Romolo Botrugno romolo.botrugno @copanitalia.com

Evaluation of Copan FecalSwab™ for Collection, Transportation and Preservation of enteric pathogenic bacteria with

manual and WASP™ automation plate seeding.

BACKGROUND

To complete the LBM devices, compatible with WASP™ automation, Copan developed the FecalSwab™ (a flocked swab and a tube with 2 ml of semi-liquid medium) for the collection, transportation and preservation of stool or rectal swab specimens.

OBJECTIVE

The objective of this study was to evaluate Copan FecalSwab™ to support the viability of ATCC entero-pathogenic strains according to the current CLSI M40-A2 standards.

METHODS

ATCC strains, listed in the table 1, were used for this study. Bacterial suspensions for each strains were prepared starting from a 0.5 McF, diluted tenfold in physiological saline to provide working suspensions of approximately 1.5x104 CFU/mL, 1.5x103 CFU/mL and 1.5x102 CFU/mL. FecalSwabs™ tubes were inoculated with 100ul of inoculum of each dilution in order to obtain a valid time 0 colony count (25-250 CFU/plate) with the above concentrations. All testing was performed in triplicate, three FecalSwabs™ for zero-time and three FecalSwabs™ for each subsequent incubation/storage time (6/24/48 hours RT, 6/24/48/72 hours refrigerated). At each interval, all inoculated FecalSwabs™ tubes were plated by direct swabbing on TSA+5% sheep blood agar plates and incubated under optimum conditions. Simulated stool samples in FecalSwabs™, seeded with each bacteria strain listed above, were plated with a 10ul loop both manually and by the WASP™ on selective agar and incubated under optimum conditions. After 24 hours plates, seeded with both methods, were assed for the presence of separate colonies.

RESULTS

All ATCC bacteria strains tested, were recovered at each storage time according to CLSI M40-A2 guidelines, except for C. difficile (T24 hours only). The simulated stool specimens had better streaking patterns and colonies separation in the WASP™ seeded plates compared to the manual.

ASM 2015 Poster Board Number: 1092 Romolo Botrugno, Arnalda Giambra, Laura Navarria, Roberto Paroni, Santina Castriciano

Copan Italia Spa, Brescia, Italy

V. parahaemolyticus pure colture on TCBS Wasp plated

Salmonella spp. positive sample on Hektoen Agar WASP plated

Salmonella spp. positive sample on XLD Wasp plated

C. Jejuni positive sample on CampyBap Wasp plated

TABLE 1: ATCC strains tested and Δlog

C. d

iffic

ile

ATC

C 9

689

C. j

ejun

i

AT

CC

332

91

C. u

tilis

AT

CC

995

0

E. c

oli

ATC

C 2

5922

E. c

oli

ATC

C 7

0072

8

E. fa

ecal

is

AT

CC

292

12

E. fa

ecal

is

AT

CC

512

99

E. fa

eciu

m

AT

CC

700

221

E. g

allin

arum

ATC

C 7

0042

5

S. s

onne

i

ATC

C 9

290

S. ty

phim

uriu

m

AT

CC

140

28

V. p

arah

eam

olyt

icus

AT

CC

178

02

Y. e

nter

ocol

itica

AT

CC

961

0

After 48h at RT -1,92* -1,70 -0,02 1,73 1,66 -0,13 1,13 0,86 1,29 1,88 1,85 1,90 1,92

After 72h at 4°C -1,85** -0,80 -0,15 -0,17 0,06 -0,04 -0,16 -0,05 -0,02 -0,04 -0,15 -0,11 -0,05

Results of bacterial performance at 4°C

Results of bacterial performance at RT

*= Result after 24h at RT; **= Results after 48h at 4°C.

CONCLUSIONS

Copan FecalSwab™: •  Proved to be efficient to maintain the viability of all the enteric

pathogens as per CLSI M40-A2 requirements; •  In combination with WASP™ automation allows better colonies

separation and identification; •  Is a device for transferring, preserving and transporting stool

samples that supports the viability of all relevant enteric pathogens. •  The FLOQSwab™ included in the FecalSwab™ device can be used

for collection of rectal swab or as transfer tool for feces. •  Eliminates the risk of overloading the device with excess stool

sample by the improved labelling. •  FecalSwab™ has been validated with molecular assays and rapid

antigen and toxin kits as per ECCMID 2015 poster P0334. •  Further studies with clinical specimens are in progress for validation

of Copan FecalSwab™ with culture and molecular assays and rapid antigen and toxin kits.

0,00E+00

2,00E+03

4,00E+03

6,00E+03

8,00E+03

1,00E+04

1,20E+04

1,40E+04

1,60E+04

0 hrs

6 hrs

24 hrs

48 hrs