Corkwing Research Articles

Protein carbonyls and antioxidant defenses in corkwing wrasse ( Symphodus melops) from a heavy metal polluted and a PAH polluted site

The use of fish in environmental monitoring has become increasingly important in recent years as anthropogenic substances, many of which function as prooxidants, are accumulating in aquatic environments. We have measured a battery of antioxidant defenses as a measure of oxidative status, as well as protein carbonylation as a measure of oxidative damage, in corkwing wrasse ( Symphodus melops) captured near a disused copper mine, where water and sediment are contaminated with heavy metals, and an aluminum smelter, a site contaminated with PAHs. Results were compared to two different reference sites. Fish at the heavy metal site had lower glucose-6-phosphate dehydrogenase activity and elevated protein carbonyls (1.8 times) compared to fish from the reference site. At the PAH site, EROD was increased 2-fold, while total glutathione and methemoglobin reductase concentration, were decreased. No differences were seen in protein carbonyl levels at the PAH site. Measures of both antioxidant defenses and oxidative damage should be used when assessing effects of xenobiotics on oxidative stress in fish species.

The liver of wrasse – Morphology and function as a mirror of point source chemical impact

Corkwing wrasse ( Symphodus melops L.), a protogynous, non-migratory lipfish species, living close to rocky shores was chosen as an indicator species for the monitoring of biological effects of contaminants. Fish were caught by local fisherman at the Norwegian west coast in fjord sites within the framework of the EU BEEP project. The sites represented different point source impacts of (I) copper (a former copper mine), (II) polycyclic aromatic hydrocarbons (PAHs, aluminium smelter discharge), (III) formaldehyde plus PAHs (kelp-factory and influence of the aluminium smelter). Livers of wrasse were studied for histopathological alterations and compared to healthy livers of fish from a reference location. Besides liver morphology, different functional and metabolic parameters were measured to link pathological alterations to functional disorders. The integrity of the lysosomal compartment was tested by the assessment of lysosomal membrane stability (lys), and the accumulation of neutral lipids and lipofuscin. Activity and intracellular localisation of the NADPH-producing enzymes in the liver were assessed histochemically and measured by computer-assisted image analysis. Histopathological alterations were most severe at the site impacted by formaldehyde and PAHs. These findings were associated with highest tumor prevalence, lowest membrane stabilities in hepatocytes and highest accumulation rates of lipofuscin in the liver. The activities of the NADPH-producing enzymes phosphogluconate dehydrogenase (PGDH) and glucose-6-phosphate dehydrogenase (G6PDH) were significantly lower compared to unimpacted reference fish. Histopathological alterations showed clear differences dependent on the input source. Potential links between specific contaminant impact and functional and morphological disorders are discussed.

Environmental Genotoxicity Studies in Mussels and Fish from the Göteborg Area of the North Sea

The frequency of micronuclei (MN) was analysed in gill cells of 97 specimens of blue mussels (Mytilus edulis), in peripheral blood of 61 specimens of flounder (Platychthys flesus) and 40 specimens of wrasse (Symphodus melops), which were collected from seven study locations in the Goteborg area of the North Sea. The reference level of micronuclei was 0.04 MN/1,000 erythrocytes in flounder from the uncontaminated Nordre Alvsborg location and 0.05 MN/1,000 erythrocytes from the site at Fjallbacka. In mussels from the Stenungsund location, the reference level of micronuclei was 0.71 MN/1,000 cells. Ten times higher frequencies of micronuclei (0.45 MN/1,000 erythrocytes) were found in flounder collected from the contaminated zone at Jordhammarvik and eight times higher micronuclei levels (0.33 MN/1,000 erythrocytes) were recorded in fish from Nya Alvsborg, the inner part of the Goteborg harbour. In mussels, the highest response (a 2.8-fold increase compared to the reference level of MN) was established in spe...

