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Enhitreje su bliski srodnici gujavica te modelni organizmi u ekotoksikološkim istraživnjima tla. Iako imaju važnu ulogu u ekološkim sustavima tla, djelovanje pojedinih zagađivala, posebice strobilurinskih fungicida nedovoljno je istraženo. Stoga je cilj ovog doktorskog rada nadopuniti postojeća istraživanja novim znanjima o utjecaju strobilurinskih fungicida na enhitreje vrsta E. crypticus i E. albidus. Razvijena je metoda za mjerenje promjena aktivnosti mehanizma multiksenobiotske rezistentnosti čime se dokazala prisutnost ovog mehanizma kod enhitreja i mogućnost njegova korištenja kao biomarkera zagađenja. Nadalje, uočeno je negativno djelovanje strobilurinskih fungicida na preživljavanje, reprodukciju i uspjeh izlijeganja enhitreja. Prikupljena inicijalna znanja o djelovanju aktivne tvari i različitih pripravaka na biokemijske i stanične biomarkere ukazuju na moguće mehanizme djelovanja strobilurinskih fungicida naglašavajući važnost sveobuhvatnog pristupa. Enchytraeids are close relatives of earthworms and model organisms of soil ecotoxicology. Although they play an important role in soil ecosystems, the effects of certain pollutants, especially strobilurin fungicides, have not been sufficiently investigated. Therefore, the main objective of this doctoral thesis is to supplement existing research with new knowledge about the influence of strobilurin fungicides on the enchytraeid species E. crypticus and E. albidus. The development of a method for measuring changes in the activity of the multixenobiotic resistance mechanism proved the presence of this mechanism in enchytraeids and the possibility of its use as a biomarker. Furthermore, a negative effect of strobilurin fungicides was observed on the survival, reproduction, and hatching success of enchytraeids. Initial knowledge collected on the effect of the active ingredient and different formulations on biochemical and cellular biomarkers indicates the possible mechanisms of action of strobilurin fungicides, emphasizing the importance of a comprehensive approach.

U ovom radu promatramo kako svjetski pokazatelji upravljanja (WGI) utječu na transparentnost proračuna. Kao razinu transparentnosti proračuna uzimamo indeks otvorenosti proračuna (OBI) te ga promatramo kao zavisnu varijablu. Nezavisne varijable predstavljaju WGI-jevi glas i odgovornost (VA), politička stabilnost i odsutnost nasilja/terorizma (PS), učinkovitost vlade (GE), kvaliteta regulatornog okvira (RQ), vladavina prava (RL) te kontrola korupcije (CC). Za uzorak uzimamo 117 zemalja za koje je izračunat OBI u 2019. WGI-jeve promatramo za istih 117 zemalja, no uz 2019., promatramo i prosjek svakog pokazatelja u razdoblju 2017.-2019. Metodom linearne regresije dolazimo do zaključka da su svi WGI statistički značajni i to pozitivno utječu na OBI. Uvodimo pojam agregiranog WGI pokazatelja kao prosjek svih šest svjetskih pokazatelja upravljanja. Zaključujemo da je agregirani WGI takoder statistički značajan za OBI i pozitivno utječe na njega. Također vidimo da su WGI pokazatelji medusobno snažno korelirani. Na kraju zaključujemo da bolje upravljanje ukazuje na veću transparentnost državnog proračuna. In this paper, we consider how Worldwide Governance Indicators (WGIs) affect budget transparency. The Open Budget Index (OBI) is taken as the measure of budget transparency and viewed as a dependent variable. WGIs are used as independent variables, 6 in total: Voice and Accountability (VA), Political Stability and Absence of Violence / Terrorism (PS), Government Effectiveness (GE), Regulatory Quality (RQ), Rule of Law (RL) and Control of Corruption (CC). We take as a sample 117 countries for which OBI was calculated in 2019. Regarding WGIs, we observe them for the same 117 countries, but except for 2019, we also observe the average of each indicator in the period 2017-2019 Using the linear regression method, we determined a positive and statistically significant effect of each WGI on OBI. We introduce the concept of an aggregate WGI indicator as an average of all six Worldwide Governance Indicators. Aggregated WGI is also statistically significant for OBI and positively affects it. Strong correlation between all WGIs is also observed. Finally, we conclude that better governance indicates greater budget transparency for a country.

