Parasitism may alter functional response comparisons: a case study on the killer shrimp <Emphasis Type="Italic">Dikerogammarus villosus</Emphasis> and two non-invasive gammarids

The Ponto-Caspian freshwater amphipod Dikerogammarus villosus has colonized most of the water bodies of continental Europe where it causes strong structural alterations in recipient communities that can lead to changes in ecosystem-level processes, mainly because of a strong predatory behaviour. Most of the D. villosus populations from the invaded range have been found infected with the co-introduced microsporidian parasite Cucumispora dikerogammari, known to decrease the predation rate of its host. Infection might thus mitigate the ecological impact of D. villosus and we wanted to test this assumption using the comparative functional response approach. We compared the relationship between resource use and resource availability (i.e. the functional response, FR) of D. villosus, either with infected individuals or not, to that of two non-invasive gammarids: Gammarus pulex and Echinogammarus berilloni. With infected individuals included, D. villosus displayed a higher FR than the two non-invasive gammarids. Although this effect was not significant, C. dikerogammari infection tended to alter the FR of D. villosus with a slight decrease in attack rate and handling time, resulting in a less steep initial slope and a higher asymptote, respectively. Removing infected D. villosus from the dataset did not affect the FR comparison with G. pulex but suppressed the difference in FR with E. berilloni. Although we cannot exclude the role of sample size reduction in this effect, this suggests that C. dikerogammari infection might increase the predation pressure on local prey populations in case of species replacement between D. villosus and E. berilloni. From a more general perspective, our study illustrates how parasites may alter our capacity to predict invasive species impacts from FR comparisons.     

Food resource partitioning of the <Emphasis Type="Italic">Mustelus lunulatus</Emphasis> and <Emphasis Type="Italic">Mustelus henlei</Emphasis> (Elasmobranchii: Carcharhiniformes)

The feeding habits and trophic ecology of Mustelus lunulatus and Mustelus henlei in the central coast of the Colombian Pacific were evaluated and compared to determine whether there was trophic niche overlap or resource partitioning between these two sympatric shark species. A total of 59 prey items were identified and grouped into 10 taxonomic categories. Mustelus lunulatus fed in large proportion on Stomatopoda and Brachyura, whereas M. henlei fed almost exclusively on Teleostei. Dendrobranchiata, Cephalopoda, Anomura and Polychaeta complemented the diets of both species. There were significant differences in diet between the two species, and there was an ontogenetic diet shift in Mustelus lunulatus, with Stomatopoda being the main prey item of juveniles and Brachyura the main prey item of adults. Dietary overlap (by sex and size) was low between species, but it was high for each species, with significant overlap between juveniles and adults of M. lunulatus and M. henlei, and between males and females of M. henlei. There were differences in the trophic levels of the species, sexes and sizes. This study suggests there is food resource partitioning, and differing ecological function of the two Mustelus species in the food web of the study area.     

Substrate preferences of coexisting invasive amphipods, <Emphasis Type="Italic">Dikerogammarus villosus</Emphasis> and <Emphasis Type="Italic">Dikerogammarus haemobaphes</Emphasis>, under field and laboratory conditions

Two Ponto-Caspian amphipods, Dikerogammarus villosus and Dikerogammarus haemobaphes, have expanded their geographical ranges from eastern Europe into Great Britain in recent years. This study represents one of the first examining the distribution and habitat preferences of coexisting populations of D. haemobaphes and D. villosus via field and laboratory experiments in the UK. Field surveys of a recently invaded lowland reservoir in the UK are complimented with ex situ laboratory mesocosm experiments examining the substrate preferences of coexisting populations of D. villosus and D. haemobaphes. Results from the field study indicated that D. haemobaphes dominated the macroinvertebrate community within the reservoir and demonstrated a strong affinity for large cobble and artificial substrates. D. villosus occurred at lower abundances but displayed a strong preference for coarse cobble substrates. A third invasive amphipod, Crangonyx pseudogracilis, was largely confined to sand/silt habitats. Laboratory mesocosm experiments clearly supported the field observations of D. villosus and D. haemobaphes with both species demonstrating a preference for cobble substrates. Results from the study highlight the importance of characterising physical habitat when investigating biological invasions and suggest that habitat availability may influence the extent and speed at which range expansion of new amphipod invaders occurs.     

