Secretory tissues

0

SECRETORY

m

-

TISSUES STORAGE

AND

CHEMICAL

ESSENTIAL

OILS

OF

PLANTS

HIGHER

KATERINA &

IN

VARIATION

SECRETORY AND

TISSUES

THEIR

SVOBODA’,

TONV

OF

BIOACTIVITY

JANICE

HAMPSONI

HUNTER*

’ De@-tment

ofPlantBiology, Scottish Agricultural College,Auchinwu&,

Ayr; Scotland i&S 5HW 2 Biomnthematicsand StatisticsScotland, The Universityof Edinburgh,James Clerk Maxwell Building The King’s Buildcng, Edinburgh, ScotlandEH9 3JZ

E

ssential

oils are found in a widl

The

annual,

takes

herbaceous

biennial

plants,

deciduous

and

of

composition

evergreen

species,

by

and

trees

quantity

ant

and are influence<

the

environment

ontogenic

trends

in relation

particular

environment.

essential

o

of the oils vary betwee

individual both

or perennia

shrubs

Parameters

am to tha

In

studyin!

oils it is useful to know when

in the plant biosynthesis

and storage o

the oil takes place, and the subsequcn chemical

changes

the

oils

undergo

within these structures. Several depth,

systematically

mostly

interest.

Our article

description

were

and

of

ir

commercia

presents

the basil

of the sile of oil synthesi some

storage,

ontogenic an

those

changes

outline

of

determining

data

regardin!

in composition,

a useful

biological

disintegrate

cells

most

simple

the form

containing

of a single

secretion-

where

it is only

that distinguish

adjacent

However, the

structure

cell

actual contents the

secretory

non-secretory

other

cells,

or

lining.

have

tissues:

in

the

(Laurus nob&),

a

ant

bioassay

fo

activity.

thick

leaf

types of plant

parenchyma

(Androfiogon

lemongrass

than

This cell type is

found in many different

of

spp.),

citronella

bay

(Cymbopogon

Cymbopogon winterianus)

nardus/

and

(Pogostemon patchouli), in the

patchouli seed

cells.

it may also be larger

cuticularized

the

it from

coat

of

(Elettaria

cardamom

cardomomurrq),in the rhizome

species and varieties

investigated

and

Secretory

range of plant species, including

(Zingiber (Curcuma

of ginger

and

offiicinale)

turmeric

Zongu), in the fruit

wall of

pepper

(Piper nigrum), capsicum

and

chillies

(Caps&m

the

perisperm

and

annuum), embryo

(Myristica fragruns), cassia

(Cassia

cinnamon

of

in

the

@cinalis)

oils,

accompanying most

with resins

commonly

secretory

or and

found

structures,

withou

(Valeriana

& Miyake

1997;

gums, in

art

specia

eilher

on

thl

Osmophores Osmophores with

are areas of flower tissues cells

secretory

structurally

from

(e.g. isodiametric

the

differing

adjacent

cells

cells in orchids).

tissues. The

type of structure

in verif;/ing the authenticity source

in

the

adulteration 1990).

There

most important

is famik

which can be usefu case

(Jackson

of the plan

of &

suspectec SnowdoI

follows an outline

of thl

of these structures.

of

these

secretory cavities

folded

Secretory

produced

from

Included

cells

spherical formed cells

are

structures

leaving

more

or

less

that

can

be

in two ways: the parenchyma

can separate intercellular

one

some

with

also store

within

thin

the oil

their

in this group

are are

and

with

walls, which

plastids.

are fruits

and

leaves of plants in the Citrus Dmily; C.

sinensis,

C.

Zimon)

as

Eucalyptus spp. and buchu are also present cloves

(C.

