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Additional file 3. Table S3 Procrustes ANOVAs of shape. % total—percentage of the total shape variation. Age categories: A1—first age category, A2—second age category, A3—third age category

Additional file 1. Table S1 Anatomical definitions of landmarks recorded on the labial view of the mandible and the ventral surface of the cranium of the yellow-necked field mouse (Apodemus flavicollis)

Additional file 2. Table S2 ANOVAs of centroid size (CS). % total—percentage of the total size variation. Age categories: A1—first age category, A2—second age category, A3—third age category

Additional file 4. Table S4 ANOVAs of centroid size (CS). % total—percentage of the total size variation. Categories regarding parasitism: P0—non-parasitized animals, P1—animals parasitized by one nematode species, P2—animals parasitized by two nematode species, P3—animals parasitized by three to five nematode species

We conducted a comparative (2D landmark-based geometric and traditional) morphometric analysis on tadpoles at early developmental stages. Two species of brown frog (Rana dalmatina and R. temporaria) and the common toad (Bufo bufo) were involved, all raised in the laboratory from fertilized eggs collected in their natural habitat. Taxonomic identification was confirmed by the DNA barcoding method with the 16S rRNA sequence as the gene marker. Interested to compare the methodologies for quantification and description of morphological differences among tadpoles of mentioned species, we aimed to: 1) calculate interspecies genetic distances as the most relevant measurement for species differentiation, 2) determine and describe size and shape variation, 3) identify relationships among the analyzed species at the morphological level and 4) assess their classification accuracy. Within the framework of the specified aims, both methodologies produced very similar results, i.e., the smallest divergence was between R. dalmatina and R. temporaria, while the most discriminative were B. bufo and R. temporaria. However, we observed subtle shape variation of the distal region of the tail that was detected only by the geometric morphometrics. Our findings support the following. Geometric morphometric method captures more subtle shape differences that were unable to be recovered from linear measurements. It performs slightly better in classification rate. Although it was not quantified, it stands to reason that there is no difference in time investment between the two approaches. Geometric morphometrics provides more information that can be leveraged to answer further questions and it has a clear advantage in visualizing.

We report the first evaluation of mandibular and cranial modularity in the greater horseshoe bat (Rhinolophus ferrumequinum). Although some authors found no modular pattern of these morphological structures in mammals, we discovered that traits integration in R. ferrumequinum is not uniform throughout the mandible and cranium, but structured into two distinct modules. Allometry does not affect mandibular and cranial modularity in R. ferrumequinum probably as a result of the low fraction of shape variation explained by size. Observed at the static level, mandible and cranium modularity in this species could be either functional or developmental or both, but we cannot rule out a genetic nature for underlying mandibular and cranial trait interactions. Evolutionary modularity among related taxa is influenced by genetic and functional modularity. Therefore, future studies on cranial integration and modularity at multiple levels of variation may shed more light on these important features of morphological variability in bats, including R. ferrumequinum. Ministry of Education, Science and Technological Development of Serbia {[}173003]

As a basis for future conservation activities, the genetic and external body morphology variability of the European mudminnow Umbra krameri, a highly endangered fish species in Serbia and in Bosnia and Herzegovina, was determined for existing populations with the use of molecular markers (mitochondrial and microsatellite DNA) and geometric morphometric methods. Mitochondrial DNA cytochrome b gene analysis revealed two previously undescribed haplotypes: Da1 (the Lugomir population from the Danube River basin) and Sa1 (the Bakreni Batar and the Gromielj populations from the Sava River system), with a corresponding genetic distance of 07\%. Paired values of F-ST and D-AS distances for microsatellite marker data show that the difference between the Danube and the Sava populations is seven to nine times higher than the difference between the populations within the Sava River system. Geometric morphometric analyses also support a clear separation of the Lugomir population from the Bakreni Batar and the Gromielj populations. The analysis of the body shape variation, however, indicates a significant difference between the two genetically indistinguishable Sava populations. The observed genetic and phenetic relationships of the analysed mudminnow populations most probably represent a consequence of historical, geographical and ecological factors. These results will offer guidelines for future protection, conservation and sustainable management of this species in the region. Ministry of Education, Science and Technological Development of the Republic of Serbia {[}173045]; Ministry of Culture of the Czech Republic {[}DKRVO 2014/14]; Ministry of Culture of the Czech Republic (National Museum) {[}00023272]

