Development of triplex assay for simultaneous detection of Escherichia coli, methicillin resistant and sensitive Staphylococcus aureus in raw pork samples of retail markets.

Escherichia coli and Staphylococcus aureus are the most important food borne pathogen transmitting from animal meat and meat products. Therefore, it is vital to design an accurate and specific diagnostic tool for identifying those food-borne pathogens in animal meat and meat products. In the current study, E. coli, methicillin-resistant and sensitive S. aureus (MRSA and MSSA) were simultaneously detected using a developed triplex PCR-based technique. To obtain an optimal reaction parameter, the multiplex assay was optimised by changing just one parameter while holding the others constant. Specificity of the assay was assessed using several porcine bacterial template DNA. The plasmid DNA was used to test the multiplex PCR assay's sensitivity and interference in spiked pork samples. E. coli, MRSA, and MSSA each have PCR amplified products with sizes of 335, 533, and 209 bp, respectively. The assay detects a minimum microbial load of 102 CFU/µl for all the three pathogens and can identify bacterial DNA as low as 10-2 ng/µl. The assay was validated employing 210 pork samples obtained from retail meat shops and slaughter houses, with MRSA, E. coli, and MSSA with the occurrence rate of 1.9%, 42.38%, and 18.1%, respectively. The rate of mixed bacterial contamination in pork meat samples examined with the developed method was 6.19%, 1.43%, 1.90%, and 1.43% for MSSA & E. coli, MRSA & E. coli, MSSA & MRSA, and E. coli, MSSA & MRSA, respectively. The developed multiplex PCR assay is quick and efficient, and it can distinguish between different bacterial pathogens in a single reaction tube.

Brute forcing on secured shell servers emphasising the role of cyber forensics - a quali-quantitative study.

BACKGROUND: Increasing numbers of cyber attacks threaten us personally and professionally. Cyber crimes include obtaining sensitive information (medical or financial) but may extend to organising heinous crimes including murders and aggravated sexual assaults. A major vector of cyber crimes is brute force attacks on secured shell servers. AIM OF STUDY: This research highlights the prevalence of the intensity of brute force attacks on secured shell servers via quali-quantitative analysis of cyber attacks. METHODOLOGY: The brute force attacks were recorded over a period of 20 days with the help of logs taken from five dedicated servers installed in a production environment. RESULTS: There were a minimum of 6470 and maximum of 22,715 attacks on a server per day. The total number of attacks on all the servers during the study period was 1,065,920. The brute force attacks were mainly targeted at the service network accounts. CONCLUSION: Growth of the field of cyber forensics is the optimal solution to prevent the malicious use of internet services and the commissioning of crimes by this means.

Characterization of piggery farm waste-borne bacterial transposable elements associated with antimicrobial resistance phenotypes.

Even though there is a link between antibiotic resistance and the presence of transposable elements few research has looked at the prevalence and distribution of transposable elements/ integrons in piggery farm samples. Present study identified the presence of six transposable elements namely Tn6763 (Accession number: OQ565300), Tn6764, (Accession number: OQ565299), Tn6765 (Accession number: OQ409902), Tn2003 (Accession number: OQ503494), Tn6072 (Accession number: OQ565298) and Tn6020 (Accession number: OQ503493) in piggery farm waste from India which are belongs to Enterobacteriaceae family. In a conjugative experiment, Klebsiella isolates carrying Tn6020 having the resistant phenotypes for nalidixic acid was used as donor cells while Escherichia coli DH5α Cells carrying chloramphenicol resistant plasmid was employed as recipient cells. Transconjugant bacterial colonies were shown to carry the Tn6020 transposable elements with both nalidixic acid (donor cell origin) and chloramphenicol (recipient cell origin) resistant antibiotic phenotypes. Given the presence of transposable elements in 21.4% of resistant Enterobacteriaceae strains, preventative measures are vital for avoiding the spread of mobile genetic resistance determinants in the piggery sector and to monitor their emergence.

Increased recognition of Chryseobacterium species as an emerging cause of nosocomial urinary tract infection following introduction of matrix-assisted laser desorption/ionisation-time of flight for bacterial identification.

Chryseobacterium species are rarely reported as aetiological agents of nosocomial urinary tract infection. Here, we evaluated the clinical significance of 19 isolates of Chryseobacterium species (15 Chryseobacterium indologenes and 4 Chryseobacterium gleum; identified by matrix-assisted laser desorption/ionisation-time of flight [MALDI-TOF]) obtained from urine or percutaneous nephrostomy drainage of 16 patients with urological complaints. The strains possessed drug resistance to multiple antibiotics. 14 isolates showed the presence of carbapenemases. Both MALDI-TOF and repetitive sequence-based-polymerase chain reaction grouped them into three clusters (Kappa 1.000). They may colonise the urinary tract acting as a reservoir for dissemination of drug resistance within hospital environment.

Rising trends of syphilis in a tertiary care center in North India.

BACKGROUND AND OBJECTIVES: Syphilis is a classical sexually transmitted disease (STD), caused by Treponema pallidum subsp. pallidum. In this retrospective study, we analyzed trends of syphilis prevalence in patient groups attending our tertiary care center. MATERIALS AND METHODS: The data was obtained by reviewing laboratory records of the STD laboratory from January 1, 2006 to December 31, 2011. Cases positive by both Venereal Disease Research Laboratory (VDRL) and Treponema pallidum particle agglutination (TPPA) tests were analyzed for seroprevalence of syphilis in different groups, and to analyze the rising or falling trends, if any. RESULTS: A total of 28,920 serum samples were received in the 6-year study period for VDRL testing, of which 972 (3.4%) were found to be reactive. Of these, 1722 sera were also submitted for TPPA testing, 374 (21.7%) of which were positive. A total of 375 samples were submitted for both tests, indicating biological false positivity of 0.27%. A rising trend, though not statistically significant, was observed in pregnant women, drug users and patients from wards/out-patient departments, while a statistically significant rise in prevalence of syphilis was found in HIV-positive individuals. A falling trend (not statistically significant) was observed in STD clinic attendees. CONCLUSION: An increasing trend of syphilis was observed during the study period when all groups were analyzed together, especially in HIV-seropositive individuals, which calls for continued and sustained efforts for case detection, treatment, and preventive measures to contain the disease.

Enlarging head size as the only presentation of Salmonella Typhimurium brain abscess in an infant: a short report.

The authors describe a 5-month-old male harbouring a giant Salmonella Typhimurium cerebral abscess who presented with progressively enlarging head size, without any features of septicemia. The report details the radiological findings, differential diagnoses and management problems regarding the index case.