Conservation implications of small population size and habitat fragmentation in an endangered lupine

Conservation measures to preserve critically endangered species aim to maintain healthy and self-sustaining populations and often involve reintroductions. Effective introductions must take into account the genetic structure and diversity remaining in the species to inform choices of germplasm for introduction strategies. Lupinus aridorum (McFarlin ex Beckner; Fabaceae) is an endangered plant found on two disjunct ridge systems in central Florida. All plants are found in areas that are favored for human development and agriculture. Few options exist for introduction locations, so the most informed strategies must be used to preserve this species. We used ten microsatellite loci, developed for this species, to determine genetic diversity and genetic differentiation among populations, and to compare L. aridorum with two closely related congeners. Our results show a low level of diversity remaining in the species in comparison with a more abundant congener and similar levels of variability among each L. aridorum population. At least three L. aridorum populations exhibit a genetic signal of partial selfing. Hybridization with a closely-related congener was not apparent; however, additional crossing trials are needed to fully assess the possibility of hybridization. Although some sources list the taxonomy of L. aridorum as a variety of a Florida panhandle species, L. westianus (Small), the genetic differences revealed by our data support listing these two species as separate entities. Our findings can be used to guide sampling strategies for ex situ conservation such as providing a mixture of source populations to improve the probability of successful introductions.     

Towards the conservation of a critically endangered species, <Emphasis Type="Italic">Aphanius farsicus</Emphasis>: embryogenesis and development

Human-induced disturbance or anthropogenic activities, natural disturbance and also limited distribution, have forced Aphanius farsicus populations to the edge of extinction, during the past few years. As knowledge of fish ontogeny is fundamental for accurate stock assessment, and reproductive potential determination, thus for future conservation and management of the critically endangered species, we describe the first detailed normal embryonic development of this cyprinodontid fish from fertilization to post-hatching. Aphanius farsicus spontaneously spawned at 24°C. The newly-laid eggs were hyaline and spherical in shape (1.40 ± 0.02 mm, n = 11), sticky and full of egg yolk (telolecithal/macrolecithal) with oil droplets. We documented the main developmental stages at 24°C to zygote (0 h), cleavage (2 h), blastula (2 h), gastrula (17 h), segmentation, (34 h), pharyngula (100 h) and hatching (162 h).     

Disentangling the stream community impacts of <Emphasis Type="Italic">Didymosphenia geminata</Emphasis>: How are higher trophic levels affected?

Human activities frequently result in either intentional or unintentional introductions of species to new locations, and freshwater environments worldwide are particularly vulnerable to species invasions. An introduced freshwater diatom, Didymosphenia geminata, was first discovered in New Zealand in 2004 but there was limited research available to predict the drivers of D. geminata biomass and how biomass variability might influence higher trophic levels (e.g. invertebrates and fish). We examined the effect of D. geminata biomass on benthic invertebrates, invertebrate drift and fish communities in 20 rivers in New Zealand with variable hydrology, physical habitat and water chemistry. Variation in D. geminata biomass was best explained by a model that showed D. geminata biomass increased with time since the last flow event exceeding three times the median annual discharge and decreasing concentration of dissolved reactive phosphorus. Analyses of biotic responses showed that high D. geminata biomass did not affect either invertebrate or fish diversity but altered the structure of benthic communities, changed the composition of drifting invertebrate communities and reduced fish biomass by 90 %, particularly trout. A partial least squares path model was used to disentangle both direct and indirect effects of D. geminata on fish communities and showed D. geminata had a significant negative direct effect on fish communities. This is the first study to show how the potential effects of the introduced diatom D. geminata can impact fish communities and has shown that D. geminata impacts fish both directly and indirectly through changes in their invertebrate prey community.     

Novel fungi from an ancient niche: lachnoid and chalara-like fungi on ferns

A survey was conducted in Brazil to collect fungi on ferns. Based on morphology and inferred phylogeny from DNA sequences of two loci, namely the internal transcribed spacer (ITS) regions and the large subunit nuclear ribosomal RNA gene (LSU), several species belonging to chalara-like genera and lachnoid fungi were recognized. Eighteen fungal isolates, collected from five host species, representing 10 different localities were studied. Three novel genera (Lachnopsis, Scolecolachnum and Zymochalara), and six novel species (Bloxamia cyatheicola, Lachnopsis catarinensis, Lachnopsis dicksoniae, Scolecolachnum pteridii, Zymochalara lygodii and Zymochalara cyatheae) are introduced. Furthermore, two new combinations (Erioscyphella euterpes and Erioscyphella lushanensis) are proposed. Two novel taxa (Lachnopsis catarinensis and Lachnopsis dicksoniae) may be included in the list of potentially endangered fungal species in Brazil, if proven to be restricted to their tree-fern host, Dicksonia sellowiana, which is included in the official list of endangered plant species in Brazil.     

