© MENNO1

MENNO Hygienemanagement

AtAtAtAt firstfirstfirstfirst

disinfectiondisinfectiondisinfectiondisinfection, , , ,

thenthenthenthen

productionproductionproductionproduction!!!!

© MENNO2

Disinfectants – active ingredients in horticulture

compound main indication handicap�Alcohol (e.g. Ethanol 70 %, etc.)

Disinfection of knives by dipping

Only working against bacteria and fungi. No effect against viruses!

�Peroxis and other oxidants (Hydrogen peroxide, Peracetic acid, Potassium permanganate etc.)

Disinfection of hard surfaces, standing areas, equipment, etc.

By conditions of horticulture reduced effectiveness. Reaction with peat and mats by loosing activity. Peracetic acid – Protection of Workers! Intensive specific smell.

�Halogenes(Chlorine-, Iodine- and Bromine-compounds)

Disinfection in hospital Phytotoxicity vapour pase. Plant damages! Under conditions of horticulture reduced effectiveness. Reaction with peat and mats by loosing activity.

�Aldehydes (Formaldehyde, Paraformaldehyde, Glutaraldehyde etc.)

Disinfection in animal houses Phytotoxicity vapour pase. Plant damages! Protection of Workers! Cancerogenic!

�Quatanery ammonia compounds(Benzalconiumchlorid, Alkyltrimethylam., etc.)

Disinfection in food industry Effective against bactericid and fungicid. Phytotoxicity in contact with plant material.

�Phenols (e.g. 2-Phenylphenol, 4-Chloro-3-phenol)

Disinfection in animal houses and in hospital

Phytotoxicity vapour pase. Plant damages!

�Didecyl-dimethylammoniachloridM&ENNO™ TER forte with registrations

Disinfection in Horticulture for use on hard surfaces, standing areas, pots, knives, equipment, etc.

Effective bactericid, fungicid (includingdurable forms), algicid and herbicid.Phytotoxicity in direct contact with plant material.

�Benzoic-acid MENNO Florades™ with registrations

Disinfection in Horticulture for use on hard surfaces, standing areas, pots, knives, equipment, etc.

Effective against viruses, viroids, bacteria andfungi (including durable forms).Exceptionell good plant tolerance.

© MENNO3

What should be disinfected – How long ?

dip-disinfection� pots, containers,

trays etc.� clay

� polypropylen� polyethylen

� styrofoam

� wood etc.

� tools� knives

� machinery

� pushcarts

� grids� sticks etc..

drench-disinfection� greenhouse structures

� glass� path ways

� metal constructions etc.

� standing grounds� concrete

� irrigation mats

� aluminum

� plastic (sheets, fabric) etc.

special applications� hands, feet etc.� seed, e.g. potato� culturs, mother plants,

young plants

Continouslypreventive and repeated andparticularly after symptoms

Everythingequipment, tyres, glass- and carrierconstructions, watering systems, ventilatiors,...

Completelyall areas and surfaces,...

Thouroughlyavoid dilution by „rest“ water in concrete and mats,...

The exposure time must be longer if adhering dirt, roots and plant remains and that may be contained permanent forms of fungi to be fight. The soaking and penetration of the disinfectant solution takes time!Short exposure times are sufficient for bacteria, vegetative forms of fungi, or viruses in the knife disinfection.

© MENNO4

Activity decreasing factors different for various fields of application

� pathogen (e.g. resting spores)

� min. contact time

� min. exposition time

� min. concentration

� dirt load, algae etc.

� plant rest, e.g. roots

� pH value

� temperature

� water quality

� others

© MENNO5

Cleaning

•Pre-cleansing (dry) - with hoover, shovel, broom, – mechanically cleansing ofsurfaces from plant rests, sand, dust, adheres of earth, etc.•Wet-cleansing with high-pressure machines, but avoiding blow up effectsof dirt rests and their spreading out! Reduced presurized water stream, bigvolumes of water and use of qualified surfactants, consideringsoaking up period.

Before starting any disinfection, all areas and equipmentmust be thoroughly cleaned with pressure, with a lot ofwater and active substances (surfactants) by applying ahigh-pressure machine.The disinfecting efficiency is ensured by a thoroughcleaning, i.e. plant rests obturating surface pores andcapillaries must be removed, to uncover the occlusionsunder the coating, to a very large extent.

The rest coatings, normally hardly water soluble, are partiallydissolved by the lipophile surfactant behaviour, andbecomes water-soluble by the action of the hydrophileportion, and is, then, rinsed off together with the detergentsolution / waste water.

Concret- resp. wooden surfaces with mit poresand capillaries

organi-calcoating

Surfac-tants

Concret- resp. wooden surfaces with mit poresand capillaries

© MENNO6

Cleaning step wise

1. Mechanically pre-cleansing

Taking away removable plant rests

3. Main cleansing

Taking away non removable plant rests,

adhasions, inorganic and organical dirt loads

2. Algicidal / herbicidal treatment

Control of spread out with wash

water or personal later on.

