Rearing Mosquitoes - [PDF Document] (2024)

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    Pubiished III.


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    Insect Control and Research. Inc.

    IIII Sorth Roiling

    Road, Baltimore, Jlcl. zrzz8

    Bulletin 30. 3

    Bulletin SO. 5

    is a\.ailable from:

    T. G.


    Executive Secretnry, ;IMCX


    Box 278, S&m, California 93662

    Price 53. j0

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    FOREWORD-KESUETH i_. KTISHT. . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . , , , . . , .



    . . . . . . . . . . . . . . . . . . . . . . . . . . .


    ITTRODL7CTIOT . . . . . . . . . . . . . .


    I. THE ITSECT_ \R1-



    Construction of room>.



    Temperature and humldlt equipment. . . . . . .



    SeiuritJ- and safet>- medrure>.

    . .

    -1. Faslllt> requirement< .

    I. Building . . . . . . . .

    2. Room layout . . .

    . .


    B. Labor&tory

    and rearing prcxedures.


    I. Eggs ...........................


    Larl-at and



    3. _klults .........................


    . .


    C. SIiscellaneow requireme nts an d recom menda tion\




    Equipm ent for handling adult>. . . . . . . . . . . .


    ;Ispxators or wition deliies.

    . . .

    2. cage> . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . .

    .Adult food . . . . . . . . . , . I. .


    _irtificial feeding techniques. . .


    Induced rndtmp . . . . . . .

    ....................... . .

    Colleiting and handling eggs .. . . . . . . . . . . . . . ..


    Larl al rearing ttihnia~ueh dncl tquipnmt


    Counting larvae


    Rearing tra -s ..................................

    Handling larl ae ...............................

    Equipm ent for feecling 1arTde.

    Lxval toocf ancl ieedm:

    ....... ............

    _ .

    c ..............................


    counting. separating and sexing pupa e.



    . . .

    . .


    culture . . . . . . . . . . . . . . . . . . . . . . . . . . . ..

    2 j


    . .

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    ....... qj









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    . . . . . . . . . . . . . . . . -. . . +. e .. . . . . . - . . .. . . . . . . . . . . . . . . . . . . . . . . . .



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    . . . . . . . . . . . . . . . . . . . . . .

    . , .

    . * . ,



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    . . . . . . . . . . . . . . . .

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    . . . . . . . . 8 . . . . , . . . . .


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    Hcienzngogm . ...,,..*.. . *. . . . . . .a

    . . . .


    Opifes . . . . .

    . . . . . *

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    . . . . . .

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    . . .

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    * . , .


    APP END IX A. List of mosq uitoes and mosquito coloniesmaintained b>- laboratories, . . . . . . . . ig

    .IPPEN DIX B. Tables of measu res and equivalents.

    . . . . . . . . . . . . . . . . . . . . . . . . . . . . , , .85

    APP END IX C. Relative humidity tables.

    . . . . . . . . . . . . . . . . . . . . . , . . . . . . . , . .. . . . . . . . . . , .


    REFE RENC ES . . . . . . . . . . . . . . . ...~...~............................................ gI


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    FIG. I.-Interior of insectary at V-alter R eed _k-m - Instituteof Resea rch--US. _km y.

    FIG. Z.-Interior or insectary at Insect Control k Rese arch .Inc.-R. Stacks. Baltimore

    FIG. 3.--Mo rlan s

    colony cage..-CDC.


    FIG. J.-Gerbergs collapsible cage-R. C. Ruehl. Jr.: Cornell C sE Co.


    FIG. 5.-Barraud cage-P. S. Barraud.


    FIG. 6.-Gillett cag e-J . D. Gillett


    FIG. 7.-Plastic cage-L7 .S. *Arm.............................................

    FIG. 8.--hicCra -s escape-proof colon , cage -CDC .


    FIG. 9.-Pollard cage -D. G. Pollard.


    FIG. I o.-_knimal restrainers in

    more Sun . . . . . . . . .

    mass feeding of

    . . . . . . . . . .

    mosquitoes on

    . . .



    . . .



    . .

    chicks-R. Stacks.

    . . . . . . . . . . . . .


    .Co\-er I

    . . .


    . .


    . . .


    . . .



    . . . . . 12

    FIG. II.--Apparatus for mem bran e feeding of mosqu itoes--U.S.;irmy.

    . . . . . . . . . . . . .


    FIG. I z.--_\ppara tus for mem bran e feeding. detail--US. -Arm>-.

    . . . . . . . . . . . . . . .

    I 5

    FIG. r3.-*kliquot container for mosquito larvae-R. Stacks.Baltimore Sun.


    FIG. I _I.--\Vhite enam el tra s for rearing larvae-L.S. _Lrmy.


    FIG. IT.----SYhite en amel dishes for rearm g larvae-K.S.D._L.

    .................... .

    . .


    . . 20

    . . .


    FIG. 16.-Construc tion of gall-anized metal rearing tra--Morlan. . . . . . . . . . . . . . . . . . .


    FIG. r;.-Mass rearing of msoqu itoes in

    galvanized tray s-R. Stacks. Baltimore Sun. . . . . . 22

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    All Metal Collapsible Cage

    (See Fig. 4, Page 9, This Bulletin)

    The Standard Cage in Major Laboratories

    All aluminum construction with plastic ham mock

    Metal construction eliminates mold and contamination

    Hinged for easy collapsing and storage

    Cages easily disassem bled for cleaning

    Escap e-proof feeding ham moc k enables e xposu re of animal forblood feeding with-

    out mosquito escapage

    Sucrose solutions on cotton balls can be fed mosquitos throughhammock ; sugar

    does not com e in contact with metal parts.

    Surgical stockinet sleeve provides easy access inside cage

    Econom ical in cost - efficient in operation

    Available in three

    convenient sizes

    Gl 12 x 12 x 12

    G2 24 x 24 x 24

    G3 177/8x 18~ 22-1116

    For further information and prices write-


    Whe ther for mosqu ito-rearing or experimen tal conditions,there

    is a Standard Humidifier available to provide accura te,auto-

    matic humidification for your needs . Famo us Standard units

    are engineere d for long, trouble-free service an d easyinstalla-

    tion, and are priced surprisingly low. All units are fully


    Wfite for 4-page

    free brochure


    Model 42L


    State de tails of your

    project for promp t




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    On behalf of the members of the American Mosquito ControlAssociation and

    of mosquito workers everywhere, I first thankfully add this finepublication to my

    library of mosquito literature and secondly extend sincerestthanks to its autho r

    for his devotion to its preparation.





    Department of Entomology

    N. C. State University

    Raleigh, N. C. 27607;

    Chairman, Editorial Board of

    Mosquito News, Journal of the

    American Mosquito Control Association


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    This manu al is dedicated to Mrs. Helen Louise TrembleyDurkee,

    who devoted a large part of her time and effort to producingthe

    original B ulletin 3, entitled Mosquito Cu lture Techn iques andExperi-

    mental Procedures. Her work has served for many years as aguide

    to entom ologists and other w orkers in the field ofculicidology.


    The preparation of the present manual was originally intended tobe the work of a

    committee. However, committees perform best when they can meet,discussand work

    together. Because distance and individual research problems madeeffective commit-

    tee work very difficult and subject to long delays, I decided totackle the problem

    alone, with whatever local assistancecould be obtained. I wasfortunate in having

    Capt. James W. Gentry, U.S.A. (Ret.), join our staff in 1967. Hespent many eve-

    nings gathering data. Dr. Ronald A. Ward, at the Walter ReedArmy Institute of

    Research, was extremely helpful in securing literature andlocating mosquito rearing

    laboratories. This, then is the working Committee to whom Ioffer my sincere thanks.

    Dr. Ross H. Arnett, Jr. kindly reviewed this manuscript and usedhis grammati-

    cally minded red pencil with justifiable abandon. My sincerethanks and appreciation

    are offered-any errors remaining are mine, not his.

    To the many other culicidologists that assisted n one way oranother, I give my

    grateful acknowledgment for help and advice.



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    The need for information on mosquito rearing has beenintensified by the increased

    interest in mo squito biology, control, chemoltherapy ofmosquito borne diseases, and

    in other investigations in which mosquitoes are used asexperimental animals. Since

    the publication of Helen Louise Trembleys Mosquito CultureTechniques and

    Experimental Procedures

    in 1955 , published literature in this field has increasedlike

    salt marsh mosquitoes after a high tide.

    The acquisition elf knowledge concerning proper con trol ofmosquitoes and mos-

    quito borne diseasesrequires that studies be made of thebiology, p hysiology, anatomy,

    genetics, taxonomy and ecology of the insect. Insecticideresistance, biological, chemi-

    cal, and integrated control must be investigated.

    Research on mosquito genetics

    may have far-reaching consequences in the control of m osquitoesand m osquito-borne

    diseases. Th e use of m osquitoes as screening agents forpesticides or chemothera-

    peutic compounds requires large numbers of mosquitoes. Allphases of mosquito

    research usually use individual or quantity rearings ofmosquitoes.

    Mass rearing of

    mosquitoes for control purposes may become, in the near future,as common as mass

    production of pesticides.

    The purpose of this man ual is to provide information on therearing of mosquitoes.

    A thorough search of the literature p rovided many usefulreferences. The great

    amou nt of literature made this no easy task. Undoubted ly,articles were overlooked,

    but it is hoped that no serious omissions have occurred. Anapology is offered to

    those authors so neglected.

    Author credit-lines omitted in the text are included in

    the list of references at the end of the rearing description forthe species.

    For some

    species, particularly those reared in the authors laboratory,the technique is described

    in more detail. Often this sam e technique will apply forrelated species as well.

    No method for rearing mo squitoes is guaranteed. Perhaps themost important

    requirem ent for successful rearing o f mosquitoes is attentionto detail. Mosquito

    rearing to be successful requires attention 24 h ours a day, 7days a week.

