USDA/GIPSA Proficiency Program
Testing for the Presence of Biotechnology Events in Corn and Soybeans
January 2004 Sample Distribution
Purpose of USDA/GIPSA
Proficiency Program
Through the USDA/GIPSA Proficiency Program, USDA seeks to improve the overall performance of testing for biotechnology-derived grains and oil seeds. The USDA/GIPSA Proficiency Program helps organizations identify areas of concern and take corrective actions to improve testing accuracy, capability and reliability.
Program Description
In February 2003, USDA/GIPSA’s Technical Services Division
expanded the program to offer samples for qualitative or quantitative analysis. Participants could request samples for qualitative
analysis or quantitative analysis. In this round of the USDA/GIPSA Proficiency Program one set
of samples was used for both qualitative and quantitative analyses. The samples were fortified with various combinations
and concentrations of transgenic events, and participants had the choice of
providing qualitative or quantitative results.
Scoring of the participant’s results was done by computing the
“percentage of correctly reported transgenic events” in the samples. Two new
biotechnology corn events commercialized in the
Sample Composition
The corn
samples contained various combinations and concentrations of the following transgenic
events: T25, CBH351, MON810, GA21, E176, Bt11, NK603, TC1507, and MON863; or,
no events (i.e., negative corn sample). To
produce the various concentration levels the appropriate amount of ground
transgenic corn was mixed with non-transgenic corn. Fortified corn samples were prepared with concentration
of 0.1 %, 0.4 % and 0.8 % of the event. The
soybean samples were either non-transgenic soybeans, or fortified soybeans samples
containing 0.1 % or 1.5 % of the transgenic glyphosate-tolerant soybeans.
Each
participant received six corn samples and three soybean samples. Each sample contained approximately 20 grams
of ground material.
Program Participants
GIPSA mailed samples to seventy-six
participants and sixty-seven participants returned results. Participants included organizations from
Sixty-seven organizations returned results by the deadline
date:
·
Twenty-nine
participants submitted qualitative results
only,
·
Nine participants
submitted quantitative results only,
and
·
Twenty-nine
participants submitted a combination of qualitative
and quantitative results.
In this report, participating
organizations are identified by either a confidential “Participant Identification
Number”, or by name. Appendix I
identifies those organizations who gave GIPSA permission to list them as
participants in the USDA/GIPSA Proficiency Program.
Data Summary Results
Data submitted by the participants are summarized in this report primarily in tables and charts. Participants reported their results on a qualitative basis, quantitative basis, or a combination of both qualitative and quantitative bases. Qualitative results were reported as the presence or absence of a particular event in each sample. Quantitative results were reported as the concentration of a particular event in the sample. Due to the complexity of the data, this report summarizes the data as follows:
Qualitative Data Summaries. This section summarizes qualitative sample analysis data:
·
Table One: Percentage correct scores for all participants
by event (DNA-based assays).
·
Figure One: Summary data of all participants for each
event combined with the number of results submitted for that particular event
(DNA-based assays).
·
Table Two: Percentage correct scores for all individual
participants by event (Lateral Flow Strip testing; Protein-based assays).
·
Table Three: Percentage correct scores for all individual
participants by event (Enzyme-Linked Immunosorbent Assay testing; Protein-based
assays).
Quantitative Data Summaries. This section summarizes quantitative sample analysis data:
·
Table Four: Percentage
correct scores for all participants by event (DNA-based assays).
·
Figure Two: Summary data of all participants for each
event combined with the number of results submitted for that particular event (DNA-based
assays).
·
Figure Three: Summary (chart) of participant’s results
relative to GIPSA’s fortification level.
·
Table Five: Summary (table) of participant’s results
relative to GIPSA’s foritification level.
·
Figure Five: Percentage of false negative and false
positive results in combined qualitative and quantitative data.
·
Table Six: Percentage correct
scores for all individual participants by event (Enzyme-Linked Immunosorbent
Assay testing; Protein-based assays).
·
Table Seven. Comparison of qualitative
and quantitative sample analysis results.
Table One. Percentage correct scores for all participants reporting Qualitative
results using DNA-based testing. Events labeled as 35S through MON863 were
assayed in corn samples. The transgenic
soybean samples contained the glyphosate tolerant event.
NR = Participants
who did not report a result
QUANT
= Participants who reported a quantitative value (refer to Table Four for those
results).