Avoiding

backwater

in mats,

because

of flowobstruction

© MENNO7

Activity decreasing factor - restwater

After cleansing wooden surfaces resp. concrete surfaces must dry of (concrete tillgrey shining).These rough materials contend cleansing- and rainwater.Otherwise, the product-solution will be diluted by this rest-water, or flowes away.

Cleansing water and rest moisture from rainExaminations at rough, untreated wooden boxes:● Water content 8 mass-% after drying● Afterwards dipping of wood pieces into water⇒ Increasing up to 30 mass-% water content

© MENNO8

© MENNO9

Profile of MENNO disinfectants

MENNO® TER forte MENNO Florades®

Active ingredients in 100 gms: 32,5 gms Didecyl-dimethylammoniumchlorido

9,0 gms Benzoic acid

Effectiveness spectrum: bactericid, fungicid, algicid, herbicid (seeds of weed)

bactericid, fungicid, virucid (enveloped/unenveloped)

When to be Applicated(presence of plants and workers)

no gas phase no gas phase

Plant tolerance in case of contactwith the diluted product (leaves)

plant damages* exceptional good plant tolerance *bactericid

Monitoring of active substancein solutions resp. dipping bath

indicatorpaper for measuring theactive substance in %

pH-IndicatorpaperApply: pH 3,0 – 4,5

Disinfection of hard surfaces matting systems, tables, standingareas (concrete surfaces)bactericid, fungicid, algicid

ebb-and-flow benches, mattingsystems, recirculating irrigation, bactericid, fungicid, virucid

Immersion of pots and trays bactericid, fungicid, algicid bactericid, fungicid, virucid

Immersion of cutting-knives bactericid bactericid, virucid

* Please pay attention to the directions for use on the labels and in the leaflets refering to the plant tolerance

© MENNO10

Disinfectants how to evaluate?

1. broad spectrum of

effectiveness

2. applicable

in many fields

3. low phytotoxicity

4. proper human-

and eco-

toxicological values

5. reasonable price

© MENNO11

Toxicology and Ecotoxicology

� Active ingredient in the concentrate: 9 % Benzoic acid⇒⇒⇒⇒ 1 % ige ready solution: 0,09 % Benzoic acid

� Acut toxicity of product concentrate:� acute oral toxicity � „non-toxic“

Rat (24 hours or 14 days): LD50 > 2000 mg/kg

� acute dermal toxicity � „non-toxic“Rat (24 hours or 14 days): LD50 > 2000 mg/kg

� acute dermale irritation � „non-irritating“Rabbit - skin (exposition 4 hours)

� acute Eye irritation � „corrosive“ (R 41 Risk of serious damage to eyes)Rabbit - (0,1 g directly into the conjunctival sac, 24hours exposition)

� Good biological degradability

� Water dangerous classification WGK1

Benzoic acid is for instance allowded as preservative

in food stuffs with 9 g/kg, as e.g. in Sausage and Mayonnaise.

