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SPS Capacity Building Program (SPS CBP) presents:


TRAINING WORKSHOP ON ARTHROPOD PRESERVATION,

CURATION AND DATA MANAGEMENT

Museum Zoologicum Bogoriense, Cibinong, Indonesia, 26-30 September 2005


MITE (ACARI: ARACHNIDA)

COLLECTION, PREPARATION, MOUNTING AND STORAGE

Sri Hartini and Ahmad Saim


L I P I ASEANET
Widyasatwaloka

Reseacrh Center for Biology – LIPI, Cibinong



INDONESIA

MITE (ACARI: ARACHNIDA)

COLLECTION, PREPARATION, MOUNTING AND STORAGE

Sri Hartini and A. Saim

Zoology Division, Center Research for Biology-LIPI, Bogor, Indonesia.



INTRODUCTION
More than 60,000 species of mites have been described from various regions of the world, the number of undescribed species is estimated to exceed 600,000 (Evans, 1992), and the majority of such undiscribed species are expected to live in tropical regions. Mites are successful colonizers of terrestrial and aquatic habitats, and are found in leaf-litter and soil, on leaf, on bark and invertebrate animals, in salt and fresh water, and so on.

Habitat of mites are free living forms and parasitic form. Some of free living mites as Predaceous and Phytophagous ( suborder Gamasida and Actinedida), most of this group mite is important as pest plants. Aside from their feeding injury, some species have been found to transmit plant viruses. Many mites are predaceous on other pest and some species have been used in biological control programmes.

In this paper we will explain of morphology, life stage, habitat, how to collect, preserve, preparation, mounting, storage and packing for carrying/sending mite specimens.

MORPHOLOGY

The Acari, comprising mites and ticks, are one of the largest and most biologically diverse groups of the Arachnida. The mites and ticks differ from most arachnids in that somatic segmentation generally is inconspicuous or absent. Abdominal segmentation is a primary attribute in all of the other subclasses except the Aranae-the spiders. The mite, ticks and spider may easily be separated from other arachnids. Mouthparts of mite and tick, contained in a discrete anterior gnathosoma; portion of the body on which the legs are inserted (the podosoma) broadly joined to the prortion of the body behind the legs (the opisthosoma) to form the idiosoma. Gnathosoma or capitulum resembles the head of the generalized arthropod. The roof of the gnathosomal tube is termed the epistome (tectume capituli of Evans and Till, 1965), and the lateral walls are made up of the enlarged coxae of palpi. Paired of chelicerae in gnathosoma is primary organs of food acquisition and may differ considerably between taxa, but are generally adapted for piercing, sucking, or chewing. Generally chelicerae are three segmented (two segmented in the suborder Ixodida) (Krantz, 1978).



Idiosoma functions parallel to those of the abdomen, thorax, and portions of the head of insect. Ventrally, the idiosoma a variety of shields (ventral, sternal, metasternal, epigynial, metapodal, and ventrianal shield). On the idiosoma found external structures are locomotory, respiratory, copulatory, sensory and secretory in function.

Locomotion. Legs of adults and nymphs four pairs, the larva has three pairs. The legs are divided into seven primary segments (coxa, trochanter, femur, genu, tibia, tarsus and pretarsus).

Respiration. Stigmata or spiracles is system respiratory in the Acari for exchange of carbon dioxide and oxygen. The presence and position or absence of stigmata for identifying the acari suborder

HABITAT

Two major habitat of mites: I) free living form and II) parasitic form.


I) Free-Living form:

A. Predaceous Mites

1. Ground species

2. Aerial species

3. Storage species

4. Littoral-intertidal-marine species

5. Aquatic species

B. Phytophagous Mites

1. Ground species

2. Aerial species

3. Storage species

C. Mycophagous Mites

D. Saprophagous Mites

E. Other Microphytophagous

F. Coprophagous and Necrophagous Mites

G. Phoretic Mites


II. Parasitic Form

A. Ectoparasitic Mites

1. Vertebrate ectoparasites

2. Invertebrate ectoparasites


B. Endoparasitic Mites

1. Vertebrate endoparasites

2. Invertebrate endoparasites

LIFE STAGE

The acari generally have four life stage: egg, larva, nymph and adult. The typical larva have six legged form, some family in suborder have prelarva. Nymph and adult have 8 legged. Nymph form are known single, two or three individual nymphal stages and depend of the suborder. Two nymphal stages (protonymph and deutonymph) occur in most of the suborder Gamasida and three nymphal stages (protonymph, deutonymph and tritonymph) occur in the suborder Oribatida.