    The basic

    rules are: avoid over-crowding mosquitoes, and overfeedinglarvae; a void pesticide

    contamination, observe temperature and humidity requirements,standardize rearing

    methods and avoid nonstandardized food.

    Observance of these principles should

    produce uniform animals.

    The following guide to mosquito insectary construction andrearing practices is

    presented step by step with no elaboration of theory. Much ofthis theory m ay be

    yearned by reference to the literature cited.



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    the room.

    Whenever possible use coved wall floor junctures and caulk andseal the

    cracks and crevices n the walls.

    The best insectariesare windowless with low ceilings

    (not more than 8 also painted white or light colored. Ifpossible, use recessed

    lighting fixtures that are flush with the ceiling.

    Smooth floors without a floor covering are recommended. A lightcolored epoxy

    floor paint can be used to improve the appearance. The doors ofthe rearing area are

    best constructed of metal and painted white or light colored.Weatherstripped door

    frames and jambs are necessary for a tight fit. Overlappingmarquisette curtains

    hung on the inside of the door frame provide additional mosquitobarriers (see sec-

    tion on mosquito security).

    Hot and cold running water and a sink with ample drainingsurfaces are required

    in the rearing room. A water blending tank r installed above thesink facilitates

    the filling of trays, separation of pupae and other tasks thatrequire water to be a

    certain temperature. Several water outlets around the room willavoid the clutter of

    hoseson the floor.

    Movable tables, benches, chairs, storage cabinets, etc., arepreferable, since, in

    general, permanently installed furniture prevents newarrangements to meet changing

    conditions. Some laboratories have all furniture mounted oncasters or wheels so

    that it can be readily moved.


    Temperature and humidity controls are probably the mostimportant factors in the

    successful earing of mosquitoes. The simplest method ofproviding regulated heat

    and humidity is the use of a small electric light bulb and a wettowel draped over

    the cage. Th

    e most elaborate is a complex environmental chamber withprogrammed

    electronic controls of temperature, humidity, andphotoperiod.

    The size of the

    insectary will regulate the type of temperature and humiditycontrol system required.

    The proper design of equipment for the production, control, andrecording of tem-

    perature and humidity may require the services of anengineer.

    If a large installation is planned, it is better to have


    smaller heating and cooling

    units than one large one.

    For example if it is determined that 6 tons of aircondition-

    ing is

    required, use two S-ton units rather than one 6-ton unit.

    Install the heating

    and air conditioning system of the peripheral area so that ifall the inner area systems

    fail, the peripheral system can be shunted to the inner area.When there is a possi-

    bility of contamination from pesticides from adjoininglaboratories, install air condi-

    tioning equipment to insure that the outside air intake does notpick up contaminated

    exhaust air.

    Automatic thermostat controls for the heating and airconditioning

    system are best.

    Humidity regulation is usually provided by comparatively smallcommercially avail-

    able units.2

    If a source of clean compressed air is available, the air supplyand water

    supply can be connected and regulated by solenoids andhumidistats. The water is

    forced out through fine nozzles installed in a hose and producesa fine mist. Steam

    units are available which will provide warm moisture. Variousimprovised humidi-

    1 Sarco Co., Inc., Allentown, Pa. Blender Type DB.

    2 Walton Laboratories, Inc., Union, N.J., Standard EngineeringWorks, Pawtucket, R.I., Bete Fog

    Nozzle, Inc., Greenfield, Massachusetts.


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    fying methods have been described, including such methods asusing wet towels,

    blowing a fan through layers of wet excelsior or gauze; steamfrom boiling water or

    from radiators; wet sand; chemicals in water.

    In determining the type of humidifier unit that should be usedin a rearing room,

    certain factors must be taken into consideration:

    (I) The relative humidity desired and what variation would bepermissible; (2)

    Room size-length, width and height; (3) A

    verage temperature; (4) Heat factors,

    i.e., number of people or animals in the room; number of lightsand wattage, other

    sources of heat, and heat loss; (5) Construction of room-walls,floors, ceiling, parti-

    tions, windows; (6) Openings to other rooms; (7) Exhaustfans-capacity. As in

    temperature control, it is better to have two smaller units thanone large one.

    Use a recording hygrothermograph

    for a permanent, continuous record of tem-

    perature and humidity conditions.

    Humidity can be checked with a sling psychrom-

    eter, a bulb psychrometer, or a Honeywell hygrometer withelectronic sensors which

    will show the relative humidity at the exact location desiredwithout disturbing air

    movements. Although it is recognized that the critical factor isthe rate of evapora-

    tion, usually an indication of the relative humidity issufficient for successful earings.

    A maximum-minimum thermometer will indicate the total range oftemperature

    change for any desired period.


    Barlow (1961); Bertram and Gordon (1939); Buxton (1931);Flitters (1964); Haufe (1964);

    MacPhee and Patterson (1958)



    rovost et al. (1965); Wharton and Kniille (1966); Winston

    and Bates (1960).


    The photoperiod and light intensity affect the development ofthe various stages

    in the life cycle of the mosquito and are the subject of a greatmany papers (see

    references). In the insectary, a cycle of 14 hours of light andIO hours of darkness

    appears to allow the best and most uniform development, withoutinterfering with

    normal working hours of the staff.

    Some speciesmay require a crepuscular period to swarm and mate.The automa-

    tion system described by Levin, Kugler and Barnett (1958)coordinates the numerous

    pieces of electrical equipment that regulate temperature,humidity, and light. This

    type of equipment can be readily modified to dim lightsgradually for dusk effect,

    turn off all lights for darkness, then turn up lights graduallyfor dawn and turn on

    full lighting for daylight.

    The standard fluorescent lights usually supply adequate lightfor daylight.


    especially made fluorescent lamps generate light at wavelengthsapproaching day-

    light, when required by exacting experimental work.

    Incandescent lights connected

    to a variable transformer can be used for gradually diminishingor increasing light

    intensity. A bl

    ue or blue-green light stimulates swarming of some species(Bates

    949; TheOdOr and ParSOnS 1945).

    Commercial light meters are used to measure light intensity.Evans (1961) de-

    scribes a portable visual photometer for measuring very lowlight intensities. The

    1 Bendix Corp., Environmental Science Division,


    Model NO. 594,

    Belfort Instrument Company, Baltimore, Maryland 21202.


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    scale of a regular Weston light meter used for photograph y canbe converted e asily

    to foot-candles by multiplying the reading by


    References- I

    Bates (1941)


    Belton et al. (1967) ; Brennan and Harwood (1935)


    Callot (1965)



    (1963); Evans (1961); Flitters (1964); Fowler et ctl. (1958);Haufe (1962); Hubert et al.

    (1954); Jobling (rg35), (1937); Killough and Weidhaas (1963);Levin et al. (1958); Nielsen

    and Haeger (1955); Omardeen (1958); Parker and Rozeboom (1960);Seaton and Lumsden

    (1941) ; Tate and Vincent (1932) ; Theodor and Parsons(1945).


    The escape of possible insect vectors of disease, like therelease of pathogenic

    organisms cannot be tolerated.

    The responsibility of the entomologist rearing mos-

    quitoes is similar to the responsibility of the bacteriologistcultivating pathogenic

    bacteria. Certain routine sec urity measu res are practiced bylaboratories involved in

    the rearing of medically important arthropods.

    The Comm ittee l on Experimental Use of A&s aegypti of TheEn tomolog ical So-

    ciety of America has developed a set of recomm endations onmosquito security and

    surveillance that serves as a guide for mosq uito rearinglaboratories. The follolwing

    recomm endations closely follow, but are not an exact copy ofthe ESA report.





    Window less construction, if possible, is preferred. If window sexist, keep locked.

    A sign perman ently fastened near the lock indicating that thewindows always should

    be locked, will help assu re this protection.

    Glass brick can som etimes be used to

    replace windows.

    Screened windows norm ally are not to be used, but if forsome

    specific reason they are required, double screens with


    mesh screening and an addi-

    tional outside protective screen of heavy hardw are cloth willprovide the required


    2. Room layout

    a. When the rearing room and laboratory are not one and thesame, then the

    rearing room should be an annex of the laboratory room and openinto it,

    If another

    door exists or is required for safety reasons it shou ld be marked for emergency use

    only .

    Plan exits from the laboratory roo m o r rearing room (if not anannex to the

    laboratory room ) through an ante-room , i.e. a small vestibulewith entry or exit doors

    or through another small room.

    A com pletely empty ante-room is best, with white

    or light colored walls and ceiling.

    Post signs to be sure one door is closed before the

    other is opened.

    Protect the entrance to the ante-room from the laboratorywith

    some protective barrier such as close weave nylon netting or anair curtain.a

    b. Paint the walls and ceiling of the laboratory, rearing room,ante-room and other

    work areas with a light shade of paint (gloss ename l, epoxy,etc.), preferably white.

    Light colored ceramic tile may be substituted for paint.


    Committee members: G. Craig, Jr., E. J. Gerberg, C. Judson, D.W. Micks, M. W. Provost, I-I.

    F. &hoof, C. N. Smith.

    2 Ecco Aire, Inc., New Castle, Pa.; Dynaforce Corp., QueensVillage, N.Y. 11428; Penn Ventilator

    Co., Inc., Philadelphia, Pa., 19140; Neico Products, Inc., SanFrancisco, Calif. 94124.


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    c. Screen all floor drains even though they enter a closedsystem. It is best to

    drain into a tank or dry well in which insecticide is regularlyintroduced.

    d. Screen or filter all air vents to the outside.


    I. Eggs

    (a) Store eggs in a container in the rearing room.

    (b) Treat surplus unhatched eggs with hot water or chemicals tokill them. If

    possibleburn the old egg papers.

    2. L.uruae and Pupae

    a. Larvae

    ( I) In laboratories that do not maintain

    containers with tight fitting lids to

    adults. In mass-rearing laboratories,

    several larval rearing racks are housed

    b. Pupae

    ( I) Collect pupae routinely, so that no


    T-day-week operations, rear larvae in

    prevent the escape of any emerging

    normally on a T-day-week operation,

    in enclosures nside the rearing rooms.

    adults emerge in the larval rearing

    (2) Place pupae in special containers inside cages. Provide eachcontainer with

    a lid so that the container can be closed after the pupae haveemerged. It

    should be removed from the cage through a sleeve.