Non-indigenous fish in the northern branch of the central European invasion corridor

The assessment of risks associated with non-indigenous species implies a detailed knowledge of their taxonomical composition and distribution within a certain region. The northern branch of the central European ‘invasion corridor’—a series of canals connecting different watersheds—has formed an important migratory route for Ponto-Caspian fish (i.e. fish from the Black Sea, the Sea of Azov and the Caspian Sea). However, the current status of non-indigenous fish species in this region is very scarce. This article presents a comprehensive overview of recent distribution data of non-indigenous fish species in the northern branch of the central European invasion corridor. Here, extensive data are integrated based on studies performed during 2003–2014 comprising reliable published and unpublished records from the last 12 years. Altogether, ten non-indigenous fish species were currently found in the northern branch of the central European invasion corridor, constituting 19 % of its freshwater fish diversity. Three species, including the Prussian carp (Carassius gibelio), the round goby (Neogobius melanostomus), and the Chinese sleeper (Perccottus glenii), are considered invasive species. Eight species may potentially invade this region in the near future. A comparison of the history of non-indigenous fish species introduction in the inland waters of the northern branch and other countries of the central European invasion corridor revealed similar introduction trends. Potential expansion of non-indigenous fish species across the central European invasion corridor has international implications that require awareness, cooperation, and government support from each individual country. Disclosure of recently operating vectors for non-indigenous fish introductions within the central European invasion corridor will help predict and prevent their further spread and establishment in this region.     

Fish assemblages in deep Italian subalpine lakes: history and present status with an emphasis on non-native species

We provide a comprehensive overview of the status of fish assemblages in eight Italian subalpine lakes using multiple data sources: historical records, standardised sampling and fishery catch statistics. Thirty-four fish species were recorded, twenty of which are native, seven introduced historically and another seven introduced recently. The number of fish species in individual lakes has increased by between 25 and 65% from 1855 to the present, with the increase being higher in the larger lakes. Non-native species contributed between 4.0 and 71.5% to standardised catches by numbers and between 4.8 and 65.1% by biomass. Roach Rutilus rutilus and pumpkinseed Lepomis gibbosus dominated electrofishing and benthic catches by number, while open-water catches were dominated by the coregonids European whitefish Coregonus lavaretus and gangfish Coregonus macrophthalmus. In terms of biomass, electrofishing catches were dominated by crucian carp Carassius carassius, largemouth bass Micropterus salmoides and wels catfish Silurus glanis, benthic catches were dominated by roach, European whitefish and crucian carp, and open-water catches by coregonids. Commercial fisheries are important ecosystem services provided by subalpine lakes, with their harvests now dominated by non-native species. To deliver effective biodiversity conservation and ecosystem services provision, the prioritisation of targets and interventions of management of non-native species is needed.     

<Emphasis Type="Italic">Spartina versicolor</Emphasis> Fabre: Another case of <Emphasis Type="Italic">Spartina</Emphasis> trans-Atlantic introduction?

Intercontinental introductions are widespread in the genus Spartina, with important ecological and evolutionary consequences. The native or introduced status of Spartina species is then critical with regard to biodiversity assessment, especially for vulnerable Mediterranean coastline ecosystems. Spartina versicolor was first recorded in southern France in 1849, then successively in various places on the European and North-African Mediterranean and Atlantic coasts. This species is considered to be either a European native or an invasive species introduced from North America which has a high morphological similarity to the Atlantic American species Spartina patens. We performed extensive sampling of S. versicolor in Europe and North Africa (from natural populations and herbarium collections) and compared these samples to other European and American Spartina species (including S. patens). Chromosome counts were reported for the first time and revealed that S. versicolor is tetraploid (2n = 4x = 40). Phylogenetic analyses based on chloroplast and nuclear ribosomal DNA sequences did not reveal any molecular variation within S. versicolor. In this species, a single haplotype, that is identical to one haplotype of S. patens, was found in the four chloroplast and the nuclear ribosomal ITS regions investigated. In addition, simple sequence repeat markers were used and revealed a low level of genetic diversity within S. versicolor, suggesting that the introduction of S. versicolor occurred from a narrow genetic pool of S. patens from North America.     

