Objective. Unravelling the determinants of parasite life-history traits in natural settings is complex. Here, we deciphered the relationships between biotic, abiotic factors and the variation in 4 life-history traits (body size, egg presence, egg number and egg size) in the fish ectoparasite Tracheliastes polycolpus. We then determined the factors affecting the strength of the trade-off between egg number and egg size. Methods. To do so, we used 4-level (parasite, microhabitat, host and environment) hierarchical models coupled to a field database. Results. Variation in life-history traits was mostly due to individual characteristics measured at the parasite level. At the microhabitat level (fins of fish hosts), parasite number was positively related to body size, egg presence and egg number. Higher parasite number on fins was positively associated with individual parasite fitness. At the host level, host body size was positively related to the individual fitness of the parasite; parasites were bigger and more fecund on bigger hosts. In contrast, factors measured at the environmental level had a weak influence on life-history traits. Finally, a site-dependent trade-off between egg number and egg size existed in this population. Conclusion. Our study illustrates the importance of considering parasite life-history traits in a hierarchical framework to decipher complex links between biotic, abiotic factors and parasite life-history traits.

Over the past decades, pollution, overfishing, and habitat degradation have driven the population size of Taiwan shoveljaw carp down markedly in Taiwan. Cryopreservation is a useful tool which could be used to maintain genetic resources to protect and preserve this endemic species. Four cryoprotectants [dimethyl sulphoxide (DMSO), dimethylacetamide (DMA), glycerol and methanol] and six freezing rates (0.5, 1, 2, 4, 8, 16 °C min-1) were tested in order to develop an optimal controlled slow-freezing protocol for Taiwan shoveljaw carp spermatozoa. Samples were subsequently examined under the scanning electron microscope to reveal whether cryopreservation had affected their ultrastructural morphology. The highest survival rate (50.1 ± 2.0%) was observed with a freezing rate of 8 °C min-1 in 1M DMSO, using SYBR-14 + PI staining. Fertility and hatching rate results using frozen-thawed spermatozoa (90.2 ± 2.2% and 22.3 ± 2.5%, respectively) were not significantly different from results with fresh spermatozoa. After cryopreservation, 21.0 ± 1.6% of frozen-thawed spermatozoa had mid-piece swelling and rupture of the head. Cryopreservation might, therefore, slightly affect Taiwan shoveljaw carp spermatozoa in terms of morphological change. However, these alterations could be compensated by using large enough numbers of normally functioning frozen-thawed spermatozoa to achieve a standard equal to fresh spermatozoa. This is the first report of successful cryopreservation of Taiwan shoveljaw carp spermatozoa using a controlled slow-cooling method.

To address whether a fish host is involved in the life cycles of malacosporeans of the genus Buddenbrockia, cohabitation experiments with different bryozoan and fish species were conducted. Samples were analysed by malacosporean-specific PCR, partial sequencing of the 18S rDNA, and light and electron microscopy. Co-habitation challenges with bryozoans resulted in malacosporean infections detected mainly in the kidney of common carp (Cyprinus carpio) and minnow (Phoxinus phoxinus). Sequences of the minnow parasite and of worm-like Buddenbrockia stages in Plumatella repens were identical and showed 99·5% similarity to Buddenbrockia plumatellae and 96·3% similarity to the sequence obtained from carp. One sample, comprising 4–5 zooids of statoblast-raised bryozoans cohabitated with infected carp was PCR-positive, but no overt infection could be observed in the remaining colony. Light and electron-microscopy of kidney samples of infected minnows revealed single cells within kidney tubules, whereas in carp, sporogonic stages were found in kidney tubules. Phylogenetic analysis of the Buddenbrockia spp. known to date placed the carp-infecting species at the base of the B. plumatellae clade, but low posterior probability makes this node questionable. The present study showed that Buddenbrockia spp. were able to infect cyprinid fish, showing stages in kidney-tubules strikingly similar to those of T. bryosalmonae.

