Water samples (100 mL) were either processed directly or diluted when necessary into IDEXX (Westport, ME, United States) Colilert or Enterolert substrate for enumeration of total coliforms/E. coli and enterococci, respectively, as recommended by the manufacturer. In brief, the mixtures were then transferred to an IDEXX Quanti-Tray/2000, sealed using the IDEXX Quanti-Tray Sealer PLUS, and then, respectively, incubated at 35 and 41°C for 24 h as recommended by the manufacturer. Positive control organisms, consisting of Klebsiella pneumoniae subsp. pneumoniae (ATCC [American Type Culture Collection] 13883) and Escherichia coli (ATCC 25922) for Colilert and E. faecalis (ATCC 29212) for Enterolert, were utilized to confirm the effectiveness of each new substrate lot. After the 24-h incubation period, the Colilert and Enterolert trays were viewed under a 365-nm UV light, with blue fluorescing wells marked positive for E. coli and enterococci. In Colilert trays, yellow wells under normal lighting were marked positive for total coliforms. The number of positive wells were converted to most probable number (MPN) using the IDEXX result interpretation table and the dilution factor.
Quanti tray sealer plus
Manufactured by IDEXX
Sourced in United States
The Quanti-Tray Sealer PLUS is a laboratory equipment designed for sealing Quanti-Tray sample trays. It provides a consistent and secure seal to facilitate the most accurate count of microbiological samples.
Automatically generated - may contain errors
4 protocols using quanti tray sealer plus
1
Enumeration of Waterborne Coliforms and Enterococci
2
Quantification of L. pneumophila in Water
3
Quantification of L. pneumophila in Water
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The Legiolert® test kit by IDEXX was used to quantify L. pneumophila (detects 1 organism in 100 mL within 7 days) in non-potable water following manufacturer recommended methods, briefly described here [22 ]. A volume of 100 mL of deionized water and the content of a Legiolert® blister pack were added to a specimen container. Using a sterile microtube, 0.2 mL of Legiolert® Pretreatment and 0.2 mL of non-potable water sample were mixed and incubated at room temperature for 60 s (±5 s). Approximately 0.2 mL of the mix contained in the microtube was transferred to the specimen container. After mixing, the contents of the specimen containers were poured into Quanti-Tray/Legiolert® trays. The trays were immediately sealed in an IDEXX Quanti-Tray Sealer PLUS (IDEXX Laboratories, Inc., Westbrook ME, USA), and incubated paper-side down with the wells facing up at 37 ± 0.5 °C, 85% humidity for 7 days. For comparison, a negative control blank containing Legiolert® reagent and deionized water was incubated. Any sample that exhibited a brown color or a turbidity higher than the negative control was positive for L. pneumophila. A lack of brown color change and turbidity less than the negative control indicated a negative result. All the positive wells were counted to obtain the concentration in most probable number (MPN) as indicated by the Legiolert® MPN table (dilution factor = 0.0001).
4
Quantifying Culturable L. pneumophila via MPN
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The quantification of culturable L. pneumophila through the MPN method was performed using the 100 mL potable water Legiolert/Quanti-Tray kit (IDEXX Laboratories Canada Corp., Markham, ON, Canada) according to the manufacturer’s instructions. In short, aliquots of 100 mL of water were transferred into sterile polypropylene vessels and first analyzed for water hardness using Aquadur® test strips (Macherey-Nagel, Düren, Germany). Due to overall low water hardness (zero to two positive pads on test strips), 0.33 mL of Legiolert Supplement were added to each vessel, as well as one Legiolert reagent blister pack. Finally, Legiolert water sample mixtures were transferred to 96-well plates and sealed with the IDEXX Quanti-Tray Sealer PLUS. Plates were then incubated at 39 ± 0.5 °C for seven days and results were read by counting any brown or turbid wells. The MPN method ranged from 10 to 22,726 MPN/L.
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