    (3) Treat unneeded larvae, pupae, and discarded rearing water byheating or

    with chemicals so that only dead waste materials enter thesewage system.

    3. Adults

    a. Use sturdy cages, preferably of metal, fitted with a surgicalstockinette sleeve.

    When used properly the tight-fitting sleeve will allowmanipulation of objects within

    the cage. Th

    e mosquitoes obtain their blood meals by feeding on the hostthrough

    the screening of a nylon mesh hammock.

    This is preferable to introducing the verte-

    brate host animal into the cage.

    b. Keep the rearing room neat and clean.

    Remove excess sugar pads, fruits, etc.

    c. Hosts for blood feeding.

    (I) Provide animal rooms separate from the rearing room. Checkhost animals

    to be sure they are free of mosquitoes.

    d. Transfer of adults.

    All insects ransported from one secure laboratory to anothermust be housed within

    a screened, securely closed container.


    I. Train employees to handle mosquitoes and make them aware ofthe potential

    hazard of escapees.

    Loose mosquitoes endanger the entire project. Even one

    loose mosquito must be killed or caught immediately.

    A sign in the rearing room

    indicating the name of the individual responsible for themosquito security helps

    to maintain vigilance.

    Replace careless workers or technicians with competent,

    conscientious personnel.

    2. Do not remove any living stage of the insect from securityconditions without

    taking proper precautions.


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    3. Place surveillance oviposition-traps (Fay and Perry, 1965 ;Fay and Eliason, 1 966)

    in the laboratory, rearing rooms and adjoining rooms. Numb ereach trap and record

    on a map. Keep a record of the weekly check.

    Place additional surveillance oviposi-

    tion-traps outside within 25 feet of the laboratory bu ilding,at approx imately r5o-foot

    intervals. Rem ove all potential breeding containers from thesurveillance area. (For

    detailed information on preparation of surveillanceoviposition-traps, and selection of

    sites, see Ovipos ition trap reference handbook , CD C, Aedesaegyptt Handboo k series

    No. 6, 1967.)

    4. Personnel

    Limit access o rearing and laboratory facilities to authorizedpersonnel who under-

    stand the need to retain all live s tages under securityconditions.



    (1958) ; Fay

    and Eliason (1966);


    and Perry (1965); Hocking,

    B. (1960)



    and Jakob (1967).

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    this is done the significance of the DR S can be ascertained forany species of m os-

    quitoes. Preliminary research indicates that a DR S of 1.8 givesgood results.

    Many cages for holding and/or mating mosquitoes have beendescribed. McKiel

    (1957 ) describes a cage 30 wide x


    long x


    high (RS= 12,90 0 sq. cm.) for

    4000 Aedes aegypti adults.

    This would provide a DR S of 3.2. Weathersby (1962 )

    used a cage 18x18~18 for

    Aedes togoi



    sp. Blakeslee et

    al., (1962)

    used a cage 24x24~24 for Culex erythrothorQx. Kitzmiller andMicks (1954) cage

    is 12X12x12

    and 24X24X24

    cages for Culex


    Hayes and Morlan ( 1957)

    used a similar size cage for Aedes


    Williams ( 196 2) employed a 12x9s x

    14 cage for the same species.

    None of these authorrs mention the density per cage.

    McLintocks (1952 ) cage is 12x12~2 2 (RS= 6812 )

    f or 1500-2000 Culiseta inornata.

    This gives a DR S of approximately 3.4-4.5. Casanges et al.(1949 ) describe a

    22x.36x24 cage (RS-17 957) f



    000-5000 Anopheles quadrimaculatus, or a DRS

    of 3.6-6. Aedes aegypti have been kept in many types ofcages.

    Christophers (1960)

    descriibes a numb er of cages for this species including onetype 8 2x8 2x8 1/z and

    another 14x8~/~~12.

    Liles et aZ. (1960) employed a cage 12x8~9 for IOO A.aegypti.

    Wall is (1954) cage is 36x36x30

    for 10-350 female A. aegypti and concludes that

    the density of ovipositing females did not increase thepercentage of eggs produced.

    Roth (1948 ) used an 11x11~15 cage for his work on aegypti.

    Morlan et al. (1963)

    indicate that their colony cages (Fig. 3)

    consist of approximately 10,00 0 adults in



    FIG. 3.-Mm-hs colony cage.


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    FIG. q.-C &be rgs collapsible cag e.



    cage, (RS-12,487 ) or a DRS of 1.2. Thus, there is nostandardized

    size, type or construction material or population density.

    A standard size cage may be of value for rearings for researchpurposes.

    To this

    end a light weight aluminum cage has been developed 1 that isnow widely used

    (Fig. 4). This cage is constructed of pressed-form .032 inchalum inum frame 7s

    thick by 3/4

    wide and is screened with 18 x 22 mesh aluminum screening.The

    screening is held in place by a thin rubbe r sp line, for easyreplacem ent of screening

    when necessary.

    The floor of the cage is constructed of either .090 or .03 2inch

    aluminum sheeting depending upon the model of the cage. A hammock constructed

    of a fine mesh nylon is an integral part of the top of the cage.

    Anesthetized animals

    such as guinea pigs with shaved backs can be used for bloodfeeding.


    cotton balls, fruit, and su gar, can be placed in the hamm ockfor o ther types of feed-

    ings. The

    cage is fitted w ith a 24-inch long su rgical stockinette sleevewhich fits

    a 9l/2x97/s opening.

    The size of the cages may be


    (3ox3ox3ocm) or

  • 8/10/2019 Rearing Mosquitoes


    ; 1

    I 1





    ;, I




    ,._. _----

    _ -q


    CAG E


    FIG. 5.-Barraud cage.


    Gillet, J. G.(1962)

    PIG. 6.-Gillett cage







    Pollard, D.G,(1960)

    FIG, 8,-&kCrays escape-proof colony cage,

    FIG. g.-Pollarcl cage.


  • 8/10/2019 Rearing Mosquitoes



    (6ox6ox6ocm ) ( h

    own in Fig. 4 as G-I, G-2, and G-3, respectively), or

    other combinations.

    All cages are collapsible for ease of storage and can readilybe

    cleaned by steam or hot water.

    These cages have been successfully used with many

    species of mosquitoes in most of the major mosquitolaboratories.

    Many kinds of cages for special purposes have been described.Perhaps one of

    the most useful cages for field work is the Barraud Cage, Fig.5, (Barraud, Igag),

    a cage with a wire frame over which is hung netting. Lanternglobe cages, made

    from the glass globes of kerosene lanterns have been widely used(Fig. 6) (Trembley

    195 5; Gillett 1962 ). Others have modified glass cylinders(Burgess and Young

    rg46), glass jars (Eldridge and Gould 196 0) and plasticcontainers (Fig. 7) (Yolles


    7.-Plastic cage.

    and Knigin 1943; Young and Burgess 1946; Barnett 1955; Gerberget al.



    1968 ). Escape proof cages have been described by Bar-Zeev andGa lun (1960 )

    and McC ray (Fig. 8) (1963 ), b u are not practical for massrearings. Pollard (1960 )

    describes a cage (Fig. 9) that minimizes the escape ofmosquitoes and is still easy to


    Alger (1968); Barnett ( 1955) ; Barraud ( Igag) ; Bar-Zeev andGalun ( 1960) ; Blakeslee et al.



    Burgess and Young (1946); Casanges et al. (1949); Christophers(1960); Eastwood

    and Jamieson (1965); Eldridge and Gould (1960) ; Gerberg et al.(1967) ; Gillett (1962) ;

    Hayes and Morlan (1957); Kitzmiller and Micks (1954); Lennox(1959); Liles et al. (1960);

    McCray (1963); McKiel (1957); McLintock (1952); Morlan et al.(1963); Pollard (1960);

    Reynolds (1960); Roth (1948); Trembley (1955); Wallis (1954);Weathersby (1962);

    Williams (1962); Yolles and Knigin (1943);

    Youngand Burgess1946).

  • 8/10/2019 Rearing Mosquitoes



    Although the female mosquito usually must ingest a blood mealfor ovarian devel-

    opment, adu lts of both sexes require carbohydrate foods inaddition. Carbohy drates

    are generally supplied as a sugar solution.

    Although sucrose and g lucose in concen-

    trations of from 37/o to 20% have been used,


    sucrose, made by dissolving


    gms of ordinary household white sugar in one liter of waterappears to provide

    the best results.

    Other forms of suga r, such as corn syru p, honey, variousfruit

    juices, raisins, apple slices, and bananas , have also beenused.

    FIG. Io.--Animal restrainers in use; mass feeding of mosquitoeson chicks.

    Soake d cotton balls are the easiest method of providing thesuga r solution to ad ult

    mosquitoes. Th

    e cotton balls are soaked in the


    sugar solution, the moisture is

    squeezed out and the balls are then placed on the top of thecage. Usually 4 cotton

    balls, changed daily, are sufficient for a 30x30~30 cm


    cage. The cotton balls

    must be changed daily. Coluzzi (1964) used a tank and a side cuptrough. The

    liquid in the cup maintains its level and keeps a piece offilter paper wet continuously.

    Porter et al. ( 196 1) describe a small solution feeder foradult mosquitoes m ade from

    a ro-dram plastic vial in which is inserted a wick made of asmall roll of blotting


    The vial is filled with the feeding solution and the mosquitoesfeed through


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    small holes punched through the wall of a plastic vial. Eliason(1963) used a tech-

    nique of feeding mosquitoes on solid sugar.

    Blood meals are provided by supplying animals that have beenanesthetized or

    restrained, and placed on the adult cage, rather than in thecage.

