Tau-fluvalinate content of Apistan® strips

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Tau-fluvalinate content of Apistan® strips

Mark F. Feldlaufer

To cite this version:

Original

article

Tau-fluvalinate

content

of

_Apistan®

_strips

Mark F. Feldlaufer

USDA,

Agricultural

Research _Service, Bee Research

_{Laboratory, Bldg}

476, BARC-East, Beltsville, MD 20705, USA

(Received

1 July

1998;

accepted

12 November ₁₉₉₈₎

Abstract - The tau-fluvalinate content of 13 lot numbers of

_{Apistan® strips}

was determined

_{by high}

performance liquid chromatography

(HPLC). The average percent tau-fluvalinate in

_strips

from each lot number _ranged from 9.95 to 10.64 %, which falls within the range

specified

in the Confidential Statement of Formula _(CSF) on file with the Environmental Protection

_Agency.

It is not

_likely

that the reduced

_efficacy

in

_parasitic

mite (Varroa jacobsoni) control

_reported

in certain areas of the US

is due to the tau-fluvalinate content of

_{Apistan® strips.}

© _{Inra/DIB/AGIB/Elsevier,} Paris tau-fluvalinate content /

_Apistan®

/

_{liquid chromatography}

/

_Apistan®

resistance

1. INTRODUCTION

Apistan®

(Wellmark International,

Bensenville, IL)

is the trade name for

plas-tic

_{strips impregnated}

with the

_synthetic

pyrethroid

tau-fluvalinate

(10

%

_w/w)

that are used to control the

_honey

bee

_parasitic

mite

_{Varroa jacobsoni}

Oudemans. Tau-flu-valinate is considered

_relatively

non-toxic to

_honey

_bees,

and

_Apistan®

_strips

are the

sole

_legal

treatment available to

beekeep-ers in the US for the control of V.

jacob-soni.

_During

the summer of

_1997,

anecdotal

reports surfaced that

_beekeepers

in certain

geographic

areas of the US were

experi-encing

reduced mite control when

_using

Apistan®

[8].

While the

_possibility

of mite resistance to tau-fluvalinate was

_being

inves-tigated,

concerns were raised about the

pos-sibility

of formulation and/or release rate

problems

associated with

_Apistan®

_strips.

In this

_study,

we determined the tau-flu-valinate content of

_Apistan®

_strips

from a

variety

of years and lot numbers in an effort to

_explain

the reduced

_efficacy

of

_Apistan®

strips

in certain situations.

E-mail:

_{mfeldlau@asrr.arsusda.gov}

Disclaimer: Mention of trade names or commercial

_products

in this article is

_solely

for the purpose of

providing specific

information and does not

_imply

recommendation or endorsement

_by

2. MATERIALS AND METHODS 2.1.

_Strips

Apistan®

strips

for this

_study

were obtained from several sources: _i)

_existing

Bee Research

Laboratory stocks, which were _{used both}

exper-imentally

and for routine

_{(twice-yearly)}

control of V.

_jacobsoni;

ii) from

_unopened

_boxes

sup-plied by

the manufacturer _{(Wellmark Intl.,}

Bensenville, IL); and iii) from several

_beekeepers

experiencing

control

_problems.

In addition, we

obtained and

_analyzed

a number of ’Section 18

emergency

strips’

that were used when V.

jacob-soni was first encountered in the US. _These

par-ticular

_strips

were formulated with a

_phthalate

plasticizer

not

_currently

used. All

_strips

exam-ined in this

_study

were unused.

2.2. Extraction

_procedure

Three

_Apistan®

_strips

from each lot number

were taken for

_analysis.

Three disks (∼4 mm in

diameter) were

_punched

from different areas

(top,

middle, bottom) of each

_{strip, weighed,}

and

placed

in standard scintillation vials to which 10 mL

_methylene

chloride was _{added. After}

cap-ping, samples

were shaken on an orbital shaker

(Bellco; Vineland, NJ) for 15 min and

subse-quently

left to stand at room _temperature for a

minimum of 72 h. Prior to

_analyses,

all

_samples

were re-shaken as above. Previous tests indicated this

_protocol

was sufficient in

_extracting

all of the tau-fluvalinate in the disks.

2.3.

