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ECU.',ITV CLASSIFICATION OF THIS PAGE (When Dei Entered)REPORT DOCUMENTATION PAGE READ INSTRUCTIONS

BEFORE COMPLETING FORMI. REPORT NUMBER 2. GOVT ACCESSION NO. 3. RECIPIENT'S CATALOG NUMBER

ANNUAL-REPORT No.2 1TITLE(endSubtitle) Evaluation of Insecticides, TYPE OF REPORT & PERIOD COVERED

0clothing repellents, and other approaches to the Annual-Oct. 1, 1980 tocontrol of coastal sand flies, CuLicoides spp Sept.30, 1981

6. PERFORMING ORG. REPORT NUMBER

7. AUTHOR(,) 8. CONTRACT OR GRANT NUMBER(e)

Daniel L. Kline and R. H. RobertsP.O. Box 14565 N0 1-79-F-0070Gainesville, Florida 32604

9. PERFORMING ORGANIZATION NAME AND ADDRESS 10. PROGRAM ELEMENT. PROJECT. TASKAREA 8 WORK UNIT NUMBERS

Insects Affecting Man and Animals ResearchLaboratory NR 133-997

II. CONTROLLING OFFICE NAME AND ADDRESS 12. REPORT DATEOffice of Naval Research Code 443 01 November 1981

800 N. Quincy Street t3 NUMER OF PAGES

Arlinthnr Vionin ~~714. MONITORING-AGENCY NAME & ADDRESS(If different from Controlling Office) IS. SECURITY CLASS. (of thl report)

UnclassifiedSame as 11. IS.. DECLASSIFICATION/DOWNGRADING

SCHEDULE

16. DISTRIBUTION STATEMENT (of this Report)

Approved for public release; distribution unlimited.

17. DISTRIBUTION STATEMENT (of the abstract entered In Block 20, it different from Report)

IS. SUPPLEMENTARY NOTES

19. KEY WORDS (Continue on feee* ide If . e... eayand Identify by block number)

Seasonal population dynamics, CuZ{icoides spp., treated screens, habitat0characterization, laboratory developmental studies, artificial membrane,

field ground and aerial ULV.LIJ

-J 20. ABSTRACT (Conti.ue on reverse ade if nece.ay and identify by block nub.er)U-,

DD IJAR? 1473 EDITION OF I NOV GS 1OBSOLETES/N 0102-LF-0144601t

FSECURITY CLASSIFICATION O7 TNIS PAGE (l1he. Date SniEseo)

... . . . . . . ..-"--7

20. ABSTRACT

Population dynamics and control studies on CuZicoides sand flies were continuedat Parris Island, South Carolina, and Yankeetown, Florida.

Adult seasonal patterns were monitored by NJ light traps. Five species, C.barbosaiWirth and Blanton, C. furens (Poey), C. hoZlensis Melander and Brues, C. meZleus(Coquillett), and C. mississippiensis Hoffman are considered abundant. C. barbosai,C. furens and C. melleus are present from mid-April through late October; C.hollensis and C. mississippiensis peak in the spring and fall of the year. Acomparison of several population survey techniques indicated that CDC-type trapsbaited with CO2 gas and used in conjunction with New Jersey traps should give areliable estimate of CulZicoides activity during chemical control tests.

Twenty-one different larval habitats were identified at Parris Island. Areas alongthe upland edge of the salt marshes and areas underneath large logs which hadfallen into the marsh consistently yielded larvae, while Salicornia and denudedpanne areas seldom yielded any larvae. At Yankeetown fluctuations in larvaldensity were correlated with plant cover. Based on mean number larvae/sample,substrate samples taken from Distichlis areas yielded the greatest number oflarvae followed in decreasing order by Spartina and Juncus areas. These datasuggest that a properly timed effective larvicide could reduce adult emergence by53% by treating 30% of the marsh. 'Remote sensing techniques were used to locateand quantify breeding sites.

In laboratory studies, 20*C was the optimum temperature for development (57 days)of C. mississippiensis. Reared females were 100% autogenous. Techniques were de-veloped to blood-feed C. mississippiensis through a cloth-reinforced siliconemembrane on bovine blood; there was no significant decrease in egg production*compared to that of females fed on a human host.