    This prevents loss

    of mosquitoes, hazards to the technician, and destruction ofmosquitoes by the blood


    Animals may be anesthetized by the use of I grain nembutal (seeappendix)

    administered intraperitoneally at the rate of


    ml for each I lb (0.5 kg) of body

    weight. Various types of restrainers are available commerciallyand are described

    by Porter et al. (1961); Gerberg et al. (1967) (Fig. IO); Morlanet al. (1963);

    Trembley (1955); Dunn (1932); Jones and Scheltema (1952).

    The more commonly used sources of blood meals, other than atechnicians bared

    arm, are guinea pigs,

    rabbits, mice, rats, hamsters, monkeys, and chicks.


    meals may also be supplied by artificial means (see artificialfeeding techniques).

    Shave the animal prior to presentation to the mosquitoes.

    Some prefer shaving or

    clipping the hair from the back of animals, and others theunderside. Animal clip-

    pers have proved very useful for this purpose.


    Dimondet al. (1955); Dunn (1932); Eliason 1963); Gerberg t

    al. (1967);



    Knierim et al. (1955); Lang and Wallis


    L 1es


    al. (1960); Morlan et



    Porter et al. (1961); Roy (1936);

    T rembley

    1955); Woke (1937);

    Woke et al. (1956).


    Xmembrane feeding technique is one approach for artificiallyproviding a blood

    meal to mosquitoes.

    This technique, used with other haematophagous arthropods

    prior to the successfulfeeding of mosquitoes, consists of amembrane-covered con-

    tainer, such as a glass tube or test tube, containing the liquidmosquito food. The

    container is placed so the membrane side is accessible o themosquitoes for feeding.

    The liquid should b

    e several degrees warmer than the ambient temperature toinduce

    certain species to feed. Tarshis (1958)

    review of artificial feeding techniques

    includes the various types of membranes used, and describesvarious techniques and


    Knowlton and Rowe (1935)

    used animal mesentery bags to feed mos-

    quitoes on

    suspensionsof diseased (equine encephalitis) brains. Woke (1937)fed



    on whole blood through rat skin membranes. Yoeli (1938) gave

    Anop&des elutus blood through rabbit skin membranes. Bishoppand Gilchrist

    (1944, 1946) induced feeding by Aedes aegypti on blood throughchicken skins.

    Greenberg (1949,


    reports the feeding of Aedes aegypti on a variety ofartificial

    fluids through Baudruche (a bovine

    intestinal preparation) capping membranes.

    Eyles ( 1952)

    used hog-gut sausage casings, and Trembley (1952) usedBaudruche

    capping membranes

    and chicken-skin membranes.

    Kartman (1953) fed Aedes


    blood through hog-gut sausage casings.

    Bar-Zeev and Smith (1959) in

    studying the action of repellents, gave A.

    aegypti titrated blood through a membrane

    made from the outermost layer of ox caecum.

    Tarshis (1959) discusses ive different

    1 Oster Mfg. Co., Milwaukee, Wise. Model A2 Clipper, No. 40blade.

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    memb ranes and an apparatus for feeding Culex


    Collins et al. (1965 ) used

    the Baud ruche (untreated) memb rane to feed Anophelesmosquitoes Plajmodium


    infected blood.

    FIG. I I.-Apparatus for me mb rane feeding of mosq uitoes.

    Rutledge et

    al. (



    escribe in detail their techniques for mem brane feeding

    of mosquitoes, Figs. I I, 12.

    The feeder is mad e of heat res istant glass or stainless


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    Mosquito Membrane Feeder


    FIG. I x--Apparatus for


    membrane feeding,

    steel. Circulating warm water controls

    the temperature of the blood. Accord-

    ing to them, chick skin is superior to

    the Baudruche membrane, and chick

    blood is more acceptable than erythro-

    cyte extract or serum.

    Blood meals can be supplied by means

    of preserved blood absorbed on a cotton


    Russell (

    193 I )

    reports feeding

    mosquitoes blood distributed over net-


    Others (MacGregor and Lee

    1929; Knowlton and Rowe 1935; Shiavi

    and Franc0 1949; McL,intock 1952;

    Dimond et al. 1955; Trembley 1952;

    Liles et al. 1960) mix blood with honey,

    sucrose or glycerine.

    Knierim et al.

    (1955) used frozen titrated beef blood plus 10% honey (or

    without honey) heated

    to 35-38 c.



    and Smith (1959)


    Behin (1967)


    Bishop and Gilchrist (1944, 1946)


    Collins (1963)


    Collins et al. (1965, 1966); Co11

    ns et al. (1964)


    Collins et a/. (1964)


    Dimond et al. (1955) ;

    Eyles (1952) ; Galun (1965) ; Greenberg (1949) ; Grifliths andGordon (1952) ; Jones (1956) ;

    Kartman (1953) ;


    and Wallis (1956) ;

    Liles et al. (I

    960) ; MacGregor (I

    930) ; MacGregor

    and Lee (1929); Mattingly (1964); McConnell (1956); Msangi(1956); Ogden (1961);

    Ogunba (1967); R

    ass (1956) ; Russell (1931) ; Rutledge et al. (1964) ; Schiaviand Franc0

    (1949) ; Shambaugh (1954); St. J h n et d. (1930); Tarshis(1957, 1958, 1959); Trembley

    (1952); Willis (1958); Woke (1937); Yoeli (1938).


    Aedine mosquitoes are normally difficult to maintain in thelaboratory but Mc-

    Daniel and Horsfall (1947)

    used efficient procedures for stimulating copulation and

    obtaining Aedes stimulans and Aedes vexans eggs. Horsfall andTaylor (1967)

    modified the technique and used it successfully to mate over 40species of mosquitoes.

    Copulation is induced by manipulating an immobilized female intocontact with a

    decapitated male.

    This technique consists of using at least 3 day oldmosquitoes.

    The female is anesthetized lightly with chloroform, and ispicked up with a suction

    pipette and manipulated to make contact with the male. Thedorsum of the thorax

    of living decapitated males is cemented with a non-toxic glue toa small piece of

    glass slide. Contact between the two immobilized mosquitoes isaccomplished

    manually under a



    The female is placed in the mating position

    with the ventral side up and the head directed away from themale.

    Apexes of the

    two abdomens are then brought together, venters uppermost. Themale is stimu-

    lated by touching the claspers several times with the female.Insemination is usually

    completed within a 5-50 second period.

    Usually 3-4 males are thus prepared so

    that if one male is unresponsive,

    others are immediately available. Frizzi (1958)

    used the induced copulation technique on Anoptieles macdipenniswith variable

  • 8/10/2019 Rearing Mosquitoes


    results, and Carvaglios (1961 ) using the technique withoutdecapitation of males

    maintained a colony of Anopheles Zabranchiae.

    After mating, the females are transferred to a feeding cage forrecovery from


    A blood meal is provided after 12 hours. Bake r et L$ (1962 )obtained

    IOO O/~ fficiency in single pair matings of A.


    by induced copulation.

    Anopheles punctipennis, A. quadrimaculatus, A. freeborni, and A.albimanus were

    maintained in the laboratory by the above authors, using theinduced copulation


    Wheeler (1962 ) modified the technique by gluingcold-anesthetized mosquitoes to

    the heads of insect pins.

    The female-bearing pin is inserted into a cork glued to a

    microscope slide so that the mosqu itos body axis is in ahorizontal plane. The

    male-bea ring pin is inserted into an adjustable pinned insectholder an d then

    oriented until the genitalia meet. Ow Yang et al.


    simplified the technique

    by using 3-6 day old males caught w ith a fine pipette attachedto a suction device, and

    pinned latera lly through the thorax using a minuten pin fixedinto the end of a 6-

    long soft wooden stick. The females are blood-fed 2 days afteremergence, and are

    collected in individu al tubes the next m orning.

    The female is anesthetized with

    ether and tipped out on her back onto a clean white table. Copulation is achieved by

    bringing the pinned male down at about a 45 angle to the female.When copula-

    tion is affected the pair can be lifted together, with the female firmly clasped by the

    inale, and gently dropped into a container, by removing the malefrom the pin.


    mating procedure is carried ou t under a microscope at a roomtemperature of 26-28

    C. and RH of about 80%. The authors were successful inmaintaining a laboratory

    colony of

    Anopheles maculatus

    by using this technique.

    Coluzzi (1962) maintained Culex


    and C.


    by induced mating.

    Hors fall (1964 ) states that species of Aedes, Anopheles,Culex, Culiseta and Psoro-

    phora have been effectively insemina ted in his laboratory. Herecomm ends that

    adults of the same age be held for a 60-72 h ours prematinginterval at 80 + 7; RH .

    They are fed on a carbohydrate diet (honey diluted I :I w ithwater). Mating is

    carried out in a room with a temperature of 26-28 C. and about85% RH. One

    technician prepares the males, the second prepares the femalesand joins the two in

    copula, at the rate of 40 matings per hour.

    The males are collected into individual

    tubes and the open vial is inverted over a Buchn er funnelthrough which flows a

    gentle stream of CQ2.

    Immediately upon knockdow n the male is plucked from

    the funne l with forceps and is glued ventral side upw ard to aglass slide (casein

    glue is used).

    Four or five males are placed in a row. Decapitation mayprecede

    gluing, or delayed for longer surviva l until just prior to theact of copulation, w hich

    will not take place unless the male is decapitated. The malesurvives longer if

    decapitation is delayed.

    Initiate the mating procedure only after full recovery ofthe

    males (2-5 minutes).

    The females are anesthetized with chloroform only to the

    point of relaxation and are then picked up with a vacuu m pipette made from a 5-mm

    glass tube, bent at right angles and d rawn to an open tip smallenough to apply to

    the anterior slope of the mesonotum.

    A vacuu m pum p with a bleeder tube or valve

    is used. Th


    emale is held ventral side uppermost and broug ht into contactwith

    the claspettes.

    Manipulate the female so that her genital opening is broughtproximal

    to the phallosome of the male.