The correlation between frequency of B chromosomes and quality of habitat was analysed in 680 specimens of yellow-necked mice, Apodemus flavicollis, collected at six localities in Serbia. Animals with B chromosomes were present at all localities in frequencies ranging from 0.23 to 0.40. Quality of habitat was judged from an index of overall body size (BSI). At the studied localities, the frequency of animals with Bs was negatively correlated with the index of overall body size (r = -0.93), indicating that B chromosomes in this species are mentioned due to the effects that they exert at the level of populations. This finding supports the heterotic model of maintenance of B chromosomes. null

Rod Aceria obuhvata preko 1000 vrsta, parazita raznih monokotiledonih i dikotiledonih biljaka, i smatra se taksonomski problematičnim, usled nejasnog taksonomsog statusa velikog broja vrsta. Šest vrsta roda Aceria je do sada opisano sa biljnih vrsta iz 16 rodova familije Brassicaceae: Aceria drabae (Nal.), Aceria capsellae (Nal.), Aceria cardaminesbellidifoliae (Liro), Aceria cardaminis (Cotte), Aceria ciansensis (Cotte) i Aceria longior (Nal.). Većina pomenutih vrsta se smatra varijetetima ili sinonimima A. drabae, tako de je nerazjašnjeno da li se radi o jednoj oligofagnoj vrsti ili je u pitanju kompleks kriptičkih monofagnih vrsta. Među navedenim vrstama ističe se A. drabae, usled moguće primene u klasičnoj biološkoj kontroli korova Lepidium draba L. Ciljevi istraživanja su bili ispitivanje interspecijske morfološke i genetičke varijabilnosti populacija Aceria spp. koje su u asocijaciji sa različitim domaćinima iz familije Brassicaceae, ispitivanje intraspecijske varijabilnosti populacija Aceria spp. koje su u asocijaciji sa jednim domaćinom, identifikovanje i opis eventualnih kriptičkih vrsta, revidiranje liste domaćina za A. drabae kao i prikaz anatomsko-morfoloških promena koje na vrstama iz famiije Brassicaceae formiraju vrste roda Aceria. Tokom istraživanja pregledano je 355 uzoraka biljnih vrsta iz familije Brassicaceae, sakupljenih sa 162 lokaliteta. Od 26 identifikovanih, na osam biljnih vrsta registrovane su vrste roda Aceria. Prilikom opisa ili dopune opisa taksona mereno je 65 morfoloških karaktera ženki, mužjaka i juvenilnih jedinki, a za uporedne kvantitativne morfometrijske analize merena su 23 karaktera protoginih ženki. Utvrđivanje vаrijаbilnosti populacija Aceria spp. nа molekulаrnom nivou izvršena je аnаlizom sekvenci COI mtDNK. Morfološka i genetička varijabilnost analizirana je za šest populacija Aceria spp. sa različitih biljaka domaćina (Aurinia petraea (Ard.) Schur, Berteroa incana (L.) DC., Capsella bursa - pastoris (L.) Medik., Cardamine hirsuta L., Lepidium draba i Sisymbrium orientale L.)... The genus Aceria includes over 1000 species of parasites of various monocotyledonous and dicotyledonous plants and it is considered taxonomically problematic due to undefined taxonomic status of these numerous species. Six species of the genus Aceria from 16 plant genera belonging to the Brassicaceae family have been described so far: Aceria drabae (Nal.), Aceria capsellae (Nal.), Aceria cardaminesbellidifoliae (Liro), Aceria cardaminis (Cotte), Aceria ciansensis (Cotte) and Aceria longior (Nal.). Most of the mentioned species are considered to be varieties or synonyms of A. drabae, so it is still not clarified whether this is an oligophagous species or it is a complex of cryptic monophagous species. A. drabae stands out among the listed species because of its potential use in classical biological control of the weed Lepidium draba L. The objectives of the study were to investigate interspecific morphological and genetic variability of populations of Aceria spp. which are in association with different hosts from the family Brassicaceae, to study the intraspecific variability of Aceria spp. which are in association with one host, to identify and describe possible cryptic species and revise the list of hosts for A. drabae, as well as to present anatomical and/or morphological alterations on the species from the family Brassicaceae caused by species of the genus Aceria. During the research, 355 samples of plant species from the family Brassicaceae, collected from 162 localities, were examined. Out of 26 identified plant species, eight were associated with the species from the genus Aceria. Descriptions or additional descriptions of certain taxa were based on 65 morphological characters of females, males and juveniles, while for the comparative quantitative morphometric analysis 23 characters of protogyne females were measured. Population variability of Aceria spp. at the molecular level was performed by analysing the COI mtDNA sequences...