Development from egg to adult may require as little as 4-5 days in the Acari, but more often extends to several weeks or month depend of each suborder and condition.

Classifications of the subclass Acari by Krantz, 1978:
I. Order Parasitiformes

A. Suborder Opilioacarida

B. Suborder Holothyrida

C. Suborder Gamasida

D. Suborder Ixodida
II.Order Acariformes

A. Suborder Actinedida

B. Suborder Acaridida

C. Suborder Oribatida


I. SUBORDER GAMASIDA (ORDER PARASITIFORMES)
Gamasida is one of suborder in order Parasitiformes. Important of characteristic this group is in the ventral side have shield. Member of sub order Gamasida are 66 family (Krantz, 1978), some of family have economic important. This family are long legged and rapid in movement, colored in shades of red, yellow or green. Widely distribution of this family and most in the tropic. Habitat of this group mites on leave and some in soil. The life stage of this group from eggs, larva, protonymph, deutonymph and adult. Eggs put on the leave. Member of suborder Gamasida, family Phytoseiidae, two species Phytoseiulus persimilis (AthHenr) and P. macropilis (Banks) have already been used very successfully in greenhouse to control the red spider mite in Europe; these species multiply much more rapidly than the tetrachyds and are already produced commercially. These species also as predaceous areal mites, preying primarily on phytophagous mites or their eggs (spider mites, Eriophyd mites, Tarsonomid mites, trips and soon), as predators in pest management programs or biological control.


II. SUBORDER ACTINEDIDA (ORDER ACARIFORMES)
Actinedida is one of suborder in order Acariformes have variation of habitat. Suborder Actinedida have 127 family and some of family have economic important are:

1. Family Tetranychidae, Red Spider Mites.

Habitat is this family on the plant and widely distribution and aerial species mite. This species are slow-moving or sedentary mites which are weakly sclerotized. The majority of species are red, yellow, or green in color while some appear white or transculent. They feed by inserting stylet-like chelicerae into the cells of the plant host and sucking up the contents and they life undersurface of the leaves. The life stage from eggs, prelarva, larva, protonymph, deutonymph and adult. This group are some of most important arthropod pest plants. In Indonesia, this family pest on the cassava, and tea plantation. This family of spider mites have been known as being considerably injurious to sugar cane in eastern Asia (Ehara, 1969). Aside from their feeding injury, some species have been found to transmit plant viruses. Species of the family Tetranychidae on plant are:



1. 1. T. cinnabarinus (Boisd) (= telarius, bimaculatus), carmine or Red Spider

Mites
This widely distributed species, is the most common species in the tropics and it also is a pest of greenhouse in temperate regions. In Java, Indonesia the species was first found on cassava (Leefmans, 1915). It feeds on many hosts including crops, such as cotton, legume, greenhouse, nursery, citrus and ornamental plants, as well as weeds. Other host plants recorded in Bogor are castor, rubber, papaya, green manures, dadap trees, peas, beans, and tomato (Kalshoven,1950).

This spider mite is variable in color, usually with shades of green, yellow, or red, and two darker pigmented spots. The first instar is pale yellow and has 3 pairs of legs, the mature females being the larger of the two sexes and showing more pigmentation. Male are smaller size and narrow, more pointed abdomen. Tiny, pale green spherical eggs are deposited usually on the undersides of the leaves, where feeding and spinning of delicate webs take place. All active stages remove plant sap, usually from the undersurface of the leaves, resulting in tiny light spots. This gives the foliage a speckled appearance, and with severe infestation the entire plant may be killed. Dry hot weather favors the rapid development of spider mites. At normal greenhouse temperatures approximately, 20 days are required for development from egg to adult.