*
Figure One. Group average of percentage correct for
Qualitative reports on each event combined with the total number of results
reported using DNA-based testing. Events labeled as 35S through MON863 were assayed in
corn samples. The transgenic soybean
samples contained the glyphosate tolerant event producing the CP4 EPSPS protein. Data are shown on a composite basis
(i.e., all participants combined).
Tables Two and Three show the percentage correct scores for all participants reporting qualitative
results using Lateral Flow Strip (LFS) testing and Enzyme-Linked Immunosorbent
Assay (ELISA) testing (protein-based testing).
Events labeled as 35S through MON863 were assayed in corn samples. The transgenic soybean samples contained the
glyphosate tolerant event producing the CP4 EPSPS protein.
Table Two. Lateral Flow Strip (LFS) Testing (Protein-based testing)
Table
Three. Enzyme-Lined Immunosorbent Assay (ELISA)
Testing (Protein-based testing)
Table Four. Percentage correct scores for all participants reporting Quantitative
results using DNA-based testing. Events labeled as 35S through MON863 were
assayed in corn samples. The transgenic
soybean samples contained the glyphosate tolerant event.
NR =
Participants who did not report a result
QUAL =
Participants who reported a qualitative value (refer to Table One for those
results).
Figure Two. Group average of percentage correct for Quantitative reports on each event combined with the total number of results reported using DNA-based testing. Events labeled as 35S through MON863 were assayed in corn samples. The transgenic soybean samples contained the glyphosate tolerant event. Data are shown on a composite basis (i.e., all participants combined).
Figure Three. Accuracy of reported transgenic event quantifications. The composite mean of participants’ reported quantifications (observed value) of the event compared to the GIPSA target concentration determined gravimetrically. Corn samples were prepared to contain events at the concentrations of 0%, 0.1%, 0.4%, and 0.8%. Soy samples were prepared to contain events at the concentrations of 0%, 0.1%, and 1.5%. The relative error calculation was used to evaluate how close participant’s reported value agreed with the .
Relative error =
reported conc. – target conc. x 100 =
percentage relative error
target
conc.
Table Five:
Summary Quantitative Data
|
Transgenic Event |
Fortification level |
Observed level |
Standard dev'n |
Number of |
Relative Error |
Coeff. Var'n |
|
|
(%) |
(%) |
(%) |
Results |
(%) |
(%) |
|
T-25 |
|
|
|
|
|
|
|
|
0 |
0 |
0 |
28 |
0 |
0 |
|
|
0.1 |
0.67 |
0.48 |
32 |
570 |
72.7 |
|
|
0.4 |
0.73 |
0.54 |
31 |
83.7 |
71.4 |
|
CBH 351 |
|
|
|
|
|
|
|
|
0 |
0.004 |
0.02 |
24 |
0 |
500 |
|
|
0.1 |
0.14 |
0.13 |
22 |
40 |
92.1 |
|
|
0.4 |
0.35 |
0.16 |
22 |
(-)12.5 |
44.5 |
|
MON 810 |
|
|
|
|
|
|
|
|
0 |
0.02 |
0.07 |
48 |
0 |
380 |
|
|
0.1 |
0.13 |
0.15 |
25 |
34 |
111 |
|
|
0.4 |
0.3 |
0.2 |
25 |
24.3 |
66.6 |
|
|
0.8 |
0.8 |
0.43 |
50 |
0 |
53.7 |
|
GA 21 |
|
|
|
|
|
|
|
|
0 |
0 |
0 |
32 |
0 |
0 |
|
|
0.1 |
0.15 |
0.22 |
32 |
50 |
146 |
|
|
0.4 |
0.41 |
0.18 |
34 |
2.5 |
45.6 |
|
Ev
176 |
|
|
|
|
|
|
|
|
0 |
0 |
0 |
34 |
0 |
0 |
|
|
0.1 |
0.34 |
0.88 |
42 |
240 |
258.2 |
|
|
0.4 |
0.71 |
0.95 |
44 |
77.3 |
134 |
|
Bt 11 |
|
|
|
|
|
|
|
|
0 |
0 |
0 |
41 |
0 |
0 |
|
|
0.1 |
0.16 |
0.29 |
40 |
66 |
174 |
|
|
0.4 |
0.55 |
0.29 |
40 |
39 |
53.1 |
|
NK 603 |
|
|
|
|
|
|
|
|
0 |
0 |
0 |
22 |