Prekomjerna primjena fungicida na poljoprivrednim površinama može imati negativan učinak na organizme tla, poput enhitreja, te posljedično utjecati na kvalitetu tla. Enhitreje su prepoznate kao pokazatelji kvalitete tla [1], a u ekosustavima tla obavljaju istu funkcionalnu ulogu kao gujavice [2]. Na površinama s intenziviranom obradom dolazi do smanjenja brojnosti ili potpunog izostanka gujavica, dok se aktivnost enhitreja povećava [3]. Međutim, njihova uloga u tlu ih čini lakim metama onečišćenja okoliša. Postojeća znanja o učincima strobilurinskih fungicida stečena su prvenstveno na temelju ispitivanja toksičnosti aktivnih sastojaka, dok su učinci komercijalnih preparata i dalje nejasni [4]. Stoga je cilj ovog rada pridonijeti novim saznanjima o učincima tri komercijalna preparata strobilurinskih fungicida na vrstu Enchytraeus albidus. Testirani su fungicidi Retengo® (piraklostrobin - PYR), Zato WG 50® (trifloksistrobin - TRI) i Stroby WG® (kresoksim- metil - KM). Dobiveni rezultati ukazali su kako na preživljavanje i reprodukciju enhitreja najviše djeluje PYR (LC50=7, 57 mga.i./kgtla, EC50=0, 98 mga.i./kgtla), a zatim TRI (LC50=72, 98 mga.i./kgtla, EC50 =16, 93 mga.i./kgtla) i KM (LC50=73, 12 mga.i./kgtla, EC50=>30 mga.i./kgtla). Nakon sedam dana izlaganja, aktivnost mehanizma multiksenobiotske rezistentnosti (MXR) je inhibirana pri najvišim koncentracijama svih testiranih fungicida. Nadalje, pojava oksidativnog stresa (indukcija enzima SOD, CAT i GST) uočena je nakon sedam dana izlaganja, dok je povećanje lipidne peroksidacije uočeno nakon 21. dana. Izloženost fungicidima dovela je do promjena u raspoloživoj energiji, te izazvala promjene u omjeru glavnih metabolita (ugljikohidrata, lipida i proteina). Nakon izlaganja KM smanjenje raspoložive energije bilo je posljedica prekomjernog trošenja energije zbog istovremene obrane od oksidativnog stresa i reprodukcije, dok su enhitreje izložene TRI nedostatak energije kompenzirale izostankom reprodukcije. Visoka toksičnost PYR uzrokovala je smanjenje energetskih zaliha bez obzira na izostanak reprodukcije. Iako su svi testirani komercijalni proizvodi pokazali negativan učinak na enhitreje, ozbiljnost i način djelovanja su se razlikovali, naglašavajući važnost temeljite procjene više krajnjih točaka u različitim vremenskim razdobljima.

Due to their high efficiency, broad spectrum, and security to crops, strobilurin fungicides are among the most widely used agricultural fungicides. With extensive and continuous use, strobilurin fungicide residues can occur in soil, air, and water, leading to ecological impact and potentially adverse impact on nontarget organisms. Commercial preparations of fungicides often get applied directly in soil ecosystems through agricultural practices, even though only the effects of the active substance are included in common EU regulations. Thus, the toxicity of commercial fungicide preparations should be investigated in more detail to better understand their effects on e.g. enchytraeids that are known to play an important role in soil fertility and quality. In addition, enchytraeids are often used as a model organism in soil ecotoxicology. To fill the knowledge gaps about the impact of strobilurin fungicides on enchytraeids, this study evaluated the impact of three commercial products on E. albidus. To obtain a broader picture of the mode of action, multiple endpoints were assessed. The test species E. albidus was exposed to commercial fungicide products based on the active ingredients pyraclostrobin (Retengo), trifloxystrobin (Zato WG) and kresoxim-methyl (Stroby WG). Standard guidelines for the Enchytraeid Reproduction Test were followed. Standard enchytraeid reproduction test (ERT) showed that, in terms of survival, pyraclostrobin (LC50=7.57 mg/kg) was the most toxic, followed by trifloxystrobin (LC50 = 72.98 mg/kg) and kresoxim-methyl (LC50=73.12 mg/kg). Reproduction was affected in the same order (pyraclostrobin EC50=0.98mg/kg, trifloxystrobin EC50=9.39 mg/kg and kresoxim-methyl EC50=>30 mg/kg). Furthermore, tested fungicides exhibited adverse effects on MXR activity, biochemical parameters of oxidative stress (superoxide dismutase (SOD), catalase (CAT), glutathione S- transferase (GST) activity, and lipid peroxidation - malondialdehyde (MDA) content), and available energy (protein, lipid and carbohydrate content). Results of this study show a broad spectrum of negative effects of the investigated fungicides on enchytraeids, and also indicate the need to investigate, not only the active substances, but also the commercial preparations. Otherwise, environmental risks could be significantly underestimated.