Symptoms of the species on cassava, the leaves initially show yellow blotches, then coalesce and the leaf tissue becomes entirely yellow and finally redish (hama merah). The mites are visible as red spots, the size of a leaves wither and fall, but the plants do not die.

The damage to cassava plantation may be considerable especially in Central and East Java areas have been totally defoliated with considerable loss of yield (Kashoven, 1950).

Predaceous mites in the genus Typhlodromus are very important in keeping mite populations checked. Insecticides are highly toxic to the predaceous but not to the plant-feeding species. Frequent syringing of plants with a strong stream of water is of some value in spider mite control.

1. 2. T. urticae Koch (=telarius, bimaculatus), two spotted spider mites

The mites are a major pest in orchards, cotton, castor, apple, papaya. Female yellowish green with two large dark spots dorso-laterally; in older females these spots extend backwards to the end of abdomen.


1. 3. T. pacificus McGregor, Pacific Spider Mite
This species is one of the most pest of deciduous fruits. It also attacks many fruit, legume, and vegetable crops, as well as ornamental plants and weeds. Adult females are pale green with dorsal dark spots medially and a pair of dark spot near the posterior end of the body.

1. 4. T. schoenei McGregor, Schoene Spider Mite

This species is widely distributed, attacking deciduous fruit and ornamental trees and shrubs, as well as field crops. Adult females are pale green with 4 dorsal darkened spots.



1. 5. T. canadensis (McGregor), Four-Spotted Spider Mite

T. canadensis attacked in apple, cotton, rose, elm, linden, plum, horse chestnut, and osageorange has been found in scattered areas of the eastern half of the United States and in soutern Canada.

1. 6. T. atlanticus McGregor, Atlantic Spider Mite/strawberry spider mite

This species commonly feed on cotton, strawberries, legumes, ornamental scrubs, and fruit trees in United State.



1. 7. Metatetranychus ulmi (Koch), European red mite
This spider mites has become one of the most important pest of fruit trees in northern United States and adjacent regions of Canada introduced from Europe. The mite attacks elm, apple, pear, peach, plum, and prune to an injurious extent, and may be found on other deciduous trees and shrubs as well. Sap removeal by the piercing sucking active stages results in bronzing and off-colored foliage and, under severe infestations, defoliation and undersized, poorly colored fruits.

The mites are usually rusty in color, but newly emerged females are bright velvety red, changing in time to dark red-brown. Males are dull green to vulvous. The red orange eggs of the European red mite are easily recognized by the prominent hair like projection on the upper side. Development from egg to adult 2 to 3 weeks.




1. 8. M. citri (McGregor), citrus red mite

This mite is known as the purple mite in Florida and as the citrus red spider in California. Both nymphs and adults extract the sap from foliage, fruits, and tender branches with their piercing-sucking mouthparts, producing tiny gray or silvery spots on the leaves and fruits. When leaf damage is severe, the normal photosynthetic processes of the plant are greatly inhibited, resulting in leaf drop, decreased plant vigor, and smaller, poorer quality fruits.




1. 9. Bryobia praetiosa Koch, Clover Mite
This species called the brown or almond mite in the pacific Coast, is a world wide pest of fruit trees, clover, and other legumes, as well as a number of garden and field crops, including cotton. The adult is flattened, dark brown to dull green, with rather long front legs.

1. 10. Genus of Oligonychus
This genus recorded four species in Eastern Asia are assigned to the Pratensis group (Ehara, 1966). Mites of this group feed on the undersides of host leaves.

Some of the species of Oligonychus are:


1. 10. 1. O. orthius Rimando
O. orthius was originally described from sugar cane and Imperata in the Philippines. Recently this mite was recorded from Okinawa Island, Japan on sugar cane (Ehara, 1966) and para grass, and from Taiwan on sugar cane and banana (Lo, 1968).