0 |
0 |
|
|
0.1 |
0.17 |
0.15 |
11 |
70 |
88.2 |
|
|
0.4 |
0.36 |
0.23 |
11 |
(-)10 |
62.5 |
|
|
0.8 |
0.96 |
0.04 |
22 |
20 |
4.58 |
|
TC 1507 |
|
|
|
|
|
|
|
|
0 |
0 |
0 |
10 |
0 |
0 |
|
|
0.1 |
0.08 |
0.04 |
5 |
(-)20 |
55 |
|
|
0.4 |
0.6 |
0.79 |
5 |
50 |
132.6 |
|
|
0.8 |
0.43 |
0.32 |
10 |
(-)46 |
75.2 |
|
MON 863 |
|
|
|
|
|
|
|
|
0 |
0 |
0 |
18 |
0 |
0 |
|
|
0.1 |
0.37 |
0.69 |
9 |
272 |
185 |
|
|
0.4 |
0.57 |
0.34 |
9 |
42.5 |
60.5 |
|
|
0.8 |
0.89 |
0.46 |
18 |
12.3 |
51.6 |
|
Soy |
|
|
|
|
|
|
|
Glyphosate |
0 |
0 |
0 |
14 |
0 |
0 |
|
Tolerant |
0.1 |
0.21 |
0.28 |
30 |
113 |
130 |
|
|
1.5 |
1.86 |
0.52 |
46 |
24 |
28 |
Figure Four. Percentage
of false negatives and false positives observed in combined qualitative and
quantitative participant data reports. Results were classified as false negatives
when a participant reported the absence of a transgenic event in a sample that
contained the event. Results were
classified as false positives when a participant reported the presence of a
transgenic event in a sample that did not contain the event.
Table Six. Percentage correct scores for all
participants reporting Quantitative results using ELISA (Protein-based
testing). Events labeled as T25 through MON863 were assayed in
corn samples. The transgenic soybean
samples contained the glyphosate tolerant event producing the CP4 EPSPS protein.
NR= no result reported.
Table Seven. Comparison of performance between qualitative and quantitative
methods of sample analysis. The mean percentage correct for each event on both
types of sample analyses were compared as differentials (Qualitative mean %
correct minus Quantitative mean % correct).
|
|
T25 |
CBH351 |
Mon810 |
GA21 |
Ev176 |
Bt11 |
NK603 |
TC1507 |
MON863 |
RUR |
|
QUAL |
92.1 |
92.7 |
96 |
96 |
96 |
95 |
97.8 |
95.1 |
93.6 |
95.2 |
|
QUANT |
99 |
93.9 |
93.9 |
95.1 |
97 |
95.2 |
95.8 |
93.3 |
90 |
100 |
|
Difference |
+6.9 |
+1.2 |
-2.1 |
-0.9 |
+1.0 |
+0.2 |
-2.0 |
-1.8 |
-3.6 |
+4.8 |
Summary of Findings
Qualitative Sample Analysis
1.
The group
average “percentage correct scores” was above 95%
correct for all events except T25 and CBH351.
The group average “percentage correct” for T25 and CBH351 were
approximately 92% (See Table One). The
participants’ lower performance on T25 and CBH351, as compared to their
performance on other events, was consistent with the trend observed in the
previous round of samples (September 2003).
This differential in percentage correct was largely due to the higher
number of “false negative” results reported for T25 and CBH351 (See Figure
Four).
2.
Interestingly,
although the events TC1507 and MON863 were new transgenic events, the participants
did well in identifying their presence or absence. This is likely due to a smaller number of
participants reporting results for these events.
3.
The results
show that Lateral Flow Strip test methods can be used to correctly identify a number
of events; however, it proved less reliable for identifying the marker protein
associated with the T25 event. This is
likely due to the lowest fortification level (0.1 %) being below the Limit of
Detection.
4.
Generally, the
ELISA test methods were able to correctly identify the presence or absence of
the marker protein associated with an event (See Table Three). However, the performance of the ELISA test for the
glyphosate tolerant soybean marker protein (CP4 EPSPS) was surprisingly low (67%).
Quantitative Sample Analysis
1.
