Evaluation of an alternative method for the enumeration and confirmation of Clostridium perfringens from treated and untreated sewages. 2006

T Wohlsen, and J Bayliss, and B Gray, and J Bates, and M Katouli
Public Health Microbiology, Queensland Health Scientific Services, Queensland, Australia. tracy_wohlsen@health.qld.gov.au

OBJECTIVE Clostridium perfringens is recommended as a suitable indicator bacterium for human enteric viruses, Giardia cysts and Cryptosporidium oocysts in finished water and in the assessment and evaluation of water treatment. Several agars and confirmation procedures were evaluated in parallel with the Australian/New Zealand Standard (AS/NZ) Method for the enumeration of Cl. perfringens from treated and untreated sewage samples. RESULTS The current AS/NZ method utilizes tryptose sulfite cycloserine agar (TSC), lactose gelatin medium (LG) and nitrate motility medium (NM) at an incubation temperature of 37 degrees C. Sixty treated and untreated sewage samples were used to evaluate TSC agar, membrane Cl. perfringens agar (mCP), Perfringens agar (OPSP) and Perfringens agar with 4-methylumbelliferyl phosphate (OPSP-MUP) for enumeration of Clostridium. An incubation temperature of 44 degrees C for 24 h was used for comparison. Confirmation procedures were also evaluated using 103 isolates and included LG and NM, ortho-nitrophenyl-beta-D-galactopyranoside (ONPG) with MUP (ONPG-MUP) and phosphatase reagent (PR). OPSP compared favourably with TSC agar. One false negative result was obtained from each of the LG/NM and ONPG-MUP procedures. No false results were obtained using the PR confirmation procedure. CONCLUSIONS OPSP agar and PR were determined as suitable replacements for the AS/NZ Standard procedure with no interference from spreading organisms. CONCLUSIONS This is a simple and rapid method for isolating and enumerating Cl. perfringens from sewage samples and confirmed results can be reported more quickly due to shorter analytical turnaround times.

UI MeSH Term Description Entries
D010744 Phosphoric Monoester Hydrolases A group of hydrolases which catalyze the hydrolysis of monophosphoric esters with the production of one mole of orthophosphate. Phosphatase,Phosphatases,Phosphohydrolase,Phosphohydrolases,Phosphomonoesterase,Phosphomonoesterases,Phosphoric Monoester Hydrolase,Hydrolase, Phosphoric Monoester,Hydrolases, Phosphoric Monoester,Monoester Hydrolase, Phosphoric
D003016 Clostridium perfringens The most common etiologic agent of GAS GANGRENE. It is differentiable into several distinct types based on the distribution of twelve different toxins. Clostridium welchii
D003470 Culture Media Any liquid or solid preparation made specifically for the growth, storage, or transport of microorganisms or other types of cells. The variety of media that exist allow for the culturing of specific microorganisms and cell types, such as differential media, selective media, test media, and defined media. Solid media consist of liquid media that have been solidified with an agent such as AGAR or GELATIN. Media, Culture
D001616 beta-Galactosidase A group of enzymes that catalyzes the hydrolysis of terminal, non-reducing beta-D-galactose residues in beta-galactosides. Deficiency of beta-Galactosidase A1 may cause GANGLIOSIDOSIS, GM1. Lactases,Dairyaid,Lactaid,Lactogest,Lactrase,beta-D-Galactosidase,beta-Galactosidase A1,beta-Galactosidase A2,beta-Galactosidase A3,beta-Galactosidases,lac Z Protein,Protein, lac Z,beta D Galactosidase,beta Galactosidase,beta Galactosidase A1,beta Galactosidase A2,beta Galactosidase A3,beta Galactosidases
D012680 Sensitivity and Specificity Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed) Specificity,Sensitivity,Specificity and Sensitivity
D012722 Sewage Refuse liquid or waste matter carried off by sewers. Sludge,Sludge Flocs
D013696 Temperature The property of objects that determines the direction of heat flow when they are placed in direct thermal contact. The temperature is the energy of microscopic motions (vibrational and translational) of the particles of atoms. Temperatures
D015169 Colony Count, Microbial Enumeration by direct count of viable, isolated bacterial, archaeal, or fungal CELLS or SPORES capable of growth on solid CULTURE MEDIA. The method is used routinely by environmental microbiologists for quantifying organisms in AIR; FOOD; and WATER; by clinicians for measuring patients' microbial load; and in antimicrobial drug testing. Agar Dilution Count,Colony-Forming Units Assay, Microbial,Fungal Count,Pour Plate Count,Spore Count,Spread Plate Count,Streak Plate Count,Colony Forming Units Assay, Microbial,Colony Forming Units Assays, Microbial,Agar Dilution Counts,Colony Counts, Microbial,Count, Agar Dilution,Count, Fungal,Count, Microbial Colony,Count, Pour Plate,Count, Spore,Count, Spread Plate,Count, Streak Plate,Counts, Agar Dilution,Counts, Fungal,Counts, Microbial Colony,Counts, Pour Plate,Counts, Spore,Counts, Spread Plate,Counts, Streak Plate,Dilution Count, Agar,Dilution Counts, Agar,Fungal Counts,Microbial Colony Count,Microbial Colony Counts,Pour Plate Counts,Spore Counts,Spread Plate Counts,Streak Plate Counts
D018508 Water Purification Any of several processes in which undesirable impurities in water are removed or neutralized; for example, chlorination, filtration, primary treatment, ion exchange, and distillation. It includes treatment of WASTEWATER to provide potable and hygienic water in a controlled or closed environment as well as provision of public drinking water supplies. Waste Water Purification,Waste Water Treatment,Wastewater Purification,Wastewater Treatment,Water Treatment,Purification, Waste Water,Purification, Wastewater,Purification, Water,Treatment, Waste Water,Treatment, Wastewater,Treatment, Water,Waste Water Purifications,Waste Water Treatments,Water Purification, Waste

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