1. 10. 2. O. kadarsani Ehara
This species described by Ehara (1968) from sugar cane in Pasuruan, Java, Indonesia.

1. 10. 3. O. shinkajii Ehara
O. shinkajii was previously known from corn and rice in greenhouses of Kanagawa Prefecture, Honshu, Japan and sugar cane in Taiwan.

1. 10. 4. O. exsiccator (Zehyntner)
This species was originally described from Java on sugar cane in 1897 and also this species has been recorded from sugar cane in Hawaii. Greenish yellow patches with a few red spots indicate the presence of colonies on the leaf underside. Later more red spots appear and oblong rust-coloured patches. The leaves wither and die prematurely but this damage is normally confined to the secondary shoots and no harm is done.

Natural enemies is small coccinellids and gall midges and keep the population of mites.



1. 10. 5. O. ununguis (Jacobi), Spruce Spider Mite
O. ununguis is an important pest of arbour vitae, hemlock, cedar, pine, and spruce, especially blue spruce.
1. 10. 6. O. coffeae (Nietn.), Red Tea Mite
This species are mostly found in tea and coffee, and also occur on castor and many trees and shrubs. They are found on the upper surface of mature tea leaves which then turn into yellowish-brown rusty or purple colour.

1. 11. Schizotetranychus celarius (Banks)
S. celarius is readily recognized by the dosoventrally depressed body, and by having the dorsal body setae of various lengths. The adult mite are pale greenish yellow to pale green. Mites of this species form restricted colonies and live between the under surfaces of host leaves and dense spinning canopy. Within the scope of the canopy the mites feed on the under surfaces of the leaves, and the withist feeding scars can be found on the upper surfaces of the leaves. S. celarius is known from Japan and USA and it is a common pest of bamboo and sasa bamboo in Japan. Recently it was recorded from sugar cane, Ficus, and Miscanthus in Okinawa Island, and rice in Honshu, Japan (Ehara, 1966). Also this species recorded from rice in Nagano Prefecture, Honshu, Jepan (Ehara and Miyashita, 1962).

1. 12. Brevipalpus (=Tenuipalpus) phoenicis (Geijsk.) and B. obovatus (Donn.),

Scarlet Tea Mite
These two closely related species are widely distributed in the tropics and subtropics of the Old and New Wold. In Java they are the most destructive mite species on tea. This mite also live on fruit trees, crops, sweet potatoes, Hibiscus etc.

1. 13. Tenuipalpus orchidarum Parf., red mite of orchids
This species attacked leaves of orchids Phalaenopsis become silvery then yellow. Damage can be considerable, even to seedling.

2. Family Tarsonemidae
Few groups of mites are adapted to feeding on live plant tissues in soil,

phytophagous ground species feed on root tissue, corms or bulbs, and often are responsible for economic injury to ornamentals and vegetables. Members of this group are mostly opaque white or transculent, slow moving forms with short legs and with little or no distinctive idiosomal sclerotization, some have chelate-dentate chelicerae for grinding and macerating plant tissues. This family is most of the species parasit or predator on insect. Habitat of this family as parasit on insect (honeybee) on trachea and some of the species on plant and mycophagous mite. The life cyclus from eggs, larva and adult. Most of the genus of this family as pest on plant (Polyphagotarsonemus pest on plant of cotton, tea, rubber, coffe, citrus, garden plant, and vegetable) Steneotarsonemus as pest on rice field in Taiwan.



Steneotarsonemus pallidus (Banks), cyclamen mite is chiefly a pest of greenhouse plants, especially cyclamen gerbera, snapdragon, and African violets. At times it seriously injuries garden plants such as delphinium, dahlia, and strawberry. Symptoms of their presence, caused by removal of plant sap, are distortion and stunting of flower and foliage, with or without blackening of the areas fed upon. Attacks are confined primarily to the younger leaves.

