Variation in male mating success in the paternal nest brooder, Rhinogobius goby, along a river course

Rhinogobius sp. CB (cross band type) is an amphidromous freshwater goby which receives paternal nest care under stones on river beds. This goby is known to migrate to the upper reaches of rivers as it grows. In the present study, the relationships among male mating success of Rhinogobius sp. CB, male body size, the size frequency distribution of river bed stones and the presence of the sympatric goby Tridentiger brevispinis (which uses nest sites similar to those of Rhinogobius sp. CB) were investigated along the course of the Aizu River, Wakayama Prefecture, Japan. In the lower reach, where T. brevispinis was present and large stones were scarce, the sizes of the nest stones and the egg clusters of male Rhinogobius sp. CB were smaller than those of T. brevispinis. In the middle and the upper reaches, where T. brevispinis was absent and large stones were abundant, males of Rhinogobius sp. CB used larger nest stones than in the lower reach. In both gobies, there were positive correlations among male body size, nest size and egg cluster size in all reaches. Nest-choice experiments carried out in the laboratory, with or without a rival, showed that males of both gobies preferentially used large nest sites and that large males had an advantage in male-male competition for large nest sites. These results indicate that large male body size can increase male mating success and that male mating success increases in the upper reach in this goby. The migration pattern of Rhinogobius sp. CB was also discussed in relation to these findings.     

Flexible migration of Japanese freshwater gobies Rhinogobius spp. as revealed by otolith Sr:Ca ratios

The migratory histories of six Rhinogobius spp., the cross‐band type (R. sp. CB), the large‐dark type (R. sp. LD), the dark type (R. sp. DA), the cobalt type (R. sp. CO), the orange type (R. sp. OR) and R. flumineus, were studied by examining strontium (Sr) and calcium (Ca) concentrations in their otoliths using wavelength dispersive X‐ray spectrometry on an electron microprobe. The Sr:Ca ratios in the otoliths changed both with the ontogenetic development and with the salinity level of the habitat. Most fishes had high Sr:Ca ratios around the otolith core in spite of the fact that those fishes live most of their lives in a freshwater environment. The high ratios in the otoliths were thought to be a physiological effect in those fishes. Thereafter, the Sr:Ca ratios changed remarkably along the life‐history transect, showing intraspecies and interspecies variations. The otolith Sr:Ca ratios of Rhinogobius sp. CB, R. sp. LD and R. sp. CO collected from three rivers connected to the sea were low around the core, subsequently increased sharply to the points 180–345 μm from the core and then decreased again towards the edge. They were thought to reflect the typical amphidromous life history. The R. sp. CO, however, remain in a brackish‐water environment after migration from the sea, while the other species showed typical amphidromous lives with complete freshwater residence after migration from the sea. The five species (R. sp. CB, R. sp. LD, R. sp. CO, R. sp. DA and R. sp. OR) collected above dams had never migrated to the sea, spending their whole life in a freshwater environment, although Rhinogobius species, except for the fluvial type, were thought to have an amphidromous life history according to previous studies. These species are thought to have a landlocked life cycle. The otolith Sr:Ca ratios of R. flumineus showed consistently low ratios towards the edge except for only around the core, although they were collected from a river connected to the sea. These species could have a fluvial life history corresponding to a previous study. The present study clearly suggests that the migratory histories of Rhinogobius spp. are highly different both within and between species and that they have flexible migratory patterns allowing them to utilize the full range of salinity during their life history.     

Genetic variations estimated from PCR-RFLP analysis of two morphs of the freshwater goby <Emphasis Type="Italic">Rhinogobius</Emphasis> in the Lake Biwa water system