    Proper positioning is prerequisite to bringing abo ut

    complete union.



    an angle of about 120 between the .?bdomens will


  • 8/10/2019 Rearing Mosquitoes


    permit proper union.

    For Culex the angle must be nearer to 180 Horsfail blood-

    feeds on the day following mating


    hours between time of anesthesia and


    Baker (1964) feeds th

    e males, prior to copulation, with honey on a cotton ball,

    and the females with water on a saturated sponge. Generally heprovides a blood

    meal for the female shortly before mating. Baker does notanesthetize the males

    but collects them with an aspirator and softly blows thembetween layers of cotton

    wool. This holds them until pinned laterally through the thoraxusing a dissecting

    needle fixed into the end of wooden stick


    cm (3-6) long. If it is advan-

    tageous to keep the male alive, he can be held with a vacuumpipette.

    Baker reports

    success with Culex t~salis using this technique.

    Esah and Scanlon ( 1966) estab-

    lished a colony of Anopheles b. balabacensis by inducedmating.


    Baker (1964)


    Baker and Kitzmiller (I 961)


    Baker et al. (1962)


    Burcham (1957)



    (1961); Coluzzi (1962); Esah and Scanlon (1966); Frizzi (1958,1959); Gopal and Wattal

    (1962) ; Hayes, D. E. (1968) J Hayes, R. 0. (1953); Horsfall(1964); Horsfall and Taylor

    (I 967) ; McDaniel and Horsfall ( 1957) ; Ow Yang et al. ( 1963); Wheeler (I 962) ; Wheeler and

    Jones (1963).


    The eggs of mosquitoes are usually deposited on the surface ofthe water, or on

    surfaces that are subsequently inundated. To obtain mosquitoeggs, a reasonable

    substitute must be provided for the oviposition site.

    A plastic container approximately rgxrox5cm (6~4x2) has provedquite successful

    for anophelines.

    A layer of cotton balls is placed in the bottom, covered witha

    sheet of filter paper or paper toweling and then flooded withwater so that the water

    surface barely covers the filter paper (Gerberg et al. 1968).Some investigators use

    bowls or pans of open water as oviposition sites. Giglioli(1947) used a white glazed

    china finger bowl (II cm in diam. 7 cm in depth) half filledwith water. Others

    have lined the oviposition container with paper to prevent theanopheline eggs from

    sticking to the sides of the container. Kepler et al. (1964)employed a water-filled

    petri dish, the bottom lined with a strip of filter paper. Yoeliand Bone


    provided small cork rafts and protruding stones on which thefemales alight prior

    to laying eggs.

    Trembley (1955) reports that some Anopt2eZes preferred darkcolored

    containers or containers lined with darker paper toweling.

    Species of the genus CuZex usually lay their rafts of eggs onopen water, in pans,

    bowls, petri dishes or other type containers. Krishnan (1964)claims that more egg

    rafts were obtained in trays containing hay infusion than inthose containing tap


    Jupp and Brown (1967)

    confined individual gravid females in laying tubes

    r in. in diameter containing a little water and a strip offilter paper down the side

    which provided a suitable surface for the mosquito to rest on.The tubes are closed

    with nylon gauze on which is placed a cotton pledget soaked insugar solution.

    Takahashi (1968) bt

    ained eggs of C. portesi by placing an inverted clay flower

    Fat, with an additional I

    side entrance hole in a white enamel dish containing


    Inside the flower pot and standing in the water is ablack-painted g ass

    oviposition jar.


  • 8/10/2019 Rearing Mosquitoes


    Laurence (1960) collects Mansonia eggs from floating discs ofplastic-impregn ated

    &aft paper in an oviposition chamber.

    Usually aedine eggs can withstand desiccation and are collectedon a moist sub,


    The most comm on method for the collection of A. mgypti eggs isto use a


    wide paper toweling, lining the interior of a pint ice creamcarton o lr a glass jar.

    Fill the carton with enough water to cover approximately I in.of the paper toweling.

    The eggs are

    conditioned by leaving the egg paper in the container for anaddi-

    tional 24 hours, after removing the co,ntainer from thecage.

    The egg paper is then

    air dried (at 80 F. and 80% RH) for





    s and then stored for future use. Other

    A&es species may be induced to lay eggs on various types ofm oist surfaces.


    wrapped in cheesecloth, porous sponges, sphagnum moss wrapped incheesecloth,

    large-pored plastic materials, and other types of rough orporous materials have

    been used as oviposition sites.

    Reynolds (1960 ) describes an escape proof m osquito

    egg harvester, for use under maximum mosquito security.

    Anoplieles, Culex and

    Culiseta eggs

    are not readily stored and are usually hatched

    in the water where laid. Cond itioned aedine eggs can be stored.To hatch these

    eggs, place in water with a reduced oxygen content.

    Many methods for deoxygena-

    tion have been described, since the most uniform method ofhatching is by imm ersing

    the egg papers in deoxygenated water.

    To make deoxygenated water, submerge

    quart Mason jars in boiling water and cap under water.

    The jars can then be stored

    a t 27 C . ( 80 OF.).

    When an egg paper is to be used, the jar is uncapped and the

    egg paper is immediately placed in the w ater and the capreplaced.

    Eclosion usually

    begins in minutes.

    Some egg papers can be used over again as all the eggs donot

    always hatch after the first immersion.

    Tho irgh it is possible to count aedine eggs on the egg paper,for mass rearing

    purposes this is not feasible.

    Instead an aliquot sam pling m ethod is used on the

    newly hatched larvae.


    Burgess (1959); Elzinga (1961); Fay and Perry (1965); Giglioli(1947); Gjullin et

    al. (1941);

    Horsfall (1958); J d

    son (I 960) ; Jupp and Brown (1967) ; Keppler et al. (1964) ;Krishnan

    (I 964) ; Laurence (I 960) ; Lavoipierre (1953) ; Reynolds (I960) ; Shute (1933) ; Takahashi

    (1968); Trembley (1955); Williams (1962); Yoeli and BonC(1967).


    Counting Lmuae


    Newly hatched larvae may be counted individually using a handlens and a hand


    For large rearing trays or mass rearing purposes, the aliquotsample method,

    Fig. 13, (Gerberg et OZ. 196 8) is useful. Pour the first instarlarvae into a gallon

    battery jar and fill to 3500 ml of 27O C. (80 F.) water. Thebattery jar has 5

    random ly placed outlets (at 5 levels).

    A variable speed stirrer, mo unted on a ring

    stand, is immersed in the water.

    The stirrer is operated at approximately 250 rpm,

    to suspend the larvae

    uniformly in the water.

    Twenty-five samples of I ml each are

    withdrawn through the random outlets and the mean number oflarvae per sample

    (ml) is determined.

    It is then possible to withdraw a given qu antity of water

    containing a known number of larvae.

    Morlan et al. ( 1963)


    escribe and figure an autom atic dispenser for obtaining


  • 8/10/2019 Rearing Mosquitoes


    Fig. r3.-Aliquot container for mosquito larvae.

    large volumes of dispersed mosquito larvae. This equipment ismade from a modi-

    fied agitator type washing machine and a rain gaug e type oftripping bucket.

    Bar-Zeev (19 62) describes a simple technique for obtainingstandard numbers of

    newly hatched mosquito larvae.

    Rearing Trays

    A wide variety of containers capable of holding water may beused as rearing trays.

    Trembley (1955) suggests white enam el photographic developingtrays 16xgf/2x2,

  • 8/10/2019 Rearing Mosquitoes


    or containers of 3 quart capacity, 2-3 inches deep, preferably,but not necessarily,

    opaque . Containers of these types are shown in Figs.


    and 15. For mass rearing

    of A&es aegypti, Morlan et al. (1963)

    used a galvanized metal tra y 2x10~ 72.

    (Fig. 16). Th

    e ra was coated with a thin layer of paraffin. Each tray had aj/4


    copper-tube outlet to facilitate draining and was stacked on ametal rack mad e of

    i/sx 3/4

    3/4 I1

    angle iron. Each

    rack held 24 rearing trays. They used 7 liters of

    water in each tray to rear 7000 larvae. This system provides for1.4 larvae/sq. cm.

    of surface area and 1.4 larvae/ml of water. Fay et al. (1963)reared


    larvae per tray (2~10x 72) with no significant difference ofpercentage of yield or

    adult vitality.

    In the IC R laboratories, mosquitoes are mass reared inparaffin-coated, galvanized

    trays r38x7 6x5cm (54x20x2), (Fig. 17). Each tray is filled to adepth of 2cm

    ( g) with water and approximately 15,000 larvae are introduced.Gahan (1967)

    suggests 100 -250 Amp/&es larvae for a polyethylene pan 12in diameter.

    For small scale rearings, white polyethylene pan s 13I/4x1ox2have been used very


    Handling Larvae

    If larvae have to be moved or transferred, Trem bley (195 5)suggests a 50 ml.

    volumetric transfer pipette shortened to 8-10 and fitted with arubber bulb. We

    have found very useful a 12-18

    length of rigid clear plastic tubing covered with a

    FIG. Iq.-White enamel trays for rearing larvae.


  • 8/10/2019 Rearing Mosquitoes



    IS.-White enamel dishes for rearing larvae.




    314. LCl 1.062 WALL)





    FIG. 16.-Construction of galvanized metal rearing tray.


  • 8/10/2019 Rearing Mosquitoes


    FIG. IT.-Mass rearing of mosqu itoes in galvanized trays.

    fine mesh cloth at one end and fitted into a rubber bulb. Alsouseful for picking

    up larvae or pupae is a piece of 20-40 mesh stainless steelscreening 27x75m m


    and bent so that approximately


    inch is at a 45 angle to the remaining 2

    Equipment for Feeding Larvae


    can be premeasured into small containers and then dispensed overthe water


    Some technicians prefer a salt shaker and can judge the amountof food


  • 8/10/2019 Rearing Mosquitoes


    by the number of shakes. Heal and Peregrin (1945) recommendsheet metal spoons.