Adult mites are amber- or caramel-colored; the younger stages are white. They are too small to be seen without magnification. Tiny pearl-like eggs hatch in a week or more. Breeding is continuous in the greenhouse, and a generation may be completed in about 3 weeks. Considerable natural control results from predatory mites in the genus Typhlodromus. Sanitation measures should be practiced where infested plants are handled, fumigation with methyl bromide effective method of ridding plants of the mites.



3. Family Eriophyidae, leaf and gall mites
Most of member family Eriophydae on gymnospermae and angiospermae. The specific of this family is the body flat or vermiform and they feed by inserting stylet-like chelicerae into the cells of the plant host and sucking up the contents. The life stage from eggs, larva, protonymph and adult. Member of this family causes discoloration or russet and the formation of galls on the leaves. This family pest on the plant of citrus, wheat, corn, appel and onion. Some species of the family Eriophydae are:

3. 1. Calacarus (= Eriophyes) carinatus (Gr.), Purple Tea Mite

This species is a pest in tea plantation on the East-coast of Sumatra and also in Java. The species was first described in India. Attacked of this species on the plants become purple, and leaves are covered with fine dust, especially along the veins, formed by the white cast skins of the mites.


3. 2. Eriophyes boisi Gerb
E. boisi is gall mite cinnamon trees. E. endigofera Nal. lives on Indigofera.
3. 3. Eriophyes (=Acaphylla) theae Watt., Pink Tea Mite
This species found in Indonesia and India as a minor pest of tea. It attacked on the youngest leaves and first look whitish and later become yellow.
3. 4. Aceria (=Eriophyes) sheldoni (Ewing), Citrus Bud Mite
Widely distribution of this species, in tropics and subtropics. In Java this species is sporadically serious pest of all citrus.
3. 5. Eriophyes pyri (Pagenstecher), pear leaf blister mite
E. pyri introduced from Europe about 1870, this tiny mite is now generally distributed in North America. It attacks buds, foliage, and fruits of pear, apple, mountain ash, sevice berry, and cotonester. Its feeding causes the formation of galls of blisters on the leaves. These blisters are yellow to green at first, becoming red, and finally black on the leaves of pear. Fruits may be resseted or deformed and will often crack open. They produce their characteristic injury by feeding and burrowing into the leaf tissue.
3. 6. Phyllocoptruta oleicora (Ashmed), citrus rust mite

This mites is one of the most common and serious pest of citrus in Florida, Texas, and also found in some citrus districts in California. The mites feed on the sap of leaves, twigs, and fruits of all kinds of citrus, lemon, lime, grapefruit, and orange are most severely damaged. Leaves lose their glossy appearance, becoming bronzed and stunted under heavy infestations, often dropping prematurely. New twig growth is stunted and discoloured. Damaged fruits become light brown to black in the areas where the mites feed. This discoloration is called russeting and accounts for great losses every year owing to reduced size, quality, and poor appearance of the fruits.

Citrus rust mites are very small, averaging 0.1 mm in length

COLLECTION, PREPARATION, MOUNTING,

STORAGE AND PACKING FOR CARRYING OF MITE SPECIMENS

1. COLLECTIONS
Collections samples of mites depends of the habitat for the acari:
1. 1. Terrestrial Free Living Mite

Phytophagous, predaceous, mycophagous, saprophagous ground and soil form collected use of a Berlese Tulgren apparatus (Krantz, 1978). Mite sample from Berlese Tulgren funnels usually is made into a preservative such as 70% ethanol. Prolonged preservation in ethanol more than 3 months, mite should be collected to another preservative. A mixture of glycerine (50 parts), water (40 parts) and glacial acetic acid is recommended for this purposed (Krantz, 1978). A bulb aspirator of the Singer type used to collect mite from plant hosts directly (Singer, 1964).


Other methods are available to collect mite on plant:

1) Method for collecting mites on leaves one by one

2) Leaves with mites are cut with scisors collected in the field and fixed in 70 %

Ethanol.

3) Branch of leves with mites are set on the white paper, and beaten by

wooden bar in the field, mite collected fixed in 70% ethanol.



1. 2. Aquatic and Marine Mites
Aquatic mites (Actinedida, phalanx Hydrachnidia) are collected in a variety of ways, depending on whether the mites are free swimming, benthic or parasitic.