Displacement of native Patagonian freshwater silverside populations (<Emphasis Type="Italic">Odontesthes hatcheri</Emphasis>, Atherinopsidae) by introgressive hybridization with introduced <Emphasis Type="Italic">O. bonariensis</Emphasis>

The Patagonian silverside Odontesthes hatcheri is a native fish restricted to streams and lakes of Patagonia (Argentina and Chile). Stocking programs to enhance recreational fisheries in man-made reservoirs have introduced a nonnative, closely-related species (the pejerrey O. bonariensis) in Patagonia almost a century ago, and yet little is known about the invasiveness of this species. To evaluate the impact of these introductions we analyze genetic data (microsatellite markers and mitochondrial DNA) to quantify the incidence of hybridization between these two species and assess potential effects on native population structure. Phylogeographic analyses reveal weak geographic differentiation among populations of O. hatcheri, in agreement with previous studies for other freshwater fishes in Patagonia strongly influenced by Quaternary glaciations and hydrographic basin changes since the last glaciation. However, many populations have unique genetic pools. In several areas, introductions resulted in extensive hybridization, with high frequencies of F2 and backcrossed hybrids in natural populations, and in some cases O. bonariensis has completely displaced the native populations. The negative impact of these introductions on native populations is correlated to temperature, a critical parameter in the face of global warming, suggesting that invasiveness of O. bonariensis may increase in the future. Our results advise against continuing stocking programs to preserve the integrity of natural populations of the Patagonian silverside.     

Associations between stream habitat characteristics and native and alien crayfish occurrence

Human mediated introductions of non-indigenous crayfish species (NICS) are responsible for their rapid colonisation of European freshwaters. The introduction of North American crayfish is furthermore linked to the spread of crayfish plague and the decline of indigenous crayfish species (ICS). As the management of ICS and NICS have become necessary, a detailed knowledge on their distribution and ecological requirements is needed. We studied the current range of native noble crayfish Astacus astacus and stone crayfish Austropotamobius torrentium, as well as alien signal crayfish Pacifastacus leniusculus in Carinthia (Austria) and evaluated environmental and physical habitat features in streams with and without crayfish. Meanwhile, the loss of many ICS populations was recorded and alien P. leniusculus was found to be widespread in this region. Most of the habitat features of streams having crayfish differed significantly from sites lacking crayfish for at least one investigated native or alien species. Furthermore, multivariate and regression analyses showed specific differences in the habitat use of the investigated crayfish. Our results showed that the presence of alien P. leniusculus was associated with larger and smoother sloped lowland rivers, while the occurrence of the two native species was confined to smaller streams either at higher altitudes and with distinct physical habitat conditions (A. torrentium) or with moderate water temperatures (A. astacus). This study helps to identify potential refuge areas for the endangered native species and to predict the further spread of the most common non-native crayfish species in European streams.     

Distribution and abundance of fish in the southwest of Caspian Sea coastal waters

This study was carried out in southwestern Caspian Sea coastal area to elucidate demersal bony fishes distribution and abundance. Twenty two species were found in shore line, which belong to 6 families. In deeper waters down to 7 meters only 5 species of Gobiidae and 1 species of Syngnathidae were identified. Two species Neogobius caspius and N. pallasi were the main components of demersal fishes; however in some regions Rutilus caspicus, Rutilus kutum and Liza spp. were dominated. Most places were dominantly occupied by Atherina boyeri, which had the highest abundance among the pelagic fishes. Conversely, the Ponticola gorlap, N. melanostomus and Proterorhinus nasalis showed the lowest abundances (less than 1 ind./100 m²). Benthophilus stellatus and P. nasali had the lowest niche overlaps with other species. Both of these two species and P. gorlap are suggested to be classified as endangered or vulnerable species based on their low abundance. The distribution of Syngnathus abaster depends on algae, while algae grow up into the stony construction of the coastal line. A negative insignificant correlation was observed between pelagic and demersal fish abundances. The commercial fisheries data of 76 beach seines in our study area during 2002–2013 was compared with our results on fish abundance and species composition. More than 93% of total fish catch constituted Liza spp. and Rutilus kutum. The relationship between the abundance and distribution of the studied species, and the restocking activities performed by Iranian fisheries organization are discussed. The high abundance and distribution of small size fish in tidal zone could be related to the distribution of crustacean, which provide a good source of food for these fish species.     