The freshwater goby Rhinogobius is the most abundant fish in the shore area of Lake Biwa, Japan. Recently, it has been reported that two morphs of Rhinogobius inhabit this lake. In this study, genetic variations in Rhinogobius sp. OR (Orange-type) and Rhinogobius sp. BW (Biwa-type) in the Lake Biwa water system have been investigated using polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) analysis of mitochondrial DNA, including the variable D-loop region. Samples of Rhinogobius sp. OR were collected from the middle sites of three rivers, two outlets, and two lakeshore sites, whereas samples of Rhinogobius sp. BW were taken from two lakeshore sites. Rhinogobius sp. OR and Rhinogobius sp. BW did not share any haplotypes, suggesting that PCR-RFLP analysis is effective for distinguishing between these species of goby. The haplotype diversity of Rhinogobius sp. OR (0.214–0.543) was lower than that of Rhinogobius sp. BW (0.543–0.682). There were no significant differences in haplotype frequencies between Rhinogobius sp. BW groups from the two localities. In addition, haplotype frequencies in Rhinogobius sp. OR did not differ significantly among samples from the middle sites of rivers, the outlets, and the shores. These results indicate that in Rhinogobius sp. OR there is frequent gene flow among fish inhabiting different sites, and that this species of goby consists of a single population throughout the Lake Biwa water system.     

Genetic diversity and divergence of the endangered freshwater goby Rhinogobius sp. BB in Okinawa Island

The genetic diversity and population structure of Rhinogobius sp. BB, the fluvial landlocked endangered goby, were investigated and were compared with those of the closely related amphidromous Rhinogobius sp. MO at 11 microsatellite loci. Specimens of Rhinogobius sp. BB were collected from the Genka and Takae-A rivers, and those of Rhinogobius sp. MO were collected from the Genka and Suginda rivers in Okinawa Island. At 11 microsatellite loci, the two populations of Rhinogobius sp. BB showed lower variation than the two Rhinogobius sp. MO populations: the average number of alleles was 3.6 and 2.0 vs. 8.6 and 7.6, respectively; and the observed heterozygosity was 0.263 and 0.281 vs. 0.440 and 0.545, respectively. Pairwise F_st tests showed significant differences (P < 0.001) among the populations: F_st was 0.525 between the two Rhinogobius sp. BB populations, 0.079 between the two Rhinogobius sp. MO populations, and varied from 0.456 to 0.462 for comparisons among Rhinogobius sp. BB and MO. Nei’s genetic distance between the two Rhinogobius sp. BB populations is extremely large (0.604) compared with that between the two Rhinogobius sp. MO populations (0.126). The two populations of Rhinogobius sp. BB are genetically divergent, and they have extremely low genetic diversity. Therefore, the conservation of Rhinogobius sp. BB in Okinawa Island requires the assessment of each river’s population.     

A new lentic form of the “yoshinobori” species complex, Rhinogobius spp. from Lake Biwa, Japan, compared with lake–river migrating Rhinogobius sp. OR

 The various life history strategies seen within the “yoshinobori” species complex of the genus Rhinogobius, which differentiated from an amphidromous ancestor, have been grouped ecologically into amphidromous, fluvial, and lacustrine types. In the Lake Biwa water system, two lacustrine forms exist, a newly discovered, exclusively lentic form and the already well known Rhinogobius sp. OR, which generally undergoes lake–river migration but also includes lentic individuals that share spawning grounds with the former lentic form. Detailed morphological comparisons and allozyme analysis revealed consistently clear differences between the two forms, indicating them to be distinct species. The newly discovered lentic form has many distinctive morphological features that enable it to be distinguished from sympatric Rhinogobius sp. OR: dwarfness, short snout, longitudinally slender pelvic fin, undeveloped frenum with a low lamella, low first dorsal fin in adult males, lesser caudal peduncle depth, scaleless predorsal and ventral areas, and incomplete sensory canals. The lentic form was temporarily named Rhinogobius sp. BW. The life history patterns of the lake-inhabiting populations were separated into two categories: a lake–river migratory lifestyle and an exclusively lentic lifestyle.     

Migratory history of Rhinogobius sp. OR morphotype “Shimahire” as revealed by otolith Sr:Ca ratios

The migratory history of the Rhinogobius sp. OR morphotype “Shimahire” collected from a slower current environment in the Maruyama River, the Ibo River, and the Kako River, Japan, was studied by examining the strontium and calcium concentrations in the otoliths. The otolith Sr:Ca ratios of almost all samples showed consistently low ratios from the core to the edge, reflecting the fact that the organisms had been living in a freshwater environment their entire lives. The ratios of Rhinogobius sp. CB and Rhinogobius flumineus living in the sympatric habitats with the Rhinogobius sp. OR morphotype “Shimahire” in the Kako River showed an amphidromous and fluvial life cycle, respectively. These findings suggest that the Rhinogobius sp. OR morphotype “Shimahire” has a freshwater resident life cycle.     