C. aurantium, C. bergamia,

auranAJXa,

well

as

leaves. They

in the flower buds of

(Syzygium aromalicum),

the fruit

walls of pimento

(Pimenta dioicu), and

in the elongated

cavities in the bark of

(Cornmiphora molmol) benzoin benzoin)

(Boswellia

spp)

Secretory

ducts

Ducts

and

frankincense

(vittae)

are

elongated

predisposed

cells

parenchyma

undergo

division

cavities.

from spaces

another called

lumina or lacuna; or an actual cell can

Several

within

the

asynchronous

and in doing so, they expand

Lhe initial

space in the middle

the cells were all adjacent,

where

to form

cavity. Some of these cells forming of

the

cavity

biosynthesised

cavities cavities

cells also

enlarging filled

will

secretory/cpithelial These

the essential plants, several

The

continually become

wall

surface of the plant or within the plan or species specific,

layers formed.

(Stymx

Ming et al., 1996).

Essential

cells that produce

oils. In high oil yielding

nutmeg of

spaces are lined with

or epithelium and it is

cells

zeylanicum),

of valerian

(Maseda

these

bark

and

These

secretory

myrrh

anqustifolia)

(Cinn,amomum

and in the root

in

leaving a cavity within the

epidermis.

and

reticulum then They

within via

move

change

cells. The

a

the into

oils are

their leucoplasts

the

endoplasmic

into the cavity. These cavities

become can

Umbelliferde

be

‘joined’ found family

(Pimpinella an&m),

to form in

all

including fennel

ducts. of

the anise

(Foeniculum

very

dense

protoplasm

large central large

vacuole,

nucleus.

without

There

plasmodesmata

the

as well as a very are

numerous

across the cell walls of

the gland cells, and especially

between

the stalk cell and the collecting

cell. In

the very young gland, the intracellular organisation of

the

is nearly identical

adjacent

cells,

secretory

cells

changes

occur. The

progressively fully

as

develop, membrane

system

and in the

glands,

cytoplasm

the

complex

degenerates,

developed

granular

to that

but

only

is left

a fine

(Bourett

et

al., 1994). Cells usually

in the

have

normal

multicellular

nuclei

numbers

of

(endopolyploidy) Dracocephalum

moldavica

var. Budapest

vulgare),

dill

(Anethum

graueolens),

coriander

(Coriandrum sativum),

(Cuminurn (Angelica

archangelica)

1996).

the

In

case

angelica

(Bosabalidis,

of celery

[Apium,

graveolens) they can branch

to create

network

the

through

extending

and fruits.

They

in the Pinaceae,

Hypericaceae The

roots,

the stem to the leaves, then to

the flowers present

from

a

Coniferae

Compositae,

and Coniferae

resin

ducts

in

the

can reach 410

with between

are also families. xylem

of

cm in length

2 and 7 ducts per leaf.

of the

Glandular

trichomes

epidermal

hairs

and

are can

modified be

found

Labiatae

basil

lavender

(Lavandula

well

reticulum.

basilicurn),

the

marjoram

changes

spp.),

developed Essential

subcuticular

and there endoplasmic

oils accumulate cavity.

All

in the glands occur

(Mentha spp.) and thyme (Thymus

spp.)

the glandular

(Werker

et al.,

1985).

cells are attached

The

secretory

to a single

stem or

(Nishizawa et al., 1992). The formation and transformation

surface

is

heavily

however, takes place continuously

cuticle

usually

the

cutinired

and,

completely

glands

as the

covers the trichome,

are no pores

or perforations

there

present.

Instead, the essential oils accumulate spaces

diffuse

cuticle.

and it is thought

outwards

The glandular

normal

plant

in

cells

through

the

formation

senescence The

COMPOSITION FROM

they have

essential

OF

the

until

et al.,

1992).