Population data of rodent strictly protected species Dinaromys bogdanovi, Dryomys nitedula, Muscardinus avellanarius and Cricetus cricetus in Croatia is generally insufficiently known. Field protocols for the first two species are not available and have yet to be developed. The ‘zero state’ knowledge of populations will be established by processing the existing avail able data (habitat modelling) and collecting of new field data. This will greatly enhance the knowledge on the ecology of target species and consequently facilitate the development of effective conservation management. Some of the available methods of data collection are laborious, expensive and lack desired sensitivity to detect the target species. We are therefore seeking for improvements of the available protocols and the development of novel ones, specifically techniques which will provide unbiased information on the distribution, population size and habitat requirements. At last, we are keen to estimate the anthropogenic pressures and to assess the conservation status of the species. If appropriate, we shall propose implementation of urgent conservation measures already in duration of the project. Also, methods tested and improved for the duration of this survey will be again implemented during the next reporting periods. It is therefore crucial to standardize monitoring protocols and provide detailed instructions for the field work, sampling design, data processing and long-term storage of primary data.

The state of the populations of strictly protected species of small mammals in Croatia is generally insufficiently researched, therefore an expert assessment of the significance of the impact of planned projects and/or strategic documents on them is not possible. As part of the Operational Program Competitiveness and Cohesion 2014-2020, the Republic of Croatia is implementing a monitoring program for four species of small mammals: Dinaromys bogdanovi, Dryomys nitedula, Muscardinus avellanarius and Cricetus cricetus. By processing the existing available and collecting additional data, a ‘zero state’ of populations in Croatia will be established, which will greatly help in defining the impact of planned interventions and strategic documents on the ecology of target species. The analysis will also suggest ways of collecting data for better determination of distribution, population size, habitat quality, as well as the pressures needed to assess the conservation status of the species. Urgent conservation measures to be implemented on the basis of recorded direct threats during field research will be proposed. Also, the research will define methods for determining the presence and monitoring of populations for the next reporting periods through proposals for monitoring programs that include monitoring methodology in the form of detailed instructions for field work, a sampling plan and forms for each type of methodology and description of organization, storage and data processing.

Strobilurin fungicides (SFs) are a group of new fungicides produced by strobilurin A as a lead compound. Azoxystrobin, the best-selling SF, is a highly efficient broad-spectrum fungicide with relatively high-water solubility, which can lead to accumulation in water, sediments, and soil. Due to its wide application and impact on non-target soil organisms, azoxystrobin has attracted attention. Soil dwelling organisms are among the first in line of exposure to pesticides. However, their risks are often based on the pure active ingredient (AI) and not on the commercial products (CPs) actually applied in the field. Enchytraeus albidus is a model species in soil ecotoxicology used for the assessment of environmental risk. Due to their important role in proper soil functioning, it is essential to investigate the toxicity and adverse effects of fungicides used in agriculture on enchytraeids. Previous research has demonstrated a negative impact of azoxystrobin on the reproduction and oxidative status of earthworms, and the survival, reproduction, and hatching success in enchytraeids. However, most studies have assessed the effect of AI, while only a few use a CP. Therefore, we investigated the effects of pure AI, azoxystrobin, and CP Quadris on survival, reproduction, MXR activity and oxidative stress biomarkers in E. albidus. The results showed that CP exhibited higher toxicity in terms of survival (LC50CP = 16.7 mg/kg soil, LC50AI = 15.3 mg/kg soil), while reproduction was affected only by CP (EC50 = 3 mg/kg soil). Furthermore, while AI caused induction of the multixenobiotic resistance system, CP caused inhibition. Moreover, although both tested substances caused oxidative stress, the toxicity of the CP was higher and long term impact in form of lipid peroxidation was observed. These results suggest the importance of evaluating the effect of AI and CP. Furthermore, to obtain more accurate information on the mode of action, access is required through different endpoints. These findings highlight the need to further explore the toxicity of CPs compared to AIs, in order to predict a more realistic environmental hazard of fungicides.

