Faecal indicator organisms in frozen prawn products. Pt. 1. Incidence and general distribution

A general survey carried out on several brands of frozen prawn products has shown that along with the standard plate count (SPC), the numbers of pathogenic organisms like Escherichia coli, enterococci and coagulase positive staphylococci have also to be taken into consideration for the evaluation of the quality of these products. No correlation could be established between the total plate count and the number of E. coli, enterococci or staphylococci. Enumeration of enterococci, however, is advocated as a better index of faecal contamination of the products than E. coli.

Microbiological quality of some iced fishes of Imphal market, Manipur

Imphal is the main marketing centre of fish in Manipur. As fish production of the state is not sufficient to meet the demands, about 120 metric tons of iced fishes are annually brought from other states and sold in this market. Microbiological quality of iced Wallago attu, Labeo rohita, L. gonius and Aorichthy aor in respect of total fungal count (TFC), total plate count of bacteria (TPC), Most Probable Number (MPN) of coliforms, Streptococci, Staphylococcus, Salmonella and Escherichia coli in four tissues (skin, muscle, gill and intestine) were analysed. In all cases, the counts were highest in the gills and lowest in the muscles. The values of TFC, TPC, coliforms, Streptococci and Staphylococci were 0-10³/g 10(sup)6-10⁸/g, 2-α/g, 10-10⁵/g, 10-10⁵/g respectively. E. coli and Salmonella were not detected in any of the samples while the ice used in the preservation contained 10⁵-10⁷ of TPC per gram. The microbiological qualities of the iced fishes of Imphal market were adjudged poor. The extremely high counts of bacteria might be due to (1) poor quality and left over fishes being packed, (2) contact with contaminated ice and (3) repeated thawing and freezing during the process of marketing and transportation.

Chitosan as a water clarifying agent

Chitosan may be used to reduce the bacterial load of water. Material prepared according to the method of Radhakrishnan & Prahu described in Res. & Ind., 16(4), pp. 265, used in 1% solution in 1% acetic acid was added at 10 ppm level to contaminated water and allowed to stand for 45 min. Cultures of E. coli, Staphylococci and a mixture of the 2 were inoculated into ordinary and muddy water. Bacterial load was determined, and it is shown that chitosan has excellent qualities as a coagulant/water clarifying agent, especially for muddy waters or those contaminated with suspended matter or bacteria.

A bacteriological study of the natural flora of edible oyster, Crassostrea madrasensis

The total viable bacterial populations in the oysters and the sea water from the edible oyster farm at Tuticorin were in the range of 10 super(3) to 10 super(4) per ml and 1 super(2) to 10 super(3) per ml respectively. The maximum most probable number of faecal coliform recorded during the one year period of study of both the oysters and seawater were 33 per 100 ml. Pathogenic bacteria (Salmonella sp., Vibrio cholerae, coagulase positive staphylococci and faecal streptococci were absent in oysters and farm water. Study of 197 (98 taken from oyster liquid and 99 from oyster farm water) randomly isolated cultures indicated that gram negative asporogenus rod-like bacteria of the Vibrio, Flavobacterium, Achromobacter and Pseudomonas groups were the dominant flora of the oyster liquid as well as seawater.

Assessment of quality of fish landed at the Cochin fisheries harbour

Assessment of quality of fish and shrimps landed at the fish harbour, Cochin, was made over the period January, 1980 to December, 1982. A total of 201 samples were analysed. Nearly 75% of the samples scored between 6-10 in a 10 point hedonic scale and the percentage unacceptability based on sensory assessment was 5. 5%. In 10.1% of the samples, total volatile nitrogen was >30 mg% and in 8.3% of the samples trimethylamine- nitrogen was >10 mg%. Both, the gr Torrymeter and Intellectron Fish Tester VI readings marked significant correlation with sensory scores and chemical indices; but failed to bear any significant correlation with bacterial counts. Of the 5-1 total samples 66.7% had total plate count (TPC) ≥10 super(5) g super(-1) and 8.5% were considered unacceptable based on TPC >5x10 super(5) g super(-1); 63.2% of the samples were free from Escherichia coli; 26.4% had >20 E. coli g super(-1) and 20.4% of the samples contained faecal streptococci >10 super(3) g super(-1). Seven percent of the samples showed coagulase positive staphylococci >100 g super(-1). Salmonella could not be detected in any of the samples examined. Crushed ice samples and the platform of the harbour had high bacterial loads.

Quality of certain ready-to-eat fish products of the domestic trade

Quality of 181 samples of ready-to-eat fish products comprising fried fish, fish curry and fish/prawn pickles collected from Cochin and Calicut were studied. Salmonella was absent in all the samples. V. cholerae was tested in the samples collected at Cochin and was absent in all the cases. Coliforms, E. coli, faecal streptococci and coagulase-positive staphylococci were present in some of the samples studied. The study indicated the necessity to improve the sanitary and hygienic conditions of the hotels engaged in the preparation of these products. The study further indicated that fried fish and fish curry shall not be served after 6 hours of their preparation. Added care is to be exercised in the selection of shrimps and fish for the preparation of pickles.