precursor is, with high

synlhesised (Gershenzon

There

are

that

ESSENTIAL

two can

from

gland

oil biogenetic

probability,

THREE

VARIOUS

of

pyrophosphate

leucoplasts glands

of the essential oils,

(Ydmamura

isopenlenyl

cells differ from

in that

in

the

et al., 1992). main

types

exhibit

OILS,

SOURCES

Melissa

Melissa

Melissa

Melissa

Dracoce@zlum

Ne$eta cataria

Hose et al.,

Borne et al.,

Eire

Arulussa

SAC 1997

var. citriodora

1976-1994

1997

1996

1998

1998

1 - 37

tr-26

-

19

30 - 45

15-44

9-21

tr-10

-

4

20-29

10-29

5- 15

3-77

-

1 - 30 -

Geraniol

1 - 22

Nero1

tr-4

4

tr - 1

SAC 1995

4-8

tr-32

8 - 39

l-2

-

9 - 30

Citroneli

-

1-4

-

5

l-2

-

Citronella1

tr-40

1-52

1-2

7

1-2

tr - 3

Geranyl acetate

6 - 17

-

2 - 43

P-caryophyllene

-

2- 17

I1

Germacrene

-

tr-4

13

Numbers are percentage,s of total oil tr = trace

10 - 13

of

minor

Lawrence

Neral [n]

by

the time the leaves are about 5 mm long

The outer

the

and

hairs are frilly formed

basal cell in the epidermis. of

in

these

at a very

early stage in the leaf development

(Otigunum

most

Melissa

Citral [g + n]

D

the

(this might reflect

mint

oregano

MELISSA

[g]

These

are

organelles

spp.),

and

COMPARATIVE

Geranial

family.

(Ocimum

a

the

and in fully mature

a high energy requirement) is

heads

double

chromosomes

mitochondria

abundant

as the calyx in many

include

they

trichomes

hichome

such

subcuticular Glandular

glands,

leaves, stems, and even parts

of flowers plants

parsley and

crispurn)

covering

cumin

cyminum),

(Petroselinum

White secretory

with

Gland

on leaf surface

of Melissa

officinalis

variations.

Epidermal

(i) Peltate

glands with one basal

range

cells

Essential

oils obtained

cell, a short stalk and a large six to eight

not usually secreted

celled head.

but

(ii) Capitate

trichomes

with either

merely

cytoplasm,

diffuse

basal cell, a short a) one monocellular stalk and a two-cellular head or

low. Examples

stalk and a small

globose

oils from 0.075%,

The

these

hairs,

through

the

yield of essential

species include

acacia

and jasmine

unicellular

by glandular

the ccl1 wall and the cuticle

to the outside.

b) one basal cell, a multicellular

from flowers are

is usually very rose (Rosa spp.) spp.)

(Acacia

0.084%

In

every

distinctive shape

species,

variations

of the glands.

yarrow

(Achilles

trichomes

of

there

in the

size and

For example, floret

proazulene,

and they occur

the corolla

lobes

in

secretory

mill~folium) the

are

produce mainly in

of the ray and disk

florets,

but also on the leaves. When a

floret

reaches

mature

trichomes

developed

from

0.25

microns,

are present, protodermal

They have 10 cells including

fully having cells.

a pair of

The

composition

depends the

of

plant

material

(Hay

1993; Hose et al., 1997). proved

for various

investigated

at SAC are described.

officinalis

101

of

oil

is

minimum

0.05-0.1

%

countries

on the seed source,

yield

and

in

it reaches

0.2-

of citral depends and was found

to

The

Bavarian

52.0

%

examined

Research

a collection

provenances/cultivars

of

of

Melissa

and evaluated the best types (Bomme al., 1996). highest

The

percentage

but

favourable

growing

are very important.

month

period

content

decreases

the

et

of citral is

in young leaves and just before

plants

percentage

After a 6

of storage,

the -citral

from 84 to 50 %. As

become

older,

of citral decreases

to 12 %, and citronella1 1 to 32 %. Table components

an

the

from 37

increases

from

1 shows the main

of Melissa oil derived from

new plantations

extraction

In melissa, a reasonable

and

for Soil and Plant Production

(Germany)

is extracted

L.

0.3%. The percentage

Pais, 1994).

and

of the species

75.OY0, with

1.0

Institute

using Melissa

pairs of glandular

SC

species

from

Mediterranean

(Figueiredo

& Svoboda,

This has been

plant

some

oils

stage of

the results

basal cells, a pair of stalk cells, and 3 cells

essential

on the ontogenetic

and

I).

conditions

head.

8.7

between

(Table

flowering,

spp.) 0.04%.