-In chemical control studies, laboratory treated screens, field ground ULV andaerial ULV tests were conducted. NRDC-161 and permethrin gave ca. 100% mortality168 days after the screens were treated at .125% A.I. (wt/vol. technical in acetone)for NRDC-161 and at .5% for permethrin., Ground ULV tests against C. mississippiensiswith resmethrin gave indeterminate results, and aerial ULV tests with half thelabel dosage rate of naled at Parris Island gave a disappointing level of control.In both of these spray tests the level of control was probably due in part toinsufficient penetration of spray in vegetated areas.

Deet-treated net jackets were tested in Panama and provided ca. 90% protection.

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' I.

SUMMARY

Population dynamics and control studies on Culicoides biting midges (sand flies)were continued at Parris Island, South Carolina, and Yankeetown, Florida.

Adult seasonal patterns were monitored by modified New Jersey light traps. Threespecies (C. furens Poey, C. hollensis Melander and Brues, and C. me~leus (Coquillett)at Parris Island and 3 species (C. barbosai Wirth and Blanton, C. furens andC. mississippiensis Hoffman) at Yankeetown are considered major pests. C. barbosai,C. furens, and C. melleus have several peaks from mid-April through late October.Both C. hollensis and C. mississippiensis peak in the spring and fall of the year.At Yankeetown, fifteen other species were collected infrequently, and comprised<1% of the total trap collections.

A comparison of several trapping methods (Malaise, CDC traps baited with CO2 gas,

unbaited CDC traps, and New Jersey light traps) was done at Parris Island todetermine their reliability to assess the effects of ground and aerial ULV sprayson natural populations. From this comparison study, we concluded that baitedCDC traps used in conjunction with N.J. traps should give a reliable estimate ofany adult Culicoides activity at Parris Island.

Larval habitat characterization studies were conducted at Parris Island andYankeetown. At Parris Island the emphasis was placed on identifying all thedifferent potential habitats. Twenty-one different types of habitats wereidentified, of which 2 consistently yielded larvae (upland edge of the marshshaded by live oaks and/or water oaks; and beneath large logs which have fallenout into the marsh), and 2 areas which yielded practically no larvae (Salicorniaand completely denuded panne areas).

At Yankeetown the emphasis was placed on completing the study on population fluctua-tion of immatures within a gridded 125 m X 425 m section of typical salt marshwhich contained three main vegetative types (Juncus, Spartina, and Distichlis).Based on the mean number of larvae/sample, overall substrate samples taken fromDistichlis areas yielded the greatest number of larvae followed in order bysamples taken from Spartina and Juncus areas. The study area, however, wascomposed of ca. 70% Juncus, 23% Spartina, and 7% Distichlis. When this factor istaken into account, the overall projection for FY81 was that Juncus would accountfor 62%, Spartina 28%, and Distichlis 9% of the larvae recovered. If this studyarea is truly typical of salt marshes in the vicinity of Yankeetown, then a poten-tial 38% reduction in the adult population could be achieved by treating 30% ofthe marsh area with an effective larvicide. If the correct month for treatmentwere selected, such as February, up to 53% of the larvae could be killed by

treating the same 30% of the marsh.

Studies to determine the feasibility of using remote sensing techniques to locateand quantify breeding sites was initiated in February, 1980. Two remote sensingflights over the salt marsh grid area at Yankeetown, and one over Parris Islandwere flown by the 363rd TRW/DOO, Shaw AFB, South Carolina.

In the laboratory, C. mississippiensis was successfully reared at 150, 20, and25'C. Development times were inversely related to temperature. The optimumtemperature for rearing was 20°C. Complete development at this temperature re-quired a total of 57 days. Reared females were 100% autogenous and matured anaverage of 123 eggs. Blood-fed wild females with the same wing length as thereared females matured an average of 35 eggs.

![I

Techniques were developed for blood-feeding field-collected C. mississippiensison preservad bovine blood through a cloth-reinforced silicone membrane. Therewas no significant difference (p < 0.05) between the numbers of eggs matured byfemales which had fed on a human host (40 + 11) or through the membrane on bovineblood (36 + 13).