Većina dosadašnjih istraživanja strukture i obrazaca varijabilnosti glavenog skeleta odnosila su se na grupe sa akinetičkom lobanjom. Istraživanje kinetizma u okviru glavenog skeleta zmija može doprineti boljem razumevanju odnosa razvojne i funkcijske integracije, uticaja alometrije na varijabilnost oblika i morfološku integraciju glavenog skeleta, promena oblika tokom ontogenije, kao i morfološke divergencije između polova. Pored toga, analizama varijabilnosti pileusa može se steći uvid u opštu sliku varijabilnosti glavenog regiona. Osnovni zadatak ove disertacije je utvrđivanje obrazaca morfološke varijabilnosti i faktora koji utiču na varijabilnost glavenog skeleta i pileusa kod dve sestrinske vrste zmija iz roda Natrix (belouške, N. natrix i ribarice, N. tessellata) primenom metoda kompjuterizovane tomografije i geometrijske morfometrije. Rezultati su pokazali da se belouška i ribarica razlikuju u veličini i obliku elemenata glavenog skeleta. Uočene razlike najverovatnije odslikavaju razlike u tipu i veličini plena, sredini u kojoj love, strategijama lova i sposobnostima držanja i gutanja plena. Polni dimorfizam u veličini i obliku elemenata glavenog skeleta veoma je izražen kod belouške i ribarice. Kod obe vrste moždana čaura i elementi trofičkog aparata su visoko integrisani, dok se kosti njuške ponašaju kao zasebni moduli. Funkcija najviše doprinosi integrisanosti elemenata glavenog skeleta. Utvrđeno je da se tek rođene belouške i ribarice razlikuju u obliku pileusa, dok se unutar vrsta polni dimorfizam u obliku pileusa ispoljava tek kod adulata. Između moždane čaure i pileusa ustanovljeni su visoki nivoi korelisanosti, kako za veličinu, tako i za oblik. Most of the previous investigations on the structure and patterns of cranial skeletons variability were done in groups with akinetic skull. Research of snake’s kinetic cranial skeleton can provide a better understanding of the relationship between developmental and functional integration, influence of allometry on shape variation and morphological integration of the cranial skeleton, shape changes during ontogeny, as well as morphological divergence between the sexes. In addition, analysis of pileus variability may provide insight into general pattern of head variability. The main aim of this thesis was to define the patterns of morphological variability and factors that affect the variability of head skeleton and pileus in two sister snakes species from the genus Natrix (Grass snake, N. natrix and Dice snake, N. tessellata) using the methods of computed tomography and geometric morphometrics. The results showed that N. natrix and N. tessellata differ in size and shape of cranial elements. Observed differences probably reflect differences in the type and size of the prey, hunting environment and strategies, handling abilities and prey swallowing. Sexual dimorphism in size and shape of cranial elements are pronounced among Grass and Dice snakes. In both species, braincase and elements of the trophic apparatus are highly integrated, while the bones of the snout can be considered as separate modules. The function is the main factor of morphological integration between cranial elements. Hatchlings of Grass and Dice snakes differed in pileus shape, while sexual dimorphism in pileus shape was detected in adults only. For both size and shape, high levels of correlation between braincase and pileus were estimated.

