In this paper the results of a ten year investigation of mosquitoes using dry ice baited CDC traps in the city of Osijek, Croatia are presented. In the first year we recorded 16 species. During these ten years the total of 20 species were determined. The single method, dry ice baited CDC traps, makes it possible to determine almost the entire fauna in a given area. The estimated fauna is statistically calculated. Five non-parametric estimators were chosen for evaluation: one-factorial Jackknife (Jackknife-1); two-factorial Jackknife (Jackknife-2); one-factorial Chao (Chao-1); two-factorial Chao (Chao-2) and Bootstrap. The estimated fauna contains between 22 and 29 species. The method used turned out to be very good when used over a long period of time, because new species were registered even in years without many mosquitoes.U ovom radu govori se desetgodišnjim rezultatima istraživanja komaraca, koji su hvatani metodom CDC klopke uz suhi led kao atraktant u Osijeku. Prve istraživačke godine zabilježeno je 16 vrsta. Tijekom deset istraživačkih godina broj poznatih vrsta popeo se na 20. Predpostavka ovog rada je da se korištenjem samo jedne metode uzorkovanja može se doći do skoro kompletne faune nekog područja. Očekivana fauna izračunata je statistički. U tu svrhu korišteno je pet neparametarskih estimatora: jedanfaktorijalni Jackknife (Jackknife-1); dvofaktorijalni Jackknife (Jackknife-2); jedanfaktorijalni Chao (Chao-1); dvofaktorijalni Chao (Chao-2) i Bootstrap. Ovim statističkim metodama je određeno da se broj vrsta komaraca u Osijeku kreće od 22 do 29 vrsta. Korištena metoda se pokazuje kao dobra ako se koristi kroz duže vremensko razdoblje, a naročita joj je vrijednost u tome što su se nove vrste evidentirale u godinama kada i nije bilo puno komaraca. Do otkrivanja kompletne faune ovom metodom, potrebno je uzorkovati još neko vrijeme