Xi-Irritant

© MENNO12

Tested spectrum of effectiveness - Fungi

Agaricus bisporus*9 Didymella bryoniae*17 Phytium ultimum*10

Alternaria alternata*10 Erysiphe cichoracearum*17 Phytophthora cinnamomi*1

Alternaria solani*10 Fusarium spp.*17 Phytophthora cryptogea*1

Alternaria sp.*1 Fusarium oxysporum f.sp.cyclaminis*1 /*12

Phytophthora infestans*10/*11

Aspergillus sp.*6 Ramularia beticola*10

Botrytis cinerea*1 Fusarium oxysporum (Stamm Elatiorbegonien) *1

Rhizoctonia solani*10

Candida albicans*13 Rhizopus sp.*6

Cercospora beticola*10 Fusarium solani var. coeruleum*1 Streptomyces scabies *1

Chalara elegans*8 Helminthosporium solani *1/*10/*11 Taphrina deformans *15

Colletotrichum coccodes*10 Mucor sp.*6

Colletotrichum sp.*1 Ophiostoma quercus*1 Thielaviopsis basicola*1

Cylindrocladium scoparium*1 Peronospora tabacina*8 Trichoderma harzianum*9

Cylindrocladium spathiphylli*1 Pythium sp.*6 Trichoderma viride*1

Dactylium dendroides*1 Phythium aphanidermatum*17 Verticillium fungicola*1/*9

Effectiveness tests

against Fungi are

prepared with

vegetative forms and

also against spores

and duration forms

*1 State Research Institute Geisenheim, Special Field: Phytomedicine, D-65366 Geisenheim, Prof. Dr. Wohanka *2 University Hamburg, Institute for applied Botany, D-2000 Hamburg 36, Dr. C. Büttner*3 Eidgenössische Forschungsanstalt für Obst-, Wein- und Gartenbau, CH-8820 Wädenswil, Schweiz*4 Institute for Plantdisease and Plant Protection, University Hannover, D-30419 Hannover, Prof. Dr. Maiß*5 Albert-Ludwigs-Universität Freiburg, Institut für Forstbotanik und Baumphysiologie, D-79085 Freiburg i. Br., Priv. Doz. Dr. C. Büttner*6 Praxisgutachten über den Einsatz ... Florades (.... Einsatz im gärtnerischen Bereich), Dr. M. Wölk, D-56204 Hillscheid*7 HUMBOLDT-UNIVERSITÄT ZU BERLIN, Institut für Gartenbauwissenschaften, Fachgebiet: Phytomedizin, Frau Prof. Dr. C. Büttner*8 Landesanstalt für Pflanzenbau Forchheim, Deutschland, Dr. N. Billenkamp*9 Horticultural Research International, Dr. H. Grogan, Wellesbourne, Warwick, England*10 Institut für Pflanzenpathologie und Pflanzenschutz der Universität Göttingen, Dr. M. Benker, D-37077 Göttingen*11 Institut PPO Wagening, Applied Plant Research BV, NL-8200 AK Lelystad, Dr. H.T.A.M. Schepers, Dr. A. Veerman*12 Institut PPO Wagening, Applied Plant Research BV, NL-1431 JV Aalsmer, Dr. A. Hazendonk, Dr. J.P. Wubben*13 Technische Mikrobiologie Dr. J. Höffler GmbH, D-22045 Hamburg*14 Institut für Pflanzenschutzmitte lprüfung, Österreichische Agentur für Gesundheit und Ernährungs., Wien, Dr. M. Keck, Dr. P. Fida*15 Dienstleistungszentrum Ländlicher Raum, Rheinhessen-Nahe-Hunsrück, A. Thomas, Dr. G. Albert*16 Bretagne BiotechnologieVégétal (BBV), E. Pajot, F-29250 St. Pol de Léon, France,*17 Crop Diversification Centre South, Alberta Agriculture, Food and Rural Development, Dr. M.W. Harding, Dr. R.J. Howard, Canada*18 Wageningen UR Glastuinbouw, (untersucht MENNO CLEAN correspond M.F.) I. Stijger, R. Hamelink, Wageningen, The Netherlands

© MENNO13

Tested spectrum of effectiveness - Bacterias and Viruses

Viruses / Viroids BacteriasArMV*2 (arabis mosaicnepovirus)

PLCV*2 (pelargonium leafcurl tombusvirus)

Acidovorax avenae ssp. cattleyae*1

Pseudomonas fluorescensmarginaeis *16

BePMV*7 (bell peppermottle virus)

PLPV*2 (pelargonium line pattern virus)

Agrobacteriumtumefaciens*1

Pseudomonas lachrymans

CarMoV*4 (carnation mottle carmovirus)

PMMoV*7 (pepper mild mottle virus)

Clavibacter michiganensisssp. michiganensis*1

Pseudomonas putida

CGMMV*18 (cucumbergreen mottle mosaic virus)

PSTVd*7 (potato spindletuber viroid)

Clavibacter michiganensisssp. sepedonicus*1

Pseudomonassolanacearum*1

CMV*4 (cucumber mosaicvirus)

PVX*4 (potato virus X) Enterococcus faecium*13 Pseudomonas syringaePVY*4 (potato virus Y)

CSVd*7 (chrysanthemum stunt viroid)

RMV*4 (ribgrass mosaic tobamovirus)

Erwinia amylovora*3/*14 Ralstonia solanacearum*1

CyMV*5 (cymbidium mosaic virus)

TBRV*2 (tomato blackringnepovirus)

Erwinia carotovora ssp. atroseptica*1

Staphylococcus aureus*13

MNSV*7 (melon necrotic spot virus)

TMV*2 (tabacco mosaic virus)

Erwinia carotovora ssp. carotovora*1/*10

Xanthomonas axonopodispv poinsettiicola*1

ORSV*5 (odontoglossumringspot virus)

ToMV*2 (tomato mosaic virus) Escherichia coli*13 Xanthomonas campestrispv. begoniae*1TSV*/ (tobacco streak virus)

PepMV*7 (pepino mosaicvirus)

TSWV*2 (tomato spottedwilt tospovirus)

Proteus mirabilis*13 Xanthomonas campestrispv. campestris*1

PFBV*2 (pelargoniumflower break virus)

ZyMV*7 (zucchini yellow mosaic virus)

Pseudomonas aeruginosa*13

Xanthomonas campestrispv. pelargonii*1

© MENNO14

0

20

40

60

80

100

10

1

0,9

0,1

Myz

elw

ach

stu

m (

%)

Wir

ksto

ffko

nzen

t rat

ion

(ai g

/l )

KontrolleCc 3aCc BBA 62126Cc BBA 70879Hs BBAHs GeisenHs H9-14cHs H9-2aHs H9-5aRs NLRs BBARs Celle

Colletotrichumcoccodes Helminthosporium

solani Rhizoctoniasolani (AG 3)

Examination of effectiveness for MENNO Florades with applicationagainst economical relevant diseases at potatos and sugar-beets

The product MENNO Florades with the active ingredientBenzoic acid was at first for floriculture registered as Special-Disinfectant. In the Potato Production it is allowed amongothers for control of the quarantine diseases Clavibactermichiganensis ssp. sepedonicus and Ralstonia solanacearumfor disinfection of warehouses, boxes as well as equipmentand machines. Because of the fungicid and bactericideffectiveness and additionally the exceptional good planttolerance of MENNO Florades in vitro-tests, trials ofdisinfection and treatments of plants- and crops as well as leafapplications on potato and sugar-beetwas carried out.