    Hunt and DaveY (1947) used reversed pen nibs. Morlan et al.(1963) used a

    plastic spoon of appropriate capacity. Regardless of the methodof measurement, it

    is imperative that a rigid feeding schedule be followed and thetype and amount of

    food be standardized if uniform and consistent results aredesired.


    Bar-Zeev (1957 , 196 2); Fay et al. (1963); Galun (1967);Gerberg et al. (1968); Heal and

    Peregrin (1945); Long and Breland (1956); Morlan et al. (1963);Soderstrom and Levitt

    (1967); Trembley (1955).


    Mosquito larvae are fed everything from a highly refinedchemical diet to a witchs

    brew made from guinea pig droppings, mud, and the ground-upskins of water

    boatmen. Perhaps one of the greatest reasons for variability inthe results of rearing

    mosquitoes is the lack of uniformity or standardization oflarval food. An abundance

    of literature refers to larval foods, both natural andchemically defined. Asahina


    provides an excellent review of food material and feedingprocedures for

    mosquito larvae.

    Some of the early investigators (Bacot 1916) report thatbacteria and yeasts are

    essential items of diet.



    increased the bacterial and protozoan

    population by the addition of guinea pig feces. Boyd et al.(1932) used hay infusion

    to provide protozoan food for the larvae. Weyer (1934) addeddried blood and

    powdered calfs liver.

    Crowell (1940) was probably the first to use commercial dog

    biscuits as a complete and somewhat standardized larval food.Perhaps the most

    widely used food and feeding schedule is the one developed atthe Communicable

    Disease Center (Morlan et al. 1963). The larvae are fed dog chow(5% crude fat)

    ground to pass through a 40 mesh screen.

    This food is primarily for A&es aegyptz

    but will work for other speciesas well, with the followingschedule:

    Day o (day of hatching) o.zmg/larva







    Note that the addition of a live yeast mixture (0.7 gm/I literof water/Io,ooo

    larvae) on day 3 improves adult production.

    Trembley (1955)

    used the following procedure for Anopheles quadCmacuZatus,

    using a 16x9 l/*x2

    tray for 300 larvae.

    12 hours before day o

    25 mg powered dried brewers yeast

    and 25 mg Bacto-brain heart infusion.

    Day o


    mg of

    I :I

    powdered yeast and

    powdered dog food



    50 mg

    Day 2-3

    100 mg

    Day 4-14

    150 mg

    Gerberg et al. (1968)

    use the following schedule for Anopheles stephensi.


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    The larval food consists of a 50:5 0 mixture of dog chow a ndpig liver pow der

    ground to 40 mesh.

    Day o-Day 2 .I 3 mg/larva

    Day 3 .20 mg + .045 mg Fleischman ns

    dried brewers yeast/larva

    Day 4 .26 mg/larva

    Day 5 .40 mg/larva

    Day 6-9 .53 mg/larva

    The nutritional requiremen ts of mos quito larvae wereinvestigated by Trag er

    (1935, 1937, 1953), de M 11

    l on et al. ( 1945), Goldberg ( 1947), Goldberg and de

    Meillon ( 1948), Lea et al. ( 1956), Lea and DeLong ( 1956),Singh and Brown

    (1957) and Akov (1962).


    Akov (1962);


    (1964); Racot (1917);

    Boyd et al. (1932): Buddington (1941);

    Crowell (1940) ; de Meillon et al. (1945) ; Frost et al. (1936); Gerberg ez ai. (1968) : Golberg

    (1947); Goldberg and de Meillon (1948); Hinman (1930, 1932,1933); Lea et al. (1956);

    Lea and DeLong (1956); MacGregor (1915); Morlan et al. (1963);Singh and Brown (1957);

    Trager (1935, 1937, 1942, 1953); TrembleY (1955); WeYer(1934).


    Pupae may be picked up by means of pipettes or by small liftersmade of screening.

    The comm ercial m edicine dropp er with the open end filed offto ma ke a wide m outh,

    or Kom ps modification of a syringe (Trem bley 195 5) may beused. Plastic pipettes

    have become popular.

    The rubber bulb normally supplies adequate suction but

    others have relied on the use of vacuum pumps or water-operatedsuction pumps

    (Johnson 194 7) to increase the efficiency of operation.

    Lifters can be mad e from small pieces of metal screening, 20-40 mesh, 25x75c m

    (1x3). Individual pupae can be lifted by means of small wireloops.

    Pupae may be counted by manually picking them out with a pipetteor lifter.

    They can also be counted vo lumetrically.

    A volumetric counter can be made from

    a piece of plastic tubing with an ID of 3/s-2, approxim ately 6long with one

    end covered with fine wire screening. A know n num ber of onesize pupae are added

    to the tube and the level of pupae is marked on the tube. Fromthen on pupae

    can be added to the m ark, the tube inverted and the pupaewashed out by flushing.

    Pup ae can be separated rapidly from larvae by the ice watertechniques of Ram a-

    krishnan et QZ. 1963), Weathersby ( 1963 ) and Hazard ( 1967).Larvae and pupae

    are concentrated in a sieve and im mersed in ice water.

    The larvae sink immediately

    and the pupae float and are poured off.

    Bar-Zeev an d Ga lun ( 1961 ) used a rather

    unique method of separating larvae from pupae by magneticmeans.

    Iron dust is

    added to the rearing trays and the culture exposed to a magneticfield. Only the

    larvae, which have ingested the iron p articles, are held b ackby the mag netic field.

    Fay and Morlan (1959) describe a mechanical device not only forseparating larvae

    from pupa e, but ca pable of sexing the pupae of somespecies.

    The principle of sepa-

    ration and sexing is based on size differences.

    The culture of mixed larvae and

    pupae is

    polured through a funnel into a wedge-shaped space between twoslightly

    separated and adjustable glass plates.

    The female pupae, being largest are retained


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    in a line at the upper levels of the plates, the smaller malepupae are held below

    the females and the even smaller larvae are retained towards thebottom. The

    smallest larvae drain out. The authors claim the separation ofIOOO pupae in 20

    minutes. Grose et al. ( 1966) made improvements on Fay andMorlans design.

    McCray (1961) d escribes a simple separating device made from analuminum

    sheet with stamped-out 0.039 slits. The larvae and male pupaewash through

    and the female are retained. The author claims that 30,000individuals can be

    processed n 5 minutes. Gentry and Gerberg are developing a smallseparating and

    sexing device that uses removable comb-like plates in a trougharrangement. The

    plates are interchangeable and the proper size plate is inserteddepending upon the

    species to be separated and the size of the pupae andlarvae.

    The sorted pupae are placed in containers of convenient sizesand the containers

    placed inside cages to await adult emergence. Various deviceshave been used to

    aid the emergence of adults. Some workers use small floatingobjects, such as small

    pieces of cork, (Trembley 1955, Christophers 1960) or twigs toassist the adults in

    leaving the water.

    Smith and Whitlaw (1963) d escribe an emergence disc of wide

    mesh dacron cloth that is claimed to reduce the mortality ofemerging adults.


    Bar-Zeev and Galun (1961); Christophers (1960); Fay and Morlan(1959); Gillett (1955) ;

    Grose et al. (I 966) ; Jones (1957) ; Lowrie and Gubler (I 968);Hazard (I 967) ; Johnson (I 947)

    McCray (1961); Ramakrishnan et


    (1964): Smith and Whitlaw (1963); Trem bley (1955);

    Weathersby (1963).


    Rearing of mosquitoes under aseptic conditions may be requiredfor


    nutrition investigations for in vitro studies, and other typesof research.

    Eggs may be surface sterilized.

    Techniques described by Atkin and Bacot (1917)

    use 0.5% lysol; Barber (1927)

    dripped 80%

    alcohol for 2-3 minutes over A.


    eggsand Roubaud and Colas-Belcour (1929) surface sterilized eggsusing



    and also mercuric chloride. Whites (193 I) solution, consistingof mercuric

    chloride 0.25 gm, sodium chloride 6.5 gm, hydrochloric acid, 125ml, ethyl alcohol


    250 ml and distilled water 750 ml, was more effective againstbacterial contamination

    than against other types of contaminants.

    Hinman (1932) disinfected eggs in hexyl-


    Trager (1935, 1937)

    used 5% castile soap and then Whites solution.

    Lea et ul. ( 1956)

    and Jones and DeLong (1961) surface sterilized eggs by


    the eggs in

    70% alcohol for 5 minutes, transferring eggs by means of flamedforceps

    to a 0.87% sodium hypochlorite solution for 2 minutes, then to asecond 70% alcohol

    rinse for 4 minutes,

    and then to a final wash in autoclaved distilled water. Mar-

    tignoni and

    Milstead (1960) sterilized A. aegypti eggs on paper by placingone-

    month-old eggs in a petri dish containing an autoclaved IO:/,solution of benzalko-

    nium chloride for 30-60 minutes,

    then washing in sterile water, followed with

    immersion for


    minutes in 80/~ alcohol and then two more washings insterile

    water. Akov (1962) describes a method of obtaining sterilelarvae of uniform age

    by placing

    the sterile eggs in a desiccator and reducing the pressure for10-20 minutes.

    Trager (1935, 1937); de Meillon et al. (I945), Lea et al.(I956), Lea and DeLong

    (1956), Nayar (1966)~ Singh

    and Brown (I957), Akov (1962) reared larvae on a

    sterile chemical media.

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    Pupae can be washed to surface sterilize them, and adults may befed on sterile


    Thou gh it may be superfluous to mention, it is important to seethat all wo rk in

    rearing aseptic anim als is condu cted under com pletely asepticcond itions.


    Akov (1962, 1964) ; Atkin and Bacot (1917) ; Barber (1927) ;Boorman (1967) ; Butt and Keller

    (1961); Hinman (1930, 1932); Jonesand DeLong (1961); Lea et al.(1956); Lea and DeLong

    (1956); de M ei on1 et al. (1945)


    Lichtenstein (1948)


    Martignoni and Milstead (1960);

    Nayar (1966)


    Roubaud and Colas-Belcour (1929)


    Singh, K. R. P. and Brown (1957) ;

    Trager (1935,


    White (1931).