Med je naravni izdelek, ki ugodno vpliva na zdravje in počutje ljudi. Izdelujejo ga čebele rodu Apis mellifera. Vse lastnosti medu, vključno s hranilnimi in zdravilnimi učinki, izhajajo iz njegove edinstvene kemične sestave. Le-ta je posledica obdelave nektarja in rastlinske mane medonosnih čebel. Med je predvsem sestavljen iz ogljikovih hidratov in vode, hkrati pa vsebuje organske kisline, minerale, vitamine, aromatske snovi, beljakovine, encime, itd. Znan je po svojim antioksidativnih, protibakterijskih in protiglivičnih učinkih in se uporablja za zdravljenje različnih bolezenskih stanj. Pravilnik o medu pravi, da mora biti poreklo medu jasno označeno na izdelku. Zaželeno je, da ima med poleg geografskega porekla označeno tudi botanično poreklo. Na ta način ima med večjo komercialno vrednost, ker je zagotovljena avtentičnost izdelka in kupec naj bi bil zaščiten pred goljufijami. Vendar prihaja pri proizvodnji in prodaji medu do ponaredb. Bodisi je med napačno označen bodisi ga med procesiranjem zamenjujejo s cenejšim medom ali mu dodajajo sladkor. Posledično se v zadnjem času aktivno išče metode, s pomočjo katerih je možno določiti oziroma preveriti geografsko in botanično poreklo medu. Poleg tradicionalnih metod, kot so melisopalinološka metoda in določevanje fizikalno-kemijskih lastnosti, aktualne metode temeljijo na določitvi elementne sestave, uporabe stabilnih izotopov (H, O, C, N, S in Sr), določevanju hlapnih organskih spojin, NMR profiliranja, itd. Za namen diplomske naloge je bila določena elementna sestava in izotopsko razmerje 87Sr/86Sr v petih vzorcih medu iz Črne gore in Madžarske. Vsebnost elementov v medu je odvisna od vrste medu in lastnosti tal na izvornem območju. Elementi predvsem izhajajo iz tal, se prenašajo do rastlin skozi koreninski sistem in se posledično tudi nahajajo v nektarju. Elementna sestava ni povezana samo s sestavo tal, ampak tudi z antropogenimi dejavnostmi. Zato je med tako indikator geografskega porekla kakor tudi onesnaženosti okolja na izvornem območju. Kot dodatni indikator geografskega porekla se uporablja tudi izotopsko razmerje 87Sr/86Sr. Izotopska sestava Sr se med privzemom iz tal v rastline ne spremeni in tako odraža izotopsko sestavo okolja. Za določitev elementne sestave in 87Sr/86Sr izotopskega razmerja smo uporabili metodo masne spektrometrije. Elementno sestavo smo določili z masnim spektrometrom z induktivno sklopljeno plazmo (ICP-MS). Za določitev izotopskega razmerja Sr smo uporabili multikolektorski ICP-MS. Med ima kompleksno matrico in ga je potrebno pred vnosom v masni spektrometer ustrezno pripraviti. Popoln razkroj vzorca je ključnega pomena za natančno in ponovljivo določanje tako elementne sestave kot Sr izotopskega razmerja. Za določitev elementarne sestave medu smo vzorce pripravili z mikrovalovnim razkrojem. Za določitev sestave Sr izotopov smo primerjali med seboj metodo z mikrovalovnim razkrojem in suhim sežigom. Primerjavo smo izvedli na vzorcu madžarskega medu, ker smo ga imeli na razpolago v večjih količinah. Za točno in natančno določanje izotopskega razmerja 87Sr/86Sr je potrebno izolirati Sr iz matrice, kjer je prisoten tudi 87Rb, saj ta izotop moti določitev 87Sr. Najpogosteje se za izolacijo Sr uporablja Sr-selektivna smola, ki smo jo uporabili tudi mi. Rezultati primerjave priprave vzorcev z mikrovalovnim razkrojem in suhim sežigom so pokazali, da način priprave ne vpliva na izotopsko sestavo Sr, saj se vrednosti razlikujejo na zadnjem signifikantnem mestu . Za nadaljnje analize črnogorskega medu smo uporabili samo metodo suhega sežiga, ker je hitrejša, naenkrat smo lahko obdelali večjo količino vzorca in število korakov priprave je manjše kot pri mikrovalovnem razkroju. Izmerjene vrednosti 87Sr/86Sr v medu iz Črne gore smo primerjali z vrednostmi v tleh z istih področij, od koder je prihajal med. Ugotovili smo, da na Sr izotopsko sestavo v medu ne vplivajo samo tla, saj se vrednosti za med in tla med seboj niso ujemale za vse vzorce. Na 87Sr/86Sr izotopsko razmerje v medu najverjetneje vplivajo tudi tip tal, voda in prašni delci. Rezultate elementne analize in 87Sr/86Sr izotopskega razmerja smo obdelali z metodo glavnih osi (PCA). Namen je bil ugotoviti, ali se med z različnih lokacij razlikuje. PCA, ki je bila narejena na podlagi samo elementne sestave, je pokazala, da je ločba med črnogorskim in madžarskim medom možna. Dodatna uporaba rezultatov 87Sr/86Sr izotopskega razmerja ni dodatno vplivala na diferenciacijo vzorcev. Za boljšo diferenciacijo različnih vzorcev črnogorskega medu bi potrebovali še dodatne metode, kot na primer uporabo stabilnih izotopov bio-elementov (H, O, C, N in S). Honey is a natural product that has a beneficial effect on human health. Properties of honey, including nutritional and medical effects, result from its unique chemical composition. This is due to the processing of nectar and the plant manna by the honeybees. Elemental composition of honey depends on the type of honey and the properties of the soil in the source area. Therefore, honey can be used both as indicator of a geographical origin as well as biomarker because it reflects environmental pollution in the area of origin. This in particular applies to elements such as As, Cd, Cr, Cu, Fe, Ni, Pb and Zn. These elements can lower the quality of honey and pose a potential threat to human health. As an additional indicator of geographical origin, the 87Sr/86Sr isotopic ratio is also used. The isotopic composition of Sr does not change when absorbed from soil and water by plants through root system and thus reflects the isotopic composition of the environment. For the purpose of my thesis, the elemental composition and isotopic ratio 87Sr/86Sr was determined in 5 samples of honey from different places in Montenegro and Hungary. The mass spectrometry with an inductively coupled plasma mass spectrometer (ICP-MS) was used to determine the elemental composition. To determine the 87Sr/86Sr isotope ratio, the multi-collector ICP-MS was used. Honey is a sample with a complex matrix that should be properly prepared before further analysis. Complete digestion is crucial, otherwise the results are not accurate and reproducible. For the determination of elemental composition in honey, the samples were prepared by microwave digestion, while for Sr isotope composition determination microwave digestion and dry ashing methods were compared. The effectiveness and usability of the two methods for Sr isotope composition determination was tested on Hungarian honey, because it was available in greater quantity. In order to accurately determine the 87Sr/86Sr isotopic ratio, Sr had to be isolated from the matrix, because 87Rb is present, which can influence the results. To isolate Sr from the solution, Sr-selective resin was used. The final results were processed with Principal Component Analysis (PCA) and it was determined that samples differ, with the Hungarian honey standing out the most.