Comparison of trophic function between the globally invasive crayfishes <Emphasis Type="Italic">Pacifastacus leniusculus</Emphasis> and <Emphasis Type="Italic">Procambarus clarkii</Emphasis>

Impacts of invasive species may manifest most strongly if these organisms are highly distinct functionally from the native species they often replace. Yet, should we expect functional differences between native and invasive species of generalist organisms like freshwater crayfish? Some existing evidence has pointed to native and invasive crayfish species as ecologically equivalent. We contribute to this literature by comparing the trophic niches of the globally invasive crayfishes Pacifastacus leniusculus and Procambarus clarkii, by applying carbon and nitrogen stable isotope analyses to replicated allopatric (alone) and sympatric (together) lake populations in western Washington State, USA, where P. clarkii has been recently introduced and P. leniusculus is presumed native. Our study corrected for potential inherent differences in lake food webs as a consequence of lake abiotic or biotic characteristics using random effects in linear mixed effects models. We found that although overall trophic niche size or area of these species was not significantly different, P. leniusculus was significantly higher in trophic position than P. clarkii when also accounting for the effects of body size, sex, and lakes as random effects. This pattern of increased trophic position of P. leniusculus over P. clarkii was conserved over time in one sympatric lake for which we had data over multiple years. Cumulatively, our findings point to trophic differences between the globally cosmopolitan crayfishes P. leniusculus and P. clarkii, particularly when accounting for the ways that ecosystem context can affect food web structure of communities and the trophic resources available to these consumers.     

A functional response evaluation of pre-infestation with <Emphasis Type="Italic">Bemisia tabaci</Emphasis> cryptic species MEAM1 on predation by <Emphasis Type="Italic">Propylea japonica</Emphasis> of <Emphasis Type="Italic">Myzus persicae</Emphasis> on host plant tomatoes

Herbivore feeding on host plants may induce defense responses of the plant which influence other herbivores and interacting species in the vicinity, such as natural enemies. The present work evaluated the impact of pre-infestation with the tobacco whitefly Bemisia tabaci cryptic species MEAM 1, on the predation ability of the ladybird Propylea japonica, to the green peach aphid Myzus persicae, on tomato plants. The results show that B. tabaci pre-infestation density, duration, and leaf position, can impact prey consumed by P. japonica under various aphid densities. The aphids consumed by P. japonica in each treatment were fit using the Holling type II functional response equation. The predatory efficiency (a/T _h) of P. japonica was the highest in the treatment with 60 aphids and 48-h infestation directly on damaged leaves. The predatory efficiencies of P. japonica decreased with a reduction of pre-infestation density and duration. We also observed that pre-infestation on young and undamaged leaves increased predation by P. japonica.     

Life table demography of <Emphasis Type="Italic">Asplanchna brightwellii</Emphasis> Gosse, 1850 fed with five different prey items

We conducted life table experiments on the freshwater rotifer Asplanchna brightwellii to analyze its demography when fed with prey items from several taxonomic groups (cladocerans, protozoans, and rotifers) and under two different temperature regimes (20 and 25°C); the aim of the study was to determine the preferred prey for A. brightwellii in terms of fitness (evaluated as reproductive success) among five cladoceran, protozoan, and rotifer preys, and to test which temperature (20 or 25°C) is better for life table parameters of Asplanchna. Our analysis identified Brachionus calyciflorus as the preferred prey for A. brightwellii based on life table statistics, ingestion rate and electivity indices. The greatest values for net reproductive rate and intrinsic growth rate were achieved when A. brightwellii was fed B. calyciflorus. Greater reproductive values (R _o and r) were found at 25°C than at 20°C for A. brightwellii across the five prey species. We found significant differences in the ingestion rate and electivity index among zooplanktonic and benthic preys. The influence of temperature, the cost of predation, and how prey selection by A. brightwellii is influenced by: biomass, size, and swimming speed; they are discussed hoping to gain a better understanding of trophic transfers in zooplankton communities.     