Introduction

In vitro

In vitro-tests evaluatingeffectiveness of Benzoic acid onthe Mycel- resp. Germ growth wascarried out with several potato-and sugar-beet-germs. Startingwith a concentration of 0,9 ai g/lactive ingredient (= 1% product)Benzoic acid confirms a very goodmycel- resp. growthinhibition effect(fig. 1 + 2).

Marianne BenkerInstitut für Pflanzenpathologie und Pflanzenschutz,

Grisebachstraße 6, D-37077 Göttingen

fig. 1

Control

© MENNO15

0

20

40

60

80

100

3,6 (4%)

1,8 (2%)

0,9 (1%)

0,09 (0,3%)

Myz

el-

bzw

. Kol

oni

ewac

hstu

m (%

)

Wi r

ksto

f fko

n zen

t rat

i on

(ai g

/l)

[Auf

wan

dmen

ge in

%]

Isolate

Kartoffeln

Zuckerrüben

(0,1%)

Kontrolle

Phytophthora infestansAlternaria alternataAlternaria solaniErwinia carotovora (108 cfu/ml)

Cercospora beticola

Rhizoctonia solani (AG 2-2IIIB)Ramularia beticola

Pythium ultimum

fig. 2

Control

Examination of effectiveness for MENNO Florades with applicationagainst economical relevant diseases at potatos and sugar-beets

© MENNO16

Examination of effectiveness for MENNO Florades with applicationagainst economical relevant diseases at potatos and sugar-beets

Phytotoxicity Examinating possibly Phytotoxiciyt Seed- as well as leafapplications on potatos and sugar-beets in highconcentrations was carried out. The Seedtreatment on potato tubers with MENNO Florades before planting makesno Phytotoxicity, but stimulates the growth of the potatoplants (fig. 3).

Behandlung1 2 3 4 5 6

Aufgang in Tagen

0

20

22

24

26

KontrolleKontrolle + H2OPencycuron 2x (1200 ml/t)Mancozeb 2x (4000 g/t)Benzoesäure 2%Benzoesäure 4%

Sorte: Karlena(mit Keimen <5mm)

Benzoic acid 2 %

Benzoic acid 4 %

Control

Control + H2OGrowth in days

Treatment Sort: Karlena(with sprout <5mm)

fig. 3

But: Seed- and Plant treatmentsare not registered until now andare not allowed for use!

© MENNO17

Investigations on virus transmission of Pepino mosaic virus (PepMV) – Poster 1st of 3

Pepino mosaic virus (PepMV) has been spreading fast in European tomato cultivars within the last two years (JORDA et al., 2000). PepMVwas first reported from Peru in Solanum muricatum (JONES et al., 1980) and in 1999 from tomato in Netherlands (van de VLUGT et al.,2000). Severe yield losses in tomato have been observed repeatedly. The virus is transmissible by vegetative propagation and mechanicallyfor example by contact between infected and healthy plants, or contact of healthy plants with conta-minated hands, clothes or tools.Investigations on transmission of PepMV by seeds were carried out in this study.

Investigations on virus transmission ofPepino mosaic virus (PepMV)

I. Neven, M. Bandte, C. Obermeier and C. Büttner Humboldt-University of Berlin

Institute of Horticultural Science, Dept. PhytomedicineeMail: phytomedizin@agrar.hu-berlin.de

Jones RAC, Koenig R, Lesemann DE, 1980: Pepino mosaic virus, a new potexvirus from pepino. Annals of Applied Biology 94, 61-68.

Jordá C, Lázaro A, Font I, Lacasa A, Guerrero, MM, Cano A, 2000: Nueva enfermeded en el tomate. Phytoma-España 119, 23-28.

van der Vlugt RAA, Stijer CCMM, Verhoeven J, Lesemann DE, 2000: First report of Pepino mosaic virus on tomato. Plant Disease 84, 103.

© MENNO18

Investigations on virus transmission of Pepino mosaic virus (PepMV) – Poster 2nd of 3

Fig. 3: Negative staining of a PepMV-particle (length ~ 500 nm)

50 PepMV-infected tomato plants (var. Cal-Ace and Peto) were produced by mechanical inoculation of young seedlings. Infected tomatoplants exhibited chlorosis and necrosis; discolouration of fruits was only observed on the cultivar Cal-Ace (Fig. 1). The infection was verifiedby bioassay (Fig. 2), electronmicroscopy (Fig. 3) and ELISA (antibodies obtained from Plant Research International, Wageningen,Netherlands).Tomato plants were dissected into roots, stems, leaves, flowers and fruits; tomato fruits into fruit pulb, gelantine (columnar epithelium andplacenta), seed coat, endosperm and embryo (Fig. 4). These parts were tested by ELISA. Additionally homogenized plants sections weretested by mechanical inoculation of tobacco plants.The disinfectant MENNO-Florades (Menno-Chemie GmbH, Norderstedt, Germany) was tested to decontaminate PepMV-contaminatedseeds using various concentrations and incubation times.