Većina dosadašnjih istraživanja strukture i obrazaca varijabilnosti glavenog skeleta odnosila su se na grupe sa akinetičkom lobanjom. Istraživanje kinetizma u okviru glavenog skeleta zmija može doprineti boljem razumevanju odnosa razvojne i funkcijske integracije, uticaja alometrije na varijabilnost oblika i morfološku integraciju glavenog skeleta, promena oblika tokom ontogenije, kao i morfološke divergencije između polova. Pored toga, analizama varijabilnosti pileusa može se steći uvid u opštu sliku varijabilnosti glavenog regiona. Osnovni zadatak ove disertacije je utvrđivanje obrazaca morfološke varijabilnosti i faktora koji utiču na varijabilnost glavenog skeleta i pileusa kod dve sestrinske vrste zmija iz roda Natrix (belouške, N. natrix i ribarice, N. tessellata) primenom metoda kompjuterizovane tomografije i geometrijske morfometrije. Rezultati su pokazali da se belouška i ribarica razlikuju u veličini i obliku elemenata glavenog skeleta. Uočene razlike najverovatnije odslikavaju razlike u tipu i veličini plena, sredini u kojoj love, strategijama lova i sposobnostima držanja i gutanja plena. Polni dimorfizam u veličini i obliku elemenata glavenog skeleta veoma je izražen kod belouške i ribarice. Kod obe vrste moždana čaura i elementi trofičkog aparata su visoko integrisani, dok se kosti njuške ponašaju kao zasebni moduli. Funkcija najviše doprinosi integrisanosti elemenata glavenog skeleta. Utvrđeno je da se tek rođene belouške i ribarice razlikuju u obliku pileusa, dok se unutar vrsta polni dimorfizam u obliku pileusa ispoljava tek kod adulata. Između moždane čaure i pileusa ustanovljeni su visoki nivoi korelisanosti, kako za veličinu, tako i za oblik. Most of the previous investigations on the structure and patterns of cranial skeletons variability were done in groups with akinetic skull. Research of snake’s kinetic cranial skeleton can provide a better understanding of the relationship between developmental and functional integration, influence of allometry on shape variation and morphological integration of the cranial skeleton, shape changes during ontogeny, as well as morphological divergence between the sexes. In addition, analysis of pileus variability may provide insight into general pattern of head variability. The main aim of this thesis was to define the patterns of morphological variability and factors that affect the variability of head skeleton and pileus in two sister snakes species from the genus Natrix (Grass snake, N. natrix and Dice snake, N. tessellata) using the methods of computed tomography and geometric morphometrics. The results showed that N. natrix and N. tessellata differ in size and shape of cranial elements. Observed differences probably reflect differences in the type and size of the prey, hunting environment and strategies, handling abilities and prey swallowing. Sexual dimorphism in size and shape of cranial elements are pronounced among Grass and Dice snakes. In both species, braincase and elements of the trophic apparatus are highly integrated, while the bones of the snout can be considered as separate modules. The function is the main factor of morphological integration between cranial elements. Hatchlings of Grass and Dice snakes differed in pileus shape, while sexual dimorphism in pileus shape was detected in adults only. For both size and shape, high levels of correlation between braincase and pileus were estimated.

Кечига (Acipenser ruthenus L., 1758) представља потамодромну јесетарску врсту, која насељава реке у сливовима Црног, Азовског, Каспијског, Балтичког, Белог, Баренцовог и Кара мора. Значајан пад бројности услед излова током XX века довео је до потребе за бољом заштитом, па је на IUCN Црвеној листи угрожених врста, кечига сврстана међу рањиве врсте (VU). Ради испитивања морфолошке и генетичке диференцијације кечиге у средњем и доњем току Дунава, узорковано је 95 јединки из три популације, од којих две из Србије (Нови Кнежевац на Тиси и Бачка Паланка на Дунаву) и једна популација из Румуније (Гринду на Дунаву). Међупопулациона варијабилност спољашње морфологије (промене величине и облика тела и главеног региона) анализирана је применом метода геометријске морфометрије. Уни- и мултиваријатном анализом варијансе утврђено је постојање статистички значајних разлика у величини и облику анализираних морфолошких целина између јединки са различитих локалитета. Канонијска дискриминантна анализа (CVA) укупне варијабилности облика, код латералног и вентралног приказа тела и латералног приказа главеног региона, показала је да се по првој оси раздваја популација из Румуније од популација из Србије. На нивоу вентралне перспективе тела, као и на нивоу обе перспективе главе утврђен је статистички значајан ефекат алометрије на укупну варијабилност облика. На основу CVA неалометријске компоненте варијабилности облика тела (вентрална перспектива) и главеног региона (латерална и вентрална перспектива) утврђено је да по првој оси постоји раздвајање јединки из Румуније у односу на јединке из Србије. Такође, CVA неалометријске компоненте варијабилности облика главеног региона у обе перспективе указује на могућност да кечиге из Србије (нарочито оне из Тисе) одговарају краткорилој, док кечиге из Румуније одговарају дугорилој форми. Употребом митохондријске ДНК и микросателита као молекуларно-генетичких маркера, детектован је значајан ниво диверзитета (13 хаплотипова, просечан број алела по локусу и популацији 4.3 – 4.9 и добијена хетерозиготност 0.577 – 0.648) али не и јасна диференцијација између популација... Sterlet (Acipenser ruthenus L., 1758) is a