Coloration and ontogenetic features of fluviatile species of <Emphasis Type="Italic">Rhinogobius</Emphasis> (Gobioidei: Gobiidae) in Amami-oshima Island,Ryukyu Islands,Japan

 The fluviatile Rhinogobius sp. 1 (hereinafter R. sp. 1) (Japanese name kibara-yoshinobori or English name yellow belly medium-egg type) is broadly distributed in the Ryukyu Islands ranging from Amami-oshima Island to Iriomote Island. However, based on the molecular evidence, this species has probably had multiple origins from amphidromous species. We compared the coloration of specimens at Amami-oshima I. with that of the other specimens (islands) in the Ryukyu Is. Amami-oshima I. specimens of R. sp. 1 were different from the specimens of other populations in having five to six cross lines on the caudal fin. We examined the early life history of fluviatile Rhinogobius at Amami-oshima I., based on rearing experiments, and estimated the age of wild specimens by counting otolith increments. Spawned egg capsules of “Rhinogobius sp. 1” at Amami-oshima I. were larger than those of amphidromous species. Their developmental stage in newly hatched larvae was similar to that of Okinawan R. sp. 1 in having a straight notochord; however, it had a flexed notochord earlier than that of Rhinogobius sp. 2 (hereinafter R. sp. 2) (aobara-yoshinobori or blue belly medium-egg type). The ontogenetic trajectory of “Rhinogobius sp. 1” in Amami-oshima I. was similar to Okinawan R. sp. 1 (30 days in larval duration) but different from R. sp. 2 (15 days). In amphidromous relatives of R. sp. 2, developmental timing during larval period was earlier than in the amphidromous relatives of R. sp. 1. It has been suggested that the phenomenon of fluvial landlocking has been accompanied somewhat by egg size increase (on the basis of a few examples). However, the ontogenetic trajectories of fluviatile Rhinogobius species may be a reflection of those of ancestral amphidromous species. Received: March 18, 2002 / Revised: November 7, 2002 / Accepted: December 10, 2002     

Mitochondrial DNA divergence in yoshinobori gobies (Rhinogobius species complex) between the Bonin Islands and the Japan–Ryukyu Archipelago

To investigate the genetic differentiation between the Bonin (Ogasawara) Islands' freshwater goby Rhinogobius sp. Bonin Island (BI) form (Ogasawara-yoshinobori) and the Japan–Ryukyu Archipelago relatives, the mitochondrial DNA (mtDNA) phylogeny of Japanese Rhinogobius species was inferred from partial nucleotide sequences of the mitochondrial NADH dehydrogenase 5 subunit (ND5) gene (945 bp). The resultant tree showed that the Bonin Islands group separated first from the other Japanese lineage, and a test calculation indicated the divergence date to be approximately 3 million years BP. Although it is necessary to use a more reliable estimate to confirm the divergence date, Rhinogobius sp. BI has retained its mtDNA lineage in the islands for millions of years.     

Foraging modes of stream benthic fishes in relation to their predation effects on local prey density

Habitat use and foraging behavior of two benthic insectivorous gobies, Rhinogobius sp. CO (cobalt type) and Rhinogobius sp. DA (dark type), were examined in relation to their predation effects on local prey density in a small coastal stream in southwestern Shikoku, Japan. Correlations among the foraging range, frequency of foraging attempts and current velocity indicated that individuals using fast-current habitats had small foraging ranges and infrequently made foraging attempts while those in slow currents frequently foraged over large areas. The former and the latter were recognized as ambush and wandering foragers, respectively. Interspecific comparisons of habitat use, foraging behavior and prey preference suggested that Rhinogobius sp. CO selectively forage mobile prey by ambushing in fast currents, whereas Rhinogobius sp. DA randomly forage available prey by wandering in slow-current habitats. A cage experiment was conducted to assess prey immigration rate and the degree of predation effects on local prey density in relation to current velocity. The results of the experiment support, at least in part, our initial predictions: (1) prey immigration rates increase with current velocity and (2) the effects of fish predation on local prey density are reduced as current velocity increases. Overall results illustrated a link between the foraging modes of the stream gobies and their predation effects on local prey density: fish adopt ambush foraging in fast currents, where the decrease in prey density tends to be less, whereas fish actively forage over large areas in slow currents, where the decrease in prey is relatively large.     

Genetic identification of larvae and juveniles reveals the difference in the spawning site among Cyprininae fish species/subspecies in Lake Biwa

To understand the differences in the spawning sites among Cyprininae fishes in Lake Biwa, we conducted periodic sampling of larvae and juveniles at three sites (irrigation ditch, St. 1; river, St. 2; and satellite lake, St. 3). On the basis of species/subspecies identification by using RAPD analysis, we examined the species composition of the larvae and juveniles at these three sampling sites. The number of specimens was 616, 68, and 117 at St. 1, St. 2, and St. 3, respectively. Based on morphological and genetic identification, the specimens were found to include nine fish species/subspecies, namely, Carassius auratus grandoculis, Carassius cuvieri, Carassius auratus langsdorfii, Cyprinus carpio, Sarcocheilichthys sp., Silurus asotus, Oryzias latipes, Odontobutis obscura obscura, and Rhinogobius sp. The species composition at the three sites also differed. Among the Cyprininae fishes, C. auratus grandoculis, C. auratus langsdorfii, and Cyprinus carpio were found in abundance at St. 1; C. cuvieri was not collected from St. 1 but was found at the other two sites, particularly St. 3. Among the other fishes, Rhinogobius sp. was collected at St. 1 and St. 3, whereas the other four occurred only at St. 1. These results suggest that the selection of spawning sites by C. cuvieri differs to a certain extent from that of the other Cyprininae fishes, and the irrigation ditch in the lake is an important habitat for the larvae and juveniles of native fish species.     

Stable nitrogen and carbon isotope ratios as migration tracers of a landlocked goby, Rhinogobius sp. (the orange form), in the Lake Biwa water system

Fluvial–lacustrine migrations of a landlocked goby, Rhinogobius sp. (the orange form) in the Lake Biwa water system were traced using stable nitrogen and carbon isotope ratios. The ¹⁵N values of Rhinogobius sp. individuals in the lake were significantly higher than those of large individuals in a tributary river (1⁺ age) without overlap of the range, although there was little difference in the ¹³C values between these two locations. Because these values reflected those of the corresponding prey organisms in either location, it was revealed that none of the large individuals had migrated from one location to the other. The ¹⁵N values of the small individuals in the tributary river (0⁺ age) were too high to be fluvial residents. This result indicated that all of the small individuals studied had spent their larval periods in the lake, and that immigrant individuals can be distinguishable by measuring the ¹⁵N values retaining the effect of foraging in the alternate location. We conclude that the ¹⁵N value works as a useful tracer to clarify the fluvial–lacustrine migration pattern of the fish.     

Female mimicry in a freshwater goby <Emphasis Type="Italic">Rhinogobius</Emphasis> sp. OR

 We found female-mimic males in a freshwater goby, Rhinogobius sp. OR, from Kamo River, Ehime, Japan. Of 171 adult males collected, three had a female appearace. A discriminant analysis showed that they had a spectrum of morphology from a wholly female morph, through a neutral morph, to a somewhat malelike morph. These three males made an extremely higher gametic investment than did the normal males, suggesting that they have the potential for sneaking fertilizations. Among Rhinogobius species, female mimicry might be more common than previously known. Received: September 24, 2002 / Revised: January 23, 2003 / Accepted: February 4, 2003     

Isolation of microsatellite loci in the freshwater goby,Rhinogobius sp. (Gobiidae)

The Rhinogobius species complex (Pisces: Gobiidae) includes great variation in colour, morphology and ecological form. In particular, R. sp. OR (Orange type) exhibits some of the greatest colour variation among the species complex, although the genetic relationships among the variations in this fish remain unclear. Thirteen microsatellite loci were identified from R. sp. OR, and all loci were polymorphic with two to 17 alleles per locus. Observed heterozygosity ranged from 0.1 to 0.9, while the expected heterozygosity ranged from 0.19 to 0.96. Multiplex polymerase chain reaction reaction (PCR) were optimized. All loci except Rhi‐5 conform to Hardy–Weinberg equilibrium (P > 0.05).     

Variations in clutch and egg sizes in the amphidromous goby <Emphasis Type="Italic">Rhinogobius</Emphasis> sp. CB along a river course and within a spawning season

Rhinogobius sp. CB is a common amphidromous goby in Japan. I investigated egg and clutch sizes along a river course and within a spawning season in the Aizu River, Kii Peninsula, Japan. Clutch size within the same female size did not differ along the course of the river, but decreased from early to later in the spawning season. Egg size within the same female size decreased from early to later in the spawning season and from the upper to the lower reaches. These results mean energy resources allocated to the ovary decreased seasonally and locally. Several factors are presented to explain these variations, such as female somatic conditions, a tradeoff between reproduction and growth, food availability, water temperature, and presence of competitive species. Some factors explaining the seasonal and local variations in egg size, such as water temperature and food availability for offspring, are also presented. Egg size correlated well with female size. The trend in egg size variation with female size seems to be an adaptation for their migrating pattern, as larvae from larger female, which tended to inhabit the upper reaches, would have a greater endurance against starvation during their migration to the sea.     

Efficient synthetic signal peptides for Streptomyces

A short synthetic signal peptide (SSSP) of 26 amino acid and a long one of 35 amino acids (LSSP), having an additional ribosome binding site (RBS), were synthesized. The SSSP sequence was based on the comparison of known efficient Streptomycessignal sequences. The SSSP and the LSSP were connected to the Streptomycessp. TO1 amylase gene (amyTO1) without its signal peptide. These constructions, when cloned into Streptomycessp. TO1 and placed under the control of the ermE-up promoter of Saccharopolyspora erythrea, increased the secretion of the amylase up to six-fold when compared to the natural amyTO1 signal peptide.     

Intra-specific variations of egg size,clutch size and larval survival relatedto maternal size in amphidromous <Emphasis Type="BoldItalic">Rhinogobius</Emphasis> goby

Synopsis Newly hatched amphidromous Rhinogobius sp. CB (cross band type) larvae drift downstream to the sea to grow and develop before returning upstream as juveniles. Since larger and older individuals usually inhabit the upper reaches of rivers, larvae from larger females are more likely to suffer higher risks of starvation or predation during their longer migration to the sea. We examined the relationship between reproductive parameters (egg volume and clutch size) and maternal length. We collected adult Rhinogobius sp. CB along the course of the Aizu River (Wakayama Prefecture, Japan) and spawned them under laboratory conditions. We measured egg volumes and clutch sizes, as well as larval starvation tolerance. Both egg volume and clutch size increased with standard length or age of the maternal fish, while egg density in these clusters did not correlate with standard length. Gonad-somatic index (GSI) also tended to increase with maternal standard length. There was a significant positive correlation between egg size and the 72-h survival rate of unfed hatchlings. Intra-specific variation of egg volumes and clutch sizes in this species seems to be an adaptation for enhancing offspring survivorship during migration to the sea. Some females spawned a second time. Second spawned egg sizes were smaller than first spawned egg sizes, although there was no difference in clutch size between the two. Egg size variation between subsequent spawns may be an adjustment to the changes in seasonal environmental conditions.     