Branimir Hackenberger Kutuzović

Enrih Merdić

Ivana Vrućina

Mirta Sudarić Bogojević

HRČAK - Portal of Croatian Scientific and Professional Journals

original scientific paper / izvorni znanstveni radDYNAMICS OF MOSQUITO (DIPTERA, CULICIDAE)SPECIES RICHNESS IN SAMPLES OF DRY-ICEBAITED CDC TRAPS IN THE URBAN AREAOF OSIJEK, CROATIAENRIH MERDI]*, BRANIMIR HACKENBERGER KUTUZOVI], MIRTASUDARI] BOGOJEVI] & IVANA VRU]INADepartment of biology, J. J. Strossmayer University in Osijek, Trg LjudevitaGaja 6, HR-31000 Osijek, CroatiaMerdi}, E., Hackenberger Kutuzovi}, B., Sudari} Bogojevi}, M. & Vru}ina, I.: Dynamics ofmosquito (Diptera, Culicidae) species richness in samples of dry-ice baited CDC traps in the ur-ban area of Osijek, Croatia. Nat. Croat., Vol. 17, No. 3., 149–155, Zagreb, 2008.In this paper the results of a ten year investigation of mosquitoes using dry ice baited CDCtraps in the city of Osijek, Croatia are presented. In the first year we recorded 16 species. Duringthese ten years the total of 20 species were determined. The single method, dry ice baited CDCtraps, makes it possible to determine almost the entire fauna in a given area. The estimated faunais statistically calculated. Five non-parametric estimators were chosen for evaluation: one-factorialJackknife (Jackknife-1); two-factorial Jackknife (Jackknife-2); one-factorial Chao (Chao-1); two-facto-rial Chao (Chao-2) and Bootstrap. The estimated fauna contains between 22 and 29 species. Themethod used turned out to be very good when used over a long period of time, because new spe-cies were registered even in years without many mosquitoes.Key words: mosquitoes, dry ice baited CDC traps, species richness, non-parametric estimators,CroatiaMerdi}, E., Hackenberger Kutuzovi}, B., Sudari} Bogojevi}, M. & Vru}ina, I.: Dinamikabogatstva vrsta komaraca u uzorcima (Diptera, Culicidae) hvatanim metodom CDC klopke uzsuhi led na podru~ju grada Osijeka. Nat. Croat., Vol. 17, No. 3., 149–155, Zagreb, 2008.U ovom radu govori se desetgodi{njim rezultatima istra`ivanja komaraca, koji su hvatani meto-dom CDC klopke uz suhi led kao atraktant u Osijeku. Prve istra`iva~ke godine zabilje`eno je 16vrsta. Tijekom deset istra`iva~kih godina broj poznatih vrsta popeo se na 20. Predpostavka ovograda je da se kori{tenjem samo jedne metode uzorkovanja mo`e se do}i do skoro kompletne faunenekog podru~ja. O~ekivana fauna izra~unata je statisti~ki. U tu svrhu kori{teno je pet neparametar-NAT. CROAT. VOL. 17 No 3 149¿155 ZAGREB September 30, 2008Croatian Natural History Museum, Demetrova 1, Zagreb, Croatia* corresponding author, e-mail: enrih@biologija.unios.hrskih estimatora: jedanfaktorijali Jackknife (Jackknife-1); dvofaktorijalni Jackknife (Jackknife-2); je-danfaktorijali Chao (Chao-1); dvofaktorijalni Chao (Chao-2) i Bootstrap. Ovim statisti~kim metoda-ma je odre|eno da se broj vrsta komaraca u Osijeku kre}e od 22 do 29 vrsta. Kori{tena metoda sepokazuje kao dobra ako se koristi kroz du`e vremensko razdoblje, a naro~ita joj je vrijednost utome {to su se nove vrste evidentirale u godinama kada i nije bilo puno komaraca. Do otkrivanjakompletne faune ovom metodom, potrebno je uzorkovati jo{ neko vrijeme.Klju~ne rije~i: komarci, CDC klopka uz suhi led kao atraktant, bogatstvo vrsta, neparametarskiestimatori, HrvatskaINTRODUCTIONThe efficiency of various mosquito traps has been investigated by many re-searchers. New Jersey Light Traps (NJLT), gravid Reiter traps, bed net traps, trapsbaited with synthetic oviposition pheromone (MBOREA et al., 2000) and more re-cently the propane-powered mosquito trap (KLINE, 2002), have demonstrated differ-ent results, with dry ice-baited CDC traps providing the best results (REISEN et al.,2000, REISEN et al., 1999). Dry ice, octenol and light are among the most frequentlyused attractants (BURKETT et al., 2001, RITCHIE & KLINE 1995), of which dry iceproved to be the best for host-seeking females. The CDC trap has the best resultswhen a combination of dry ice plus octenol is offered as attractant (BECKER et al.,1995, WHITT et al., 2001).Because of their versatility, battery-operated CDC traps can be used in a varietyof situations (in inhabited or uninhabited areas, marshland, forests, and even at dif-ferent elevations in forests, (LUNDSTROM et al., 1996), making it possible to collectdata on ornithophilic species. Much research has been carried out on the