potamodromus sturgeon species, which inhabits river stretches of Black, Azov, Caspian, Baltic, White, Barents and Kara sea drainages. Sterlet experienced a decline during 20th century, which led to necessity for better protection, and inclusion on IUCN Red List of endangered species as a vulnerable species (VU). То analyse morphological and genetic differentiation of sterlet in Middle and Lower Danube sections, 95 specimens were sampled from three populations, two from Serbia (Novi Kneževac on Tisza River and Bačka Palanka on Danube River) and one from Romania (Grindu on Danube River). Variability of external morphology (size and shape variation of body and head region) was examined using geometric morphometric methods. Statistically significant interpopulation differences in size and shape of analyzed morphological structures were found. Canonical variate analysis (CVA) of overall shape variation (for lateral and ventral views of body and for lateral view of head region) showed that first CV axis separated Romanian sample from Serbian samples. Statistically significant effect of alometry was observed for ventral view of the body, as well as for lateral and ventral views of the head. CVA of non-allometric component of shape variation for body in ventral view and for head in lateral and ventral views showed separation of Romanian population from Serbian populations. Also, non-allometric shape changes of head in lateral and ventral views correspond to shape differences between blunt-snout and pointed-snout sterlet morphs. Use of mitochondrial DNA and microsatellite markers, showed significant level of diversity (13 haplotypes, allelic richness range from 4.3 to 4.9, observed heterozygosity from 0.577 to 0.648). Genetic analysis (mtDNA and microsatellite) showed lack of clear population differentiation. Additionally, five new haplotypes were detected, while eight previously described haplotypes had extended their previous range. Analysis of molecular variance indicated that the genetic variance could be attributed almost entirely to individuals from same populations and that interpopulation variability was absent. Samples from the Danube River were most divergent (FST=0.038), while lowest FST (0.018) was observed between samples from Serbia (Tisza and Danube River)...

Integracija, modularnost i razvojna ograničenja predstavljaju važne komponente fenotipske varijabilnosti. U ovoj tezi analiziran je uticaj ovih komponenti na morfološku evoluciju ekstremiteta pet vrsta velikih krestatih mrmoljaka. Veliki krestasti mrmoljci predstavljaju važne model organizme za istraživanja u oblasti morfološke integracije i razvojnih ograničenja, posebno ako se analiziraju ekstremiteti. To je grupa blisko srodnih vrsta sa poznatim filogenetskim odnosima, i izraženom vezom izmedju morfološke forme/tipa (oblika tela i dužine ekstremiteta) i preference prema akvatičnoj/terestričnoj sredini. Akvatične vrste se odlikuju izduženim telom i kratkim nogama, dok su vrste sa više terestričnim životnim stilom snažnije i robusnije građe sa dugim nogama. Takođe, kao grupa sa složenim životnim ciklusom, gde larveni, juvenilni i adultni stupanj imaju različite morfologije, ekološke sredine i preference, načine ishrane i lokomocije, veliki krestasti mrmoljci imaju posebno mesto u izučavanju ontogenetskih putanja tokom razvića. Ontogenetska promena niše i tranzicija između akvatične i terestrične sredine je povezana sa procesom metamorfoze. Iako ekstremiteti mrmoljaka ne prolaze kroz drastične promene strukture tokom tog procesa, funkcionalni zahtevi za efikasnom lokomocijom u te dve sredine su bitno različiti. Pretpostavka je da razlike između vrsta u pogledu stepena korišćenja akvatične tj. terestrične lokomocije, može biti faktor koji doprinosi razlikama u dužinama skeletnih elemenata i različitim obrascima korelacija između elemenata ekstremiteta, oslikavajući adaptacije na različite uslove sredine i način života (više ili manje akvatičan-terestričan)... Integration, modularity and developmental constraints are three related components of phenotypic variability. Their impacts on morphological evolution on limbs in five species of crested newts were analyzed in this thesis. Crested newts are valuable model or ganisms in studies of morphological integration and developmental constraints, especially when concern limbs. They are closely related group of species with known phylogenetic relations, and each characterized by a specific body-to-limb conformation associated with their respective ecologies. Aquatic species of newts have small, elongate bodies and small limbs while more terrestrial species are with large, stocky bodies and robust limbs. Also, as group with complex life cycle, where larval, juvenile and adult stage have different morphologies, ecol ogies and different diet and locomotor preferences, crested newts have special pl ace in studies of changes in ontogenetic trajectories. Ontogenetic niche change and transition between aquatic and terrestrial environment is connected with process of metamorphosis. Although limbs in newts do not undergo drastic chan ges in structure during that process, functional demands for high efficient loco motion in both environments are quit different. It is assumed that the differenc es between species in degree of aquatic and terrestrial locomotion may be the fa ctor which can attribute to changes in length of skeletal elements and differe nt patterns of correlations between limb elements, reflecting adaptations on diffe rent environments and life styles of analyzed species of crested newts...