Reproductive tables of predatory phytoseiid mites (<Emphasis Type="Italic">Phytoseiulus persimilis,Galendromus occidentalis</Emphasis>, and <Emphasis Type="Italic">Neoseiulus cucumeris</Emphasis>)

Feeding of predatory mites (Phytoseiulus persimilis, Galendromus occidentalis, and Neoseiulus cucumeris) on different life stages of Tetranychus atlanticus under optimal conditions was studied. Daily and total consumption of prey by predators and selection of prey of different life stages were studied for 5 days and for the entire feeding period. Average daily food consumption [number of individuals] for the entire life period of mature mite females constituted 0.43 females + 5.0 [nymphs and males] + 3.4 eggs of Tetranychus atlanticus in P. persimilis; 0.12 females + 3.70 [nymphs and males] + 3.10 eggs in G. occidentalis; and 0.19 females + 4.10 [nymphs and males] + 3.50 eggs in N. cucumeris. During the entire period of feeding, P. persimilis preferred large individuals and at the postembryonic stages selected prey to a greater extent than G. occidentalis and N. cucumeris (61.8 and 55.1%, respectively). The use of a 5-day express-method is possible for estimation of some biological characteristics of phytoseiids that previously consumed the same food for a long period. In other cases, analysis of characteristics for the entire life period is necessary.     

Molecular evidence for hybridization between invasive <Emphasis Type="Italic">Solidago canadensis</Emphasis> and native <Emphasis Type="Italic">S</Emphasis>. <Emphasis Type="Italic">virgaurea</Emphasis>

Hybridization between alien and native species is biologically very important and could lead to genetic erosion of native taxa. Solidago × niederederi was discovered over a century ago in Austria and described by Khek as a natural hybrid between the alien (nowadays regarded also as invasive) S. canadensis and native S. virgaurea. Although interspecific hybridization in the genus Solidago is considered to be relatively common, hybrid nature of S. × niederederi has not been independently proven using molecular tools, to date. Because proper identification of the parentage for the hybrid Solidago individuals solely based on morphological features can be misleading, in this paper we report an additive polymorphism pattern expressed in the ITS sequences obtained from individuals representing S. × niederederi, and confirm the previous hypothesis that the parental species of this hybrid are S. canadensis and S. virgaurea. Additionally, based on variability at the cpDNA rpl32-trnL locus, we showed that in natural populations hybridization occurs in both directions.     

Intraguild predation on the aphid parasitoid <Emphasis Type="Italic">Aphelinus asychis</Emphasis> by the ladybird <Emphasis Type="Italic">Harmonia axyridis</Emphasis>

We investigated intraguild predation (IGP) on an aphid parasitoid, Aphelinus asychis Walker (Hymenoptera: Aphelinidae), by the multicolored Asian ladybird, Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae), and used the green peach aphid, Myzus persicae (Sulzer) (Hemiptera: Aphididae) as the prey/host in the laboratory. The ladybirds reared on artificial diet and on aphids consumed more aphids than mummies, while those reared on parasitized aphids consumed similar numbers of aphids and mummies. The ladybirds chose more mummies in treatments when mummies were more abundant, and more aphids when numbers of aphids and mummies were equal, or when aphids were more abundant. However, at all density treatments, rejection rates of mummies (36%) were much greater than of aphids (2%). H. axyridis prey on more aphids than A. asychis mummies, which enhances biological control by the two species. However, prior feeding experience affected subsequent choice, increasing the competition between natural enemies which would reduce their combined effectiveness for biological control.     

The very hungry amphipod: the invasive <Emphasis Type="Italic">Dikerogammarus villosus</Emphasis> shows high consumption rates for two food sources and independent of predator cues

The invasion of the Ponto–Caspian amphipod Dikerogammarus villosus in European rivers is assumed to reduce macroinvertebrate diversity and to alter ecosystem functions. D. villosus shows an extraordinarily flexible feeding behavior including the ability to use various food sources. On the other hand, its response to predation risk seems to depend on environmental factors. To evaluate the ecological function of D. villosus, we estimated the daily food consumption for different food sources and analyzed potential effects of predator avoidance behavior on feeding. D. villosus consumption of willow leaves or chironomid larvae was quantified in 24-h laboratory experiments with and without kairomones of the European bullhead (Cottus gobio). Consumption rates were estimated based on gut content and gut evacuation rate under semi-natural laboratory conditions enabling the animals to feed over the whole time of the evacuation rate experiment. We observed very high evacuation rates and consequently high consumption rates up to 89% of body weight per day. Consumption rates differed significantly between food sources: D. villosus ingested more leaves than chironomid larvae. In contrast, predator cues did not affect the feeding of D. villosus. This might be explained by its strong refuge affinity and probably benefits its successful invasion. A comparison of the estimated consumption rates with results of an own consumption experiment (and other studies) under more artificial conditions indicated that more natural conditions result in higher consumption rates. Consequently, feeding rates from highly artificial experiments should be used with great caution to assess the ecosystem function of D. villosus.     