Feeding ecology of the Kızılırmak toothcarp, <Emphasis Type="Italic">Aphanius marassantensis</Emphasis>: ontogenetic shift and seasonal diet variation

We studied the feeding ecology of an endemic cyprinodontid fish, Aphanius marassantensis (K?z?l?rmak Toothcarp), with special emphasis on seasonal and ontogenetic diet shift. The dietary composition revealed an omnivorous diet with great seasonal and ontogenetic variation. Cladocera and Calanoid Copepods (Diaptomus sp.) dominated the diet in Spring and Summer, whereas the food items of plant origin e.g. filamentous algae and diatoms were the most important components in Autumn and Winter. An ontogenetic diet shift was also demonstrated. While the feeding of juveniles was based mostly on planktonic organisms, adult individuals preferred larger prey taxa, such as Gammarus sp. and Gastropods. There was no significant sex-related variation in feeding with the exception for the volume of gut content. The investigation on the feeding strategy of A. marassantensis suggested a generalised feeding pattern with some specialised individuals. This generalist feeding habits may account for the well-established population of this species in a big reservoir under coexistence of two non-native fish species, Pseudorasbora parva and Atherina boyeri.     

Non-native aquatic animals introductions have driven disease emergence in Europe

In this paper it is argued, using examples of disease emergence in aquatic animals in Europe, that the introduction of non-native species drives disease emergence by both extending the geographic range of parasites and pathogens and facilitating host-switching. Enteric red mouth disease and infectious haematopoietic necrosis of salmonids have extended their geographic range from North America to Europe with the import of live fish (Pimephales promelas) and rainbow trout eggs, respectively. Host-switching results in disease emergence when previously unidentified commensal organisms or known pathogen switch to new naïve hosts. The most serious endemic diseases of wild aquatic animals in Europe in recent years can be traced to the introduction of non-native species. Across Europe dramatic populations declines have occurred in native crayfish (e.g. Astacus astacus), oysters (Ostrea edulis) and eels (Anguilla anguilla), all which can be attributed, in varying degrees, to diseases (crayfish plague, Bonamia ostreae and Anguillicoloides crassus, respectively) introduced with non-native species. The severe adverse effects at a population level can be attributed to the lack of immunity in the new hosts. The impact of parasites more recently introduced to Europe, Sphaerothecum destruens (the rosette agent), and Batrachochytrium dendrobatidis, have yet to be fully determined. Both are generalists, with wide host ranges, and may present serious threats to native species. Aquaculture is the key driver for the introduction of non-native species. Most farming systems allow pathogen exchange between farmed and wild populations which underpins host-switching. Subsequently movements of animals between farms may result in the spread of newly emerged diseases. The introduction of non-native aquatic animals drives disease emergence, thus the ex-ante assessment of these hazards is severely limited. Generic risk mitigation measures (e.g. movement of disinfected eggs in place of live animals) and improved methods for rapid detection of new diseases are vital.     

The recent occurrence,establishment and potential impact of <Emphasis Type="Italic">Geophagus proximus</Emphasis> (Cichlidae: Perciformes) in the Tietê River reservoirs: an Amazonian fish species introduced in the Paraná Basin (Brazil)