Crnka (Umbra krameri Walbaum, 1792) je jedina autohtona i reliktna vrsta iz roda Umbra prisutna u Evropi, endemična za basene Dunava i Dnjestra. U okviru današnjeg diskontinuiranog areala, usled isušivanja i zagađivanja staništa, populacije crnke značajno su smanjene poslednjih decenija XX veka. Iz navedenih razloga crnka se nalazi na IUCN Crvenoj listi ugroženih vrsta i ima status ranjive vrste (VU). Na nacionalnom nivou ima status kritično ugrožene vrste (CR) i nalazi se u neposrednoj opasnosti od iščezavanja. U ovoj disertaciji su uz distribuciju vrste u Srbiji i Bosni i Hercegovini, sa novozabeleženom populacijom u Srbiji (lokalitet Lugomir), prikazani i rezultati analize ekoloških karakteristika i morfološko-genetičke diferencijacije istraživanih populacija, a dat je i status ugroženosti vrste na nacionalnom nivou sa ugrožavajućim faktorima i merama zaštite i očuvanja. Za potrebe rada uzorkovano je ukupno 76 jedinki iz tri populacije, dve iz Srbije (23 jedinke sa lokaliteta Lugomir i 21 jedinka sa lokaliteta Bakreni Batar) i jedna iz Bosne i Hercegovine (32 jedinke sa lokaliteta Gromiželj). Za svaki istraživani lokalitet određeni su osnovni geografski parametri (geografska dužina, širina i nadmorska visina) i fizičko-hemijske karakteristike staništa (širina vodenog basena, dubina vode i mulja, temperatura vode i vazduha, pH vrednost, elektroprovodljivost, koncentracija kiseonika, saturacija kiseonikom, koncentracije amonijaka, nitrita i fosfata). Sve jedinke u uzorku su prvo fotografisane za potrebe analiza geometrijske morfometrije, a nakon toga su im merene totalna (TL) i standardna (SL) dužina tela i težina tela za potrebe analize strukture populacija. Takođe, jedinkama su uzete gonade za utvrđivanje pola, uzorci digestivnog trakta za analizu ishrane, uzorci krljušti za određivanje starosti i uzorci tkiva za analizu genetičke varijabilnosti. Na svim istraživanim lokalitetima utvrđen je kvalitativni sastav biljaka i riba, a izvršeno je i uzorkovanje beskičmenjaka iz mulja i submerzne vegetacije u cilju utvrđivanja njihovog kvalitativnog i kvantitativnog sastava i analize ishrane crnke. Sva tri istraživana lokaliteta prema analiziranim fizičko hemijskim parametrima i sastavom zajednice biljaka i riba predstavljaju tipična staništa crnke... European mudminnow (Umbra krameri Walbaum, 1792) is the only autochthonous and relic species of the genus Umbra present in Europe, and it is endemic for the Danube and Dniester river basins. In scope of its present discontinuous range, due to habitat desiccation and pollution, the populations of European mudminnow significantly decreased during the last decades of the 20th century. For this reason it is listed on the IUCN Red List of Threatened Species as Vulnerable (VU). At the national level, it is listed as Critically Endangered (CR), with a serious threat to be soon extinct. The range of the species in Serbia and in Bosnia and Herzegovina is presented in this dissertation, including a newly recorded population in Serbia (locality Lugomir), along with the results of ecological and morphological-genetic differentiation of the analysed populations, and the threat status at the national level with threat factors and conservation measures. A total of 76 individuals from three populations, two from Serbia (23 individuals from Lugomir and 21 individuals from Bakreni Batar) and one from Bosnia and Herzegovina (32 individuals from Gromiželj), were sampled. Basic geographical parameters (longitude, latitude, altitude) were determined for each locality, along with physical and chemical habitat parameters (width of the water basin, water and silt depth, water and air temperature, pH, electroconductivity, oxygen concentration, oxygen saturation, ammonia, nitrites, phosphates). All sampled individuals were photographed for the purpose of geometric morphometrics analyses, and their total (TL) and standard (SL) body length and body weight were measured for population stucture analyses. Furthermore, gonads were taken for sex determination, digestive tract for diet analysis, scales for age determination, and tissue samples for genetic variability analysis. Species composition of aquatic plants and fishes was determined at all localities, and invertebrates from mud and submersed vegetation were sampled with the aim to determine their diversity and to analyse the diet of European mudminnow. For their physical-chemical parameters and aquatic plant and fish species composition all three studied localities represent typical habitats of European mudminnow...