_{High performance}

liquid

chromatography (HPLC)

A constaMetric® 4100 solvent

_delivery

_system

(Thermo

Separation

Products;

Piscataway,

NJ)

was used in

_conjunction

with a

_Rheodyne

7125

injector

(5

μL sample loop)

and a Waters Model 481 UV detector.

_Samples

were

_{analyzed by}

injecting

30 _{μL (six}

_loop

volumes) of the

methy-lene chloride extract onto a

_C

₈

column ₍₁₅₀ mm × 4.6 mm; 5 μm

particle

size; YMC _Inc.,

Wilm-ington,

NC)

eluting

90 % acetonitrile-water at

I mL/min. The eluant was monitored at 254 nm

and column _temperature was held constant at 33 °C. Under these conditions, tau-fluvalinate eluted at

_{approximately}

4.5 min. Tau-fluvalinate

in each

_sample

was

_quantified

with a Shimadzu CR3A

_integrator

(Columbia, MD),

by comparing

the

_peak

area in each

_sample

with the

_peak

area

obtained from a tau-fluvalinate stock solution. Percent tau-fluvalinate was calculated

_by

divid-ing

the amount of tau-fluvalinate

_by

the

_weight

of the individual disks. All

_analyses

were

_performed

in

_triplicate.

All solvents for extraction and

analysis

were from Burdick & Jackson _(Baxter

Healthcare

_Corp.,

McGaw Park, IL). The tau-fluvalinate standard was

_{supplied by}

Wellmark Intl.

3. RESULTS

Apistan®

strips

manufactured _{in years} 1994

_through

1997 were included in this

study,

as well as ’Section 18

_emergency’

strips

(approximately

10 years

old).

The per-cent tau-fluvalinate in each

_{strip analyzed}

is

_given

in table I. The _{average percent} tau-fluvalinate in each lot number

_ranged

from 9.95 %

_(±

0.06

_%)

in

_strips

obtained from a

_{Pennsylvania beekeeper}

to 10.64 %

(±

0.09

_%)

in

_strips

obtained from the

man-ufacturer. All individual

_{strips analyzed}

con-tained the amount of tau-fluvalinate

speci-fied in the Confidential Statement of Formula

_(CSF)

on file with the Environ-mental Protection

_Agency.

4. DISCUSSION

Analysis

for tau-fluvalinate residues in

honey

and other bee

_products

has been

pre-viously

described

_{[1, 6, 9 10].}

Most

_analyses

utilized gas

chromatography

(GC)

with an

electron _capture detector

_(ECD)

_owing

to its

_greater

_sensitivity

and

_selectivity

_[7].

While GC-ECD has also been used to deter-mine the tau-fluvalinate content of

Apis-tan®

_strips

[3],

we chose

_{high performance}

liquid

chromatography

(HPLC)

for our

anal-yses since accuracy, not

_sensitivity,

was our

primary

concern.

_Using

a

_{reversed-phase}

HPLC

_protocol

modified from

_[1],

we were

able to

_{sample directly}

from the extraction

vial,

thereby

eliminating

the need for

mul-tiple

dilutions and

transfers,

potential

sources

Since all of the

_Apistan®

_{strips analyzed}

nominally

contained the advertised amount of tau-fluvalinate

_{(10 %; w/w),}

it is

_unlikely

that reduced

_efficacy

with

_regard

to

V. jacobsoni

control can be attributed to the

tau-fluvalinate content of these

_strips.

Rather,

a recent _report

_[2]

_suggests that

cer-tain

_populations

of V.

_jacobsoni

in the US

are

_exhibiting

resistance to

tau-fluvalinate,

a condition that has been

_previously

docu-mented in

_Europe

[4,

5,

11].

ACKNOWLEDGEMENTS

We thank L. Cutts _{(Gainesville, FL),} C. Randall (Umatilla, FL), G. & C. Tschida (Lake

Wales, FL), D.

_VanGundy

(Dallas, TX) and K.

_Young

_(Cogan

Station, PA), for

_supplying

Apistan®

strips.

The technical

_expertise

of K.

Wilzer, Jr is

_{greatly appreciated.}

We thank D.

Liewehr and J.S. Pettis for

helpful

discussion.