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    Information gleaned from literature has been compiled to presenta rearing tech-

    nique for individual species,or in some casesa genus.

    Usually, for each genus there

    is a detailed description for at least one species hat couldserve as a guide for rearing

    other members of the genus.

    The arrangement of genera and nomenclature is based on Stone,Knight and Starke





    Subfamily Anophelinae



    Subfamily Toxorhynchitinae

    Genus Toxorhynchites

    Subfamily Culicinae

    Tribe Sabethini







    Tribe Culicini



















    Genus Opifex



    Genus Culex





    Eggs may be collected in a small container, usually r5cm (6) indiameter or



    Cotton balls, or an absorbent cotton pad, are laid insidethe

    container and then barely covered with water.

    Filter paper or paper toweling is

    then laid over the cotton pads so that the water surface is evenwith the surface of

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    coarse-ground mixture of equal parts of Kelloggs concentrate,wheat germ and live

    yeast. At the 2nd instar they are transferred to clean pans withfresh water and food

    (Keppler et al. 1964). Gilotra ( 1966)

    reared 200-250 larvae in enamel pans

    I I .5x7.5x2

    containing I l/2 liters dechlorinated tap water, and fed onpowdered

    Purina laboratory chow supplemented with Fleischmanns yeast. Thelarval period

    usually lasts


    days when reared at 27 C. (80 F.).





    pupae are collected once a day and placed

    The pupal stage lasts 30-33 hours.

    in cups of fresh water (Rozeboom

    The adults are maintained in cages goxgoxgocm (2.5x2.5x2.5. Theyare fed on a

    5% honey solution or sugar solution. The females are provided ablood meal at

    dusk. A rabbit is placed in the cage for 2 hours. The besttemperature appears to

    be 28'


    (SIF.) (D

    owns and Arizmendi 1951). Adults are also maintained in

    cages 6ox6ox6ocm


    and are fed on sugar water, and various fruits. The

    females are provided with a blood meal (human arm) once a day.Cages r2xgxg

    maintained at 65% RH are also used for adults. A guinea pigprovides a blood meal

    and adults are maintained on honey-saturated balls of cottonwool (Keppler et al.

    1964). Adult longevity is approximately 30 days.


    Coluzzi (1964) ; Dow ns and Arlzmendi (1951); Gilotra (1966) ;Ikpplcr rt 4. (1964) ; Roze-

    boom (1936).

    Anopheles albitarsis

    Lynch Arribalzaga


    Barrett0 and Coutinho (1943); Galvao and Grieco (~9~3) JGalvaoet nl. (1944).

    Anopheles at-gy&arsis



    Galvao et al.


    Anopheles axtecus Hoff mann

    See methods used for A. quadrimaculatus and A. freeborni.


    Downs et

    al. (1948);

    Dow ns and Arizmendi ( 1 g5 I) ;

    Wallis (I 955 ).

    Anopheles ba.Zabacensi.s


    Eggs are collected on filter paper discs kept in contact withmoist cotton pads.

    The mean number of eggs per female is 163.

    Eggs are removed from the filter

    paper discs to enamel trays containing 2,500 ml of seasoned tapwater over a layer

    of sterilized stream sand.

    The eggs hatch in 48-72 hours.

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    Anopheles crucians Wiedemann

    See A. quadrimacuZatus techniques.


    Boy1 (I

    926) ; Boyd et al. (1935).

    Anopheles culicifacies



    Jayewickreme ( 1952)


    Pal ( 1945).

    Anopheles darlingi Root

    Eggs are collected on moistened filter paper (Freire and Faria1947) or in white

    glazed fingerbowls (diam.

    I I

    cm, depth 7 cm) half filled with water. The eggs

    are allowed to hatch in these bowls. Eggs hatch in 24-48 hoursat 27 C. (80 F.).


    The larvae are reared in containers 49x7oxIocm or in whiteenamel basins (28 cm

    in diam, and 8 cm deep). App



    larvae are placed in the basin.


    larvae are fed on an infusion made by mixing toasted fish andtoasted bread in pro-

    portions of I :2 plus a trace of brewers yeast. This mixture issuspended in water

    4-5 days at 27O C. (80 F.), and should have a pH of 7.o-7.8.Giglioli (1947)

    feeds larvae on dried brewers yeast or poultry laying mash orcombination of both.

    No special attempt is made to regulate light, temperature (70-MF.) or relative

    humidity (66-95% RH). Th

    e water used is clean, fresh, with a pH of

    larval stage ranges from 6-12 days.

    6-7. The


    The pupal period is approximately 2 days. The pupae arecollected

    placed in containers in the adult cage.

    daily and


    The adults are kept in cages 4ox4ox5ocm in an insectarymaintained at

    and 80-90~/~ RH. The time required from egg to imago is IO days.The

    __ _

    26-2a" c.

    adults are

    fed hony on cotton pads.

    Adult longevity is 35-45 days. A cage 3 m high with

    a 1x1-m base is required for mating (Freire and Faria 1947).

    Giglioli ( 1947) main-

    tains adults in a cylindrical cage 60 cm high and 35 cm indiameter, consisting of F

    cotton mosquito gauze sleeve stretched over a IO gauge wireframe. The adults

    are supplied with cotton wads soaked in cane sugar solution andchanged daily.

    Blood meals (human) are provided daily for l/2 hour in themorning. A tempera-

    ture of 80 F. and a relative humidity of 85% seems optimal.

    The first oviposition

    usually occurs 6-8 days after emergence.


    Bates (I

    947) ;

    Freire and Faria (I

    947) ;

    Giglioli (I 947).


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    The larval stage lasts from g-10 days.


    The pupal stage lasts for




    Mating will occur in an insectary.

    A minimum of


    days after emergence is

    required by adults before mating.

    Eggs are laid


    days after a blood meal. Females

    will feed on man, and animals. Adults feed on fruit juices.Females will live for

    51 days at 85% RH when provided with blood meals.


    J&m (1945); Perry (1946).

    Anopheles fluuiatilis James


    Eggs are collected in earthen pots, lined inside with mud andcontaining water.

    The eggs are transferred to large enamel basins and dishescontaining water. The

    eggs are floated inside paraffined cork rings.


    The larvae are fed on hay infusion, plus a small quantity of amixture of 2 parts

    litmus milk and


    part dehydrated blood serum.

    Dried brewers yeast may be sub-

    stituted for the above.

    The water in the basin is aerated vigorously once each day.

    Relatively larger amounts of yeast are added to the rearingbasins when the larvae

    reach the fourth instar.


    The pupae are removed from the larval rearing basins, placed ina bowl of clean

    water and placed inside the colony cage.

    A few blades of grass are floated in the

    bowl to provide a foothold for the emerging adults.


    The colony cage measures


    and is placed inside a larger cage with solid

    wooden sides and glass top.

    Humidity is maintained by suspending pieces of cloth

    in a saturated solution of common salt. Adults are fed on


    glucose on cotton

    wool. A rabbit provides nightly blood meals. Mating takes placeonly in the

    presence of a blue


    Egg laying occurs 48-72 hours after a blood meal. Len-

    gevity is

    17-18 days at 27 C. and 60-80*/~ RH.


    Mohan (1945); Pal (1943); Sin& and M&an (1951);Viswanathan et al. (1944).

    Anopkeles ft-eeborni Aitken


    Eggs are collected in a glass crystallizing dish containingdistilled water.


    mately 100-200 eggs are laid per female. The eggs hatch in 3days at 220 C.

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    The larvae are reared in enameled iron pans 25cm (IO) indiameter in water

    about r2.5-17mm (s-3/4) deep,


    rmstrong and Bransby-Williams



    water is boiled, cooled stream water.

    Approximately 150 larvae are added to each

    pan. Powdered meat meal is provided as food. Powdered blood andyeast suspen-

    sion used by Shute (1956)

    seemed to produce heavy scum.

    Wegesa ( 1964) com-

    pares meat meal and yeast and finds adult emergence and survivalrates significantly

    increased by using meat meal.

    A fine mesh tea strainer is used for applying the


    Larvae are reared in an enamel tray 35x3ox5cm at 26.5 C. andrequire


    days to pupate.

    Two hundred newly hatched larvae are placed in 2 liters ofdistilled

    water. They are fed


    g meat meal on alternate days.


    Pupae are collected daily and 300 placed in a 7.5cm (3) diameteraluminum pot

    with a minimum of larval water.

    Distilled water is added to bring the water level


    r2.5-r7mm (l/2-3/4) of the top. Pupae are maintained at 26.5 C.& 0.5 OC.

    Adult emergence is complete in about



    Eight hundred pupae are placed

    in a cage.


    Adults are kept for the first 7 days in cages 3ox3ox3ocm

    (1x1~1') and then

    transferred to larger stock cages 75x38x4ocm (30x15~16). Theadults are main-

    tained in an insulated, temperature controlled (26.5 C.) (70-90%RH) darkened


    The adults are maintained on sugar solution or corn syrup oncotton wool and

    are offered a blood meal on the day after emergence, and onalternate days after

    that. Rabbits are used as a source of blood meals. The back ofthe rat&it

    is shaved

    and the unanesthetized rabbit is placed in a box in the cage.Mating occurs at dusk

    or in a darkened room or cage.

    Maximum mating occurs 3-5 days after emergence



    Fertilized females take a blood meal more readily

    than unfertilized


    Approximately 15.6/, survive 20 days (Wegesa 1964).


    Armstrong and Bransby-Williams (1961); Causey et al. (1943);Coluzzi (1964); Gillies

    (1961) ; Goma (1959)


    Haddow and Ssenkubuge (1962)


    Jones,M. D. R. et al. (I


    ; Mathis,

    C. (1936); Mathis, M. (1935); Moores (1953); Muirhead-Thompson(1948); Philip (1930);

    Shute, G.