1. 3. Parasitic mites
Parasite mite of vertebrates is collection of the host habitat (nest, runway litter, mammal, birds etc). Nest and litter material lends itself well to Berlese Tulgren funnel separation, for mammal (example dead rodents) to brush with using fine-combing and brushing, mites will be fallen on the white paper or pan and will be used painbrush to collect the mite and fixed in 70% ethanol. Mite of invertebrate are found on their host in a wide range of location. Collection parasite and phoretic mite associated with insect may be removed with a fine brush, a pin or a pair of forceps in the surface of body insect under magnifying glass in the field or stereoscopic microscope in the laboratory. Most of the family of mite have a specific of ways to collect.


2. PREPARATION

Preparation of preserved material for use in optical of phase contrast and interference microscopy, it necessary to prepare specimens transparency and clear. Before mites specimens is placed on the slide necessary of clearing or digesting agent or dissection.

2. 1. Clearing agent.

Various chemical (lactophenol, lactic acid, Nesbitt’s fluid, Kono’s fluid) are effective in macerating internal tissues of preserved mites with little or no damage to the exoskeleton. Lactophenol is an acid corrosive, submersion of lightly sclerotized specimens for more than 48 hours tends to weaken leg conjunctiva and shield and makes dissection difficult. Formula of lactophenol: Lactic acid (50 part), Phenol crystal (25 part) and Distilled water (25 part).

2. 2. Dissection

Spesimens mite heavily sclerotized of the suborder Gamasida and Oribatida, are difficult to study only use of clearing procedures. The thickness of the dorsal and ventral shields prevents easy observation of surface structure in the microscope, needs to separate the dorsum and venter carefully with tool (microdissection forceps, minuten pin inserted in a wood matchstick). After clearing procedure (3-4 days in 60% lactic acid or lactophenol and etc. depend of sclerotized), and mite specimens to be dissected in clearing agent (Takaku, 2000).



3. MOUNTING TECHNIQUES
Some of the mite specimens (soft specimens) directly to use mounting techniques as clearing agent (Hoyer’s medium, Berlese, Nesbitt’s fluid), usually for temporarily /semi permanent specimens. Formula of Hoyer’s medium: Distilled water (50 ml), Gum Arabic (amorphic) (30 gram), Chloral hydrate (200 gram), Glycerine (20 ml); Nesbitt’s fluid: Cloral hydrate (40 gram), Distilled water (25 ml), Concentrated hydrochloric acid (2.5 ml).

Two methode for making slide specimens of mites or mounting techniques:

1) Semi permanent specimen: whole specimen mite after clearing agent or not

and mite after dissecting, put on center of object glass and mount with

Hoyer’s medium and put coverslip on specimens mite with medium and

give label should include data, host or substrate and collector as well as the

locality and date in right side and data taxa in left side. Slide specimens

stored on oven 40-45 centigrade degree for 2-7 days and after drying and

give a ring a waterproofing paint for sealing the coverslip to the slide

surface. This way is easier and better than other method.

2) Permanent specimen :

a. PVA, for mites of the family Phytoseiidae, Macrochelidae, PVA

(Polyvinyl alcohol-lactic acid mixture) medium is also good for

permanent specimen. But for other soft mites (e.g., Tetranychidae,

and most of suborder Actinedida), PVA will cause shrinkage and it

will difficult to observe specimens.

Specimens mites after clearing agent or and dissection and put on

center of object glass with PVA medium and coverslip put on

specimens mite and then give label should include data, host or

substrate and collector as well as the locality and date in right side

and data taxa in left side and stored on oven 40-45 centigrade

degree for 2-7 days.

b. Canada balsam

Methode to make permanent specimens by Canada balsam (Saito et all., 1993) (Fig. 1.):

Fixative: MA80 Sol. (80% MA Sol. (Diluted mixture of equal volume of

Methanol and Acetic acid by distilled water))


1. Fixed materials in MA80 Sol. For 1 or 2 hours (for Tetranychidae and Phytoseiidae,

at least 1 hours; for Macrochelidae, at least 2 hours).