Two synthetic pyrethroids, permethrin and NRDC-161 (Decamethrin @) were evaluatedfor their effectiveness as residual screen treatments to prevent entry of C.mississippieneis into screened areas. NRDC-161 was the most effective with screens

treated with this insecticide still causing ca. 100% mortality at all concentrationstested (.125, .25, 15, and 1.0% A.I. (wt./vol.) 168 days after they were treated.Permethrin gave comparable results at .5 and 1.0%.

Ground ULV tests against natural C. miscissippiensis populations at Yankeetownwith resmethrin gave indeterminate results. There was some indication that thefield population was reduced, but there was high variability in the collectionnumbers and in the mortality of caged mosquitoes and biting midges. Two majorproblems in achieving control appear to be timing the spray application and penetrat-ing the vegetative cover which, when dense, appears to restrict movement of theaerosol through the area.

During the period of 1-9 April, 1981 a joint USDA-DOD research project at theMarine Corps. Recruit Depot, Parris Island, South Carolina, was conducted todevelop and evaluate aerial application of naled (Dibrom 14 @ and HAN forCulicoides biting midge control. One of the goals in this first trial was to testthe use of an application rate lower than the maximum allowable rate in order toreduce costs and the possibility of environmental damage. Valuable results andinformation were obtained even though the level of control achieved was somewhatdisappointing. Collections from surveillance traps indicated that the spray had asignificant impact on the natural population of biting midges on Parris Island whencompared to the untreated area surveyed on nearby Lemon Island.

Although the population in both areas actually increased on the day following treat-ment, the post-treatment population of biting midges on Parris Island was approxi-mately 50% lower than those on Lemon Island. The effects of the spray were alsomonitored using caged mosquitoes and biting midges. Results of this bioassay in-dicated an overall 67% kill of caged mosquitoes and 58% kill of caged biting midges.This level of control was probably due in part to insufficient penetration of thespray in vegetated areas. In support of this view the kill of caged mosquitoesplaced in open or sparsely vegetated areas was 87%, while the mortality in cagesplaced in dense vegetation was only 40%. Although the number of caged CuZicoideswas somewhat limited, the Culicoides kill was also better in open areas. The lowdose of chemical (1/2 oz/A) applied was possibly insufficient to give good pene-tration through vegetation. Other environmental factors, such as marginally highwinds and low temperatures during the application, could have adversely affected thetreatment.

Light-weight net jackets treated with N, N-diethyl-m-toluamide (deet) were field-tested in Panama against 5 species of biting midges, principally C. furens andC. barbocai. The jackets provided ca. 90% protection.

Area treatment tests with deet impregnated netting were initiated at Yankeetown.These investigations are expected to be completed in 1982.

A-

I. INTRODUCTION

This report summarizes research activities on Culicoides biting midges(sand flies) conducted at Parris Island, South Carolina, and Yankeetown,Florida during FY81. Population studies were continued to complementthe first year's studies. In the chemical control studies emphasisshifted from laboratory screening to field testing.

II. POPULATION DYNAMICS AND BIOLOGICAL STUDIES

Seasonal Patterns of Adults

Seasonal population studies of adult Culicoides were continued at ParrisIsland, South Carolina, and Yankeetown, Florida. New Jersey light traps,modified by replacing the standard delivery cone with 40-mesh brass screening,were used at both sites. Data were obtained on species composition, seasonalincidence, and relative abundance.

At Parris Island, four New Jersey light traps were operated nightly.Samples were retrieved from the field at least twice weekly (3- and 4-dayintervals). Trap catch results for the period October 1, 1980 throughSeptember 30, 1981 are listed in Table 1. Three species C. furens, C.hollensis and C. melleus are considered major pest species at this loca-tion. C. furens was abundant during October and from April through Septem-ber. This species was most abundant from the last week in April throughthe first week in June. C. hollensis was abundant in light trap collec-tions for the month of October with occasional specimens collected fromNovember through February and was then very abundant from mid-March untilthe end of May. Thereafter, C. hollensis was virtually absent from lighttrap collections until the last week in September. C. melleus was abundantduring the month of October and then was not collected again until thelast week in March. This species reached its spring peak in April, thenremained at low levels throughout the summer months and reached a fallpeak during the first week of September.