    (1956); Wall (1953); Wegesa (1964).

    Anopheles kymanus


    Anopheles jamesii Theobald




    collected in petri dishes 1% in diameter.


    Larvae are reared on a medium (pH 7.1)

    consistingof mud water and tap \lrater.

    Larvae are fed on brown bread powder.


    Pupation occurs IO days after oviposition.

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    The adults are maintained in Barraud cages 6x6~6. They are fedwater-soaked,

    split raisins, placed on top of the cage and covered with a padof dam p cotton w ool.

    The adults emerge in 24 hours.



    Anopheles labranchiae Falleroni

    Anopheles labranchiae subsp.


    Van Thiel

    Eggs are collected on damp filter paper. The filter pape r isplaced on wet cotton

    in small plastic cups or in petri dishes. Eggs are left on dampfilter p aper for at least

    24 hours but not more than 72 before hatching. The eggs areremoved with the

    filter paper for hatching purposes.


    Approximately 300-40 0 larvae are placed in a white enamel orpolyethylene bowl

    34cm (14) in diameter, containing water to a depth ofapproximately 3-4 cm (I s").

    The water temperature should be about 27 C. (80 F.). Mu d or apiece of grass

    turf is used in add ition to artificial food. The use of thesevariables leads to non-

    standardized mosquitoes.

    Ground dog biscuits, grain foods and liver powder are

    used successfully.


    Pupae are usually hand picked though mechanical means ofseparation may be



    One thousand or more adults are maintained in a 2x2~2' cage. Thepup ae are

    placed in the cage in sm all containers and the containerscovered or rem oved after

    adult emergence.

    The adults are maintained on 10% sucrose soaked cotton woolor

    cotton balls. Restrained guinea pigs are laid on top of a cage,or a human arm is

    inserted into the cage to provide a blood m eal.


    DpAlessandrot al. (1961); Bertram and Gordon (1939); Coluzzi(1964); Meller (1962);

    Shute,P. G. (19 36).

    Anopheles maculatus Theobald

    Each female lays about 80-100 eggs on moist filter paper, inindividual tubes or

    in paper cups,



    et al. 1963).

    At the ICR laboratories the fertilized females

    are placed in 1~1x1

    alum inum screened cages and lay eggs in plastic containers.


    Jayewickreme (1952) reared larvae in a medium consisting of 20parts hay infu-


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    sion to 80 parts tap water.

    75 cc of hay infusion was added daily. Larvae were

    reared in an earthenware dish with a diameter of 20 cm and asurface of 314.2 sq cm

    containing 500 cc of media.

    Larvae were fed on the brown bread powder after the

    4th day.


    Maximum pupation occurred on the 13th day after oviposition.



    Maximum adult emergence occurs on the 15th day afteroviposition. The adults

    are maintained in Barraud cages 6x6~6 and are fed water-soakedraisins placed on

    top of the cage and covered with a pad of damp cotton wool.

    Jayewickreme (1952)

    does not mention mating or egg production. Ow Yang et


    (1963) were able to

    maintain a colony by artificial mating.

    Sexed pupae (by size) emerged in separate



    They were maintained at 27 C. and 70-90% RH and fed on 5%

    glucose solution. Two days after emergence the glucose wasremoved and a guinea

    pig was placed in the cage overnight. The following morningengorged females were

    collected in individual tubes.

    Males 3-6 days old were used for the artificial mating

    (see techniques section).



    (1952); Ow

    Yang etd. (1963).

    Anopheles pharoensis Theobald

    Eggs are collected in petri dishes containing water. Theembryonic period is 2-4

    days, depending upon temperature. The percentage of eggfertility varies from

    35-95% with an emergence of 70% (Theodor and Parsons 1945).


    The optimum density of larvae is roe-150 first stage larvae and50 third or fourth

    stage larvae in a bowl of 15 cm diameter.

    The rearing technique of Theodor and

    ParSOnS 1945)

    consistsof placing IOO first stage larvae into the white enamelbowl

    containing a layer of mud I cm in depth and about 4 cm of water.The larvae are

    daily fed a few drops of bakers yeast in water.

    At the second stage, they are fed

    powdered biscuits,

    sprinkled on the surface of the water-sometimes 3 or 4 times

    a day. The larval period is I 1-12 days at 28-29 C. At 23 C. thelarval period

    is 23 days and at 27O C. about 18 days.

    The pupae are removed every morning and placed in a bowl insidethe cage.


    pupal period lasts 3-4 days at



    The adults are maintained in a large cage 6ox6oxIoocm.

    Humidity is maintained

    by covering the sides and front of the cage with a moistblanket. The adults are

    fed a sugar solution on a soaked cotton pad or sponge. A rabbitwas first used for

    blood meals, but a marked reluctance to feed was observed. Humanblood is



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    acceptable. Swarming and

    white light is turned on.

    mating takes

    place in blue light and


    when a


    Abdel-Malek et al. (1966)


    de Meillon et al. (1963); Theodor and Parsons (1945).

    Anopheles pseudopunctipennis Theobald


    Downs and Arizmendi




    Anopheles punctipennis (Say)


    Hardm an (I 947).

    Anopheles punctulatus Donitz




    MacKerras and Lemerle


    Anopheles quadrimaculatus Say


    Eggs are collected in a pan or bow l of water, or on damp filterpaper or toweling.

    Avoid desiccation of the eggs. Gah an (1967 ) states thatwhenever a surplus of eggs

    is obtained, the excess supply may be wrapped in damp filterpaper o r toweling and

    may be stored in a refrigerator for IO days to 2 weeks. Eggs areusually deposited

    at night. Rep lace the egg collecting container daily. Eggs areconcentrated and

    introduced to larval rearing trays or the eggs are left to hatchand then co unted by

    the aliquot method and placed in the rearing trays. The eggshatch in 30-40 hours.


    Trays of various sizes

    and composition are used.

    White enamel


    or white

    plastic trays are used with su

Rearing Mosquitoes - [PDF Document] (2024)


What is the protocol for mosquito rearing? ›

Eggs are collected on moist filter paper. A small bowel is lined with a 3″ wide strip of filter paper. Water is added to a depth of 2.5 cm. The container is then placed in a cage of adults, the egg collecting container is left in the cage for 48 h.

How to rear mosquito larvae? ›

To prepare the large trays for rearing, each tray should be filled with 5L of water the day before adding the larvae to allow the water reaching ambient room temperature. Trays can also be filled with 5L of water the same day as larvae (water depth is between 1.2 and 1.5 cm).

How to stop mosquitoes from reproducing? ›

Prevent Mosquito Breeding
  1. Empty, drain, or cover all things that can and may hold water. ...
  2. Maintain decorative ponds and ensure swimming pools are operating properly. ...
  3. Repair or prevent outside leaks. ...
  4. Change water collection pans for plant pots weekly. ...
  5. Clean bird baths weekly. ...
  6. Mow your lawn weekly.

How do you gather mosquitoes for research? ›

Build a mosquito trap.

By building a mosquito trap, you will be able to collect and count the eggs laid by mosquitoes over a set period of time. You can then share this data with epidemiologists, who will use the information to determine the growth and location of mosquito populations.

What is the best time to start mosquito treatment? ›

The best time to start mosquito spraying is shortly before their season begins. Mosquito activity peaks when temperatures are between 70-80 degrees Fahrenheit, but they become active at temperatures as low as 50 degrees Fahrenheit.

How many days does it take for mosquito eggs to hatch? ›

Eggs will hatch into larvae within 24 to 48 hours. Larvae soon grow to become approximately 5 mm in length. Most larvae breathe through air tubes.

What do mosquitoes hate the most? ›

What smells do mosquitoes hate? Mosquitoes hate the smell of lavender, citronella, clove, peppermint, basil, cedarwood, eucalyptus, peppermint, lemongrass and rosemary.

What to put in water to stop mosquitoes from breeding? ›

Adding Kerosene | Medicinal Paraffin to Your Water

One of the traditional ways to prevent mosquitoes breeding was to use a small amount of kerosene in your water tank.

How do farmers keep mosquitoes away? ›

Clean cattle tanks and watering troughs regularly to prevent mosquitoes from multiplying. Mosquitofish may also be added to provide control. Remove or destroy troughs or tanks that are not in active use. Good agriculture results from the efficient use of water, and efficient use of water reduces mosquito populations.

Who attracts mosquitoes most? ›

The Rules of Mosquito Attraction

That means people who have a high metabolic rate and emit more carbon dioxide, including those who are pregnant, working out, or drinking alcohol tend to be more attractive to mosquitoes. The question of whether mosquitos prefer a certain blood type is controversial.

What is the best method against mosquitoes? ›

Eliminate Standing Water

One of the most effective mosquito control methods is eliminating the insects breeding grounds. Mosquitoes lay eggs in stagnant water, so inspect your surroundings for standing water sources such as empty containers, buckets, or clogged gutters.

How much water is needed for mosquitoes to breed? ›

Some mosquitoes can lay their eggs in as little as 1/4” of water and can lay hundreds of eggs at a time—so even very small sources can become a big problem. The eggs then hatch into larvae which live in the water.

How long do you have to wait after mosquito squad? ›

Our products are mixed only by Mosquito Squad management in accordance with the product labels, our technicians apply the products per the label instructions making them non-toxic to people, pets, and wildlife. However, we do recommend people and pets wait 30 minutes before re-entry outside to enjoy the property.

What are the guidelines for mosquito repellent? ›

Use only enough repellent to cover exposed skin in a thin, even layer. Using more repellent than what is required does not make it more effective and may increase skin irritation. Do not use on cuts, wounds or irritated skin. Do not apply to areas around the eyes or mouth.

What is the method of feeding mosquitoes? ›

ADULT mosquitoes of both sexes can be persuaded to take a considerable variety of foodstuffs, including fruit juices, milk, sugar solutions and various blood preparations.

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