Feeding patterns of the black-striped pipefish <Emphasis Type="Italic">Syngnathus abaster</Emphasis> in an invaded freshwater habitat

Syngnathus abaster is an euryhaline species, which has penetrated freshwaters in Europe and its range is expanding. The goal was to investigate the feeding patterns of this species in an invaded freshwater habitat, where it first appeared at the beginning of 1990s, with implications for assessing its possible effects on invaded ecosystems. In total, 36 prey items were identified in the diet of S. abaster. The most important prey were zooplanktonic organisms, among which copepods predominated significantly. The second most important prey were cladocerans, represented by 20 species/genera. The diet composition of this species showed clear seasonal dynamics but copepods remained the predominant prey item throughout the year. Significant positive relationships between the abundance of zooplanktonic organisms in the gut contents of S. abaster and their abundance in zooplankton samples were found for copepods and Ceriodaphnia sp. S. abaster showed a significant positive electivity for copepods throughout all months. It also positively selected some large cladocerans, which probably shared the same habitat. S. abaster also consumed fish items including gobiid larvae and its own juveniles, when they were abundant in May and especially in June. The possible effect of S. abaster on native ecosystems may include the shaping of zooplanktonic communities through selective predation on copepods. This species is unlikely to be a serious food competitor for native fish inhabiting European freshwaters, the majority of which are cyprinids. Nevertheless, it can have effects on some native fish through predation on their juveniles, which might share the same habitats.     

Black bear (<Emphasis Type="Italic">Ursus americanus</Emphasis>) and wolf (<Emphasis Type="Italic">Canis</Emphasis> spp.) summer diet composition and ungulate prey selectivity in Ontario,Canada

Understanding predator-prey dynamics is an important component of management strategy development for wildlife populations that are directly affected by predation. Ungulates often serve as a significant source of prey for many large mammal predators, and patterns of predation are known to influence population dynamics. Although black bear and wolf diets have been investigated extensively, prey preference has been less commonly examined, especially in analyses that take into account age class (i.e., juvenile and adult) of the ungulate prey. We examined black bear (Ursus americanus), wolf (Canis lupus), and hybrids (Canis spp.) prey preference in Ontario based on the availability of three ungulate species—elk (Cervus elaphus), moose (Alces alces), and white-tailed deer (Odocoileus virginianus). We analyzed the presence of prey items in black bear and wolf scats collected over 3 years by examining prey hair cuticular scale patterns. We applied correction factors to frequencies of occurrence of prey items found in predator scat and related diet composition to the availability of ungulates, determined by fecal pellet transect surveys. In addition, non-ungulate diet items were identified to obtain full diet composition profiles. We found that black bear diet consisted of more than 87% vegetation, and they were opportunistic, not selecting for any particular ungulate species in either adult or juvenile age class. Wolf diet was comprised mainly of ungulates (~?73.2%), muskrat (Ondatra zibethica; ~?8.5%), and beaver (Castor canadensis; ~?14.6%), and although moose were at least 1.5 times more abundant then each of the other ungulate prey species in the study area, wolves preferred elk, using moose less than expected. Although we found black bear diet to be opportunistic during the summer, wolves in our study heavily utilized both juvenile and adult ungulates, however, among ungulate species, displayed preference for elk. The preference displayed by wolves provides insight that wildlife managers can use to guide further investigation and assist with the development of strategies to ensure continued elk reintroduction success, and moose and white-tailed deer population sustainability.     

Screening and molecular identification of lactic acid bacteria from <Emphasis Type="Italic">gari</Emphasis> and <Emphasis Type="Italic">fufu</Emphasis> and <Emphasis Type="Italic">gari</Emphasis> effluents

Bacterial strains were isolated from cassava-derived food products and, for the first time, from cassava by-products, with a focus on gari, a flour-like product, and the effluents from the production processes for gari and fufu (a dough also made from cassava flour). A total of 47 strains were isolated, all of which were tested to determine their resistance to acidic pH and to bile salt environments. Four of the 47 isolates tested positive in both environments, and these four isolates also showed antibacterial behaviour towards both Gram-positive and Gram-negative microbial pathogens (i.e. Methicillin-resistance Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus, Salmonella enteritidis, Escherichia coli, Escherichia coli (O157), Yersinia enterocolitica). In most cases, the antibacterial activity was related to bacteriocin production. Molecular identification analysis (16S rDNA and randomly amplified polymorphic DNA-PCR) revealed that the four isolates were different strains of the same species, Lactobacillus fermentum. These results demonstrate that bacteria isolated from cassava-derived food items and cassava by-products have interesting properties and could potentially be used as probiotics.     