A population of Garra ceylonensis in a rocky, hill stream was studied by monthly sampling over a period of two years. The monthly population density, which varied from 1.28 to 4.16 m-2 with a mean of 2.37 m-2, was negatively correlated with the water speed. Asymptotic length (L∞) and the growth constant (K) were found to be 180.5 mm total length and 0.62. Maximum size observed was 174 mm, which is attained in about five years, but 99% of the population had a life expectancy of less than 3.4 years. The overall male to female ratio of the population was 0.58. The sex ratio was 1.0 in smaller size classes but decreased steadily with increasing size. There was a major spawning season following the main rainy period in contrast to the trend of most tropical freshwater fish species, which show enhanced breeding during the rainy season. The mean length and age at sexual maturity in females appear to be 109 mm and eighteen months. Fecundity varied between 740 and 4390. Average longevity and natural mortality were 4.8 years and 1.86. The mean biomass, average annual production and turnover ratio of the population were about 23.7 gm-2 wet weight, 41.2gm-2 and 1.7, respectively.

Twelve diel surveys were carried out in three reservoirs of Sri Lanka viz. Minneriya, Udawalawe and Victoria, to investigate diel feeding patterns, daily ration and relative food consumption in fish populations. Stomach content weights of different size classes of various fish species in the three reservoirs in 12 diel surveys were analysed using an iterative method, MAXIMS. Predominantly herbivorous or detritivorous fish species such as Amblypharyngodon melettinus and Oreochromis niloticus exhibited one peak in the diel feeding pattern. Two peak feeding periods were evident in predominantly insectivores and/or zooplanktivores (e.g., Puntius chola and Rasbora daniconius) and interestingly in macrophyte feeders (i.e., Etroplus suratensis, Puntius filamentosus and Tilapia rendalli). It might be possible that all species with two feeding peaks in diel feeding patterns rely on vision for feeding.Food consumption per biomass (Q/B ratio) defined as amount of food consumed per unit weight of an age-structured population of fish was estimated on the basis of the average quantities of food consumed over a long period of time by various size classes in order to minimize the bias of estimates.

This paper presents a comprehensive study of the impact of damming on the spawning migrations of Barbus barbus in the canalized River Meuse (Southern Belgium). A Denil fish pass on the Ampsin-Neuville dam was controlled 251 times in 1989-1993. The most striking feature is the almost complete absence of barbel in 1990-1993 (15 fishes in 4 years). Most captures of barbel in the fish pass in 1989 were clumped within a few days (mid-May) and related with spawning migrations. Stepwise multiple regression analyses revealed that attractivity was the major condition set allowing barbel to migrate successfully through the pass, while feasibility parameters (water velocity) were found not to be relevant. The variables involved in the attractivity condition set refer indirectly to the influence of water catchment by a hydroelectric plant and to the relative importance of the flow in the pass. The study concludes that this additional condition set significantly interferes with the natural environmental stimuli triggering spawning migrations in barbel and questions the effectiveness of the thermally-related reproductive strategy of the species in an environment with restricted longitudinal connectivity. This statement is discussed in parallel with the recent evolution of barbel populations in canalized rivers.

The toxicity of five cryoprotectants – methanol, DMSO, glycerol, ethanediol and sucrose – on different development stages of zebrafish embryos was investigated. Embryos were exposed to a range of cryoprotectant concentrations for 30 min at room temperature. Post-heart beat stage embryos were more resistant to cryoprotectants than carly embryonic stages. The maximum non-toxic concentrations of cryoprotectants on heart beat stage embryos were 2 M methanol, 2 M DMSO, 1 M glycerol, 2 M ethanediol and 0.5 M sucrose. Gradual stepwise addition did not reduce toxicity. Heart beat stage embryos survived 2 M methanol at room temperature for up to 5 hours, whilst DMSO and ethanediol were toxic after 3 and 1 hour exposure respectively. The effect of the nature and concentration of cryoprotectant, equilibrium time, and cooling rate were investigated during cooling to –30 °C. Methanol was more effective than either DMSO or ethanediol for zebrafish embryo cryopreservation and 0.3 °C/min was found to be the optimum cooling rate. Two-step addition of cryoprotectants, with the higher concentration of cryoprotectants added at 0 °C, improved the results. The best embryo survivals obtained after cooling were 94% at – 10 °C, 72% at – 15 °C, 43% at –20 °C, and 8% at –25 °C, no embryos hatched following cooling to –30 °C. Ice formation within the egg was found to be the main factor affecting survival of the embryos.