Integracija, modularnost i razvojna ograničenja predstavljaju važne komponente fenotipske varijabilnosti. U ovoj tezi analiziran je uticaj ovih komponenti na morfološku evoluciju ekstremiteta pet vrsta velikih krestatih mrmoljaka. Veliki krestasti mrmoljci predstavljaju važne model organizme za istraživanja u oblasti morfološke integracije i razvojnih ograničenja, posebno ako se analiziraju ekstremiteti. To je grupa blisko srodnih vrsta sa poznatim filogenetskim odnosima, i izraženom vezom izmedju morfološke forme/tipa (oblika tela i dužine ekstremiteta) i preference prema akvatičnoj/terestričnoj sredini. Akvatične vrste se odlikuju izduženim telom i kratkim nogama, dok su vrste sa više terestričnim životnim stilom snažnije i robusnije građe sa dugim nogama. Takođe, kao grupa sa složenim životnim ciklusom, gde larveni, juvenilni i adultni stupanj imaju različite morfologije, ekološke sredine i preference, načine ishrane i lokomocije, veliki krestasti mrmoljci imaju posebno mesto u izučavanju ontogenetskih putanja tokom razvića. Ontogenetska promena niše i tranzicija između akvatične i terestrične sredine je povezana sa procesom metamorfoze. Iako ekstremiteti mrmoljaka ne prolaze kroz drastične promene strukture tokom tog procesa, funkcionalni zahtevi za efikasnom lokomocijom u te dve sredine su bitno različiti. Pretpostavka je da razlike između vrsta u pogledu stepena korišćenja akvatične tj. terestrične lokomocije, može biti faktor koji doprinosi razlikama u dužinama skeletnih elemenata i različitim obrascima korelacija između elemenata ekstremiteta, oslikavajući adaptacije na različite uslove sredine i način života (više ili manje akvatičan-terestričan). Analiza morfološke integracije i modularnosti je utvrdila postojanje dva važna aspekta variranja elemenata ekstremiteta velikih krestastih mrmoljaka. Odgovarajući ontogenetski stupnjevi su sličniji između vrsta nego sukcesivni ontogenetski stupnjevi u okviru jedne iste vrste, sa najvećim nivoom integracije u juvenilnom stupnju. Visoki nivo integracije prisutan u juvenilnom stupnju predstavlja funkciju brzog, koordinisanog rasta skeleta ekstremiteta tokom perioda u okviru koga sistemski hormoni imaju odlučujuću ulogu, dok je niska ukupna integracija na adultnom stupnju je najverovatnije rezultat mnogih lokalnih i globalnih-sistemskih faktora koji menjaju jedan drugome efekat u vremenu i prostoru, pri čemu bifazični životni ciklus sa različitim načinima lokomocije, imaju odlučujuću ulogu u ontogenetskim promenama morfološke integracije i modularnosti. Razlike u obrascima integracije između više akvatičnih i više terestričnih vrsta mrmoljaka su predstavljene nižim stepenom korelacija između homologih elemenata ekstremiteta, i povećanjem korelacija između elemenata unutar ekstremiteta kod terestričnih vrsta velikih mrmoljaka (kao T. karelinii), dok više akvatične vrste (kao T. dobrogicus) imaju suprotan obrazac variranja sa većim korelacijama između homologih elemenata ekstremiteta nego između elemenata unutar ekstremiteta. Ova studija je pokazala važan uticaj razvojnih ograničenja u kreiranju morfološke varijabilnosti ekstremiteta velikih krestastih mrmoljaka. Uporedna analiza osifikacionih sekvenci ekstremiteta utvrdila je postojanje nekoliko sinapomorfnih heterohroničnih promena kod velikih krestastih mrmoljaka, koje uključuju kašnjenje u sekvenci drugog prsta i ubrzanje metakarpalnog elementa III i metatarzalnog elementa V. Ove heterohronične promene nisu adaptivne ali su važne za razmatranje morfološke evolucije ekstremiteta kod velikih krestastih mrmoljaka jer uključuju promenu preaksijalne ka postaksijalnoj dominaciji u razvojnoj sekvenci, koja nije poznata kod repatih vodozemaca. Analiza alometrijskih obrazaca variranja ekstremiteta velikih mrmoljaka je pokazala dva važna rezultata: osobenosti obrazaca rasta kod vrste T. dobrogicus, kao i zajednički obrazac rasta nakon završene metamorfoze kod svih vrsta velikih mrmoljaka. Ovi rezultati ukazuju na verovatni uticaj i ulogu kako prirodne selekcije, tako i unutrašnjih-razvojnih mehanizama koji regulišu variranje dužine ekstremiteta tokom diverzifikacije velikih mrmoljaka. Obrasci variranja u basipodijumu su slični medju analiziranim vrstama sa većim variranjem u broju karpalnih nego tarzalnim elemenata. Značajan podatak ove studije je da je u basipodijumu vrste T. dobrogicus došlo do gubitka jednog od karpalnih elemenata (centrale 1). Specifičnost ekstremiteta dunavskog mrmoljka se dopunjuje i posebnim planom organizacije karpusa i tarzusa koji odlikuje nizak nivo osifikacije