Revision of the <Emphasis Type="Italic">Serrulati</Emphasis> species of <Emphasis Type="Italic">Penstemon</Emphasis> section <Emphasis Type="Italic">Saccanthera</Emphasis> subsection <Emphasis Type="Italic">Serrulati</Emphasis>

A revision of Penstemon sect. Saccanthera subsect. Serrulati includes a new species (P. salmonensis), a new variety (P. triphyllus var. infernalis), and the elevation of a subspecies to species (P. curtiflorus), bringing the total number of species to eight, which are keyed and described, complete with nomenclature and type citations.     

Taxonomy and geographic distribution of <Emphasis Type="Italic">Carex lucorum</Emphasis> var. <Emphasis Type="Italic">austrolucorum</Emphasis> (section <Emphasis Type="Italic">Acrocystis</Emphasis>, Cyperaceae)

Carex lucorum currently comprises two morphologically similar varieties. We revise the taxonomy of C. lucorum, and map the distribution of the taxa. Based on multivariate analyses and analyses of variance of measured characters, allopatry, and previous studies revealing differences in achene micromorphology, chromosome differences, and flavonoid chemistry, we elevate C. lucorum var. austrolucorum to species status as C. austrolucorum. We provide an identification key to C. lucorum and its closest relatives.     

Continued feeding on <Emphasis Type="Italic">Diporeia</Emphasis> by deepwater sculpin in Lake Huron

Monitoring changes in diets of fish is essential to understanding how food web dynamics respond to changes in native prey abundances. In the Great Lakes, Diporeia, a benthic macroinvertebrate and primary food of native benthivores, declined following the introduction of invasive Dreissena mussels and these changes were reflected in fish diets. We examined the diets of deepwater sculpin Myoxocephalus thompsonii collected in bottom trawls during 2010–2014 in the main basin of Lake Huron, and compared these results to an earlier diet study (2003–2005) to assess if their diets have continued to change after a prolonged period of Dreissena mussel invasion and declined Diporeia densities. Diporeia, Mysis, Bythotrephes, and Chironomidae were consumed regularly and other diet items included ostracods, copepods, sphaerid clams, and fish eggs. The prey-specific index of relative importance calculated for each prey group indicated that Mysis importance increased at shallow (≤55 m) and mid (64–73 m) depths, while Diporeia importance increased offshore (≥82 m). The average number of Diporeia consumed per fish increased by 10.0% and Mysis decreased by 7.5%, while the frequency of occurrence of Diporeia and Mysis remained comparable between time periods. The weight of adult deepwater sculpin (80 mm and 100 mm TL bins) increased between time periods; however, the change in weight was only significant for the 80 mm TL group (p?<?0.01). Given the historical importance of Diporeia in the Great Lakes, the examination of deepwater sculpin diets provides unique insight into the trophic dynamics of the benthic community in Lake Huron.     

Assemblage-level effects of the introduced peacock hind (<Emphasis Type="Italic">Cephalopholis argus</Emphasis>) on Hawaiian reef fishes

The peacock hind Cephalopholis argus (family Serranidae), locally known as ‘roi’, was introduced from French Polynesia to Hawaii in the mid-twentieth century as a food fish. However, because of its association with ciguatera fish poisoning, it is rarely fished for food. Previous research indicates that roi could have a negative impact on native reef fish assemblages because of their high densities and prey consumption rates. However, it is unclear whether roi add to the cumulative mortality of prey (predation hypothesis), or whether predation is instead compensatory (doomed surplus hypothesis). This study experimentally assessed the effects of roi on reef fish populations through a long-term (5.5 year) predator removal experiment. A Before-After-Control-Impact study design was used to assess changes in fish assemblages following the removal of roi on 1.3 ha of patch reef. Increases in the density of prey-sized fish (<15 cm TL) were observed 18 months after roi removal. However, those effects did not translate into sustained increases in prey. While increases in potential competitors, wrasses (family Labridae), particularly the piscivorous ringtail wrasse Oxycheilinus unifasciatus, were observed on roi-free reefs, the fish assemblage did not diverge substantially in composition. Native reef fish appeared to resist the potential negative impacts of predation by roi, possibly through a refuge in size for some fish families. Management to protect intact fish assemblage size-structure could serve to bolster native resistance to invading species. In considering the threats facing coral reefs, and the possible solutions, roi removal alone will not likely replenish native fishery resources.     