Fig. 1: PepMV infected tomato left: chlorosis (arrow left) and necrosis (arrow right)right: discolouration of a tomato fruit (cultivar Cal-Ace)

Fig. 2: PepMV-induced symptoms on tobacco indicator plantsleft: Nicotiana benthamianaright: Nicotiana tabacum var. Samsun

© MENNO19

Pepino mosaic virus (PepMV) is systemic in tomato plants

PepMV is detectable in single seeds by ELISA

PepMV was found in the fruit pulb as well as in the gelantine (columnar epithelium and placenta) and contaminates the seed coat,

but was not found in the endosperm and embryo

The possibility and efficiency of transmission of PepMV from contaminated seed coats to emerging seedlings needs

to be evaluated further

The infection is systemic in tomato and the virus is readilydetectable by ELISA in all parts of the plant including roots, stems,leaves and flowers. PepMV was consistently detected by ELISA infruit pulb, gelantine and seed coat (Tab. 1). However all testedendosperms and embryos were not contaminated with thepathogen. Transmission of PepMV by contaminated tomato seeds isevaluated in current studies by testing seedlings raised from theseseeds by ELISA.Additionally it was tested, if PepMV-contaminated seeds can beeffec-tively disinfected using the disinfectant MENNO-Florades.Disinfection of seeds was successful using 3% of MENNO-Floradesfor 2 hours or 4%of the disinfectant for 30 minutes. The germinationrate is not affected by these treatments.

Investigations on virus transmission of Pepino mosaic virus (PepMV) – Poster 3rd of 3

+: detection of PepMV-: no detection of PepMV

gelantine

seed coatembryo

seed coatFig. 4: Parts of tomato seeds tested - gelantine, seed coat, endosperm, embryo -

Tab. 1: Detection of PepMV in seeds and parts of seeds of tomato(Lycopersicon esculentum L.) by ELISA (n=50)

endosperm

© MENNO20

Effectiveness against Pepino mosaic virus (PepMV)

HUMBOLDT-UNIVERSITÄT ZU BERLIN, FACULTY OF AGRICULTURE AND HORTICULTUREInstitute for Horticultural Sciences, Special Field PhytomedicineDirection: Prof. Dr. C. Büttner, Expertise dated from 02.05.00Resuming assessmentFlorades is perfectly suitable for disinfecting plants infested with Pepino mosaic virus (PepMV); this applies to knife disinfection (short exposure time) and to standing area disinfection (low concentration). For knife disinfection, a PepMV virus inactivation could be observed after applying a 3 % means concentration and a 30 seconds exposure time. To disinfect standing areas, we recommend a lower means concentration with a longer exposure time. The 1,5 % concentration leads to a virus inactivation after a 16 hours time, and the 2 % concentration after 8 hours. The other combinations of concentration and exposure time can be taken from table 4 containing the test results. According to Jones et al. (1980), PepMV has been known from solanum muricatum (coastal region of Peru) since 1980, and has been discovered in European tomatoe cultures in 1999, for the first time (Lesemann et al., 2000). The virus spreads quickly in the crops. At present, it is being discussed, if it must be classified as a quarantine pathogen. This shows the high importance of prophylactic measures to prevent the spreading of this virus. One of the steps to combat the virus prophylactically, consists in disinfecting cutting tools and standing areas. According to the PepMV tests, Menno Florades proves to have a prompt effect showing, at the same time, a highplant tolerance.

First symptoms of infection - course of disease - consequence

© MENNO21

Transmission by recirculation nutrient solution

Pic. 1: Ebb-flow-system: set up of Phalaenopsis

and herbaceous indicator plants

0

20

40

60

80

100

0 14 28 35 70 91

days after setting up the plants

viru

s in

fect

ed p

lant

s (%

)

healthy

virus infected

Fig. 4: Odontoglossum ringspot virus (ORSV): disease development by nutrient solution of the ebb-flow system detection of ORSV in leaves of Phalaenopsis

020406080

100

0 14 28 63days a fter se tting up the plants

viru

s in

fect

ed p

lant

s (%

)

hea lthy

virus infec ted

Fig. 5: Cymbidium mosaic virus (CyMV): disease development by nutrient solution of the ebb-flow system detection of CYMV in leaves of Phalaenopsis

© MENNO22

disinfection of ebb/flow benches

virus rinsing

with tap water no rinsing

Pelargonium leaf curl virus (PLCV) not tested 1 %, 10 seconds

Pelargonium line pattern virus (PLPV) not tested 2 %, 1 minute

Arabis mosaic virus (ArMV) not tested 2 %, 5 minutes

Tomato black ring virus (TBRV) not tested 4 %, 1 minute or 2 %, 5 minutes

Tobacco mosaic virus (TMV) 4 %, 16 hours only partially effective

Pelargonium flower break virus (PFBV) 1 %, 1 hour 1 %, 16 hours

Tomato spotted wilt virus (TSWV) 1 %, 1 hour 1 %, 4 hours

Disinfection of standing areas - efficiency against viruses -

concentrations and exposure timesfor complete inactivation of various viruses

TMV

reference: expertise Dr. C. Büttner

Consequence:

Application

after intensive

cleaning

© MENNO23

Disinfecting knives with MENNO Florades®

Tobacco mosaic virus -

TMV

reference: expertise Dr. M. Wölk

treatment concentration- 1 % 2 % 4 %

MMEENNNNOO FFlloorraaddeess -- 22 33 00

M&ENNO TER forte - 10 10 10

Sanosil - 10 10 9

flaming 0 - - -

control (water) 10 - - -

diseased tobacco plants (max. 10)

method:

1. dipping of contaminated knives for 3 minutes2. cutting of healthy, vigorous plants (tobacco)3. disease assessment after 3 weeks or ELISA

© MENNO24

The right way to test disinfectants BBA-guideline 16-4

testing the gaseous phase

The disinfectant is placed on a dish under a glass. Also a very sensitive plant culture is placed beside, to show, if theproduct has a gas phase.