praćen rastresitim pakovanjem skeletnih lemenata. Sve ove odlike ukazuju na postojanje heterohronične promene (pedomorfoze) kada je u pitanju skelet ekstremiteta. Rezultati ove studije su pokazali značajan uticaj morfološke integracije, modularnosti i razvojnih ograničenja u kreiranju fenotipske varijabilnosti u ekstremitetima velikih krestastih mrmoljaka. Integration, modularity and developmental constraints are three related components of phenotypic variability. Their impacts on morphological evolution on limbs in five species of crested newts were analyzed in this thesis. Crested newts are valuable model organisms in studies of morphological integration and developmental constraints, especially when concern limbs. They are closely related group of species with known phylogenetic relations, and each characterized by a specific body-to-limb conformation associated with their respective ecologies. Aquatic species of newts have small, elongate bodies and small limbs while more terrestrial species are with large, stocky bodies and robust limbs. Also, as group with complex life cycle, where larval, juvenile and adult stage have different morphologies, ecologies and different diet and locomotor preferences, crested newts have special place in studies of changes in ontogenetic trajectories. Ontogenetic niche change and transition between aquatic and terrestrial environment is connected with process of metamorphosis. Although limbs in newts do not undergo drastic changes in structure during that process, functional demands for high efficient locomotion in both environments are quit different. It is assumed that the differences between species in degree of aquatic and terrestrial locomotion may be the factor which can attribute to changes in length of skeletal elements and different patterns of correlations between limb elements, reflecting adaptations on different environments and life styles of analyzed species of crested newts. Analysis of morphological integration and modularity has found different aspects of variation patterns in limbs of crested newts. Equivalent ontogenetic stages of different species of crested newts show higher concordance in the correlation pattern than successive ontogenetic stages within species, with highest level of integration in juvenile stage. High integration in juveniles is function of the rapid, coordinated growth of the limb skeleton during this time period, in which systematic hormones are thought to play a major role. While, low integration in adult stage are probably due to many locally and globally-systematic factors which change effect one to another in time and space, although biphasic life cycle with different locomotion modes have decisive role in ontogenetic changes of morphological integration and modularity. Differences in patterns of integrations between aquatic and more terrestrial species of newts were observed, with lower correlation between homologous limb elements and higher correlation within-limb elements in more terrestrial species, the reverse pattern occurs in more aquatic species. This study showed important role of developmental constraints in creating phenotypic variability in limbs of crested newts. Comparative analyses revealed some synapomorphic heterochronic shifts specific to crested newts, including delay of the ossification in the second finger and accelerations in metacarpal III and metatarsal V. These shifts involve a change from pre-axial to post-axial dominance in a developmental sequence uncommon to caudate salamanders. No adaptive explanation of these shifts is apparent. Allometric analyses showed two important results: specific growth patterns in T.dobrogicus and similar growth pattern after metamorphosis in all species of crested newts. These results are reflecting impact of natural selection as well as internal developmental mechanisms which regulated variation in length of limbs during morphological diversification of crested newts. All studied species shared the same basipodial pattern, within which the carpus exhibited much more variation than the tarsus. Important point in this study is lost of one carpal element (centrale 1). Specific pattern observed in T. dobrogicus is followed with low ossification and loose package of carpale elements. It is suggested that the uniqueness of T. dobrogicus originates from heterochronic process. Results of this study showed important influence of morphological integration, modularity and developmental constraints in creating phenotypic variability in limbs of crested newts.