The introduction of allochthonous fish species happens constantly in large bodies of freshwater, like as the reservoirs of Paraná Basin, located in Brazilian southeast, representing a threat for local biodiversity. The fish species Plagioscion squamosissimus and Cichla ocellaris were introduced from the 1970s in several water bodies of this basin and had successfully established themselves in all six reservoirs located in the middle and lower Tietê River (SP, Brazil), particularly. After six decades from the first recorded species introduction, this hydrographic system remains open to the invasion of further fish species, owing to widespread fish-farming activity and by the channels opened between this system and other reservoirs and river basin. This study was an effort to confirm the Geophagus proximus occurrence in the six Tietê River reservoirs, verifying the actual introduction status and analyzing its potential environmental impacts on local species by the analysis of the population structure (abundance, body dimensions and feeding habits). By the results, this species was confirmed in the Ibitinga, Nova Avanhandava and Três Irmãos reservoirs. The abundance and feeding analysis shows, respectively, it is successfully established in the Três Irmãos reservoir with the same feeding habitats of local species, such as Geophagus brasiliensis. It was further shown to be very likely that G. proximus would spread throughout the reservoir system of the middle and lower Tietê River, in the manner of P. squamosissimus and C. ocellaris, and the competition pressure for food resources between G. proximus and the local species which represents a potential environmental impact system. These scientific evidences fortifies the knowledge basin for the implantation of a fish management system, to control and reduce the abundance of the invader and to prevent its becoming established in all the Tietê River Basin, avoiding the disastrous consequences for the native species of Paraná River Basin.     

Morphological and molecular taxonomy of novel species <Emphasis Type="Italic">Pleurotheciaceae</Emphasis> from freshwater habitats in Yunnan,China

The diversity of lignicolous freshwater fungi of the Greater Mekong Subregion are currently being studied. In this paper, 18 fresh collections of Pleurotheciaceae from submerged wood in freshwater are characterized based on morphological characters and analyses of ITS, LSU, SSU and RPB2 sequence data. Six new species of Pleurotheciella (P. aquatica, P. fusiformis, P. guttulata, P. lunata, P. saprophytica, P. submersa), one new Phaeoisaria species (Ph. aquatica) and one new Pleurothecium species (Pl. aquaticum) are introduced based on their distinct morphology and evidence from molecular phylogeny. Pleurotheciella uniseptata, Phaeoisaria clematidis and Pleurothecium pulneyense are also redescribed and phylogenetic relationships assessed herein, these species are the first records for China.     

Massive genetic introgression in threatened northern crested newts (<Emphasis Type="Italic">Triturus cristatus</Emphasis>) by an invasive congener (<Emphasis Type="Italic">T. carnifex</Emphasis>) in Western Switzerland

Genetic pollution through introgressive hybridization of local species by exotic relatives is a major, yet neglected aspect of biological invasions, particularly in amphibians where human introductions are frequent. In Western Switzerland, crested newts make an interesting case: the Italian species Triturus carnifex was introduced at least a century ago within the range of the native and threatened T. cristatus. To understand the genetic consequences of this introduction and inform wildlife management authorities, we conducted a genetic survey on the remaining northern crested newt populations known in the area, using newly-developed species-diagnostic nuclear (microsatellites) and mitochondrial (control region) DNA markers. We documented massive nuclear introgression by the T. carnifex genome, which has completely replaced T. cristatus in most populations, especially in the Geneva area where the introduction was originally reported. However, many of these individuals retained the ancestral T. cristatus mtDNA, which could be explained by asymmetric introgression between the two species, stemming from demographic and/or selective processes. Analyses of genetic diversity support multiple events of T. carnifex releases, most-likely of proximate North Italian origin. We pinpointed the last indigenous populations in the region and recommend to prioritize their protection. Our study demonstrates the invasive potential of introduced taxa through introgressive hybridization, alerts about the underestimated rate of illegal amphibian translocations, and emphasizes the need for genetic analyses to monitor such invasions.     

Introgression from non-native species unveils a hidden threat to the migratory Neotropical fish <Emphasis Type="Italic">Prochilodus hartii</Emphasis>

Invasive species are one of the greatest threats to biodiversity, due to competition, predation, pathogen spread, and hybridization. The latter may remain undetected and impair the survival of species, due to genetic admixture and hybrid swarming (i.e., interbreeding between hybrid individuals and backcrossing with parental species). The impact of invasive species remains poorly studied in the Neotropical ichthyofauna, particularly when considering the potential for hybridization between native and introduced species. Due to fisheries importance and its commercial value, species of the Prochilodus genus have been introduced to other catchments in Brazil. Here, we evaluate the introduction of non-native Prochilodus species and the potential effect of hybridization with the native migratory fish P. hartii. To evaluate possible introgression of Prochilodus spp. to P. hartii in the Jequitinhonha river basin (JRB), we employed a morphogenetic approach, analysing 219 specimens sampled from a broad extent of the river basin. Morphological analyses using meristic characters were incongruent with molecular identification by DNA barcoding (COI) in 22.83% of the analysed specimens. Haplotypes from three non-native species (P. argenteus, P. costatus, and P. lineatus) were recovered from specimens morphologically identified as P. hartii. Hybridization between P. hartii and introduced species was confirmed using co-dominant nuclear microsatellite markers. We observed a pronounced introgression pattern in this Neotropical basin, and paradoxically, despite being one of the most abundant migratory species native to the JRB, due to ongoing levels of introgression, P. hartii’s genetic integrity and conservation might be affected.     