Goldfish embryos of pre-morula, somite formation, optic vesicle, heart formation and pigment formation stages were tested for cryoprotectant toxicity, using DMSO and glycerol at 10, 12 and 15% concentration. Those that showed 50% or more survival were employed for cold shock test, from 24 to 4 °C at the rate of –1 °C/min. Only embryos at the heart formation stage protected with 12% DMSO showed 60% survival and 35% hatching. A group of embryos of that stage went through a freezing and thawing test to temperatures of –15, –25 and –35 °C. They showed 30, 5 and 2% survival and 12, 3 and 1% hatching respectively.

F1 hybrids were produced by crossing, in artificial conditions, female Barbus barbus and male B. meridionalis. Results from the genetic study describing enzymatic patterns produced after electrophoresis of experimental F1 hybrids confirmed a great part of the original interpretation of patterns observed in the natural population. Morphological studies which examined denticle length on the last hard ray of the dorsal fin (well developed in B. barbus and absent in B. meridionalis), showed that F1 hybrids' values were not exactly intermediate between the parental values. Up to two years old, experimental hybrids displayed identical growth rates and sexual dimorphism of length (in favour of the females) as B. barbus growing in the same conditions. At this age, in female hybrids, the ovaries matured and produced fertilizable ovules: in male hybrids, the testicles were morphologically normal but no sperm was produced. If this sterility is confirmed in the reciprocal cross (male B. barbus × female B. meridionalis), the phenomenon would be an essential feature in understanding this hybridization.

Two female barbel Rarbus barbus (L.) were tagged with implantable radio transmitters and tracked in the Ourthe River (Southern Belgium) from April 12th to July 7th, 1989 to study their movements and activity rhythms during the spring-summer period. Both barbels defined a 1.6 km home range and occupied four residence areas. Greatest average daily movements within the home range limits occurred during the prespawning and spawning periods and were associated with variations of mean daily water temperature and level. Barbel 2 made a 9.6 km excursion from its spring home range and homed a week later in its residence area. Barbels displayed dusk and dawn activity patterns, character- ized by the occupation of riffle areas. The activity timing was found to be dependent on water temperature while the size of daily activity area was more influenced by river flow conditions.

Three species in the group of "small" Barbus from West Africa were analyzed by enzyme electrophoresis to assess their genetic differentiation. Comparison with a species of "large" Barbus from the same region showed that the "small" Barbus are certainly diploid and the "large" tetraploid. They clearly form two distinct lineages. Phenetic dendrograms (Nei distances) and cladograms (compatibility networks) of the genus Barbus are proposed, based on three African diploid species, two diploid species from Saudi Arabia and Southeast Asia, one African tetraploid species, and two French tetraploid species. These trees reveal two sets of species, i.e. diploid and tetraploid. Several methods of data processing are suggested for overcoming the difficulties involved in simultaneously analyzing species with different ploidy levels.

Enzymatic polymorphism was studied in nine enzyme systems of four species of large AfricanBarbus (B. bynni occidentalis Boulenger, 1911; B. sacrutus Daget, 1963; B. petitjeani Daget 1962; and B. wurtzi Pellegrin, 1908), two species of small African Barbus (B. guineensis Pellegrin, 1913 and B. cadenati Daget, 1962), and two species of European Barbus (B. barbus Linnaeus, 1758 and B. meridionalis). Like European Barbus, the large African Barbus were found to express more than 20 enzymatic loci for these systems (21 to 23), whereas the small African Barbus only expressed 14. This result suggests that large African Barbus are tetraploid like European Barbus and that small African Barbus are diploid like the Asian species. The two groups of African Barbus do not appear to be phylogenetically closer to each other than they are to European Barbus. Consequently, tetraploidi-zation must have occurred long ago in large African Barbus, and may correspond to the same event responsible for the origin of the European species.