Impact of antibiotics on conjugational resistance gene transfer in Staphylococcus aureus in sewage.

Ohlsen K, Ternes T, Werner G, Wallner U, Löffler D, Ziebuhr W, Witte W, Hacker J. Institute for Molecular Biology of Infectious Diseases, The University of Würzburg, Röntgenring 11, D-97070 Würzburg, Germany. knut.ohlsen@mail.uni-wuerzburg.de The growing rate of microbial pathogens becoming resistant to standard antibiotics is an important threat to public health. In order to assess the role of antibiotics in the environment on the spread of resistance factors, the impact of subinhibitory concentrations of antibiotics in sewage on gene transfer was investigated using conjugative gentamicin resistance (aacA-aphD) plasmids of Staphylococcus aureus. Furthermore, the concentration of antibiotics in hospital sewage was measured by high-performance liquid chromatography (HPLC)-electrospray tandem mass spectrometry. Several antibiotics were found to be present in sewage, e.g. ciprofloxacin up to 0.051 mgl(-1) and erythromycin up to 0.027 mgl(-1). Resistance plasmid transfer occurred both on solidified (dewatered) sewage and in liquid sewage in a bioreactor with a frequency of 1.1x10(-5)-5.0x10(-8). However, low-level concentrations of antibiotics measured in sewage are below concentrations that can increase plasmid transfer frequencies of gentamicin resistance plasmids of staphylococci.

Comparison of the cidal activity of tea tree oil and terpinen-4-ol against clinical bacterial skin isolates and human fibroblast cells

Aim: The aim of this study was to compare both the antimicrobial activity of terpinen-4-ol and tea tree oil (TTO) against clinical skin isolates of meticillin-resistant Staphylococcus aureus (MRSA) and coagulase-negative staphylococci (CoNS) and their toxicity against human fibroblast cells.

Staphylococcus epidermidis device-related infections: pathogenesis and clinical management

Staphylococcus epidermidis, the most frequently isolated coagulase-negative staphylococcus, is the leading cause of infection related to implanted medical devices (IMDs). This is directly related to its capability to establish multilayered, highly structured biofilms on artificial surfaces. At present, conventional systemic therapies using standard antimicrobial agents represent the main strategy to treat and prevent medical device-associated infections. However, device-related infections are notoriously difficult to treat and bacteria within biofilm communities on the surface of IMDs frequently outlive treatment, and removal of the medical device is often required for successful therapy. Importantly, major advances in this research area have been made, leading to a greater understanding of the complexities of biofilm formation of S. epidermidis and resulting in significant developments in the treatment and prevention of infections related to this member of the coagulase-negative group of staphylococci. This review will examine the pathogenesis of the clinically significant S. epidermidis and provide an overview of the conventional and emerging antibiofilm approaches in the management of medical device-associated infections related to this important nosocomial pathogen.

Catheter-related bloodstream infection in adult haematology patients:Catheter removal practice and outcome

The aim of the present study was to describe the practice of central venous catheter (CVC) removal and outcomes of catheter-related bloodstream infection (CR-BSI) in adult haematology patients. Patients were identified retrospectively according to diagnosis coding of inpatient episodes and evaluated when, on examination of medical records, there had been evidence of sepsis with strong clinical suspicion that the source was the CVC. Demographic and bacteriological data, as well as therapeutic measures and clinical outcomes, were recorded. One hundred and three patient episodes were evaluated. The most frequent type of CVC was the Hickman catheter and the most frequently isolated pathogen was coagulase-negative staphylococci. Twenty-five percent of episodes were managed with catheter removal. Treatment failure, defined as recurrence of infection within 90 days or mortality attributed to sepsis within 30 days, occurred significantly more frequently in the group managed without catheter removal (52.5% versus 4%, P

Propionibacterium acnes and Staphylococcus lugdunensis Cause Pyogenic Osteomyelitis in an Intramedullary Nail Model in Rabbits

Propionibacterium acnes and coagulase-negative staphylococci (CoNS) are opportunistic pathogens implicated in prosthetic joint and fracture fixation device-related infections. The purpose of this study was to determine whether P. acnes and the CoNS species Staphylococcus lugdunensis, isolated from an "aseptically failed" prosthetic hip joint and a united intramedullary nail-fixed tibial fracture, respectively, could cause osteomyelitis in an established implant-related osteomyelitis model in rabbits in the absence of wear debris from the implant material. The histological features of P. acnes infection in the in vivo rabbit model were consistent with localized pyogenic osteomyelitis, and a biofilm was present on all explanted intramedullary (IM) nails. The animals displayed no outward signs of infection, such as swelling, lameness, weight loss, or elevated white blood cell count. In contrast, infection with S. lugdunensis resulted in histological features consistent with both pyogenic osteomyelitis and septic arthritis, and all S. lugdunensis-infected animals displayed weight loss and an elevated white blood cell count despite biofilm detection in only two out of six rabbits. The differences in the histological and bacteriological profiles of the two species in this rabbit model of infection are reflective of their different clinical presentations: low-grade infection in the case of P. acnes and acute infection for S. lugdunensis. These results are especially important in light of the growing recognition of chronic P. acnes biofilm infections in prosthetic joint failure and nonunion of fracture fixations, which may be currently reported as "aseptic" failure. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Dual functional ionic liquids as antimicrobials and plasticisers for medical grade PVCs