emissionof CO2 which actually imitates the presence of a host. In the past few years manyresearch papers on the standardization of CDC traps in Europe under the auspicesof EMCA have been published (BELLINI et al., 2003).It is evident that none of the methods used singly can provide absolute results,therefore, in order to collect as many species of mosquitoes as possible from an areain the shortest possible time, a combination of trapping methods is recommended.In this paper we wish to demonstrate the ability of dry ice-baited CDC traps to pro-vide data on most of the mosquito fauna, even if the method is more time-consum-ing. Statistical estimators are used to support this thesis. The only way to estimatespecies richness from the field investigation results is the use of various speciesrichness estimators. The better to describe species richness five common nonpara-metric estimators were used.MATERIAL AND METHODSThe investigation was carried out in the broader area of the city of Osijek, (north-eastern Croatia, UTM CR25 and CR26), during a period of 10 years. This study dis-cusses the data obtained from 1995 to 2004. Battery-operated, dry ice-baited CDC(Center for Disease Control) traps were set in 9 locations (Donji grad N 45 33 41,2 E150 Merdi}, E. et al.: Dynamics of mosquito species richness ...18 43 37,4; Jug II N 45 33 7,2 E 18 42 34,3; Sjenjak N 45 33 7,2 E 18 42 34,3; Iktus N 4533 44,7 E 18 41 46,7; @upanijska N 45 33 20 E 18 40 49,8; Strma N 45 33 57,9 E 18 407,2; Retfala S N 45 34 2,5 E 18 39 20,3; Retfala J N 45 33 25,4 E 18 39 13,3 andJosipovac N 45 34 23 E 18 36 16,9) and operated twice a month from May to Septem-ber each year. The traps were put in yards of houses 1 m from ground and active forover 24 hours (± 2h), with a 9 kg dry ice cube. A total of 207,136 mosquitoes belong-ing to 7 genera and 20 species were collected in 900 samplings during this period.All statistical calculations were performed using the statistical package »vegan«under the R software environment and Estimates.RESULTS AND DISCUSSIONIn the investigated area we collected a total of 20 species in 7 genera. The speciesAedes vexans, Culex pipiens c., Anopheles maculipennis c., Ochlerotatus cantans, Ochlero-Nat. Croat. Vol. 17(3), 2008 151Tab. 1. Mosquito species recorded in Osijek (1995–2004)Nr. Species/Year 1995 1996 1997 1998 1999 2000 2001 2002 2003 20041 Aedes vexans + + + + + + + + + +2 Anopheles maculipennis c. + + + + + + + + + +3 Coquillettidia richiardii + + + + + + + + + +4 Culex modestus + + + + + + + + + +5 Culex pipiens c. + + + + + + + + + +6 Culiseta annulata + + + + + + + + + +7 Ochlerotatus cantans + + + + + + + + + +9 Ochlerotatus excrucians + + + + + + + + + +8 Ochlerotatus sticticus + + + + + + + + +10 Ochlerotatus caspius + + + + + + + + +11 Aedes cinereus + + + + + + +14 Anopheles hyrcanus + + + + + + +12 Aedes rossicus + + + + + +13 Ochlerotatus rusticus + + + + + +15 Anopheles claviger + + + + +17 Ochlerotatus cataphylla +19 Uranotenia unguiculata +16 Culex territans +18 Ochlerotatus leucomelas +20 Anopheles plumbeus +No of species per year 16 15 11 13 15 15 13 14 12 10tatus excrucians, Coquillettidia richiardii, Culex modestus and Culiseta annulata werefound every year, and can therefore be referred to as »frequent species«. Ochlero-tatus sticticus and Ochlerotatus caspius were recorded in nine seasons; Aedes cinereusand Anopheles hyracanus were recorded in seven seasons; Aedes rossicus and Ochlero-tatus rusticus in six, and Anopheles claviger in five. Ochlerotatus cataphylla, Uranoteniaunguiculata, Culex territans, Ochlerotatus leucomelas, and Anopheles plumbeus (»rarespecies«) occurred in one season only.The abundance analysis demonstrated that the dominant species breeding in theflooded area were Aedes vexans and Ochlerotatus sticticus. These two species madeup 88.91% of the mosquito fauna in the investigated area, as they did in previousresearch (MERDI] & LOVAKOVI], 2001).The number of species found differs from year to year, varying from 10 to 16species (Tab. 1). In 1996, 1999, and 2000, we had a qualitatively identical composi-tion of mosquito fauna, with 15 species. The largest number of species, 16, was re-corded in 1995, the first year of investigation.The number of species trapped using this method increased to 17 the followingyear (1996, Ochlerotatus rusticus). Two years later, another species (1998, Culex terri-tans) was found, and three years later, yet another (2001, Ochlerotatus leucomelas),and finally, two years later one more rare species was found (2003, Anopheles plum-beus). The probability of recording the entire mosquito fauna was calculated basedon the dependence of each particular year and the probability of recording most ofthe mosquito fauna in the years of investigation.152 Merdi}, E. et al.: Dynamics of mosquito species richness ...151617181920210 50000 100000 150000 200000 250000Number of sampled mosquitosNr. of spec.SpeciesrichnessFig. 