Early otolith development in the critically endangered tooth-carp, <Emphasis Type="Italic">Aphanius farsicus</Emphasis> (Teleostei: Cyprinodontidae)

Otolith morphology in the tooth-carp/killifish genus Aphanius is a source of informative taxonomic characters at both the species and population level. Most work on otoliths has focused on adult specimens, while evidence of ontogenetic variation is rarely provided. In this study we describe the development of otolith morphology during the early life stages of an endangered and endemic species, the Fars tooth-carp Aphanius farsicus from southern Iran. The study material comprises 34 larvae and early juveniles representing nine different developmental stages (0–120 days post hatching), all reared under the same laboratory conditions. The results reveal (i) a significant correlation between standard length and otolith size (length) in larval and early juvenile stages, (ii) clear differences in otolith morphology between larvae/early juveniles and adults, and (iii) a temporal link between the appearance of the sulcus on the otolith’s inner face and the emergence of the dorsal and anal fins. Our results indicate that otoliths of Aphanius can be recognized as originating from larval or early juvenile fish based on their short rostrum and antirostrum lengths and wide excisura, in addition to their small size. These immature otoliths are, however, not diagnostic at the species level in A. farsicus, nor most probably in other species of tooth-carp. The outcome of our study is also of interest to palaeontologists working with fossil killifish otoliths, as it can help avoid misinterpretation of ancient species diversity.     

Records of Bat Flies (Diptera: Nycteribiidae and Streblidae) in the Semi-Arid Caatinga in the State of Paraíba,Northeastern Brazil

One of the largest gaps in the knowledge of ectoparasitic flies of the families Nycteribiidae and Streblidae in Brazil is the northeastern region, where most states do not have any record. Here, we present the first records of those two bat fly families for the state of Paraíba. We recorded a total of 10 species of five genera parasitizing eight bat species of four families. Trichobius diphyllae Wenzel (Streblidae) was the most abundant species, found parasitizing Diphylla ecaudata (Phyllostomidae), and T. dugesioides dugesioides Wenzel, the second, found on Trachops cirrhosus (Phyllostomidae). Three species were recorded for the first time in northeastern Brazil and seven species are new for the semi-arid Caatinga. We collected T. galei Wenzel and T. pallidus (Curran) on Natalus macrourus (Natalidae) and Furipterus horrens (Furipteridae), respectively, two endangered bat species, and the species-specific relationship with their hosts points out to some degree of vulnerability. In addition, we present information on host-parasite relationship, and data that extend the known geographic distribution of some species.     

Spatial distribution shifts in two temperate fish species associated to a newly-introduced tropical seaweed invasion

The establishment of non-native habitat-forming seaweeds into new areas may trigger important changes in ecosystem functioning, yet their community and ecosystem-level effects remain largely understudied. Here we studied the spatial distribution of two common fish species (Xyrichtys novacula and Bothus podas) which are key components of communities in unconsolidated bottoms of temperate areas regarding the colonization of the newly-introduced tropical seaweed Halimeda incrassata in the Mediterranean Sea. We used a spatially-explicit before-after-control-impact model and a unique data-set formed by 6 years of fine-scale spatial information of fish and seaweed distribution and abundance. We demonstrate a long-term alteration on the spatial distribution of X. novacula characterized by a shift towards non-native H. incrassata beds, while no effect on B. podas. The introduction of the tropical seaweed H. incrassata has led to the re-distribution of X. novacula, potentially by harbouring a greater biodiversity of species at the base of the food-web through adding biogenic structure to an otherwise bare sediment. Our work demonstrates that non-native tropical habitat-forming species have the potential to maintain or even enhance fish abundance in unconsolidated bottoms in temperate areas potentially altering the functioning of native habitats.