Two ionic liquids, 1-ethylpyridinium docusate (IL1) and tri-n-butyl(2-hydroxyethyl)phosphonium docusate (IL2), were designed and synthesized with the explicit intention of imparting a combination of plasticization and antimicrobial efficacy when incorporated into medical grade poly(vinyl chloride)s (PVCs). The glass transition (T-g) of PVC can be reduced by >20 degrees C on addition of 15 wt% IL2. Both IL1 and IL2 leached to varying extents from the base PVC resins rendering the surface of the PVCs hydrophilic. The antimicrobial activity of both ILs is related to the presence and concentration of both cationic and anionic component of the ILs leached from the PVC and inversely proportional to the extent of PVC gelation. Blends of the PVCs with IL1 displayed antibacterial activity against almost all Gram-positive bacteria tested, including coagulase-negative Staphylococci (CoNS) and methicillin-resistant Staphylococcus aureus (MRSA), but not with IL2 at low concentration in contrast to our previous study when high concentrations of IL2 were used. The more hydrophilic IL1 when added to PVC retards biofilm formation.

Novel particulate antibiotic-loaded platforms as sustained drug delivery systems for bone infection treatment

Tese de doutoramento, Farmácia (Tecnologia Farmacêutica), Universidade de Lisboa, Faculdade de Farmácia, 2015

ß-lactamases in the biochemistry and molecular biology laboratory

β-lactamases are hydrolytic enzymes that inactivate the β-lactam ring of antibiotics such as penicillins and cephalosporins. The major diversity of studies carried out until now have mainly focused on the characterization of β-lactamases recovered among clinical isolates of Gram-positive staphylococci and Gram-negative enterobacteria, amongst others. However, only some studies refer to the detection and development of β-lactamases carriers in healthy humans, sick animals, or even in strains isolated from environmental stocks such as food, water, or soils. Considering this, we proposed a 10-week laboratory programme for the Biochemistry and Molecular Biology laboratory for majors in the health, environmental, and agronomical sciences. During those weeks, students would be dealing with some basic techniques such as DNA extraction, bacterial transformation, polymerase chain reaction (PCR), gel electrophoresis, and the use of several bioinformatics tools. These laboratory exercises would be conducted as a mini research project in which all the classes would be connected with the previous ones. This curriculum was compared in an experiment involving two groups of students from two different majors. The new curriculum, with classes linked together as a mini research project, was taught to a major in Pharmacy and an old curriculum was taught to students from environmental health. The results showed that students who were enrolled in the new curriculum obtained better results in the final exam than the students who were enrolled in the former curriculum. Likewise, these students were found to be more enthusiastic during the laboratory classes than those from the former curriculum.

Sonication of removed breast implants for improved detection of subclinical infection.

BACKGROUND: Capsular fibrosis is a severe complication after breast implantation with an uncertain etiology. Microbial colonization of the prosthesis is hypothesized as a possible reason for the low-grade infection and subsequent capsular fibrosis. Current diagnostic tests consist of intraoperative swabs and tissue biopsies. Sonication of removed implants may improve the diagnosis of implant infection by detachment of biofilms from the implant surface. METHODS: Breast implants removed from patients with Baker grades 3 and 4 capsular contracture were analyzed by sonication, and the resulting sonication fluid was quantitatively cultured. RESULTS: This study investigated 22 breast implants (6 implants with Baker 3 and 16 implants with Baker 4 capsular fibrosis) from 13 patients. The mean age of the patients was 49 years (range, 31-76 years). The mean implant indwelling time was 10.4 years (range, 3 months to 30 years). Of the 22 implants, 12 were used for breast reconstruction and 10 for aesthetic procedures. The implants were located subglandularly (n = 12), submuscularly (n = 6), and subcutaneously (n = 4). Coagulase-negative staphylococci, Propionibacterium acnes, or both were detected in the sonication fluid cultures of nine implants (41%), eight of which grew significant numbers of microorganisms (>100 colonies/ml of sonication fluid). CONCLUSIONS: Sonication detected bacteria in 41% of removed breast implants. The identified bacteria belonged to normal skin flora. Further investigation is needed to determine any causal relation between biofilms and capsular fibrosis.