CylindrocladiumFor testing practicale conditionsas good as possible, nylon germcarriers are contaminated withfungi. Afterwards they store solong, until all forms have built, including spores and durationforms. If completed it will bedisinfected. So the product has to show all it‘s power of effectiveness.

test organism (standard)

►Xanthomonas campestris pv. Pelargonii

► Phytophthora spp.

► Thielaviopsis basicola

► Cylindrocladium spp.

► Fusarium oxysporum

lab test (with and without dirt load)

► bactericidal activity (suspension test)

► fungicidal activity (carrier test)

in practice (with susceptible test plants)

► disinfection of knives

► dipping pots, trays etc.

► application on standing grounds (e.g. capillary mats)

► gaseous phase (lettuce, Begonia semperflorens etc.)

© MENNO25

Fungicid effectiveness of MENNO Florades - exposure times

Survival of test germs (number germ carriers with growth of fungi after treatment)

conc. Phytophthora Colletotrichum Fusarium oxysporum Fusarium solani Cylindrocladium

[% ] cinnamomi sp. f.sp. cyclaminis var. coeruleum scoparium

without peat load1 h 4 h 16 h 1 h 4 h 16 h 1 h 4 h 16 h 1 h 4 h 16 h 1 h 4 h 16 h

0,50 0 0 0 0 0 0 not tested not tested 2 1 01,00 0 0 0 0 0 0 3 2 0 0 0 0 2 0 02,00 0 0 0 0 0 0 0 0 0 0 0 0 2 0 0

with peat load1 h 4 h 16 h 1 h 4 h 16 h 1 h 4 h 16 h 1 h 4 h 16 h 1 h 4 h 16 h

0,50 0 0 0 1 0 0 not tested not tested 2 1 01,00 0 0 0 0 0 0 3 2 0 0 0 0 0 0 02,00 0 0 0 0 0 0 0 0 0 0 0 0 2 0 0

0 = complete effectiveness (n = 3)

Results in vitro – laboratorial trials

Recommendation: 1 % and 16 hours, respective 2 % and 4 hAt this concentration and exposure time all fungi (includ ing the mostresistent fungi) and their duration forms are killed, al so with load ofdirtyness.

© MENNO26

Bactericid effectiveness of MENNO Florades - exposure times

conc. Ralstonia Pseudomonas Claviabacter mich. Claviabacter mich. Xanthomonas camp.

[%] solanacearum solanacearum ssp. sepedonicus ssp. Michiganensis pv. pelargonii

with germ carriers

metal surface without peat load

15 min 30 min 60 min 5 min 15 min 30 min 15 min 30 min 60 min 5 min 15 min 30 min 1 min 2 min 3 min

0,25 n.t. n.t. n.t. 0 0 0 0 0 0 0 0 0 n.t. n.t. n.t.0,50 n.t. n.t. n.t. 0 0 0 0 0 0 0 0 0 0 0 01,00 0 0 0 0 0 0 0 0 0 0 0 0 0 0 02,00 0 0 0 n.t. n.t. n.t. n.t. n.t. n.t. n.t. n.t. n.t. 0 0 0

with peat load

15 min 30 min 60 min 5 min 15 min 30 min 15 min 30 min 60 min 5 min 15 min 30 min 1 min 2 min 3 min

0,25 n.t. n.t. n.t. 0 0 0 0 0 0 0 0 0 n.t. n.t. n.t.0,50 n.t. n.t. n.t. 0 0 0 0 0 0 0 0 0 0 0 01,00 n.t. n.t. n.t. 0 0 0 0 0 0 0 0 0 0 0 02,00 n.t. n.t. n.t. n.t. n.t. n.t. n.t. n.t. n.t. n.t. n.t. n.t. 0 0 0

Survival of test germs (number germ carriers with growth of bacteria after treatment)

Ralstonia = Pseudomonas

n.t. = not tested

© MENNO27

Disinfecting Knives with MENNO Florades®

- Xanthomonas campestris pv. pelargonii -

treatmentXcp-positive

(n=100)trial 1 trial 2

control "healthy" (untreated, not contaminated) 0 0untreated (contaminated) 32 92ethanol (70%; contaminated) 0 0MENNO Florades (1,5%; contaminated) 0 1MENNO Florades (3 %; contaminated) 0 1

method:

1. dipping of contaminated knives for 2 minutes2. cutting of healthy, vigorous plants (geraniums)3. samples tested on special agar plates after 5 or 7 weeks