2. After fixation, materials can be preserved in 70% ethanol or should be moved to

MA80 Sol. Before next procedure.

3. Equal volume of lactic acid added in MA80 Sol. Containing materials, and stored

in oven at 55-60 centigrade degree for 2 – 7 days (for Tetranychidae, 3-7

days; for Phytoseiidae, 2-3 days; for Macrochelidae, 5-7 days).

4. If materials are too large to macerate completely, it’s necessary to make minute

hole at any part of body which is not important taxonomically. If it’s impossible,

materials should be moved to 70% ethanol, and moved to 10% KOH and stored in

it for a few hours at 55-60 centigrade degree. After this, materials are moved to

70% ethanol.

5. In 70% ethanol with materials, a few drop of filtered 0.1% Evans Blue sol. Or 1%

Acid Fuchsine sol. Added for staining for 2-24 hours.

6. Staining solution removed from vial and 70% ethanol added.

7. Stored in 70% ethanol for 10 minutes, 70% ethanol removed and 75% ethanol

added.

8. 75%, 80%, 85%, 90%, 95% ethanol for 10 minutes, respectively.



9. 99.5% ethanol for at least 1 hour.

10.In different vial, 1/3 volume of alpha terpineol prepared and 99.5% ethanol added.

In this time, ethanol should be added gradually to prevent for mixing ethanol and

terpineol.

11.Materials gradually gone down to boundary of ethanol and terpineol, and after a

few hours, materials gone down to bottom of terpineol layer.

13.Ethanol layer removed and small amount of terpineol added.

14.Material with small amount of alpha –terpineol moved on glass slide.

15.Cover glass set on material. Canada balsam added from margin of cover glass, and

alpha-terpineol absorbed and removed by a piece of filter paper from opposite

margin of cover glass. These procedure done again 2 or 3 times, and terpineol

removed completely.

16. To remove air in balsam and material, and to remove alpha-terpineol from inside

of material, air and material warmed by soldering iron from back slide of glass

slide.

17. Slide specimens stored in oven 50 centigrade for 1 or 2 days in order to dry the



Canada balsam.


Fig. 1. Method permanent specimens mites by Canada balsam (Saito et all., 1993).



CATALOGING, STORAGE AND PACKING FOR CARRYING /SENDING OF MITE SPECIMENS

Cataloging

Cataloging is record of data mite specimen collected or exchange/voucher specimens. Process of cataloging from record of data mite specimens from field book/list, Register book/ collection number of mite, Book/list of genus/species, species, hospes and distribution catalog. All collections number of each species entered into database or catalog card.(Saim and Hartini, 1999).




Storage

Wet specimens of mite storage in ethanol 70%-80% in vial to big bottle and every big bottle storage on the rack of bottle collection. Mite specimens of slide/dry storage on the slide box and every box slide put on the rack of box slide with vertical position.



Packing for carrying /sending of mite specimens
Collection of mite specimens (wet or dry/slide) are easy to broken, so need carefully to pack for carrying/sending mite specimens.

Two ways of packing specimens:


A. Wet specimen:

Mite specimens in the vial 70% ethanol with label data, cover with tissue and

put in the center of box and contained with stereofoarm/plastic.
B. Slide/dry specimens:

Slide specimens with label data put on the small box and cover with tissue or plastic and then put in center of box contained with small stereofoarm/plastic.





REFERENCES

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Indonesia)
Ehara, S. 1966. The tetranychoid mites of Okinawa Islan (Acarina: Prostigmata). J.

Fac. Sci. Hokkaido Univ. Ser. 6 Zool 16: 1-22.
_______ 1969. Spider Mites of Sugar Cane in Eastern Asia (Acarina: Tetranychidae)

J. Fac. Educ. Tottori Univ. Nat. Sci. 20(1): 19-25.
_______ and T. Miyashita. 1962. A spider mite infesting rice in Japan. Plant Protect.

16: 55-56.


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