The group average
“percentage correct scores” for all events was greater than or equal to
94%, with the exception of TC1507 and MON863. The
composite “percentage correct” scores for TC1507 and MON863 were 93% and 90%
respectively (See Table Two).
2.
The quantitative
results were scored on a “qualitative” basis, and an analysis of the
“Accuracy”
of the reported concentration of an event versus the target concentration of an
event in the samples was conducted. The
quantification accuracy was evaluated by computing the relative error for each of the means of the reported values (See Figure
Three and Table Five). The trend
observed was that at the lower fortification levels, the results were less
accurate and more variable.(i.e., higher relative
error and greater coefficients of variation).
In contrast, at the higher fortification levels, the reported
quantifications were more accurate and less variable. This inverse relationship between fortification
level and the accuracy of reported quantifications has been observed in the
previous rounds and is consistent with observations of other quantitative
analytical methods. The most dramatic dispersions in quantification accuracy were
observed for events T25, E176, and MON863.
3.
Results from
this study indicate that accurate and precise quantitation
at the 0.1% fortification level is very difficult for many organizations, and
therefore, actual results reported at this level should be very carefully
considered.
4.
When the
performance between qualitative and quantitative methods of sample analysis was
examined, there was no marked difference in performance (Table Seven). This is in contrast to the September 2003
results that showed a significant difference between organizations that
reported qualitative and quantitative results.
Note: It is important to understand that there are no internationally recognized standard reference materials for all transgenic events. The transgenic seed or grain used to prepare these samples was made available to GIPSA by the Life Science Organizations. Care was taken to ensure the transgenic material was either essentially 100 % positive for the event, or adjusted accordingly. The fortified samples were prepared using a process that has been verified to produce homogenous mixes, and representative samples were analyzed to ensure proper fortification and homogeneity.
To obtain additional information on the
USDA/GIPSA Proficiency Program, contact Dr. Ron Jenkins, USDA/GIPSA Proficiency
Program Manager, at US 816-891-04442, or by e-mail at biotech-lab@usda.gov.
Appendix I. Organizations who participanted in the January GIPSA 2004 Proficiency Program.
A. Bio. C – Molecular Biology Division
Route de Samadet
64410 ARZACQ
Contact Dr. F. Bois
Telephone 33 5
59 04 49 20
Fax 33 5 59 04 49 30
E-mail bio.moleculaire@labo-abioc.fr
AINIA (Instituto Tecnologico Agroalimentario)
Benjamin Franklin 5-11
Parque Tecnologico
46980 Paterna
Contact David Tomas
Telephone +34961366090
Fax +34961318008
E-mail dtomas@ainia.es
Behoerde fur Umwelt und Gesundheit
Institut fur Hygiene und Umwelt
Marckmannstr.129b, Haus 6
D20539
Contact Norbert Hess, Ph.D.
Telephone +49 40 42845 3629
Fax +49 40 42845 3840
E-mail norbert.hess@bug.hamburg.de
Biogenetic Services, Inc.
Contact Alex Kahler
Telephone 800-423-4263/605-697-8500
Fax 605-697-8507
E-mail biogene@brookings.net
Biolytix AG
Benkenstrasse 254
CH-4108 Witterswil
Contact Peter Brodmann
Telephone 41 (0)61 723 20 70
Fax 41 (0)61 723 20 71
E-mail peter.brodmann@biolytix.ch
Biotechnology Research and Development
Office of Biotechnology Research and Development
Tunyabury District
Contact Mr. Kittisak Kiratiya-angul
Telephone 66 2 9046885-95
Fax 66 2 9046885-95, ext. 555
E-mail nudee@ksc.th.com
Bureau of Food and
Drug Analysis (BFDA), DOH,
161-2,
Nankang
Contact Dr. Lih-Ching
Chiueh
Telephone 02-26531273
Fax 02-26531268
E-mail clc1025@nlfd.gov.tw
7877 Pleasant
Contact Parm Randhawa
Telephone 916-655-1581
Fax 916-655-1582
E-mail randhawa@calspl.com
Canadian Food
Inspection Agency
K2H 8P9
Contact Cheryl Dollard
Phone 613 228-6698, ext 5960
Fax 613-228-6669
Canadian Food Inspection Agency
Center for Plant Health
Molecular Analysis Laboratory
Contact Ms. Margaret Green
Telephone (250) 363-6650 ext.