Strobilurin fungicides (SFs) are a group of new fungicides produced by strobilurin A as a lead compound. Azoxystrobin, best-selling SF, is a highly efficient broad-spectrum fungicide with relatively high-water solubility, which may lead to accumulation in water, sediments, and soil. Due to its wide application and impact on non-target soil organisms azoxystrobin have attracted attention. Soil dwelling organisms are among the first in line of exposure to pesticides, however their risks are often based on the pure active ingredient (AI) and not on the commercial products (CPs) actually applied in the field. Enchytraeus albidus is a model species in soil ecotoxicology used for environmental risk assessment. Due to their important role in proper soil functioning, it is essential to investigate the toxicity and adverse effects of fungicides used in agriculture on enchytraeids. Previous research has demonstrated a negative impact of azoxystrobin on the reproduction and oxidative status of earthworms, and survival, reproduction, and hatching success in enchytraeids. However, most studies have assessed the effect of AI, while only a few used a CP. Therefore, we investigated the effects of pure AI, azoxystrobin, and CP Quadris on the survival, reproduction, oxidative stress biomarkers, energy metabolism, and MXR activity in E. albidus. Results showed that CP exhibited higher toxicity in terms of survival (LC50= 16.7 mg CP/kg soil, LC50= 15.3 mg AI/kg soil), while reproduction was affected only by CP (EC50= 3 mg CP/kg soil). Furthermore, although both tested substances caused oxidative stress, the toxicity of the CP was higher. Exposure to CP and AI affected energy metabolism, causing changes in lipid and carbohydrate content. Moreover, while AI caused induction of the multixenobiotic resistance system, CP caused inhibition. These results suggest the importance of evaluating the effect of AI and CP. Furthermore, to obtain more accurate information on the mode of action, access through different endpoints is required. These findings highlight the need to further explore the toxicity of CPs compared to AIs, aiming to predict a more ealistic environmental hazard of fungicides.