1. Increase of found species according to number of specimensThe increase in the number of rare species is directly connected with the numberof sampled specimens (Fig. 1). A new species appears at regular intervals (everytwo years). The value of the method used is additionally proved by the fact that anew species appeared even in the dry season.The estimation of total species richness in a biological community using a lim-ited number of survey units has drawn a great deal of attention (BUNGE & FITZ-PATRICK, 1993; COLWELL & CODDINGTON, 1994; WILLIAMS et al., 2002; CAO et al.,2004). Recent evaluations have suggested that non-parametric estimators performbetter than methods based on either species accumulation curves or species-abun-dance distributions (PALMER, 1990; BALTANAS, 1992; COLWELL & CODDINGTON, 1994;WALTHER & MORAND, 1998; KREBS, 1988; WALTHER & MARTIN, 2001, CAO et al.,2004) For some insect species Chao-2 was the most accurate and precise estimator,the Bootstrap estimator performed only slightly better than the number of observedspecies. Jacknife-1 and 2 performed intermediately. For such populations only Chao2performed more precisely than the observed number of species. Since, however, formosquito populations it is still unknown which estimator is best to describe speciesrichness, for this paper five estimators have been used.Nonparametric estimators are sampling theoretical extrapolation methods thatonly require the number of samples in which each species is found rather than anyparametric information about their abundance. In this paper five non-parametricestimators that are often recommended in the literature were chosen for our evalua-tion: one-factorial Jackknife (Jackknife-1); two-factorial Jackknife (Jackknife-2);Nat. Croat. Vol. 17(3), 2008 1531618202224262830Chao 1 Chao 2 Jackknife 1 Jackknife 2 Bootstrap SampledEstimatednumberodspeciesFig. 2. Number of predicted species using different estimatorsone-factorial Chao (Chao-1); two-factorial Chao (Chao-2) and Bootstrap. The resultsof these analysis show that the estimated fauna is between 22 (Bootstrap) and 29(Chao-2 and Jackknife-2) (Fig. 2, Tab. 2). A good basis for this hypothesis is the datathat in the neighbouring countries Hungary (MIHALY, 1963) and Serbia, in Vojvodi-na, 42 and 35 species, respectively (PETRI], 1989) were found.ACKNOWLEDGMENTSWe would like to thank the Osijek City Government for the long-term financialsupport of the project Monitoring and investigation of mosquitoes in Osijek, the resultsof which were used in this paper.Received December 3, 2007REFERENCESBALTANAS, A., 1992: On the use of some methods for the estimation of species richness. Oikos65, 484–492.BECKER, N., ZGOMBA, M., PETRIC, D. & LUDWIG, M., 1995: Comparison of the carbon dioxide,octenol and a host odour as mosquito attractants in the upper Rhine valley. 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L., 1995: Comparison of CDC and EVS light traps baited with car-bon dioxide and octenol for trapping mosquitoes in Brisbane Queensland (Diptera, Culi-cidae): Journal of Australian Entomological Society 31–34(3), 215–218WALTHER, B. A. & MARTIN, J. L., 2001: Species richness estimation of bird communities: Howto control for sampling effort?. Ibis 143, 413–419.WITT, P. B., HARRISON, R. L. & GARDNER, R. C., 2001: Evaluation of 1-octen-3-ol, carbon diox-ide, and light as attractants for mosquitoes associated with two distinct habitats in NorthCarolina. J. Am. Mosq. Contr. Assoc 17 (1), 61–66.WILLIAMS, R. J. & MARTINEZ, N. D., 2000: Simple rules yield complex food web dynamics. Na-ture 404, 180–183Nat. Croat. Vol. 17(3), 2008 155

Dinamika bogatstva vrsta komaraca u uzorcima (Diptera, Culicidae) hvatanim metodom CDC klopke uz suhi led na području grada Osijeka

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