(@m&urn

between

citronella1

in Africa, where the oil

directly from fresh plants, advanced

method

of

with 1,1,1,2-tetrafluorethane.

The very low oil yield of Melissa, and comparative

difficulties

with the distillation

connected

of fresh

material

together

with quick oil decomposition,

initiated

a quest for other

plants with

high

percentages

monoterpenes.

of

subsp.

and l)racocep,halum moldauica

citriodom

were investigated content

technology.

lemon-scented

Nepetu cataria

for their essential

oil

and quality.

included

Recent

composition (both

fresh

cataria

large

subs@. citriodora

Nepeta cataria can be grown successfully as an annual

in our northern

and in areas with a more climate,

as a perennial.

material

climate,

The

and during

were

flowers plant

between fresh

the

oil content with

is between

citral,

geraniol

at full flowering.

and

dried

and of leaves and flowers. All distinct,

light lemon scent.

et al., 1994).

Dracocephalum

molclavica throughout Siberia

antispasmodic

Geranial,

neral,

geraniol,

account

essential

oil (Table

time

and

IL has antibacterial,

and

the

is an annual

moldauicu

southern

geographical of

harvest

chamomile

herbaceous

is

a

perennial,

creeping native

Europe

and

Reaching

a height

of about 0.3 metres,

geranyl

north

plant is characterised

I), and

a long period, fresh hours,

depending

source,

on

genotype, postharvest

borne01

(tr-5.0%),

white-ray

be

the gathering

repeated

flowers

takes place over

several

become

changing

mouldy

teas,

the

flowers

must

dried

as

possible.

For

be

oil distillation,

during

drying

and a special

CHAMOMILE

ROMAN

oil content

OILS

and

on

the

during

quantity

depends

for

of chamomile

oils

British

was

chemical

market

composition,

with samples from established

new growers

(Table

2).

Due

oil from

Suffolk

Starke & Son) being of excellent and well established

soon

used as a blueprint

quality

for comparison.

cut

increases aroma

is

FROM

Cedarwood

oils

Cedarwood

oil is an important

source

of aromachemicals

VARIOUS

natural

used either

SOURCES

1

2

3

4

5

6

7

8

ol-pinene

6.3

4.8

3.3

16.2

12.2

2.2

-

5.4

10.2

A

9.5

9.5

10.8

14.7

14.5

38.4

17.3

11.8

-

9

B

18.3

17.1

13.3

7.9

10.9

-

11.5

18.5

-

c

6.6

5.9

6.7

5.7

5.3

8.8

4.8

7.9

-

D

5.3

5.0

4.0

7.1

7.9

15.2

-

5.4

-

E

15.2

14.2

15.0

8.2

10.1

1.0

-

14.7

-

F

5.8

5.0

‘7.8

-

2.8

-

4.4

-

are percentages

(R.W.B.

on the market, was

Oil Component

Numbers

to

in the GC analysis of this oil,

gathered as

is formed

precursor the

selection

available

chamomile

the

(tr-l.O%),

and pinocarvone

upon the origin and age of the flowers. A wide

within

can be left in the field for 1-3

days. The essential

and

alpha-

(1.0%))

myrcene

Chamazulene

difficulties

So

beta-pinene

natural

The

to a brown colour.

for use in herbal

material

of heads

times.

include

(tr-1.0%)

distillation

together

eastern

and

by

angelate

(l-5%),

analysed

antiviral

acetate

Africa.

and isoamyl

pinene

a

the

n-butyl

Monoterpenes

early July. As flowering can

for about 90% of the

(35%)

esters,

being

(15%).

from

of Roman

aliphatic

component

angelate

flowers that appear from late spring to

central

properties.

principal

are

(1.0.4.0%).

to

western

this aromatic

chamomile

camphene

nobile

downy stems and yellow-disc,

distributed

Europe.

1

of this

species.