At Yankeetown, two modified New Jersey light traps were operated nightly.Samples were retrieved from the field at approximately weekly intervals.Three species, C. mississippiensis, C. furens and C. barbosai were abun-dant, and trap catch results for these species for October 2, 1980 throughSeptember 30, 1981 are listed in Table 2. Fifteen other CuZicoidces species(Table 3) were collected infrequently; these comprised < 1% of the totaltrap collections. Trap downtime due to malfuntion, vandalism, and othercauses was less than 10%.

C. mississippiensis was present throughout the year, but abundant onlyin spring and fall. Data from Table 2 show a sharp rise in abundance duringNovember-early December and a second rise beginning in February and peakingin May-early June. Compared with the previous year's data, the fall peakoccurred slightly later and the spring peak was slightly more protracted.C. furens adult abundance peaked in May-June and August-September. Thesespecies Seasonal patterns are very similar to last year's. Abundance ofC. bartosai paralleled that of C. furens but on a lesser scale.

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Comparison of Techniques for Trapping Adult Culicoides spp.

In order to evaluate the effectiveness of any control procedure there is aneed to establish a survey method that gives reliable estimates of popula-tions. Since many survey techniques are available to collect adult Culicoides,we decided to evaluate several at Parris Island to determine their reliabilityin monitoring the effectiveness of ground and aerial ULV sprays.

In fall 1980 we compared unbaited U.S. Army miniature solid state type CDClight traps with our standard adult population assessment trap, the modifiedNew Jersey traps mentioned above. A complete description of this CDC trapcan be found in Driggers et al. (1980). The trap is automatically activatedat dusk in response to decreasing ambient light conditions. Once activated,the motor and lamp operate continuously until the power supply is disconnect-ed. Twelve CDC traps were used. The New Jersey traps were controlled byautomatic timers which were adjusted so that these traps were active forthe same time period as the CDC traps.

Trap locations for the New Jersey traps remained the same as in our seasonal

population assessment program. The CDC traps were placed in similar areaswhere CuZicoides were known to be a problem, based either on personal ex-perience or complaints by recruits, and were distributed in such a way thatthe entire base was surveyed. The holding device in both type traps was thekill jar, which consisted of a glass pint mason jar containing a small stripof Shell No-Pest Strip ®.

All three major pest species, C. furens, C. hollensis, and C. melleus, werecollected (Table 4). It was observed that within reasonable limits, thetrends were similar for both types of traps. The New Jersey traps generallycaught more specimens, which was expected since it used a 40-watt light bulbas opposed to the miniature 6 volt lamp used in the CDC trap. For C.furens and C. meleus, the collections in both traps accurately reflectedbiting complaints. For C. hoZlensis, however, in the latter part of Octoberthroughout the rest of the sampling period light trap collections were in-consistent with biting complaints. A possible explanation was that sincethese traps were activated at dusk and ceased activity at dawn, the ambienttemperature was too low for flight to occur during this time period. Wehave observed, as have other investigators, that Culicoides flight activity

ceases below 55*F.

Further studies were conducted in spring 1981. In this study four types oftraps were used: 4 modified New Jersey traps, 6 unbaited and 6 C02 gas-baited (200m /min) army-type CDC traps, and a fine mesh Malaise trap.A different strategy from the fall study was employed, in that all thetraps were operated continuously during any 24 hr period. From mid-Marchto mid-April, when C. holensis was the predominant species, the temperature,at dusk, was < 60*F, usually below 550 F. Within this time interval thebaited CDC trap collected more than any of the other traps with a few ex-ceptions when the Malaise trap collected more. Once the temperature re-mained consistently greater than 550 F. at dusk, the collections in theunbaited CDC and New Jersey traps increased and similar trends were noted

for C. hollen is by all 4 trapping methods.

From mid-April through June, C. furens was the predominant species. Gener-ally, this species is present during the time of year when warmer eveningtemperatures occur. Peak activity normally occurs around dusk and dawn.Consequently, the baited CDC and New Jersey traps showed similar trends.The unbaited CDC and malaise trap collections showed inconsistent trends;based on the collections in the fall, 1980 test, we are unable to explainthese poor collections.

P a .

From these data we conclude that baited CDC traps used in conjunction withour New Jersey traps should give a reliable estimate of any adult Culicoidesactivity at Parris Island.

Studies of Immature Populations

At Parris Island a search for larval habitats was initiated in December withemphasis on identifying all possible breeding sites. To date, twenty-onedifferent types of potential habitats have been investigated (Table 5).Two habitats that consistently yielded larvae were the upland edge of themarsh shaded by live oaks or water oaks, and beneath large logs which hadfallen out into the marsh. Samples taken from these types of areas inpast years, had also consistently yielded larvae. Areas covered withSalicorni, or completely denuded (panne and mudflats within tidal creeks)did not produce many larvae.

At Yankeetown the larval habitat characterization studies initiated lastyear were continued within a 125m X 425m section of typical salt marsh.This study area was subdivided into eighty-five 625m X 425m plots.Each plot was characterized according to the different vegetative typesfound DistichZis, Juncus, and Spartina and various mixtures of thesegrasses. Sod samples (ca. 10cm diam. X 8cm deep) were taken weekly fromeach major vegetative cover by means of post hole diggers. The locationof each sample was determined through random selecion of the plots inwhich the desired vegetation type is found.

The data collected at Yankeetown between October 1, 1980, and September 30,1981 are shown for the major vegetative types in Table 6. Fifty sampledates yielded 1486 samples and 15,207 larvae. Compared to FY80 results,larvae were more abundant in all three vegetative areas. For 7 months ofthe year DistichLio samples yielded the highest mean number larvae/sample,and for the remaining 5 months Spartina areas yielded the highest meannur'ber larvae/sample. With the exception of July, samples taken from ;Uncusdomitnated areas always yielded the least number of larvae.

Overall, based on the mean number larvae/sample, Juncus areas yielded 26%,Spartina areas 36% and Distichlis areas 38% of the larvae recovered. Thesefigures would imply that Spartina and DistichZis account for 74% of thelarvae produced at this marsh site. These figures, however, need to beadjusted to reflect the relative abundance of these 3 vegetative typeswithin our study area. From aerial photographs, followed by groundtruthing, the study area was calculated to be composed of ca. 70% Jun,-us,23% Spartina, and 7% Distichlis. Therefore, a readjustment of the meannumber of larvae recovered from each vegetative type acquired through

sampling must be made to account for the disportionate area each typeoccupies within the marsh. When this adjustment is made, the overallprojection for the entire year shows that Junauo would account for 62%,Spartina 29% and Distichlis 9% of the larvae recovered from this marsh inFY81. If these population projections, based on our grid sampling, aretypical for all the marshes in the Yankeetown area, then a potential 38%reduction in the subsequent adult population could be achieved by treating30% of the marsh area with an effective larvicide. An even greater re-duction can be achieved if these same areas are treated in the proper month.For example, a glance at Table 6 would indicate that Febrauary would be agood month to treat. If an effective larvicide were applied to these areasin February, 53% reduction could be achieved.

Remote Sensing Techniques for Population Distribution

Our studies on the population dynamics and habitat characterization forCulicoides included aerial surveys during the last year. Two remote sensingflights of our salt marsh grid area at Yankeetown, and one over ParrisIsland were flown by the 363rd TRW/DOO, Shaw AFB, South Carolina. Theobjective was to determine the feasibility of this technique to locate andquantify the various vegetative types. These flights indicate that aerialphotography, especially infrared techniques, can be used to quantify theamounts of different types of plants in an area. Therefore, we feel thatremote sensing techniques, coupled with reliable ground-truth data, willprovide a rational approach for assessment of biting midge production in agiven geographic area and therefore aid in their control.

Laboratory Studies on Life Cycle of C. mississirpiensis

Development of C. mississippiensis was successful at 150, 200, and 25*C,but not at 100 or 300C. Development times were inversely related totemperature. The optimum temperature for rearing was 20*C at which com-plete development required a total of 57 days. The egg stage lasted 8 days,the larval stage 40 days, the pupal stage 5 days, and the adult pre-ovi-position time 4 days. Reared females were 100% autogenous and matured anaverage of 123 eggs. Blood-fed wild females with the same wing length asthe rear&ed females matured an average of 35 eggs. Engorgement with bloodrequired 4 min 19s (S.D. = 19s).

Use of Artificial Membrane for Feeding Adults

Techniques were developed for the preparation and use of a reinforcedsilicone membrane in laboratory blood-feeding of C. mississippiensison preserved bovine blood. The membrane was made from Sears Clear SiliconeGlue and Seal ® # 9-80672 rolled over a piece of polyester organdy cloth.In 12 feeding trials conducted on 4 dates, the mean feeding percentage was77.6% (S.D.= 9.2%). Production and hatch of eggs from females fed onbovine blood through the membrane were compared to that of females capturedon the same dates but allowed to feed to repletion on a human forearm.There were no significant differences (p < 0.05) between numbers of eggsmatured by females which had fed on a human host (40 +11) or through themembrane on bivine blood (36 +13).

III. CHEMICAL CONTROL STUDIES

Evaluation of Treated Window Screens

Two synthetic pyrethroids, permethrin and NRDC-161 (Decamethrin ®), weretested at .125, .25, .5 and 1.0% A.I. (wt./vol. technical in acetone) inevaluations of their effectiveness as residual screen treatments to retardor prevent entry of (. mississippiensis adults into screened areas. Discs(9cm diameter) of standard aluminum screen (16 by 18 mesh) were treated byimmersion for ca. 10 seconds in the solution. After treatment, the screenswere hung under a shelter, designed to produce the effects as if they werehung beneath the eaves of a building to dry. Control tests were conductedwith screens immersed in acetone only. Exposure techniques were identicalto those described in the previous Annual Report (1980) NRDC-161 was themost effective with screens treated with this insecticide still causing ca.

100% mortality at all concentrations tested 168 days after they were treated.After 168 days, we had difficulty trapping flies for testing. Based onthese results, a single treatment of this chemical should give completeprotection for an entire C. missinsippiencio season. Permethrin treatedscreens at .125% gave < 50% mortality 1 day after treatment. At .25%ca. 90% mortality was produced for at least 56 days post-treatment. At

S.5 and 1.0% ca. 100% mortality was achieved for the entire 168 day period

that the screens were tested.

Ground ULV Tests

Ground ULV tests were initiated at Yankeetown, Florida, against naturalC. mississippiensi! populations with resmethrin. Effectiveness of theground ULV sprays was assessed by population monitoring with CDC lighttraps baited with CO2, caged mosquitoes, caged biting midges, and cageswith both mosquitoes and biting midges. The combination was necessary inorder to test spray penetration of the small mesh screen used to confinethe biting midges. Two tests were conducted. The resmethrin was sprayedat the rate of 0.007 lb/acre. In the first test, the spray was deliveredat 4 oz/min at 10 mph and in the second test at 9.2 oz/min at 10 mph.

The results were indeterminate. In test one, high mortality was obtainedin all cages at two locations, and slight to no mortality at the otherlocations. In test two, high mortality in the control cages negated thetest cage results. While there was some indication that the field popula-tion was reduced, high variability in collection numbers prevented accurateassessment.

One major problem in achieving control appears to be the timing of sprayapplication. C. mississippiensis is active for several hours before sun-set, therefore, treatments were made between 5 and 6 PM, ca. 2 hours be-fore sunset. At this time of day, thermal air currents, especially overthose sections of the road exposed to the sun, prevented proper aerosoldispersion. A second factor in obtaining adequate coverage with groundULV equipment is the influence of the vegetative cover which, when dense,appears to restrict movement of the aerosol through the area.

Aerial ULV Tests

An aerial application test for control of Czdicoidas at Parris Island wasconducted on April 7, 1981 using the conventional insecticide naled (Dibrom-14). Two Air Force UC-123-K aircraft applied the insecticide using techni-ques normally employed for mosquito adulticiding. However, the insecticidewas applied in two treatements spaced ca. 20 minutes apart using 2 aircraftto take advantage of any "flushing action" on the insect population causedby the first application. Each of the two treatments applied a 1:5 mix-ture of Dibrom-14:HAN (Heavy Aromatic Naptha) at a rate of 1.5 oz/A whichincluded 1/4 oz /A HAN. This resulted in a total application rate of 1/2oz/A of Dibrom-14 which is the minimum dose recommended for adult mosquitocontrol. The entire land and marsh area of Parris Island (ca.8000 acres)was treated. The aircraft were equipped with Tee Jet nozzles oriented450 forward and were flown at 140 kts airspeed, 2,000 ft swath width and200 to 250 ft. altitude. Meteorlogical data during the insecticide applica-tions were collected by a weather team from the Marine Corp. Air Station,Beufort, South Carolina. (Table7 ). Winds were generally easterly with aspeed of ca. 7 kts on the ground. Wind speed aloft increased somewhat during

_ _ _ _ _ _ _

the spray operation from ca. 8 kts, at 200' 30 minutes prior to the start ofspraying to ca. 14 kts at 200' 30 minutes after the start of spraying.The effectiveness of the insecticide treatment was determined by measuringthe population by surveillance traps and by bioassays with caged insects.

Several trapping methods were employed at different locations on ParrisIsland and in an untreated control area on nearby Lemon Island to monitorthe natural population. CDC light traps baited with CO2 and operated con-tinuosly (day and night) provided the best population indicator for C.hollensis, which was the predominant species present during this study.This species had peak activity periods that included some daylight hoursduring the morning and afternoon. Trap collections were collected andcounted twice daily (early morning and afternoon) to quantitate activitypeaks. Six CO2-baited CDC traps were operated on Parris Island and 2 onLemon Island. The traps were placed 4 ft. above ground on metal stakes.CO2 gas cylinders at a flowrate of 200 cc/min, which was measured by afloating ball flowmeter on each trap. The traps were operated for 10 dayspre-treatment and 1 day post-treatment.

Caged Aedes taeniorhynchus females (25/cage) were used in the treated areaas an indicator of insecticide distribution and effectiveness. A totalof 65 cages of mosquitoes were placed on 4 ft stakes throughout ParrisIsland with 56 cages along roadways at 0.1 mile intervals and the re-mainder placed at trapping stations. A limited number of caged sand flies(11 cages with ca. 100/cage) was also placed in the treated area. Thesand flies were field-collected using a modified CDC trap baited with CO2gas and then released onto a lighted window and aspirated into cages con-sisting of 1/2 pint cylindrical paper cartons with 40 ga screen wire onthe top and bottom.

Results of the natural population surveys and caged insect bioassays in-dicated that only a low level of control resulted from the aerial applica-tion. The surveillance traps indicated that the population level actuallyincreased in both the treatment and control areas; however, the popula-tion level in the treated area was ca. 50% lower than the level in thecontrol area. Since the pre-treatment population levels for both areashad been nearly the same this suggests that the insecticide achieved a-round 50% mortality in the treated area. This agrees with the results ofthe caged insect bioassays where the ove.rall mortality in caged mosquitoeswas 67% and in caged sand flies it was 58%.

A number of factors probably contributed to the low level of control. Wefeel that one important factor was insufficient penetration of the sprayin vegetated areas. This was indicated since the kill of caged mosquitoesplaced in open or sparsely vegetated areas was 87%, while only 40% killwas noted in cages protected by dense vegetation. Although the numberof caged sand flies was limited, their mortality was also greater in openareas. The low dose of chemical applied in this test was possibly in-sufficient to give good penetration of the vegetation. Other environmentalfactors such as marginally high winds and low temperatures during theapplication could have adversley affected the treatment.

IV. PERSONAL PROTECTION

Light-weight net jackets treated with "W,N-diethyl-m-toluamide (deet) werefield-tested in Panama against 5 species of biting midges, principally

CuZicoides furens (Poey) and C. barbosai Wirth and Blanton. The deet-treatedjacket provided 87 to 93% protection. Morning and evening tests as well astime of year appeared to influence the proportionate numbers of speciespresent. The mean coefficient of protection was slightly lower during

morning tests when C. barbosai was most abundant and higher during evening

tests when C. furens was most abundant.

Area treatment tests with deet impregnated netting were initiated atYankeetoyn, Florida. PVC pipe (1" diameter) was used to construct 2 units,each 2 m and 2 high. The sides of the units were covered with netting and

the tops were left open. The netting of one unit was treated with deetand the other left untreated. A sticky trap baited with CO2 gas (.51/min)was set up in the center of each unit and daily counts were made of thebiting midges trapped. Periodically, I or 2 subjects made biting countsinside and outside the units. These units were tested daily until they

failed to provide any protection from blood-seeking female biting midges.An outside sticky trap was also set up ca. 75 m away from either cage.These investigations are expected to be completed in 1982.

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REFERENCES CITED

Driggers, D.* P., O'Connor, R. J., Kardatzke, J. T., Stup, J. L. and Schiefer, B. A.,

1980. The U.S. Army Miniature Solid State Mosquito Light Trap. MosquitoNews 40:172-178.

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BIBLIOGRAPHY OF PUBLICATIONS/MANUSCRIPTSPREPARED UNDER THIS CONTRACT FY 1981

PUBLICATIONS

Kline, D. L. and R. H. Roberts. 1981. Effectiveness of clyorpyrifos, fenthion,malathion, and propoxur as screen treatments for control of CuZicoidesmississippiensis. J. Econ. Entomol. 74:331-333.

Kline, D. L., R. H. Roberts, and D. A. Focks. 1981. Extraction of larvae of theceratopogonid biting midge, CuZicoides mississippiensis, from salt marsh soilswith a new agar technique. Mosquito News 41:94-98.

Schreck, C. E. and D. L. Kline. 1981. Repellency determinations of four commercialproducts against six species of ceratopogonid biting midges. Mosquito News41:7-12.

Roberts, R. H. and D. L. Kline. 1981. Efficacy of insecticide-treated screensagainst biting midges (Diptera:Ceratopogonidae), Culiaoides spp. Proc. Fla.Anti-Mosq. Assn. 51(2):41-43.

Kline, D. L. 1981. Current status of Culicoides research at the USDA Laboratory inGainesville, Florida. Proc. Fla. Anti-Mosq. Assn. 51(2):71-75.

MANUSCRIPTS

Kline, D. L. and R. H. Roberts. Daily and seasonal abundance of Culicoides spp.biting midges in selected mangrove areas in Lee County, Florida, (submittedto Florida Entomologist).

Kline, D. L., D. G. Haile and K. F. Baldwin. Wind tunnel tests with seven insecti-cides against adult Culicoides mississippiensis Hoffman (accepted by MosquitoNews).

Harlan, Ii., C. E. Schreck and D. L. Kline. Insect repellent jacket tests againstbiting midges (Culicoides spp.) in Panama. (submitted to American Journ.Trop. Medicine and Hygiene).

Davis, E. L., J. F. Butler, R. H. Roberts, J. F. Reinert, and D. L. Kline.Laboratory blood-feeding of CuZicoides mississippiensis Hoffman througha reinforced silicone membrane. (Submitted to J. Med. Entomol.).

THESIS

Davis, E. L. 1981. Laboratory studies on life cycle development and adult blood-feeding of Culicoides missisippiensis (Diptera:Ceratopogonidae). M. S.University of Florida, 107 pp.

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Table 3.--Minor Culi.ooides spp.-A recorded from Yankeetown, Florida during October,1980-

September, 1981.

C.arboricola Root and Hoffman

C. baueri Hoffman

C. bermudensis Williams

C. edeni Wirth and Blanton

C. flori-densis Beck

C. gut tipennis (Coquillett)

C. haematopotus M'allock

C. hinmani Khalaf

C. insignia Lutz

C. loughnani Edwards

C. niger Root and Hoffman

C. ousairani Khalaf

C. paraensis (Goeldi)

C. steliifer (Coquillett)

C. vilZ~osipennis Root and Hoffman

a! These species comprise <1% of total trap catches

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Table 7.--Meteorlogical Data for Spray Operations Over Parris Island, South

Carolina, April 7, 1981. a

SURFACE OBSERVATIONS

1700L 1730L 1800L

CloudCover 10/10/Cl/CS 25,000? lO/l0/CIICS 25,000' 10/10/Cl/CS 25,000'

8/10 opaque 8/10 opaque 7/10 opaque

Temp 620F 61OF 60OF

Wet Bulb 520F 520F 520F

Dew Point 490F 50OF 51OF

RelativeHumidity 50% 51% 58%

Wind Drctn 0900 0800 0900

Speed (kts) 07 07 07

Winds Start of

Aloft Spray Ops (H-30) H+00 H+30

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Sfc-200' 1010/7.8KTs 095*/13.OKTs 102 0/14KTs

Sfc-300' 100*/8.8KTs 095*/14.OKTs 101 0/16KTs

Pasquill 1700L 17301, 1800L

E E E

Time of

Sunset 1847L

a! Data observed and compiled by MGySgt K. L. Hicks

Marine Corps Air Station

Beaufort, South Carolina 29904

II-DATF .