Construction of an expression vector for Lactococcus lactis based on an indigenous cryptic plasmid

To construct an expression vector for Lactococcus lactis, the EmPMT fragment which contained the erythromycin resistance gene, P32 promoter, multiple cloning site (MCS) and terminator (T) was subcloned into the small cryptic plasmid pAR141. The resulting vector, designated as pAR1411, wasfound to be stably maintained in L. lactis MG1363 after transformation for at least 100 generations under non-selective conditions. The vector was also demonstrated to be able to express the gene coding for chloramphenicol acetyltransferase (cat) in L. lactis

Evaluation of in vitro Vibrio static activity of Shewanella algae isolated from healthy Penaeus monodon

To conquer disease problem in shrimp industries, probiotic biocontrol is a well known remedy now. The antagonistic ability of separated isolates from different parts of juvenile Penaeus monodon werescreened against shrimp Vibrio pathogens; Vibrio parahaemolyticus and Vibrio alginolyticus. The most antagonistic effect was observed for an isolate that was primarily identified as Shewanella algae usingconventional methods followed by Biolog microlog software. Since production of antagonistic agents rely on cultural conditions, antagonistic ability of candidate probioic against the mentioned Vibrios was assessed using Response Surface Methodology, with central composite design in which four independents variables were assumed: temperature (10 - 50°C), pH (6 -10), NaCl concentration (0 - 50%)and time (12 – 60 h). The coefficients of multiple determinations (R2), for the responses of antagonistic effect of S. algae against V. parahaemolyticus and V. alginolyticus values were 0.807 and 0.805,respectively. Concentration of the NaCl exhibited least influence on the antibacterial effect of candidate probiotic while the other independent variables exhibited different degree of affect. The candidateprobiotic revealed a reasonable antibacterial response in quite a wide range of temperature and pH in which the maximum levels were in the same range of optimum shrimp culture

Predicting youth participation in urban agriculture in Malaysia: insights from the theory of planned behavior and the functional approach to volunteer motivation

This study examines factors associated with the decision of Malaysian youth to participate in a voluntary urban agriculture program. Urban agriculture has generated significant interest in developing countries to address concerns over food security, growing urbanization and employment. While an abundance of data shows attracting the participation of young people in traditional agriculture has become a challenge for many countries, few empirical studies have been conducted on youth motivation to participate in urban agriculture programs, particularly in non-Western settings. Drawing on the theories of planned behavior and the functional approach to volunteer motivation, we surveyed 890 students from a public university in Malaysia about their intention to join a new urban agriculture program. Hierarchical regression findings indicated that the strongest predictor of participation was students’ attitude toward urban agriculture, followed by subjective norms, career motives and perceived barriers to participation. The findings from this study may provide useful information to the university program planners in Malaysia in identifying mechanisms for future students’ involvement in the program

A simplified multiplex PCR assay for fast and easy discrimination of globally distributed staphylococcal cassette chromosome mec types in meticillin-resistant Staphylococcus aureus

A multiplex PCR assay was developed for the identification of major types and subtypes of staphylococcal cassette chromosome mec (SCCmec) in meticillin-resistant Staphylococcus aureus (MRSA) strains. The method uses a novel 9 valent multiplex FOR plus two primer pairs for S. aureus identification and detection of meticillin resistance. All 389 clinical MRSA isolates from Malaysia and 18 European isolates from the Harmony collection harbouring different SCCmec types that we tested were correctly characterized by our PCR assay SCCmec type III and V were by far the most common types among both hospital- and community-acquired Malaysian MRSA isolates, with an apparent emergence of MRSA harbouring the IVh type

Methicillin-susceptible Staphylococcus aureus from clinical and community sources are genetically diverse

Despite the association of methicillin-susceptible S. aureus (MSSA) with several life-threatening diseases, relatively little is known about their clinical epidemiology in Malaysia. We characterized MSSA isolates (n = 252) obtained from clinical and community (carriage) sources based on spa sequencing and multilocus sequence typing (MLST). The prevalence of several important virulence genes was determined to further define the molecular characteristics of MSSA clones circulating in Malaysia. Among the 142 clinical and 110 community-acquired MSSA isolates, 98 different spa types were identified, corresponding to 8 different spa clonal clusters (spa-CCs). In addition, MLST analysis revealed 22 sequence types (STs) with 5 singletons corresponding to 12 MLST-CCs. Interestingly, spa-CC084/085 (MLST-CC15) (p = 0.038), spa-non-founder 2 (MLST-ST188) (p = 0.002), and spa-CC127 (MLST-CC1) (p = 0.049) were identified significantly more often among clinical isolates. spa-CC3204 (MLST-CC121) (p = 0.02) and spa-CC015 (MLST-CC45) (p = 0.0002) were more common among community isolates. Five dominant MLST-CCs (CC8, CC121, CC1, CC45, and CC5) having clear counterparts among the major MRSA clones were also identified in this study. While the MSSA strains are usually genetically heterogeneous, a relatively high frequency (19/7.5%) of ST188 (t189) strains was found, with 57.8% of these strains carrying the Panton-Valentine leukocidin (PVL). Analysis of additional virulence genes showed a frequency of 36.5% and 36.9% for seg and sei and 0.8% and 6.3% for etb and tst genes, respectively. Arginine catabolic mobile element (ACME) was detected in 4 community isolates only. These represent the first isolates harbouring this gene in an Asian region. In conclusion, MSSA from the Malaysian community and their clinical counterparts are genetically diverse, but certain clones occur more often among clinical isolates than among carriage isolates and vice versa. (C) 2010 Elsevier GmbH. All rights reserved

Nasal carriers are more likely to acquire exogenous Staphylococcus aureus strains than non-carriers

We performed a prospective observational study in a clinical setting to test the hypothesis that prior colonization by a Staphylococcus aureus strain would protect, by colonization interference or other processes, against de novo colonization and, hence, possible endo-infections by newly acquired S. aureus strains. Three hundred and six patients hospitalized for >7 days were enrolled. For every patient, four nasal swabs (days 1, 3, 5, and 7) were taken, and patients were identified as carriers when a positive nasal culture for S. aureus was obtained on day 1 of hospitalization. For all patients who acquired methicillin-resistant S. aureus (MRSA) or methicillin-susceptible S. aureus via colonization and/or infection during hospitalization, strains were collected. We note that our study may suffer from false-negative cultures, local problems with infection control and hospital hygiene, or staphylococcal carriage at alternative anatomical sites. Among all patients, 22% were prior carriers of S. aureus, including 1.9% whom carried MRSA upon admission. The overall nasal staphylococcal carriage rate among dermatology patients was significantly higher than that among neurosurgery patients (n = 25 (55.5%) vs. n = 42 (16.1%), p 0.005). This conclusion held when the carriage definition included individuals who were nasal culture positive on day 1 and day 3 of hospitalization (p 0.0001). All MRSA carriers were dermatology patients. There was significantly less S. aureus acquisition among non-carriers than among carriers during hospitalization (p 0.005). The mean number of days spent in the hospital before experiencing MRSA acquisition in nasal carriers was 5.1, which was significantly lower than the score among non-carriers (22 days, p 0.012). In conclusion, we found that nasal carriage of S. aureus predisposes to rather than protects against staphylococcal acquisition in the nose, thereby refuting our null hypothesis. Clinical Microbiology and Infection (C) 2015 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved