Wah vtboun pe ris < “euater. oY pert . LECTURE #1 TECHNIQUES AND APPAKATUS EMPLOYED IN ENTOMOLOGICAL RESEARCH 1, Collection & Preservation Collection net Killing bottle Forceps Setting board Insect collection box Alcohol Phenyl tablets ‘Camphor Propeacges 2. Identification roscope 2D ‘Microscope 3D Glycerin jelly | Glycerin . Slides Hot plate Needles: blunt and pointed Genitalia vial Pemeapep 3. Raring in Laboratory a. Cage or Incubator b. Required Plant or food ~ c. Brushes d. Humidifier e. Air Conditioner Or Heater 4, Raring in Field a. Field Cage for plant to be studied 5. Temporary slides ‘6. Permanent slides 7. Sampling methods 8. Pesticide calibration |V ntomoleyy Bteitction LECTURE #2 COLLECTION AND PRESERVATION OF INSECTS The behavior of insects can be observed most easily in their natural environments. lowenel many species, especially the smaller ones, must be collected and properly preserve before they can be identified. Because correct identification seldom is easy, it is important specimens be preserved in the best condition possible. The identification of a particular insect or mite usually requires examination of minute details of its anatomy with the aid of a hand lens or microscope. Some specimens may require dissection or even study with the electron ‘microscope. If these details on a specimen are concealed, missing, or destroyed because of improper handling or preservation, identification is made difficult or impossible, and information about the species to which it belongs cannot be made available. Therefore, adequate preservation and proper labeling of specimens are essential to their identification. The methods used to collect insects and mites are dictated by the ultimate goal of the samples collected. Insects may be collected as a hobby for personal enjoyment of their diversity and beauty. They may be collected in conjunction with school courses on biology or entomology. Specific insects groups may be sampled to assess or measure biodiversity to help identify ehbropniate arcas to be included in reserves. Aquatic species may be used to detect changes in Water quality, Pest species may be sampled to assess presence/absence or abundance in order to determine whether control measures are necessary. Specific groups or species may be collected to acquire material for biological, physiological, ecological, molecular, and * systematic studies. Equipment and Collecting Methods (1) Forceps. Fine, lightweight forceps are recommended; if sharp-pointed forceps are used, care must be taken not to puncture Possible, grasp specimens with specimens. If the part of the forceps slightly behind the points, ‘ (2) Vials containing alcohol or other preservatives @) Killing bottles of various sizes {& Small boxes or containers for storing val from killing (6) One or more aspirators, (7) Absorbent tissue for Use in killing bottles and aspirators,Killing Jars or Bottles To make a cyanide killing jar or bottle, place a layer (about 15 mm) of cyanide erystals in the bottom. Potassium cyanide is best; sodium | cyanide is as effective but is hygroscopic, that is, it absorbs water and makes the jar wet; and calcium cyanide is seldom available. Cover the crystals with about 10 mm of sawdust and then add about 7 mm of plaster of paris mixed with water to form a thick paste, working quickly before the plaster solidifies. Then add crumpled absorbent paper to prevent water condensation on the inside glass surface. Instead of the plaster of paris, a plug of paper or cardboard may be pressed on top of the sawdust. Be sure that it fits tightly. When ready to use after a few hours, place several drops of water on the plaster or paper plug. In an hour or so, enough fumes of hydrocyenic acid will ave Been produced to make the jar operative. Do not test this by sniffing the open jar. Liquid Killing Agents . ‘Among the liquid killing agents are ethyl acetate (CH3CO2 * C2HS), ether (diethyl ether, C2HS * O + C2HS), chloroform (CHCI3), and ammonia water (NHSOH solution). Ethyl ‘most widely used. All of these chemicals are extremely volatile and flammable and acetate should never be used near fire. Children should only use them under adult supervision. Ethyl 2 Sos are Kaihas os Emomtagy 2 hol ‘ ling agent. Ethyl Alcol satisfactory liguid kill tera, and sate is regarded by many as the most s ‘adalts, small Hymenoptera. acetate 18 reg Kl small COLEOpIE!® 240 ed at TL-o% idely used to J sofi-bodied insects. It is most (ethanol or ETOH) is wi many immature insects ani concentration Aspirators and Suction Devices . jent and effective device for ‘The aspirator, known in England as a ‘pooter.” is @ convenient a od effete euboct an collecting small insects and mites. The following materials aspirator: () Vial 2.5-5 em in diameter and about 12.em long. (2) Two pieces of glass or copper tubing about 7 mm in diameter, cone piece about 8 cm long and the other about 13 cm long. (3) Rubber stopper with two holes in which the tubing will Gt snugly. (4) Piece of flexible rubber or plastic tubing about 1 meter long, with diameter just large enough to fit snugly over one end of shorter piece of stiff tubing (5) Small piece of cloth mesh, such as cheesecloth, and rubberband. Malaise Traps One of the most widely used insect traps was developed by the ‘Swedish entomologist René Malaise and that now bears his name. Several modifications of his original design have been published, and at least one is available commercially. The trap, as originally designed, consists of a vertical net serving as a baffle, end nets, and a sloping canopy leading up to a collecting device. The collecting device may be a jar with either a solid or evaporating killing agent or a liquid in which the insects drown. The original design is unidirectional or bidirectional with the baffle in the middle, but more recent types include a nondirectional type with cross baffles and with the collecting device in the center. Malaise traps have been phenomenally successful, sometimes collecting large numbers of species that could not be obtained otherwise. Attractants may be used to increase the efficiency of the traps for special purposes. Pitfall Traps Another simple but very effective and useful type of interception trap consists of a jar, can, or dish sunk in the earth (fig. 10). A cover must be placed over the open top of the jar to exclude rain and small vertebrates while allowing insects and mites to enter. A piece of bark, ‘wood, or flat stone will serve this purpose. Pitfall traps may be baited ‘with various substances, depending on the kind of insects or mites the collector hopes to capture. Although most that fall into the trap will remain there, it should be inspected daily, if possible, and desired PVA NSE 8s Nana Rte £et E ig bottle while they are in their best Specimens removed and placed in alcohol or in a condition, Screen light trap Light Trap Mounting specimens Material Required for Mounting Specimens Pinning of Different Insects mason dB ‘A,B. Grasshoppers, C. Bugs, D. Bees, E. Beetles , F,G. Butterflies ee 1 toe kt1 Late mot A point mounted specimen Method for Shipment of Specimens Specimen to send other countries. ‘Must write on box. Specimen for Study Purpose and No Commercial Value Data Label Information: Locality Date of Collection Name of Collector Host plant Technical nameyr ? , LECTURE HED RARING OF INSECTS 1, FOOD: For phytophagous insects, the host should be found ut and the plant material Tequired by the insect should be placed in the cage. Food supply should be fresh and changed everyday to prevent decaying. Unused food material as well as the fatcal miatter should be semoved everyday and the cage should be cleaned. Different species Tequire different types of food material, which should be provided in sufficient Quantity throughout the period of rearing. For example, Papilio demoleus can be reared on citrus leaves, Erias vitella on lady fingers and Leucinodes by providing it with brinjal frit, In some cases an artificial diet, if available, works very well if natural food is not available in certain season Polyphagous peeties can’ generally be reared on cruciferous or leguminous vegetables as for example Heliothis armigera and Spodoptera litura can be reared on a variety of vegetables and pulses. 2. TEMPERATURE: Majority of insects breed at temperatures ranging from 25-350C and undergo diapause at lower temperatures. However, there are species, such as Heliothis armigera that can be reared at room temperature even in winter months. Successful rearing can be done during the natural breeding season of a particular species! when conditions are optimal. But on small scale insects can be reared in rooms having temperature control devices like BODs. Special care has to be taken while rearing temperature-sensitive species. Stored grain pests can generally be reared in small spaces where temperature can easily be maintained. For some species lower temperatures and high humidity has to be maintained in summer. 3. HUMIDITY: A vast majority of insects prefer high humidity (80-90% RH) for their natural development. Humidity can be increased in cages by spreading water-soaked sponge or wet sand at the bottom of the cage, particularly in summer. A large umber of insects are easier to rear in rainy season due to naturally available optimum temperature and humidity. 4. SPACE: Space requirements differ according to the species to be reared. For example, while butterflies ‘and moths require plenty of space for nuptial flying, stored grain insects can breed in small chambers over even in glass jars. Cages are, therefore, designed keeping in mind the specific space requirements of the insects to be reared. Generally larger and flying 1 wan Kiaieingots require me anu! must be reared ore space J in separate char| ; i i a ae aS Or pe “ot Y of eS SF oon el 23 © PE Pret) Tg 22 For gst iss Pros i 2 Wt 9X, DS se oe 243 A sk sod wt ee & g - & ace as compared to the smalle Separate chambers, insects require more g and must be reared inLECTURE #64 AN ACCOUNT OF REARING METHODS GF SOME INSECTS OF ECONOMIC IMPORTANCE Rearing field-collected larvae to adult stage seule fetal ieee ee ant feeding insects can be placed in a jar or wide-mouthed : m which they were collected. The opening of the jar should be closed with a muslin cloth for sufficient aeration. Feeding material should be changed everyday and the container cleaned to avoid fungus growth and other infections. The larvae will grow and enter pre-pupal stage, when they will stop accepting food and will restlessly look for suitable place for pupation. No feeding material should be provided after that and the container should be left clean. Sometimes bits of crumpled paper or corrugated cardboard placed inside the container will help the searching caterpillars to find suitable places for spinning cocoons. Pupae should be kept at suitable temperature for proper development. Adults will emerge’ after completion of the pupal period. Adults may be fed on 30% honey and water solution if required to be Kept alive for egg-laying. Many species can survive and mate in captivity and lay eggs on the host plants provided in the cages. Exopterygotes are easier to rear as their nymphs as well as adults require same food and habitat and need lesser care. Gregarious insects such as aphids, hoppers and bugs can easily be maintained in large numbers in cages. \ Multipurpose Rearing Cage Multipurpose rearing cage is designed to breed various kinds of insects in laboratory. It contains a cuboid wooden frame with a firm wooden bottom and a sliding glass door in front as shown in the diagram. Wire mesh is fixed on the back and the top sides, while on the two lateral sides, muslin cloth with a sleeve attached to it in the middle is attached, so that hands could be inserted through the cloth sleeves and insects could be easily handled inside the cage without danger of their escaping out. Sand or sponge could be spread on the floor ‘and moistened periodically to provide optimum humidity to insects. A clay pot with host plant can be kept inside the cage by opening the front sliding door. Even twigs dipped in a water container can be used as food for larvae. Feeding material should be changed everyday and faecal material and dried plant material should be removed from the cage. Many types of insects, particularly the phytophagous species can be bred in this type of cage for several generations and a culture can easily be maintained for experimental purposes. Rearing insects on potted plants Some insects do not require much caring such as aphids and hoppers, and can be easily maintained for several generations by holding them on potted plants. Three or four sticks can be inserted in the soil of the clay pot vertically and a muslin cloth should be wrapped around these sticks, so that the insects are confined to the plant and are not allowed 1ieee =. ° je afcttonts rm Etoneligy insects can be Only small insec them from predators. mall 7 ee atta rs, scale insects, mealy bugs and paar tic can be used in place imes Mylar plast on ee This method is also commonly to escape. Also a cloth barrier wi reared by this method, such as aphids, is minimal beetles, whose space requirement i of muslin cloth but there should be provision for geratio! used to set upfield experiments under natural conditidns. Rearing stored grain insects . Sitophilus etc. are Insects infesting stored grains, e.g. Trogoderma, Ephestia, Corcyra, comparatively easy to breed in captivity, as their requirements for food, space and humidity are not as rigid. Almost all of them can be easily reared through successive generations in wide glass jars in which food material has been filled to about half and the opening covered with muslin cloth. Almost no further caring is required till the food supply is exhausted. However, low temperature induces diapause in stored grain insects and therefore suitable temperature should be maintained in winter months. The culture jars should be placed inside the water-filled trays to keep ants away from contaminating the cultures. Stored grain insects can be maintained in large numbers with minimum of efforts and are ideal experimental animals. Rearing houseflies In nature houseflies breed in cow dung or other decaying matter but in laboratory they an be reared on a paste made of milk powder or agar powder with a little yeast added to it as mixture should be allowed to ferment for at least 24 hours. A tray containing this type of Sa elle ES be placed in any type of cage in which adult flies can be confined for . (iene Aaa ay be collected from the field with a sweep net and released inside Wit rn il readily lay eggs inthe rearing medium, after which they can be released, 'ge0%s will hatch from the eggs within 26 hours and will move about actively in ge 345 gs EP =e ge 38 32 Ae ge ge = zh Sok 5 e as se a3 ae 7” ge as sf B= BS = 2 seI be fed on blood before they can lay eggs. Males can be fed on 30% honey and water solution so that they can survive and mate with the females. In order to provide blood meals to the female adults, either rats or pigeons are commonly used which should be shaven or defeathered on some body parts for easy feeding by mosquitoes. A separate cage can be used to provide blood-feed or if the cage is sufficiently large, the animal can be placed within the same cage for about an hour. Care should be taken to tie the legs and wings of the animal before keeping it in the cage so that it is not able to run around in the cage and disturb or destroy things or cut holes in the cage. gels 1 ema Field-collected adults can be used to start the culture, But if pure cultures of a particular species are required, then the field-collected adults should be identified and sorted to species level before introducing them into separate cages. In winter higher temperature has to be maintained for the sitccess of the culture.LECTURE# 5 REPOR WRITING Writing a report depends on the type of experiment but generally report may contain: (a) summary or abstract () introduction (c) materials and methods @ results and discussion <> Qanchssien (e) recommendations (f) references. For the better presentation any reputed journal of related filed could be followed.lectur>G 8 ‘Taxonomic Study of Spilomelinae (Lepidoptera: Pyraloidea : Crambidae) from China Chapter 3 General Morphology 3.1 Body form and sotiraton/ Head, thorax, patagium, labial and maxillary palpi, frons, wings and abdomen color and shape were presented and described. Wing size is described in millimeter (mm), the patterns, antemedial line, antemedial spot, discodial spot, postmedial line and subterminal lin and fringe color and structures were briefly described. discodial spot antenna Jabial palpis antemedial line subterminal antemedial 7 line fringes discoidal spot4 yw Pyraloidea : Crambidae) from China 16 ‘Taxonomic Study of Spilomelinae (Lepidoptera: 3.2 Wing venation ~~ Position of different veins (Radial, Medial, cubitus and anal veins) with proportion and position of discal cell were described. Fig. 3-1 C: Costa, Sc: Sub costa, Ri: Radial vein 1. Ra: Radial vein 2, Rs: Radial vein 3, Ra: Radial vein 4, ‘Ret Radial vein 5, My: Medial vein 1, Ma: Medial vein 2, Ms: Medial vein 3, Cuy: cubitus Vein 1, Cuz: cubitus vein 2, Avs: Anal vein 1 and 2, As: Anal vein 3 3.3 Male genitalia we Different parts of male genitalia and anal tube, including aedeagus, uncus, valva, gnathus, transtilla, harpe, juxta and saceulus are described. Fig, 3-2 A: Anal tube, U: Uncus, V: Valva, G: Gnathos, T: Transtilla, H: Harpe, J: Juxta, S: Saccus, C: Comuti,Error: Insufficient ory opkd Remale Senitalja | ror Peale ial pats anteri ior and posterior epophysis, luctus birsae, corpus bursae and Signum were described in detail dani, CB: Corpus bars 1 sae, S: Signum, CB: Corpus i is, DB: Ducts burse,S: Si ; is, A: Anterior apophysis, ‘J : Posterior apophysis, Fig. 3-3 PA: