Home-range fidelity and use of historic habitat by adult Colorado pikeminnow ( Ptychocheilus lucius ) in the White River, Colorado and Utah

Twelve wild adult Colorado pikeminnow ( Ptychocheilus lucius ), captured in the tailwaters of Taylor Draw Dam on the White River, Colorado, were implanted with radio transmitters and their movement patterns monitored from 1992 to 1994. The spawning migration of these fish was extensive. In 1993, the only full year of the study, the fish migrated an average of 658 km from the White River to spawning sites in the Yampa or Green rivers and back to the White River. Eight of these fish were translocated in the river upstream of the dam in April 1993. These fish and the 4 others below the dam remained in the river until May 1993. All 12 had migrated down the White River to spawning sites in the Green and Yampa rivers by July 1993. The fish that were located above the dam successfully passed over the dam during their downstream migration. Seven fish migrated upstream toward the Yampa River Canyon spawning site and 5 migrated downstream toward the Green River Desolation/Gray Canyon spawning site. Five of 7 Yampa River fish were found at the spawning site. The other 2 were found 5-8 km downstream of the site. One of 5 Green River fish was found at the spawning site, the other 4 between 16 and 62 km upstream of the site. All fish migrated back to the White River by August 1993 and were found near the dam by October 1993. Two fish were recaptured and translocated above the dam in September 1993. Five fish were located below the dam and 2 above the dam in April 1994. By July 1994 seven of the same fish that had migrated toward the Yampa River in 1993 were found at the Yampa Canyon spawning site. At the same time, 3 of 5 fish that migrated toward the Green River in 1993 were found at the Desolation/Gray Canyon spawning site. This included 2 fish that had been found upstream of the site in 1993. The 12 fish traveled an average of 6 km d -1 (range: 4-10 km d -1 ) during the migration period from May through October 1993. Generally, fish moved faster to the spawning site than back from the site to the White River. These fish moved very little within their home ranges in the White River. Six fish tagged in 1992 moved only 0.1-2.3 km in the tailwater reach below Taylor Draw Dam from September 1992 through April 1993. All fish, after their spawning runs, had moved up to or near the dam by October 1993. These fish were not tracked again until April 1994. Their movement patterns in April 1994 were similar to those observed in April 1993. The greatest amount of fish movement in the White River was displayed by the 8 fish placed above Taylor Draw Dam in April 1993 and the 2 placed in Kenney Reservoir in September 1993. They moved 1.1-40.6 km in the river before and after their spawning migration in spring and autumn 1993. These spawning migrations suggest that adult Colorado pikeminnow in the White River were recruited from both Green and Yampa river spawning populations and were presumably imprinted to these respective spawning sites. Those fish placed above Taylor Draw Dam established home ranges in habitats previously occupied by Colorado pikeminnow before the dam was completed. They remained there until they migrated downstream during the spawning period. Although we did not study fish passage, our study demonstrates that adult Colorado pikeminnow will use habitat if access is provided. Translocation of wild adult fish into historic but unoccupied habitats may be a valuable recovery option.     

Ecological investigation of a suspected spawning site of Colorado squawfish on the Yampa River, Utah

On 5 July 1981, 13 adult Colorado squawfish were found in spawning condition at river mile 16.5 in the Yampa River, a major tributary to the Green River. An investigation was undertaken to quantitatively describe this section of the river to gain insights on the spawning requirements of this endangered species. The substrate at the suspected spawning site was cobble with large interstitial spaces devoid of organics, silts, or clays. It appeared that larvae of several fish species utilized these cobble areas and the associated voids. Diurnal sampling indicated that larval drift occurred between 0100 and 0125 hours. Substrate size also appeared to be a dominant factor in fish distribution. Feeding intensities of these fish corresponded to macroinvertebrate drift.         

Esox lucius (Esocidae) and Stizostedion vitreum (Percidae) in the Green River basin, Colorado and Utah

Northern pike, Esox lucius , stocked in the Yampa River in 1977, invaded the mainstream Green River by 1981 and subsequently increased in range and abundance. The speed of this invasion is indicated by two recaptured pike that moved 78 and 110 km, respectively, downstream in about one year. Pike stomachs (n = 123) were usually empty (54.5%), but some contained fish (43%) and nonfish items (2.4%). Red shiner, Notropis lutrensis , and fathead minnow, Pimephales promelas , predominated among the 12 fish species eaten. Walleye, Stizostedion vitreum , presumably introduced to the Green River drainage in the 1960s, was widely distributed but low in abundance. Most of 61 adult walleye stomachs contained food (60.7%) ; of 6 fish species eaten, channel catfish, lctalurus punctatus , and fathead minnow were most frequently consumed. Northern pike and walleye were captured in habitats occupied by endangered Colorado River fishes, particularly Colorado squawfish, Ptychocheilus lucius . Predation on endangered fishes was not detected, but northern pike and walleye consumed at least three other native fishes. The northern pike may pose a threat to endangered fishes due to its population expansion, piscivory, and resource sharing. Diets of northern pike and walleye species should be further evaluated if their abundance increases.     

Aspects of Sacramento pikeminnow biology in nearshore habitats of the Sacramento-San Joaquin Delta, California

We documented distribution, relative abundance, diet composition, and body condition of Sacramento pikeminnow Ptychocheilus grandis during 2001 and 2003 at 5 sites in the Sacramento-San Joaquin Delta, California. Sacramento pikeminnow densities in nearshore habitats were higher in 2003 than 2001. In both years, spatial distribution of beach seine densities was similar. There were no significant differences in density among sampling sites except for the southernmost site where the catch was near zero. Based on rotary screw-trap data from a 6th site, we found relative abundance of Sacramento pikeminnow entering the Delta via an artificial floodplain was positively correlated with flow. Most individuals collected using all 3 gear types were age 1 or older, and appeared to grow quickly based on data from previous studies. Sacramento pikeminnow had diverse diets composed of freshwater and estuarine invertebrate and fish taxa. Incidence of piscivory was only 2% of the diet of individuals P. oregonensis , but in seeming contrast to endangered Colorado pikeminnow P. lucius , the present study showed that Sacramento pikeminnow can be successful in altered habitats. Sacramento pikeminnow; Ptychocheilus grandis; Sacramento-San Joaquin Delta; habitat use; diet composition; condition factor; Cyprinidae.     

Migrating Mormon Crickets, Anabrus simplex (Orthoptera: Tettigoniidae), as food for stream fishes

Migrating bands Mormon crickets ( Ananbrus simplex ) were observed crossing the Green and Yampa rivers in Dinosaur National Monument, Colorado and Utah, in 1986 and 1987. Swimming crickets were swept downstream and eaten by four- endemic and seven introduced fish species. Included were two endangered fishes, Colorado squawfish ( Ptychocheilus lucius ) and humpback chub ( Gila cypha ). Direct and indirect effects to aquatic food webs associated with application of pesticides for Mormon cricket control may pose a threat to these fishes and to man.     

Archaeological record of native fishes of the lower Colorado River: how to identify their remains

Archaeological sites in the Salton Basin of southeastern California and along the lower Colorado River provided opportunities to determine which fish species were present prior to extirpations, environmental degradation, and the recession of Lake Cahuilla. These remains also represent the fishes exploited by Native Americans. Bonytail ( Gila elegans ), razorback sucker ( Xyrauchen texanus ), Colorado pikeminnow ( Ptychocheilus lucius ), striped mullet ( Mugil cephalus ), and machete ( Elops affinis ) have been recovered from 117 sites in the Salton Basin, once filled by the Colorado River forming Lake Cahuilla. Bonytail and razorback sucker comprise nearly 99% of the remains. Along the lower Colorado River itself, fragmentary elements of bonytail, razorback sucker, Colorado pikeminnow, and roundtail chub ( G. robusta ) have been recovered, documenting a disappearing native fish fauna. Anatomical details are described that permit identification of diagnostic materials commonly recovered during archaeological excavations.     

Diets of young Colorado squawfish and other small fish in backwaters of the Green River, Colorado and Utah

We compared diet of young-of-year Colorado squawfish ( Ptychocheilus lucius ), an endangered cyprinid, with diets of other fish Rhinichthys osculus, Catostomus discobolus, and C. latipinnts , and nonnative Cyprinella lutrensis, Notropis stramineus, Pimephales promelas, Ictalurus punctatus, and Lepomis cyanellus. For each species, diet varied with size and between upper and lower river reaches but not between seasons for fish of similar size. Larval chironomids and ccratopogonids were principal foods of most fishes. Copepods and cladocerans were important in diets of P. lucius L. cyanellus Catostomus discobolus was the only species that ate moderate amounts of algae. Fish (all larvae) were in digestive tracts of only 10 P. lucius (21-73 mm TL), about 1% of P. lucius analyzed. High diet overlap occurred between some size-reach groups of P. lucius and C. lutrensis, R. osculus, C. latipinnis, I. punctatus, and L. cyanellus . Potential for food competition between young-of-year P. lucius and other fishes in backwaters appeared greatest with the very abundant C. lutrensis .     

Reproductive ecology of a Tibetan frog Nanorana parkeri (Anura: Ranidae)

Endemic to the Tibetan Plateau, Nanorana parkeri figures among the highest altitude amphibians in the world. The present work, which was conducted in an alpine marsh at 4300 m altitude, is the first report of the species’ reproductive ecology. The breeding season, as indicated by the presence of amplectant pairs and fresh eggs in ponds, lasted from early Mary to early July. The ratios of adult male to female were 1.3 in hibernation ponds and 2.3 in spawning ones. Males were smaller than females. Amplexus was typically axillary (81.7%), with a few exceptional amplexus in which a male clasped another male or object rather than gravid females. Scramble competition occurred in 6.8% of the normal amplexus located. The body sizes of amplexed couples correlated positively. Deposited eggs were attached to submerged plants solitarily (60.5%) or in small clusters (39.5%), and the scattered arrangement of individual eggs could be a strategy to ensure embryonic development in oxygen-poor environments. Clutch size averaged 189 [standard deviation (SD) = 64], and correlated positively with female body size; egg size averaged 2.18 mm (SD = 0.10), independent of female body size; both traits within a clutch were negatively related with each other even controlling for female body size. Relatively few and large eggs laid by the Tibetan frogs, compared with those by lowland aquatic-spawning anuran species, should be adaptive to the harsh high-altitude conditions. Larvae hatched from eggs after 16.4 days on average (SD = 5.8). Froglets emerged between late July and early August, but some tadpoles were present in winter. These findings may help to understand the evolution of amphibians in extreme environments and to protect these remote species.     

Large mixed-species dispersal flights for predatory and scavenging aquatic Heteroptera and Coleoptera, northern Arizona, USA

We report 4 incidents of large (hundreds to hundreds of thousands of individuals) mixed-species flights of predatory and scavenging aquatic Heteroptera and Coleoptera. The events occurred on normal (mostly clear, calm) autumn 2005 and spring 2006 late afternoons near Flagstaff, Arizona. Flight days were either near the full moon or in advanced waning lunar periods. At least 18 species were involved in the flights, with as many as 16 species (7 species of Heteroptera in 3 families, 9 species of Coleoptera in 2 families) in a single flight. Heteroptera (especially from the family Corixidae) were 2–4 orders of magnitude more numerous than Coleoptera. The diasporas plummeted, with much direct mortality, onto green metal roofs, and the largest event lasted more than 2 hours. Even after 2 days, uninjured individuals failed to resume their flights. The literature suggests that such flights occur for autumn movement to winter habitats or for location of springtime habitats for reproduction. The relative proportions of species in flocks were similar neither to the relative proportions in the region nor to those in nearby livestock watering tanks, indicating that the flights consisted of nonrandom assemblages of species. These events are rare or unique observations of coordinated movement of mixed predatory and scavenging invertebrate species.     

Chironomidae (Diptera) species distribution related to environmental characteristics of the metal-polluted Arkasas River, Colorado

Mining in the Upper Arkansas catchment has polluted the river with heavy metals for 140 yr. Pupal and adult chironomid species distribution and sedimentary metal concentrations are provided for 22 stations along 259 km of main river during 1984-85. Complete species identification was achieved only recently. This has produced an unprecedented record of chironomid species distribution for a comparable length of river in the USA. Chemically or physically perturbed sites had poor species richness compared with the next site downstream, suggesting that larvae may drift through unfavorable habitats to benign ones. Using canonical correspondence analysis, we found species composition to be most strongly related to variables expressing the longitudinal axis of the river (distance/altitude, temperature, latitude), while toxicity to zinc was a significant secondary correlate. These river-related environmental variables accounted for a greater proportion of pupal species variation than for adults. This was considered to result from a proportion of adults emerging from habitats beyond the main river. Multivariate analysis identified metal-tolerant and -intolerant species. Generic data revealed the same major trends but indicator taxa were lost. The study provides a disturbed-state reference for monitoring effects of remedial actions begun in 1991, and for comparisons with other Colorado rivers.     

Exclusion experiments with backwater invertebrate communities of the Green River, Utah

The role of biotic interactions in structuring freshwater invertebrate communities has been extensively studied but with mixed results. For example, fish effects on invertebrates are most pronounced in pelagic and soft-sediment benthic habitats that lack structural complexity, yet appear insignificant in benthic rubble habitats. Backwaters of the Green River, Utah, are shallow, structurally simple, quiet-water embayments adjacent to the river. These habitats form in middle to late summer and are colonized by benthic and epibenthic invertebrates that produce standing crops significantly higher than the river. Backwaters are also utilized by a large number of fish species. We used cages to determine if selective exclusion of backwater organisms could significantly change invertebrate community structure. Results showed that backwater invertebrate community components changed significantly in response to exclusion treatments. Two taxa (both predators), the chironomid genus Tanypus (Diptera: Chironomidae) and the corixid genus Trichocorixa (Hemiptera: Corixidae), increased in density in exclusion cages while cladocerans, immature copepods, the cyclopoid copepod Eucyclops speratus , and the chironomid genus Procladius all decreased in density. Diversity of adult copepods was reduced by exclusion treatments, though density of only a single species changed significantly.     

Species composition, emergence, and habitat preferences of Trichoptera of the Sagehen Creek Basin, California, USA

An extensive study of larvae and adult Trichoptera of the Sagehen Creek basin. Sierra Nevada, California, USA, revealed 77 species representing 14 families and 41 genera. Twenty-six species were restricted to small water bodies (spring sources, seeps, spring streams, temporary ponds, and intermittent streams); 27 were restricted to Sagehen Creek, a second-order stream, and the mouths of two large spring streams. Similarity between species caught at black lights and those in emergence traps was 43%. There were two major peaks in adult emergence by species, midsummer and late summer-fall. A few species emerged during winter or throughout most of the year. The species composition of the community showed affinity with Oregon, the Great Basin, and the Rocky Mountains but very little similarity with the California Coast Range. Four of the most abundant species in the basin have very restricted distributions. Ecological separation of several groups of closely related species could be explained by major differences in larval habitats or by different emergence periods.     

New approaches for sampling and modeling native and exotic plant species richness

We demonstrate new multi-phase, multi-scale approaches for sampling and modeling native and exotic plant species to predict the spread of invasive species and aid in control efforts. Our test site is a 54,000-ha portion of Rocky Mountain National Park, Colorado, USA. This work is based on previous research wherein we developed vegetation sampling techniques to identify hot spots of diversity, important rare habitats, and locations of invasive plant species. Here we demonstrate statistical modeling tools to rapidly assess current patterns of native and exotic plant species to determine which habitats are most vulnerable to invasion by exotic species. We use stepwise multiple regression and modified residual kriging to estimate numbers of native species and exotic species, as well as probability of observing an exotic species in 30 × 30-m cells. Final models accounted for 62% of the variability observed in number of native species, 51% of the variability observed in number of exotic species, and 47% of the variability associated with observing an exotic species. Important independent variables used in developing the models include geographical location, elevation, slope, aspect, and Landsat TM bands 1-7. These models can direct resource managers to areas in need of further inventory, monitoring, and exotic species control efforts.     

Natural history of the African riverine tiger beetle Chaetodera regalis (Dejean) (Coleoptera: Cicindelidae)

The tiger beetle Chaetodera regalis (Dejean) (Coleoptera: Cicindelidae) is widely distributed throughout sub-Saharan Africa. We studied activity patterns, ecological interactions, and behaviour of this species along four major rivers in Kruger National Park, South Africa. During the dry season, small numbers of adult beetles are found on sand bars along perennial rivers. In the rainy season, adults are found in large numbers on a broad spectrum of substrates (including clays, coarse and fine sands and gravels, and black organic soils) and geomorphological features (sand flats, mud flats, sand bars, beaches, riverbanks, and dry and wet sandy streambeds) in riverine areas. Predatory, defensive, thermoregulatory, and reproductive behaviours are described. This species may prove to be a useful indicator of the health of African riverine systems: adults are abundant in high-quality riverine areas; adults and larvae may be adversely affected by human activities; and adults are easily detected, even by novice surveyors.     

Effects of temperature on the survival and growth of age-0 least chub ( Iotichthys phlegethontis )

Larval and juvenile stages of many fishes require nursery habitats that provide optimal conditions for growth. Loss or degradation of these habitats limits recruitment, causing population and species declines. Least chub ( Iotichthys phlegethontis ), an endemic cyprinid in the Bonneville Basin, is currently restricted to a few spring complexes in Utah. This species utilizes the warm shallow spring margins as spawning and rearing habitat throughout the summer. This study was conducted to determine effects of temperature on survival and growth of age-0 least chub to understand the importance of temperature in selection of spring margins as rearing habitat. Age-0 least chub were exposed to 5 temperatures (14°C, 17°C, 21°C, 24°C, and 27°C) for 112 days. Growth rates varied significantly with temperature except at 17°C and 27°C, and growth rates were highest at 21°C ( P < 0.001). Maximum growth rate for age-0 least chub, estimated from a 2nd-order polynomial regression, would occur at 22.3°C. These thermal requirements indicate the importance of warm rearing habitats in producing strong year classes and viable populations. However, warm spring margins also enable western mosquitofish, a nonnative competitor and predator of least chub, to thrive in these spring habitats. Broad thermal limits of the least chub suggest that it could persist at cooler temperatures, which would reduce the viability of western mosquitofish populations.     

Biogeography and physiological adaptations of the brine fly genus Ephydra (Diptera: Ephydridae) in saline waters of the Great Basin

Four species of the genus Ephydra are commonly found in saline waters within the hydrologic Great Basin: E. hians, E. gracilis, E. packardi , and E. auripes . Though none of these brine flies is endemic (distributions also occur outside the Great Basin), they all inhabit distinctive habitat types and form the characteristic benthic insect fauna of inland saline-water habitats. The affinities of each species for different salinity levels and chemical compositions, and ephemeral to perennial habitats, appear to form the basis for Biogeographic distribution patterns. Within any habitat, changing salinity conditions over time may impose physiological or ecological constraints and further alter patterns of population productivity and the relative abundance of co-inhabiting species. Based on the physiology of salt tolerance known for these species, high salinity conditions favor E. hians in alkaline water and E. gracilis in chloride water. At lower salinities, based on limited habitat data, E. auripes and E. packardi are often more common, again showing respective preferences for alkaline and chloride chemical conditions. Specialized adaptations for alkaline carbonate waters are found in the larval Malpighian tubule lime gland of the alkali fly E. hians , while high salt tolerance in E. gracilis appears to be conferred by high hemolymph osmolality. Adaptation to ephemeral and low salinity conditions may be accomplished by swift adult colonizing ability and rapid larval development rates. It is hypothesized that adaptive specialization in both physiology and life history and varied geochemistry of saline water habitats across the Great Basin produce the Biogeographic pattern of distributions for species in this genus. This perspective on the genus Ephydra , and possibly other biota of mineral-rich Great Basin waters, suggests that interconnections among pluvial lakes may be less relevant to aquatic biogeography than chemical profiles developing in remnant lakes and ponds with the progression of arid post-pluvial climatic conditions.     

Drumming behavior and life history notes of a high-altitude Colorado population of the stonefly Isoperla petersoni Needham

Late instar nymphs and adults of a Colorado Hudsonian zone population of Isoperla petersoni Needham and Christenson were studied during the summer and early fall months of 1998, when stream temperature ranged from 3.3° C to 8.9° C. Early and middle instar nymphs were absent from July to October, and nymphs attained maximum size in mid-August during the 2nd week of adult field presence, suggesting a univoltine-slow life cycle. Nymphs were carnivorous and fed primarily on chironomid larvae. Adults were present from August to mid-October, with peak numbers of adult males and females occurring in mid-September and late September, respectively. Fecundity of field-collected females averaged 94.1 ± 45.15 eggs per female in September ( N = 81) and decreased to 85.2 ± 57.6 in October ( N = 12). Drumming duets were 2-way (male-female), and female answers followed male calls (nonoverlapped) or began before completion of the male call (overlapped). Male calls averaged 11.1 ± 2.09 beats and female answers averaged 6.2 ± 2.99, with mean intervals of 118 and 58 ms, respectively. The average number of female answer beats increased from 5.3 ± 2.56 when duets were nonoverlapped, to 6.6 ± 3.08 when overlapped.     

The reproductive ecology of the Tahoe sucker, Catostomus tahoensis, in Pyramid Lake, Nevada

The Tahoe sucker spawns in Pyramid Lake from April to August at lake temperatures of 11.7 to 22.7 C. The spawning population is comprised of a large lake spawning group and a numerically smaller river running group. The river running group is smaller in length and was not considered during this study. The sex ratio of sampled suckers significantly favored the females. This is the result of the longer life of females and greater mortality of males during spawning. &nbsp; Pyramid Lake Tahoe suckers reach sexual maturity at two to three years of age; however, those in Lake Tahoe do not mature until four or five years of age. The size at sexual maturity is different in both populations, which suggests that size or rate of growth rather than age determines sexual maturity. &nbsp; The fecundity of Tahoe suckers is positively correlated with fork length, weight, and age. Additional analysis showed that a better correlation occurred between fish size (either length or net weight) and total ovary weight. We believe that fish size is primarily correlated with total reproductive tissue produced and secondarily with fecundity. A comparison of the Pyramid Lake population and the Lake Tahoe population demonstrated that size, not age, is the most important determinant of Tahoe sucker fecundity.     

Distribution, movements, and habitat use of razorback sucker ( Xyrauchen texanus ) in a lower Colorado River reservoir, Arizona-Nevada

Distribution, movements, and habitat use of 10 wild adult razorback suckers ( Xyrauchen texanus ) were examined in Lake Mohave, Arizona-Nevada, from November 1994 through July 1997. Movement rates (0.00-17.35 km d -1 ) and ranges ( x = 39 km) were similar to those for riverine populations. All study fish returned to spawning sites used in previous years, but they also visited other spawning areas. Spawning females were significantly ( P = 0.031) more active than males (480 vs. 87 m d -1 ) and moved substantial distances between spawning sites during peak reproduction (1-28 February). Fish became most active (m d -1 , km month -1 ) after spawning and moved to areas known to support higher algal production. Fish were typically within 50 m ( P 30.0 m). Adults were detected throughout the available thermal gradient (12°-30°C), but during summer typically had body temperatures between 18° and 22°C. Vertical movements within the water column showed no correlation with depth or time of day, but seasonal shifts suggest fish may regulate body temperature by seeking specific temperatures during reservoir stratification.     

Fossil history of Notiosorex (Soricomorpha: Soricidae) shrews with descriptions of new fossil species

Notiosorex shrews (Soricomorpha: Soricidae) have a fossil history that extends to the Miocene (i.e., mid-Hemphillian). In the intervening 6.69 million years, N. repenningi, N. jacksoni, and what heretofore was considered N. crawfordi existed with varying degrees of overlap in occurrence timewise and in geographic distribution in areas of the southwestern United States and northern Mexico. Examination of unusually small fossil specimens of Notiosorex indicated the presence of another species referable to this genus present in late Pleistocene- (i.e., early Wisconsinan) to Holocene-aged deposits. Based on statistical analyses, these specimens were as distinct from any of the other fossil or extant species of Notiosorex as any other pair of species within the genus. I referred these specimens to the new species N. harrisi. Analyses also indicted fossils heretofore considered N. crawfordi were equally distinct from other forms of Notiosorex. I referred them to the new species N. dalquesti. Individuals referred to the fossil species of Notiosorex range in size from the smaller N. harrisi, to N. dalquesti, to N. jacksoni, to the larger N. repenningi. This size differential could be related to changing climatic conditions with the concomitant ecological succession that occurred, which provided a driving force for speciation within the genus. The changing climate and unique morphological and physiological adaptations allowed fossil taxa of Notiosorex to expand their distributions into areas subsequently less suitable for other forms of shrews. Present-day notiosoricini shrews occur throughout much of the southwestern United States and northern Mexico in a variety of habitats in association with rough, rocky terrain. Thus, if climate, even indirectly, was the driving force for speciation among fossil taxa of Notiosorex, other forces must have acted on the extant species N. cockrumi, N. crawfordi, N. evotis, and N. villai.