Dracocephalum plant,

et

Table

Chamaemelum Roman

and

(Collins

the main components

promising

Asia,

nerol

in quantity

al., 1993; Collins outlines

0.8 and 2.2%,

citronellal,

found

The

of

oil, of a light blue or greenish

The major constituents

oil

yield with the best quality and quantity of oil is achieved

0.7 and 1.75%

to yellow-brown.

significantly

the oil samples had a pleasant,

highest

heads and stems

between

blue colour, which can change with age

leaves

of

Flowering

can contain essential

dried)

differences

developed.

oil yield and

in

There

SAC

analysis

and

composition

favourable

at

growth

and yield, and identified

ontogenesis. Nepeta

studies

preliminary

of total oil

1. R.W.B. Starke & Son, field distillation,

whole plant, July 1997

2. RW.B. Starke & Son, British Pharmacopoeia Standard laboratory diit%h&on, flowers July 1997 3. R.W.B. Starke & Son, British Pharmacopoeia Standard laboratory distillation, leaves July 1997 4. EngEsh grower @} (anon.) experimental plots, British Pharmacopoeia Standard laboratory distillation,whole plant,July 1997 5. English grower IlI] (anon.) experimentalplots, Bri&sh Pharmacopoeia Standard laboratory distiJ&ation, whole plant,July 1997 6. Commercial sample of essential oil [a] origin unknown 7. Commercial sample of essential [b] origin unknown 8. Commercial sample of essential oil [cl origin unknown Components A-F are all esters, and they include isobutyl angelate, butyl butyrate, isoamyl-2-butyrate, isoamyl angelate, and two unidentified esters. However, which percentages correlate with which esters has not been ascertained. GCMS was done in Japan and New Zealand and has incomplete identification due to the lack of pure chemical standards. An effort is being made towards further elucidation. The main author, (Dr KP Svoboda) personally collected samples 1, 2 and 3, and these are now used as a ‘blueprint’ for other acquired samples.

directly

in fragrances

extract

further

(Adams,

1986; Adams,

al., 1988).

1991; Adams et

Juniperus

(Virginia

cedarwood).

components cuparene,

cedrol cedarwood

include

major

interest

and

distilled

Other

vir@niana are

beta-cedrene, thujopsene,

Commercial fractionally

widdrol. oil

may

be market

to remove cedrol.

sources

of cedarwood

oil

Cedrus

Cupressus spp. (China),

atlantica

(India),

(Morocco)

and Cedrusfunebris (China).

Young 3.5%.

Cedrus

trees yield less than

compared

cedarwood

to Texas and

is different.

the end product, take into

seasonal

obtained

samples

also to

were

not

and the source

of material:

whether

was heartwood,

it

sapwood,

bark, young

none

thujene

of the oils were

The

variability

min to 8 hours); boiled

how was it distilled

in water or suspended

(boiling

changes

decomposition

-

in steam

the pH of the water

consequently

and

causes

of environmental genetic

For our experiments,

fresh plant

from C. atlantica,

C. deodoru, C. brevz]olia and C. libani all grown in Scotland Material

consisted

twigs and cones from

(MacDonald, of fresh

needles, collected

C. atlantica and oil

Essential

was

hydrodistillation

C. deodoru only. obtained

of fresh material

by and

subjected samples

to GC-MS. of

cedarwood labels,

commercially oils,

were

In addition,

sold

obtained

available

under from

24

various the

UK

COMPOSITION

New pharmaceuticals

out for

activities.

products

initial

source

are important

in the

as the

development

of

anticancer

agents

harvest and the

and

alone,

and

over

developed

their

products

in the

of the increase

has

been

essential

and hope.

assays are

specialised

experiments

which

concentration

of a key substance

determine

by measuring

a biological

in

for new anti-

drugs

oils offer real potential

preparation

of

infection

prototype

Biological

still

treatment

scaled up, and in this regard

system.

the in a

its activity in

The

system may be human,

biological

animal, plant or

and

the

primary

are to show any effect and to relative

potencies.