Susceptibility of corkwing wrasse Symphodus melops, goldsinny wrasse Ctenolabrus rupestis, and Atlantic salmon Salmo salar smolt, to experimental challenge with Vibrio tapetis and Vibrio splendidus isolated from corkwing wrasse

The virulence of two Vibrio strains, previously isolated from diseased corkwing wrasse Symphodus melops and identified as V. tapetis and V. splendidus, to corkwing and goldsinny wrasse Ctenolabrus rupestris and to Atlantic salmon Salmo salar, was studied under laboratory conditions. Both bacteria were shown to be opportunistically pathogenic to corkwing wrasse, causing significantly higher mortality in the challenged groups than in the controls. Bacterial cultivation of kidney samples and re-isolation of V. tapetis and V. splendidus from most mortalities confirmed the two strains as the probable cause of mortality in the challenged groups. The control group also suffered relatively high mortality, but no specific pathogens that were suspected to be the main cause of death were isolated, other than a mixture of Vibrio spp. and, in the case of one individual, atypical Aeromonas salmonicida. Following injection challenge with both bacterial strains, no mortality was recorded in Atlantic salmon. In bath challenge trials with goldsinny wrasse, only slight mortality was observed in the challenged groups and the unchallenged control group. Bacterial examination showed that atypical Aeromonas salmonicida was the probable cause of death in both bath challenged and control groups of goldsinny wrasse, and no indication of infection by any Vibrio sp. was found.

Larval fish distribution in shallow coastal waters off North Western Iberia (NE Atlantic)

Monthly sampling for ichthyoplankton was conducted at three stations very near to the coast (near to shore shallow stations before the surf zone in direction to the shoreline) of Atlantic northwestern Portugal within contrasting localities: rocky shore, in front of a sandy beach receiving an estuary and a fishing port with heavy anthropogenic impact. Sampling was conducted from August 2002 to October 2003, always at full moon, at low tide and high tide in daylight hours, at the water column using a 48 cm diameter plankton net with 335 μm mesh. Analysis of the physico-chemical parameters pointed out the spatial (horizontal) homogeneity of the sampling area. Fish larvae from 41 taxa belonging to 17 families were identified; Blenniidae, Labridae, Ammodytidae, Clupeidae, Gobiidae, Soleidae and Gobiesocidae were the most representative during the study period. Parablennius gattorugine, Ammodytes tobianus, Symphodus melops, Sardina pilchardus, Lipophrys pholis and Coryphoblennius galerita were the most representative species (percentage contribution to total abundance). Peak abundance of fish eggs occurred during May, June and August 2003 and fish larvae occurred during May and July 2003 and August 2002 and 2003; there was a pronounced winter/early spring (March 2003) peak in larval abundance dominated by the small sandeel A. tobianus. This study identifies the occurrence of a conspicuous assemblage of larval fishes at very nearshore shallow environments of a variety of species with different adult habitats: the fish larvae assemblage was dominated by intertidal species. The present study has shown that temporal and spatial variations in the larval fish assemblage are related to environmental conditions and biological dynamics: the results suggest that abiotic conditions mediate biotic parameters, and that both abiotic and biotic characteristics regulated the larval fishes at very nearshore shallow environments.

Temporal and spatial distributions of larval fish assemblages in the Lima estuary (Portugal)