Habitat use and diet of two stream gobies of the genus Rhinogobius in south-western Shikoku,Japan

The importance of interspecific competition to habitat use by two congeneric stream gobies, Rhinogobius sp. LD (large-dark type) and CB (cross-band type), was studied by: (i) examining differences in habitat use by each species along the course of the stream; and (ii) comparing microhabitat use and the diet of each species between in allopatry and in sympatry in tributaries of the Shimanto River, south-western Shikoku, Japan. Rhinogobius sp. LD mainly used riffles through the course of the stream. Although CB also mainly used riffles in the lower reaches where CB was numerically dominant, the greater use of riffles by CB was not observed in upper reaches where LD was dominant. Microhabitat analysis revealed that both LD and CB preferred lower bottom velocity in faster current in both allopatry and sympatry. In sympatry, however, LD used coarser substrate and faster current than CB, and both LD and CB used narrower ranges of microhabitat conditions than in allopatry. Dietary analysis indicated that both allopatric and sympatric LD preferred baetids, which are apt to drift, suggesting they adopted ambush foraging. Allopatric CB preferred leptophlebiids and chironomids, which are also apt to drift, whereas sympatric CB did not prefer such invertebrates. Overall results of this study suggest that LD and CB compete for better feeding habitats, CB shift their habitat use and foraging mode under the influence of LD, and current velocity and substrate coarseness are key factors in their habitat segregation.

     

Life history and production of filter-feeding insects on stone dikes in the lower Mississippi River

Population and production dynamics of two filter-feeding macroinvertebrates, the caddisfly Hydropsyche orris and the midge Rheotanytarsus sp., were examined in the lower Mississippi River Miles 510–515. Samples were collected from September to November 1987, May to December 1988, and March to May 1989 from stone dikes that protrude into the river. Developmental synchrony was high among individuals in both populations, and seasonal changes in the relative abundance of instars indicated bivoltine and trivoltine life cycles for H. orris and Rheotanytarsus sp., respectively. The size frequency method was used to estimate production (P) and biomass (B) of each cohort in each population. P/B ratios for spring and fall cohorts of H. orris equaled 4.5 and 3.7. Spring, summer, and fall cohorts of Rheotanytarsus sp. had P/B ratios of 4.0, 4.7, and 4.1, respectively. Annual production, estimated as the sum of cohort production values, of the H. orris and Rheotanytarsus sp. population equaled 10.1 and 0.359 g m^–2 (dry weight). Annual P/B ratios equaled 10.8 and 16.1 for the H. orris and Rheotanytarsus sp. populations, closely matching values expected for bivoltine and trivoltine populations, respectively. Annual production of H. orris is in the low end of the range of published estimates for other lotic hydropsychid populations inhabiting coarse substratum in warm, lowland rivers of medium size or small, trophically enriched streams. Annual production of Rheotanytarsus sp. is the first such estimate based on a simultaneous assessment of life cycle and production. Given the lack of naturally occurring gravel and cobble substrate in the lower Mississippi River, stone dikes are an especially important habitat for H. orris and Rheotanytarsus sp.     

A new species ofRhinogobius (Teleostei: Gobiidae) from Fujian Province, China

A new freshwater goby,Rhinogobius xianshuiensis sp. nov. was collected from the upper Mulan River basin of Xianyou County, Fujian Province, eastern China. The species is distinguished from related species in the region by 3–7 predorsal scales, 27 vertebrae, cephalic lateralis canals always always with 2 derived post-orbital pores (ε1) in adults, a reddish-brown vertical line below the anterior margin of the eye, and a red netlike pattern on the opercle. A key to species ofRhinogobius with higher vertebral numbers (27–29) from eastern China, Taiwan and Japan is provided.     

Behavior and distribution of upstream-migrating juvenilerhinogobius sp. (The orange form)

Upstream-migration of juvenileRhinogobius sp. OR was studied along the Ado River, a tributary of Lake Biwa. Juvenile gobies, mainly 18–21 mm in standard length, migrated from the lake in a belt of about 3 m wide in lower reaches and about 10–20 cm wide in middler reaches. Their upstream-migration was restricted to places shallower than 20 cm with bottom current velocities less than 20 cm/sec. Individual migration speed varied between 4.2–8.9 m/min at places with bottom current velocity of 8.8–20.0 cm/sec, slowing down with increasing current velocity. Most active migration occurred between 14:00 and 16:00, number of upstream-migrating juveniles decreasing with distance from the river mouth. Riffles in the mid-and upper reaches, having few slow current areas, probably present barriers to upstream-migration of the juvenile gobies, although both adults and subadults of the species are distributed in more upper reaches.