© MENNO28

0

10

20

30

40

50

60

70

80

90

100

day 0 day 1 day 3 day 6 day 9

1 % M-Florades2 % M-Florades3 % M-Florades

efficiency (%)

days after treatment

Spraying Plants with MENNO Florades®

Xanthomonas campestris pv. begoniae on the plant surface

© MENNO29

efficiency (%)

0

10

20

30

40

50

60

70

80

90

100

1% 2% 3% 1 % prophylactic

V9801PEL01

V9801PEL02

V9801PEL03

concentration

3 experiments

Spraying Plants with MENNO Florades®

Xanthomonas campestris pv. pelargonii on the plant surface

© MENNO30

Begonia leaf and stalk bacteriosis destruction(Xanthomonas campestris pv. begoniae) on Begonia

0

10

20

30

40

50

60

70

1. week 2. week 3. week 4. week 5. week

*MENNO Florades 1 %

C.(copper sulphate) 0,03 %

Kind of infection: InoculationNumber of repetitions: 4Lot size: 10 plantsVariety: Begonia-Elatior

„Hetja dark“Stages: plant in blossom*Slight marginal blossom

necroses were observed.The treatment can only slowdown the bacterial spreading,but not stop it.

Classification: Recording the percentage of infected herbage (after infection)Grade of effectiveness

Time table of judge after weekly treatment (after 8 applications)

© MENNO31

Phytotoxicity of MENNO Florades®

after spray application on geraniums

3 %

2 %

© MENNO32

Phytotoxicity of MENNO Florades®

after spray application on begonias

1 %

3 %

1 %

3 %

© MENNO33

2,0

2,5

3,0

3,5

4,0

4,5

5,0

5,8 °dKH 11,7 °dKH 23,4 °dKH

pH

-val

ue

carbonate hardness

1 %

2 %

3 %

MENNO Florades®

monitoring of pH-value in immersion solutions!

helpful tip: 1. Renewal of dips unless they are clearly transparent !

Photo right above: Dip solution becoming dirty - throw away !2. Remove deposit of peat, soil, fertil etc. from the ground of the box, before recharging !3. Use rain water for dip solution / immersion-bath - lowering the product consumption !

© MENNO34

Personal hygiene – Hand disinfection

back of the hand

insideof

thehand

considere insufficient:

very often

less often

© MENNO35

Disinfecting Hands with MENNO Florades®

- Xanthomonas campestris pv. pelargonii -

0

1-5

6-50

>50

0

10

20

30

40

50

60

70

80

90

100

control ethanol (70 %) MENNOFlorades (1%)

MENNOFlorades (2%)

MENNOFlorades (3%)

MFD (3%) inethanol

> 50

6-50

1-5

0

severity of contamination per finger

(frequency classes in %)

exposure time

5 seconds

© MENNO36

Hand disinfection with MENNO Florades

against cucumber mosaic virus

0

5

10

15

20

Gesundkontrolle Krankkontrolle MENNO Florades (3%)

Anteil CMV-positiver Pflanzen (%)

5 ml, 5 seconds

control „healthy“ control „untreated“

Part of CMV-positve plants [%]

Results: Within serological tests of the indicationplants, CMV was only in 18 % of the plants (control “untreated”) detectable. In plants of the control "healthy" and those of the disinfectant treatment the virus could not be detected in any case.

� To study the suitability of MENNO Florades (3% )for hand disinfection under real conditions as an example cucumber mosaic virus (CMV) was tested at Surfinia hybrids.

� The contamination was done by rubbing virus containing leaf between thumb and index finger.Thereafter, the contaminated hands were disinfected by a standard procedure (5 seconds „handwashing“ with 5 ml disinfectant solution). Then the thumb surface of a hand 4 leaves of a healthy surfinia … was rubbed off. (40 plants per test group).

© MENNO37

Instant test for determinationof concentration of MENNO Florades

+ MENNO

Florades

1.) Titration determines deficit

2.) Adding of product, till solution is

adjusted correctly.

10 ml

sample + 5 drops reagent 1(Phenolphthalein 0,1 % solution in Ethanol)

sample

sample

+ reagent 2 (Sodium hydroxide 0,30 mol/L)

⇒⇒⇒⇒consumption of ml equals existing

concentration as given in the table.

A distinct change of color into pink

resp. red becomes visible.

„used“ solution can be

added with fresh product

to sharpen the

solution / dipfor renewal use

Tray washing machinewith recirculation of

solution of disinfetant

washing- / immersionsolution

© MENNO38

Instant test for determinationof concentration of MENNO Florades

Method:Analytical determination ofconcentrations of MENNO Florades insolutions by titration with Sodiumhydroxide, using Phenolphthalein(0,1 % solution in Ethanol) as indicator.

Procedure:1.) Rinse plastic-glass twice with the test

solution and empty it.2.) Rinse 10 ml plastic syringe twice with

the test solution and empty it.3.) Fill 10 ml plastic syringe with test

solution, avoiding bubbles, up to themark and fill this volume into theplastic-glass.

4.) Into this sample of solution, give 5droplets of reagent 1 from plastic-bottlewith drop-nozzle. Mix by stirring glass.