A method

using brine shrimp is a simple bioassay for natural

OF

pharmacological

conditions,

years, the search

infective

of

may then be carried

specific

Taking

antibiotic-resistant

estimate

relative

new drugs (Cragg et al., 1997).

many diseases. Because

objectives

samples

the

sophisticated

on the final product

and natural

with

representative

once more

Natural

oils indicates

of plants

play a vital role

micro-organism,

few

and

is known,

the fresh

in the search for new drugs is

all the oil samples were analysed by GC a

is reliable,

antiinfective

60%

of approved

for the treatment

drugs of these

diseases are of natural origin. There is a

increasing,

in

inexpensive toxicity

as toxicity

The method

groups

origin of the plant material.

extracts

1997).

with wood

both

are manifested

to the shrimp.

synthetic.

technology,

importance

recent

of plant material).

material was collected

within

compounds

bioassays

samples

or pinene

and commercial

The

(1

representatives.

of the tested

contained

whether

it was fresh or dried material;

and clearly

and none

Bioactivity

of distillation

authentic

However,

or old trees, twigs and needles or cones; what was the length

in this study. Several mislabelled

and post-harvest

variations

by GC and GC-

3 and 4 show some of the

the huge influence

In considering

it is important

account

results

samples

oil, but is less valued because

its fragrance

and analysed

MS. Tables

were

1% of oil

oil is almost

in composition

Virginia

deodora

with older trees which yield

Chinese

identical

of material

Source

(Texas

The

of commercial

alpha-cedrene,

of cedar

ashei

and Juniperus

cedarwood)

to

compounds

The major sources

are

oils

or processed

primary

product

a broad

range

CEDRUS

research. of

Activities

known

ATLANTICA

active

0

diverse structural

array of compounds

that

considered

may

be

chemopreventive, may

be

considered

(Pezzuto,

in

1997).

this

aromatic

flavones,

diallyl disulfide,

antioxidants,

ellagic

carotenes,

calcium have

unmodified synthetic

(Ca)

and nerolidol

Drugs of natural

been

classified products,

products

derived

products,

based

on

(Dewick,

acid,

retinoids,

natural

natural

between

include

compounds,

(Wrigley et al., 1997). origin

category

Examples

phenols,

terpenes,

cancer

and over 600 agents

or synthetic

natural

product

1997).

Data

as semifrom

products models published

1983 and 1994 show that from

a total of 520 new products, biological

origin,

unmodified

natural

semisynthetics, product

purely

synthetic. role

rave played,

are

products,

from 127 are

46 are modelled

natural

important

28 are of

30

molecule These that

on the

and 289 are data show the

natural

and continue

products to play, in

hug development.

or-pinene p-pinene

myrcene

ru-atlantone& himachalc

Cut twigs

68.9

-

7.3

-

15%

Female

71.0

4.4

29.5 / 16.8

;ysternatically

cones

Wood Twigs/needles

79.9

-

Source: Royal Botanic Gardens, Edinburgh Numbers are percentages of total oil

It has been estimated of

species n-esence

the

of higher of

that only 5-

approximately plants

investigated bioactive

250,000 have for

been the

compounds.

Examples of drugs derived from plants nclude: ;lycosides

steroids, (lligitalis),

cardiotonic anticholigernics

(Relladana

type

tropane

analgesics

and

antitussives

alkaloids),

alkaloids), (opium

antihypertensives

(reserpine) , cholinergics antigout

(pilocarpine),

agents

anaesthetics

(Colchicinej, muscle

(cocaine),

relaxants

and

(tubocurarine),

anticancer

drugs

current

50

available, derived

(see above).

or

so

Of the

anticancer

drugs

the most important ones are from

Catharanthus

naturally

Tuxus Drevz@lia, Podofihyllum

?-OS&L&

peltatwm, and Camptheca acuminata. Antimicrobials

& antioxidants

Many essential

oils possess

antibacterial, antioxidant

activities

1990;

Deans

years,

attempts

isolate

1992).

have

for of

the

these

and

al.,

made

to

compounds

activities.

essential

temperate

et

In recent

been

identify

responsible

and

(Clarke

et al.,

and

number

significant

antifungal

oils

tropical

A

from

both

plants

were

tested at SAC for their activity against a range of bacteria, public

health

plant

including

animal

spoilage/poisoning In

addition,

tested

for

their

antifeedant

activities

promotion

activity of some plants

essential to

be

oils is meaningful exploited

for

be

transformed

activity into more as

superoxide,

singlet

such

metabolic

reactive

forms

hydrogen

oxygen

and

Other

active oxygen:

toxic since it by

factors

can

dioxide.

oxygens

are

All

generate pollutants

nitrogen

sulphur

radicals

known as active

UV radiation,

as ozone,

such

peroxide,

hydroxyl

which are collectively oxygen.

the

of human well-being.