The Pannonian island mountains is a term for isolated mountains scattered across the Pannonian Plain, particularly its western and southern parts, in Hungary, Serbia and Croatia. These mountains were islands of the ancient Pannonian Sea that disappeared about 600, 000 years ago. In Croatia they encompass several mountains: Dilj, Krndija, Papuk, Psunj, Požeška Gora and Medvednica. The majority of area is covered with beech forest. Within this area several endemic earthworm species were identified (Aporrectodea papukiana, Dendrobaena papukiana). In this research a comprehensive sampling was carried out as well as the use of barcoding for confirmation of earthworm identification. Particular focus was on the protected areas of Papuk mountain.

Strobilurins, a commonly used class of fungicides, are known to impact an embryonic development of aquatic organisms and show toxicity to different soil organisms. Standard tests for soil invertebrates are usually limited to the assessment of endpoints like survival and number of juveniles, while hatching and embryotoxicity tests have emerged recently. Besides earthworms, enchytraeids are the most commonly used bioindicators of soil contamination. Their size and short life cycle makes them suitable for this type of research. The main aim of the study was to assess the impact of strobilurins on reproduction and hatching delay of enchytraeid Enchytreus crypticus. Standardised OECD reproduction tests with azoxystrobin (AS), pyraclostrobin (PS) and trifloxystrobin (TS) were conducted. A decreased number of juveniles and an increased number of unhatched cocoons in strobilurin treatments were recorded. TS was the most toxic in terms of reproduction (EC50 0.04 mg a.i./kg soil), followed by PS (EC50 1.85 mg a.i./kg soil) and AZ (EC50 93.10 mg a.i./kg soil). The use of synchronised cocoons enabled monitoring of hatching dynamics. After a standard 11-day exposure more than 90 % of cocoons were hatched in the control while in treated soil those values were lower. However, as hatching was monitored after that period (up to the 19th day) the percentage of hatched cocoons significantly increased with the time of exposure. Consequently, EC50 values changed. These results show that supplementary endpoints such as hatching success included in standard tests could increase tests sensitivity and accuracy

Fotomonografija Prirodna baština - Skriveno blago pleterničkog kraja kroz niz fotografija prikazuje raznolikost stanišnih tipova, ali i biljnih i životinjskih vrsta zastupljenih na području grada Pleternice.

Strobilurin su jedna od najzastupljenijih skupina fungicida, a dokazano je da utječu na embrionalni razvoj vodenih organizama, te pokazuju toksičnost za različite organizme tla [Zhang i sur., 2020]. Standardni testovi za beskralježnjake tla obično su limitirani na procjenu krajnjih točaka kao što su preživljavanje odraslih i broj juvenilnih, dok su testovi izlijeganja i embriotoksičnosti ustanovljeni tek nedavno [Gonçalves et al., 2015]. Osim gujavica, enhitreje su najčešće korišteni bioindikatori zagađenja tla [Castro-Ferreira et al., 2012]. Nadalje njihova veličina i kratak životni ciklus čine ih pogodnima za ovakvu vrstu istraživanja. Cilj ovog istraživanja bio je utvrditi utjecaj strobilurina na preživljavanje, reprodukciju i izlijeganje kod vrste Enchytraeus crypticus. Provedeni su standardizirani OEC reprodukcijski testovi s azoksistrobinom (AS), piraklostrobinom (PS) i trifloksistrobinom (TS). Na preživljavanje je najviše utjecao TS (LC50 = 2.34 mg/kg), te PS (LC50 = 4.26 mg/kg) i AS (LC50 = >150 mg/kg). Izlaganje AS i PS pokazala je negativan utjecaj na uspjeh izlijeganja, a na kraju reprodukcijskog testa zamijećen je značajan porast broja neizleženih kokona. Produljeni test izlijeganja pokazao je odgodu izlijeganja pri nižim koncentracijama AS i PS, dok je pri višim koncentracijama izlijeganje potpuno izostalo, te kokoni više nisu bili izleživi. Produljeni test izlijeganja omogućava razlikovanje odgode izlijeganja i njegovog potpunog izostanka. Osim što su dokazani negativni učinci AS, PS i TS na preživljavanje, reprodukciju i izlijeganje kod vrste E. crypticus, dobiveni rezultati ukazuju na potrebu korištenja različitih krajnjih točki. Korištenje produljenog testa izlijeganja može popuniti prazninu između reprodukcijskih i više generacijskih testova i omogućuju bolje razumijevanje štetnih učinaka testiranih tvari.