222/248
Fax (250)
363-6661
E-mail greenmg@inspection.gc.ca
Canadian Grain
Commission
R3C 3G8,
Contact Tigst Demeke
Telephone 204-984-4582
Fax 204-983-0724
E-mail tdemeke@grainscanada.gc.ca
CONGEN
Biotechnology GmbH
Robert Roessle Str.
10
13125
Telephone Fon +49-(0)30-9489 3506
Fax +49-(0)30-9489 3510
E-Mail l.grohmann@congen.de
Chemisches und Veterinäruntersuchungsamt Freiburg
(CVUA)
Bissierstrasse 5
79114
Contact Hans-Ulrich Waiblinger/Dr. Pietsch
Telephone ++49 761 8855151
Fax ++49 761 8855100
E-mail hans-ulrich.waiblinger@cvuafr.bwl.de
CNTA-Laboratorio
Ctra N-134 km 50
31570 San Adrian
Navarra
Contact Blanca Jauregui, Ph.D.
Telephone 34 948 670159
Fax 34 948 696127
E-mail bjauregui@ctncv.es
DNA Technology Laboratory
Kamphaengsaean, Nakorn Pathon 73140
Contact Apiwan Yoojinda
Telephone 66-34-355192-4
Fax 66-34-355196-7
E-mail apiwan@dnatec.kps.ku.ac.th
Eurofins Scientific, Woodson-Tenent Laboratories
Division
3507 Delaware
Des Moines , IA 50313
Contact David Pinero
Telephone 515-265-1461
Fax 515-266-5453
E-mail davidpinero@eurofinsus.com
European Commission - JRC
Via Fermi 1
I-21020 Ispra (VA)
Contact Marco Mazzara
Telephone 0039 0332 785773
Fax 0039 0332 789333
E-mail marco.mazzaro@jrc.it
FASMAC CO., LTD
Kanagawa 243-0041
Contact Dr. Satoshi Futo
Telephone +81 46-295-8787
Fax +81 46-294-3738
E-mail sfuto@fasmac.co.jp
Federal Institute for Risk Assessment
BfR
Thielallee 88-92
D-14195
Contact Jutta Zagon, Ph.D.
Telephone 49-30-8412-3876
Fax 49-30-88412-3685
E-mail j.zagon@bfr.bund.de
GeneScan Analytics
GmbH,
Engesserstr. 4
79108
Contact Dr. Castor Menendez
Telephone +49-(0)761-5038
Fax +49-(0)761-5038-111
E-mail gmoanalytics@genescan.com
Genetic ID NA
Contact Jane Pappin/Bernd Schoel
Telephone 641-472-9979, ext 124
Fax 641-472-9198
E-mail jpappin@genetic-id.com/bshoel@genetic-id.com
GeneScan do Brasil
Ltda
Gerente de Qualidade
Avenida Antonia Gazzola, 1001
3 andar
13.301-245
ITU - SP -
Contact Flavia Machado
Telephone +55 11 4023 0522
Fax +55 11 4023 0625
GeneScan USA, Inc.
Belle
Contact Dr. Frank Spiegelhalter
Telephone 504-398-0940
Fax 504-398-0945
E-mail fspiegel@gmotesting.com
JenaGen GmbH
JenaGen Diagnostik-Gentechnik-Biotechnologie
Loebstedter Str. 78
D-07749
Contact Dr. Reinhard
Baier
Telephone: +49(0)3641-464913
Fax: +49(0)3641-464991
E-mail: r.baier@jenagen.de
Indiana
Contact Huabang Chen
Telephone 765-523-2535
Fax 765-523-2536
E-mail Chen@indianacrop.org
Instituto de Biologia
Molecular de Barcelona (IBMB)
Departmento de Genetica Molecular
Consejo Superior de Investigaciones Cientificas (CSIC)
Jordi Girona Salgado 18-26
08034
Contact Marta Hernandez Perez
Telephone +34 93 400 61 00
Fax +34 93 204 59 04
Kantonales Labor Basel-Stadt
Kannenfeldstrasse 2
Postfach
CH-4012
Contact Philipp Huebner, Ph.D.
Telephone +41 61 385 25 27
Fax +41 61 385 25 09
Laboratorie IFRA
38 rue de l’industrie
BP 70192
67405 Illkirch Cedex
Contact Philippe Rousselin,
Ph.D.