Interactions between the grass shrimp <Emphasis Type="Italic">Palaemonetes pugio</Emphasis> and the salt marsh grasses <Emphasis Type="Italic">Phragmites australis</Emphasis> and <Emphasis Type="Italic">Spartina alterniflora</Emphasis>

The grass shrimp Palaemonetes pugio, a species common to Spartina alterniflora-dominated marshes, may be sensitive to the invasion of the common reed Phragmites australis in northeastern US salt marshes. We examined two questions: (1) Do grass shrimp have a preference for the native plant over the non-native plant? (2) Are grass shrimp more effective foragers on P. australis? We tested the first hypothesis by comparing the amount of time shrimp spend in physical contact with the plant types over a 1-h period. Shrimp were observed under different arrangements of vegetation to control for differences in conspicuous structural features. Additionally, the amount of time shrimp spent foraging on S. alterniflora and P. australis shoots was compared to determine if shrimp graze more often on S. alterniflora. Shrimp spent significantly more time in contact with S. alterniflora only when plant types were grouped at opposite ends of aquaria, but did not exhibit a foraging preference for this plant type. To address our second question, we investigated the effects of shrimp foraging on stem epifauna, an assemblage of semi-aquatic invertebrates associated with macrophyte shoots. Previous research showed that P. australis supports a lower density of stem-dwelling epifauna relative to S. alterniflora. We hypothesized that the primary grazer of this community, P. pugio, can forage on P. australis stems more effectively due to structural differences between the two plants, causing the lower abundance of epifauna through top-down effects. We exposed individual shoots inhabited by epifauna to shrimp and compared faunal densities on exposed shoots to densities on control shoots after 18 h. The reduction of epifauna by predation was proportional on the two plant types. Therefore, top-down effects can be ruled out as an explanation for the patchy distribution of epifauna observed in P. australis–S. alterniflora marshes.     

<Emphasis Type="Italic">Cantharellus</Emphasis> sect. <Emphasis Type="Italic">Amethystini</Emphasis> in Asia

In this contribution on the genus Cantharellus in Asia, C. subvaginatus is described from the Republic of Korea as a close relative to the Chinese C. vaginatus, which is here reported for the first time from India. Both species are here placed in Cantharellus subg. Cantharellus sect. Amethystini, together with the Indian C. pseudoformosus (syn.: C. umbonatus) and the Malayan C. subamethysteus. As such, Asia has suddenly become the continent with the highest diversity for Amethystini. Species delimitation in sect. Amethystini is molecularly supported by a combined phylogenetic analysis of rDNA sequences obtained for LSU and ITS and additionally suggests the existence of a still undescribed species in North America. Character variability is discussed for all known members of Amethystini, including atypical specimens of the North American C. lewisii that are morphologically more reminiscent of the South Korean C. subvaginatus.     

Co-invasion of three Asian earthworms, <Emphasis Type="Italic">Metaphire hilgendorfi</Emphasis>, <Emphasis Type="Italic">Amynthas agrestis</Emphasis> and <Emphasis Type="Italic">Amynthas tokioensis</Emphasis> in the USA

Earthworm invasions are one of the most serious causes of ecological deterioration in the temperate deciduous forests of North America. Non-native earthworms impact understory vegetation, leaf litter layer, carbon dynamics, nutrient availability, and the associated food webs. Here we report a significant status change and confirm expansion of known range of Amynthas agrestis, one of the most serious invasive species in North America, and two of its close relatives, A. tokioensis and Metaphire hilgendorfi. The three species have never been confirmed to co-occur in North American ecosystems. We examined 1760 earthworms collected from 30 sites across northeastern USA, and identified them using a new morphological key. Our data show that sympatric occurrence of at least two, and often all three, species is more common than having only one species. In addition, A. tokioensis was dominant in many of these earthworm communities. The status change in species composition from only one species to two or three and the shift in dominance are most likely caused by previous incorrect species identification. Our results support expansion of known range of A. tokioensis and M. hilgendorfi northward and westward into states with colder winters. This range expansion may have taken place alongside that of A. agrestis in the last 10–20 years, but has long been overlooked. Altogether, results highlight an urgent need for correct species identification. The recognition of an expanding multi-species system represents a unique opportunity to further evaluate complex interactions among co-invading and resident species, and to investigate whether interspecific interactions have unexpected non-additive impacts on ecological processes.