Oxygen is potentially can

al.,

oils were

tansy and oregano).

The antioxidant enough

et

several

melissa,

and their

food

(Graven

1992).

and

and

pathogens

(hyssop,

types with

significance,

oxide these

extremely

and active

reactive

and

short lived, and they cause damage proteins,

lipids

and

nucleic

Direct damage to membrane as peroxidation to the

breakdown

and

function.

Hydroxyl

react

with bases

mutations. inactivate

Vitamin

can

carotene

can

oxygen, radicals.

They

scavenge

E and or

pollutants

and

enhancement

aromaticurn) their

from

radiation,

antioxidant

any systems

effects.

Essential vulgaris)

UV

of

has beneficial

pressures

This

prevents

from

oils from thyme (77hymu.c

and

clove

were

assessed

capacity,

reference

liver and retina

(Syzygium

successfully

antioxidant

with

to the protection

of polyunsaturated

easy animal eggs

fatty acids

during

ageing

in the (Deans

can be stored

fa

they are returne d the eggs absor b begins,

embryogenesis

completed after

by between

developes breaks

antennae away

swimming though

lipid

crustacean

The and

from

the

and becomes nauplius.

and

16 to 36 hour

immersion.

The brine

to the subclass

When

to the saline solution,

Brine shyimp toxicity belonging

are abet

and as long as the

water,

and

(Artemia salina) is a

and study. Th

shrimp

dehydrated,

membrane, shrimp

to a wide range c makes it a relative1

brine

long periods.

It

rangin

to the ultrasalinc

to culture

of the

remain

Anostraca. in water

the brackish

0.2mm in diameter

beta-

deactivate

superoxide

order

world-wide

salinity (lo-22Og/l)

increased

et al., 1993; Deans et al., 1995).

C, vitamin

singlet hydroxyl

peroxide

enzymes.

their permeability.

With

particular

cause

Branchiopoda found

acids.

radicals and

chain-breaking the membranes

This high tolerance

for

can

a

also have the ability to bind free fatty

of structure

in DNA

Hydrogen

lipids

by

and stabilise

by decreasing

acids.

structures

of membrane

leads

to

peroxidation reaction

The

embry

Iis 0

mandible: L hatchin

gl

an active fret :larva

grow ‘S

about 15 stages and survives for

72 hours

I

‘s

on its yolk resource

alone.

I

After

about

reaches

15-35

a length

days,

the

animal

of 8.5-9.5mm

the number

and is

sexually mature. During

active life is indicated

of its growth, by movement

the

of

particles

(unicellular

towards

the

algae, yeast cells)

head.

the

The

ease

availability

of hatching

them

and the relative

ease of maintaining

population

under

simple

make

and

bioassays stages shrimp rate

the brine

the

petroleum

eggs

hours

after

older

stages

screening.

carcinogens

hours

be

used

of exposure

(the

acute

Five

oils

German

were

used

tests: Roman chamomile,

thyme,

For each

four

of 30 wells were

plates

characterise

of the shrimps

concentration

of essential

each 200,

insecticide lppm).

LD50)

between

107ppm

concentrations estimates from

a preliminary

1OOOppm. On

each

well)

of shrimps

were

33,

100,

plate

wells. The

being

and 279ppm

which

required

activities

indicates Further to

assess

of the oils and their

components.

500

there data

and were (for

into the well and

shrimp

inally

bioassay

support; Professor

melissa

Asakawa

‘harmaceutical 3unri

Arulussa

for

770,

and

(Faculty

Sciences,

University,

rokushima

Ltd,

oils

of

Tokushima

Yamashiro-cho, Japan)

analysis of essential

for

GCMS

oils.