Telephone 00 33 3 88 66 77 70
Fax 00 33 3 88 66 37 90
Laboratorie IFRA
38 rue de l’industrie
BP 70192
67405 Illkirch Cedex
Contact Philippe Rousselin,
Ph.D.
Telephone 00 33 3 88 66 77 70
Fax 00 33 3 88 66 37 90
Landesuntersuchungsanstalt fur das Gesundheits-und Veterinarwesen Sachsen
Sitz Dresden
Amtliche Lebensmitteluberwachung
Fachgebiet 6.6
Postfach 2002744
D – 01192
Contact Dr. Gerda Hempel
Telephone +49-0351-8144-474
Fax +49-0351-8144-497
Mid-West Seed
Services
Contact Kalyn Brix-Davis
Telephone 605-692-7611
Fax 605-692-7617
E-mail kalynb@mwseed.com
National Food
Institute
2008 Soi Charansanitwong
40
Contact Phattraphorn Choo-in
Telephone 66(0)2886-8088
Fax 66(0)28868106-7
E-mail phattrapornc12@hotmail.com
National
Vecna pot 111
1000
Contact Dr. Jana Zel
Telephone +386 1 4233388
Fax +386 1 2573 847
E-Mail jana.zel@nib.si
Niedersachsisches Landesamt fur Verbraucherschutz und Lebensmittelsicherheit
Lebensmittelinstitut Braunschweig
Dresdenstrasse 2 + 6
38124 Braunschweig
Contact Manuela Schulze, Ph.D.
Telephone 0531/6804 205
Fax 0531/6804 201
E-mail manuela.schulze@lua-bs.niedersachsen.de
Nippon Yuryo Kentei Kyokai
Yokohama
(Japan Oil Stuff Inspectors Corporation)
Bankokubashi Bldg
Kaigan-dori Naka-ku
231-0002,
Contact Ms Kumi Goto
Telephone 045-641-1037
Fax 045-641-1038
E-mail goto@nykk.or.jp
Ocimum Biosolutions, LLC
Contact Sujata Pammi, Ph.D.
Telephone: 317-228-0600
Fax: 317-228-0700
e-mail: sujata@ocimumbio.com
OMIC USA Inc.
Contact Dr. Farin Hajar
Telephone 503-223-1497
Fax 503-223-9436
E-mail dna.us@omicnet.com
Reading Scientific Services Ltd.
The
Lord Zuckerman Research Centre
Whiteknights
Reading RG66LA
Contact Andrew P Tingey, PhD.
Telephone +44 (0)118 986 8541
Fax +44 (0)118 986 8932
E-mail andrew.p.tingey@rssl.com
SE-230 53 Alnarp
Contact PeO Gummeson/Anders Dahlqvist
Tel: +46
40415321
Fax: +46
40415321
e-mail: per.olov.gummeson@scangene.se
Silliker, Inc.
Contact Dr. Daniel Wetsch
Telephone 319-366-3570
Fax 319-366-4018
E-mail daniel.wetsch@silliker.com
Staatliches Veterinaeruntersuchungsamt
Zur Taubeneiche 10-12
D-59821
Contact Jochen Kilwinski, Ph.D.
Telephone 49-2931-809-220
Fax 49-2031-809-290
E-mail kilwinski@svia-arnsberg.nrw.de
Staatliches Veterinaeruntersuchungsamt
Detmold
Westerfeldstrasse l
D-32758
Contact Dr. Margit Hagen
Telephone +49 (0)5231/911-740
Fax +49 (0)5231/911-503
E-mail margit.hagen@svua-detmold.nrw.de
TECAM
Rua Fabia, 59
Sao Paulo – SP – CEP: 05051-030
Contact Dr. Daniela Contri
Telephone 55 11 3873 2553
Fax 55 11 3862 8954
E-mail biomol@tecam.com.br
Thuringer
Landesamt fur Lebensmittelsicherheit und Verbraucherschutz
Sitz Jena/Winterstein
Amtliche
Lebensmitteluberwachung
Nauburger Str. 96 b
D-07743
Tobacco Research
Board
Kutsaga Station
Airport Ring Road
Contact Dr. Dahlia Garwe
Telephone 263 4 575290/4
Fax 263 4 575288
E-mail Dahlia_Garwe@kutsaga.co.zw