Résumé - Teneur en tau-fluvalinate des

lanières

_{d’Apistan®.}

On a déterminé la teneur en tau-fluvalinate des lanières

d’Apis-tan® afin

_{d’essayer d’expliquer}

la baisse d’efficacité du

_produit

contre

_Varroa

jacob-soni dans certaines

_régions

des

États-Unis.

Les lanières

_{d’Apistan®}

utilisées

_provenaient

de diverses sources, années et lots :

i)

stocks

présents

au Bee Research

_{Laboratory, qui}

étaient utilisés à des fins

_{expérimentales}

et

en routine deux _{fois par} an contre V.

jacob-soni ;

ii)

boîtes non ouvertes fournies _par le fabricant

_{(Wellmark Intl., Dallas, TX) ;}

et

iii)

lanières de divers

_apiculteurs

_ayant

ren-contré des

_problèmes

de traitement. Toutes les lanières étaient neuves. Trois

_disques

de 4 mm de diamètre environ ont été

décou-pés

dans le

_haut,

le milieu et le bas des

ajouté

10 mL de chlorure de

_méthylène.

Après

obturation des

_flacons,

les échan-tillons ont été

_placés

sur un

_agitateur

orbital

pendant

15 min

_puis

laissés à

_température

ambiante durant 72 h au minimum. Tous les échantillons ont été de nouveau

_agités

comme

_indiqué

_ci-dessus,

_{puis analysés}

en

chromatographie liquide

à haute

perfor-mance

_(HPLC)

_par

_injection

de 30

_μL

(six

fois le volume de la

_boucle)

de l’extrait de chlorure de

_méthylène

sur une colonne

_C

₈

(150

mm ×

4,6

mm; taille des

particules

5

_μm)

_puis

élué avec un

_mélange

acétoni-trile-eau à 90-10

_(ou

bien : acétonitrile à 90 %

(v/v)

dans

l’eau)

à la vitesse de 1 mL/min. L’élution a été suivie à 254 nm et la

tem-pérature

de la colonne maintenue constante à 33 °C. Dans ces

conditions,

le tau-fluva-linate est élué en 4,5 min environ. La quan-tité de tau-fluvalinate dans

_chaque

échan-tillon a été mesurée avec un

_intégrateur

Shimadzu

_CR3A,

_par

_comparaison

de la surface du

_pic

de

_chaque

échantillon avec

la surface du

_pic

obtenu à

_partir

d’une solu-tion de tau-fluvalinate _{standard. Le}

pour-centage de tau-fluvalinate a été calculé en

divisant la

_quantité

de tau-fluvalinate par la

masse de

_chaque

_disque.

Toutes les

_analyses

ont été

_répétées

trois fois.

Le taux _moyen de tau-fluvalinate a varié de

9,95

à

10,64 %,

ce

_qui

entre dans _{la gamme} des valeurs mentionnées dans le

Confiden-tial Statement

_{of Formula}

_(CSF)

de

_l’Agence

de Protection de l’Environnement des

États-Unis. L’efficacité réduite de

_{l’Apistan®}

ren-contrée dans certaines

_régions

des

États-Unis dans la lutte contre V.

_jacobsoni

n’est vraisemblablement _{pas due} à la teneur en

tau-fluvalinate des lanières

_{d’Apistan®.}

©

Inra/DIB/AGIB/Elsevier,

Paris

Varroa jacobsoni

/ résistance /

_Apistan®

/ tau-fluvalinate / HPLC

Zusammenfassung -

Gehalt von Tau-Flu-valinat in

_Apistan®

Streifen. Für eine

Mög-liche

_Erklärung

der reduzierten Wirksam-keit von

_Apistan®

Streifen bei der Varroa

jacobsoni

Kontrolle _{in bestimmten}

geogra-phischen

Bezirken der

_Vereinigten

Staaten wurde der Gehalt von Tau-Fluvalinat in

Api-stan® Streifen bestimmt. Die

_Apistan®

Strei-fen stammten aus verschiedenen Jahren und aus einer

großen

Zahl verschiedener

Quel-len:

_i)

aus den vorhandenen

_Lagern

der Ver-suchslaboratorien für

Bienen,

die sowohl für Versuche als auch für Routinekontrol-len von V.