0

1998

National

Holistic

Association

Aromatherapy

Reprinted

with

NAHA from World

for

(NAHA).

permission

from

the proceedings

of the

of

Aromatherapy

International

II

Conference

and Trade

?? Adams, R.P. (1986)

Investigation

Show.

)fJuniiperus species of the United States ‘or new

sources

of

Cedarwood

oil.

konomic Botany 41: 48-54. ?? Adams,

uniper

R.P. (1991).

and other

Analysis

forest

of

tree oils. In:

Modern method.5ofplant analysis - oils and “axes. Eds. Linskens,

H.-F. and Jackson,

?? Adams, R.P., McDaniel,

The

Scottish

Agricultural

financial

Andrew

photographs Chestnuts

Fisheries Syred

Photolibrary,

Beguildy,

Middle

Near

(R.W.B.

Knighton,

UK) for SEM

activities

(1988) in

)ark/sapwood pecies

the

heartwood, of Juniperus

and leaves

from

the

C.A. and

Termiticidal

United

States.

3ochemical Systematics and Ecology 76: 153-456. ?? Bosabalidis,

Intogenesis, norphometry

A.M.

(1996)

ultrastructure of the petiole

and oil ducts of

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:elery (Apium graveolens L.). Flauour and

& Son,

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Farm, Eye, Suffolk.

7EIN) for financial

F.L.

Starke

of essential

Starke

The

Agriculture,

Powys. Wales LD7 lUW, Richard

from

and

(Microscopix Travelly,

College

support

Office

Department;

using

the Zambia

:arter,

after

obtained

oils and

in

.F., 131-157.

Environment

consists of the total number pipetted

(fennel)

high bioactivity.

These

experiment

of

LD,,

to

oil. On each

Essentia Hinckley,

(ideal

and the

chosen

LD,,

(ideal

be

would

specific

Ltd, Street,

or for use

All five oils had LD,,

chamomile)

individual

Bond

possible

tests

Scottish

were

the

comparatively

(30, 60, 100, 150,

concentrations

each

melissa

480ppm).

of the

also two control

the

used

of

activity of compounds

(German

low

500-l 800 ppm.

of the test lies mainly

indicator

around

receives

and

and showed relatively

as an

wells were used to test

concentration 240

an

camphor,

between

the mortality

plate, 4 adjacent

tested

oils,

and citral have all

LD50 being less than 40ppm)

of the ftve oils,

the relationship

carvone,

menthone

after 6

in

essential

antitumor

chamomile,

and fennel.

there are no references

regarding

The significance as

in the

Station).

Although available

used

lsed

inancial

1997,

oils

Shirley Price (Shirley

,eics. LElO lRS, UK) for supplying

,usaka,

Rothamsted

for

makes the test rapid and simple. preliminary

Experimental

Trust,

Upper

a

is

and

Aromatherapy

-Iouse,

a

in the statistical

the

of the lethal

for 50% mortality

and

of oil) is “probit

toxicity if used between

for toxicity.

although

also

of

between

(copyright

Lawes Agricultural

been

contaminants

Identification

concentration

to

stage used is 2448

can

GENSTAT

material

shrimp bioassays; ‘rice

the

(alive/dead)

available as a procedure

terpinen-4-01,

hatching,

relationship

stimulus (concentration

limonene,

used as criteria

constitute

estimating

response

after

oil, pesticides,

have been

brine

slant

of

effects of the oils.

used: the hatching

and other environmental The most common

in

exposure

plate)

for

the

quanta1

All

of the

triples

The standard statistical method

package a

animal studies.

cycle

have been

of

test

toxicology life

a

shrimp

28

analysis” (Finney, 1971). This method

laboratory

effective

and

in the

The (per

data

analysing

of

into larvae, the rapid growth of nauplii

conditions,

dose.

toxicological

which help to sweep food

the eggs,

alive after 6 hours together

the

measurements

all stages

appendages

with

support,

UK IP23 supplying

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