5.) Fill 2 ml plastic-syringe with Reagent 2up to 2 ml mark, avoid bubbles.Add Reagent 2 drop by drop whilestirring, until permanent change of colorto a reddish pink occurs.

6.) Read consumption of Reagent 2 fromscale on the syringe.

consumptionreagent 2

(Natronlauge 0,30 mol/l)

in ml

concentration MENNO

Florades solution in %

0,1 0,25

0,2 0,50

0,3 0,75

0,4 1,00

0,5 1,25

0,6 1,50

0,7 1,75

0,8 2,00

0,9 2,25

1,0 2,50

consumption reagent 2

(Natronlauge 0,30 mol/l)

in ml

concentration MENNO

Florades solution in %

1,1 2,75

1,2 3,00

1,3 3,25

1,4 3,50

1,5 3,75

1,6 4,00

1,7 4,25

1,8 4,50

1,9 4,75

2,0 5,00

Remarks: store Reagents in a cold place (15-25 °C), pay attention to datesof expiry. After each use, close bottles of Reagents immediately.Reagents: Reagent 1: Phenolphthalein 0,1 % Lösung in Ethanol

Reagent 2: Sodium hydroxid 0,30 mol/lPrecautions: Avoid contact of Reagents with skin or eys.

© MENNO39

0

20

40

60

80

100

3,0 3,4 3,8 4,2 4,6 5,0 5,4

conc. [%]

pH

pKa

acc. HUGO & RUSSELL; 1992

Biocidal Activity of Organic Acids Impact of pH

decreasingbiocidalactivity

not dissociated (AH)

COOH

dissociated (A-

)

COO-

© MENNO40

Application technique

0,5 %Lilac

4 %Blue

3 %Orange

2 %Dark brown

1 %Aqua

ConcentrationDosage control jet

One 3 l container full of product concentrate is sufficient for 300 l ready mixed solution, enough for a surface of750 m² by using 0,4 l/m² (ready mixed solution, i. e. wet surface).

The colored dosing jets inserted into the suction tube allow precise control of concentration levels.

MENNO Florades

With the MENNO® disinfectant applicator► for connection on a hose with a snap connector► for automaticly exact dosage of disinfection-concentrate► foam - as a spray control – checking surfaces have been treated► to guarantee longer contact and reaction times

also for surfaces like plastic (e.g. boxes, trays, pots, ...), as far as concrete, metall, glass, etc.

© MENNO41

Personal hygiene – the right protective clothingfor keeping the surface disinfected after treatment

© MENNO42

© MENNO43

Foam with big air bubbles, suitable for horicontal surfaces

© MENNO44

Disinfectant foam application with low pressure up to 6 bar

aqueous solution for

absorbent surfaces Foam with

smooth surfaces

© MENNO45

„Creamy“ foam with very small air bubbles, suitable for smooth vertical surfaces and horicontal undersites

© MENNO46

Foam application - to be documened photographically digital

10.09.2010 13:30 Uhr

© MENNO47

hygitrix®

Measurement, control and documentation of

process relevant values for hygiene

© MENNO48

How to realize exceptional cleaning of plastic boxes?

After a steak was fried in a pan, it is with hot water, detergent and a sponge cleaned mechanically.What must be done to achieve an adequate result in boxes?

-Warm / hot water – recirculating the waterto reduce waste, cost of energy /product

- Presurized water- Right nozzle technique- Flushing out water- Avoiding dirt water restsentering in the disinfection unit

- Efficient Detergent:Detergents from the scope of foodindustry are made for the removal ofanimal proteins and fatty dirt loads!But, such dirt loads are not present in horticultural production! => VENNO Hortisept Clean

© MENNO49

hygitrix® for right dosage, monitoring, and documentation

For electronic measurement, automaticdosing and data recording of hygienerelevant prozess parameter formonitoring and documentation ofcleansing and disinfection processes. Concentration of product (measuring theelectronical conductivity), temperatur ofwater and application rate (as volumestream per unit of time) are measuredelectronically, recorded as data file andprovided graphically via software tool forsaving on the computer.

© MENNO50

hygitrix® facilities

© MENNO51

applications

© MENNO52

Plastic boxes – stackable, for folding, … large variety

© MENNO53

Dirt load: dust adherences ? Gradiations of white color

© MENNO54

Create box / Create box / Create box / Create box / plasticplasticplasticplastic material material material material totototo bebebebe cleanedcleanedcleanedcleaned

After After After After cleaningcleaningcleaningcleaning withwithwithwithsubsubsubsub----optimal optimal optimal optimal conditionsconditionsconditionsconditions

After After After After cleaningcleaningcleaningcleaning withwithwithwithoptimizedoptimizedoptimizedoptimized conditionsconditionsconditionsconditions

© MENNO55

Create box washing machine:- pre-washing - washing with heated water and

recirculation as far as resharpening of process water- disinfection with foam for optimizing the contact time

© MENNO56

STOP

At first disinfection,

then production !

„Plant-Hygiene“ resp. disinfection

is the essential part of the

Plant Protection Strategy and have to

be done first of all and consequently!

Thank you very much

for your kind attention