_jacobsoni

_genutzt

wurden,

ii)

aus

ungeöffneten

Paketen direkt aus der Fabrik

(Wellmark Intl., Dallas, TX);

und

iii)

von verschiedenen

Imkern,

die

_{Schwierigkeiten}

mit der

_{Varroabehandlung}

hatten. Alle Strei-fen waren noch unbenutzt. Drei Scheiben

(d =

4

_mm)

wurden aus verschiedenen Stel-len

_{(oben, Mitte, unten)}

der einzelnen

Api-stan® Streifen

_{ausgestanzt ,}

_{gewogen und} in Standard Scintillations Gefässe

überführt,

in die 10 mL

_{Methylenchlorid zugefügt}

wur-den. Nach dem

_{Verschließen}

wurden die Proben für 15 min in einem Orbitalschüttler

geschüttelt

und anschließend für mindestens 72 h bei

_{Raumtemperatur}

stehen

_gelassen.

Vor der

_Analyse

wurden alle Proben noch einmal wie oben

_{geschüttelt.}

Die Proben wurden in einem

Hochdruck-Flüssigchro-matographen

(HPLC)

analysiert.

Dazu

wur-den 30 mL

_(das

Volumen von 6

Schleifen)

des

_{Methylenchloridextraktes}

auf eine

_C

₈

Säule

(150

mm ×

_4,6

mm; 5 _μm

_{Korngröße)}

injiziert

und mit 90 % Acetonitrilwasser mit 1 mL/min eluiert. Das Eluat wurden bei 254 nm

_{kontrolliert,}

die

_{Säulentemperatur}

wurde konstant bei 33 °C

_gehalten.

Unter diesen

_Bedingungen

wurde

_jede

Probe mit einem Shimadzu CR3A

_Integrator

quanti-fiziert,

indem man die Peakfläche

_jeder

Probe mit der Peakfläche einer Tau-Fluva-linat

_{Stammlösung verglich.}

Der Anteil Tau-Fluvalinat wurde berechnet durch die Divi-sion der Tau-Fluvalinat

_Menge

durch das Gewicht der einzelnen Scheiben. Alle

Ana-lysen

wurden 3 mal wiederholt.

Im Durchschnitt

_lag

der Prozentsatz von

Tau-Fluvalinat zwischen

9,95

% und

10,64

%

Umwelt-schutzamt dokumentiert ist. Es ist

unwahr-scheinlich,

daß

die

_geringere

Wirksamkeit bei der

_{parasitischen}

Milbe

_(V.

_jacobsoni)

in verschiedenen Bezirken der USA vom Gehalt des Tau-Fluvalinat in den

_Apistan®

Streifen

_abhängig

ist. © Inra/DIB/AGIB/

Elsevier,

Paris

Tau-Fluvalinat Gehalt /

_Apistan®

/ HPLC /

Apistan®

Resistenz / Varroa

_jacobsoni

/ USA

REFERENCES

[1] Atienza J., Jimenez J.J., Bernal J.L., Martin M.T.,

Supercritical fluid extraction of fluvalinate residues in _honey. Determination _{by high} per-formance _{liquid chromatography, J. Chromatogr.} A 665 (1993) 95-99.

[2] Baxter J., Eischen F., Pettis J., Wilson W.T.,

Shimanuki H., Detection of fluvalinate-resistant varroa mites in U.S. _honey bees, Am. Bee J. 138 (1998) 291.

[3] Cabras P., Floris I., Garau V.L., Melis M., Prota R.,

Fluvalinate content of _{Apistan® strips during} treatment and _efficacy in colonies _containing sealed worker brood, _Apidologie 28 (1997) 91-96.

[4] Colin M.E., Vandame R., Jourdan P., Di Pasquale S., Fluvalinate resistance of Varroa jacobsoni Oudemans _{(Acari: Varroidae)} in

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[7] Fitch W.L., Helisten C.C., Visser I.M.,

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[8] Fore J., T. H., Florida _{beekeepers question} Apis-tan _efficacy, The _Speedy Bee 26 ₍₁₉₉₇₎ 3. [9] Kubik _M., Nowacki J., Marcinkowski J.,

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[10] Kubik M., Nowacki J., Marcinkowski J., Pidek A.,

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