The Lima estuary (NW Portugal) is at the end of an international watershed, whose potential role as a spawning and nursery habitat for local fish populations has not been previously examined. To address this knowledge gap, fortnightly plankton surveys were conducted between April 2002 and April 2004. A total of 12,903 larvae, belonging to 20 families and 50 taxa were collected, with a mean abundance of 8 individuals per 100 m 3. Gobiidae was the most abundant family comprising 71% of the total catch, followed by Clupeidae with 12% of the total. The top six abundant taxa ( Pomatoschistus spp., Sardina pilchardus, Ammodytes tobianus, unidentified Clupeidae, Symphodus melops and Solea senegalensis) represented 91% of the total catch. Fish larvae showed a seasonal trend with abundances increasing during spring and summer. Diversity was generally low ( H′ = 0.65) with high dominance of very few taxa. Near the ocean, the larval fish assemblage was more diverse due to the presence of marine species. In the lower estuary Channel zone, abundance was lower than in the two upstream salt marsh zones (North and South zones) and no statistical differences in abundance or diversity values were found within the latter zones. ANOSIM results demonstrated seasonal differences in the species composition, mainly during the second winter period which was typified by a pelagic species A. tobianus. The community in the Channel zone was more diverse in comparison with the other zones, which were highly dominated by the most abundant species. The spatial and temporal trends of the most abundant species were typical for Iberian estuaries, with the exception of the low abundance of anchovy larvae and the unusually high numbers and frequency of S. pilchardus, usually mentioned as accidental in estuarine systems. Overall results suggest that the Lima estuary larval fish assemblage has a strong seasonality and affinity to the salt marsh zones. It seems that spawning seasonality controlled the presence of temporary estuarine residents, while environmental aspects controlled the general abundance trend of the resident species.

Occurrence of leeches (Hirudinea, Piscicolidae) on some marine fishes in Norway

Marine fish from several locations along the west coast of Norway were examined for leeches. Seven leech species were detected, Calliobdella nodulifera (Malm, 1863) from codfish, flatfish and a ray, Calliobdella lophii van Beneden & Hesse, 1863 from Lophius piscatorius L., Platybdella anarrhichae (Diesing, 1859) on Anarhichas lupus L., Platybdella quadrioculata Malm, 1863 on Symphodus melops (L.), Oceanobdella sp. on Zeugopterus punctatus (Bloch), Malmiana brunnea (Johansson, 1896) on Myoxocephalus scorpius (L.) and Malmiana bubali Srivastava, 1966 on Taurulus bubalis (Euphrasen). Calliobdella nodulifera infected unrelated hosts (euryxenic); the others appeared to be host specialists (oio- or stenoxenic). Details on the occurrence of Calliobdella nodulifera on Enchelyopus cimbrius (L.) during 1 year are given, which together with other observations suggest that its life cycle exceeds a year. A brief account on the morphology of “Platybdella” quadrioculata is provided, justifying its transfer to the genus Oceanobdella Caballero, 1956 as O. quadrioculata comb. nov. With the new additions, the Norwegian marine piscicolid fauna contain 12 species. At least ten additional species, known from adjoining areas, also probably occur along the Norwegian coast.

A short-review of brown ring disease, a vibriosis affecting clams,Ruditapes philippinarumandRuditapes decussatus

Brown ring disease (BRD) in Ruditapes philippinarum and Ruditapes decussatus is a shell disease caused by Vibrio tapetis. This disease has begun in 1987 in clams beds in Brittany (France) and then has spread along the European Atlantic coast. Since about fifteen years, research on BRD has progress a lot and the purpose of this review is to give a short description of BRD in clams in retrospect and in addition of recent and pertinent results. Diagnosis including isolation, biochemical, serological and molecular characterization has been developed to identify and detect the pathogen. Therefore, Vibrio tapetis-like strains has been detected in other host specie, in cultured fishes during mortalities, such as the wrasse, Symphodus melops in Norway and the halibut, Hippoglossus hippoglossus in Scotland. Mechanisms of host-pathogen interactions modulated by environmental factors were studied using in vivo bacterial challenge and in vitro bio-assays. According to these studies, adherence and cytotoxic factors have been yet identified as virulence factors; identification and characterization of virulence genes are in progress. Field and experimental studies confirmed the significant effect of temperature on the development of BRD and on clam defense related activities. A significant increase in temperature over 21 ◦ C may have a preventive effect on the development of the disease.

Underwater television observations of Serpula vermicularis (L.) reefs and associated mobile fauna in Loch Creran, Scotland

The behaviour of Serpula vermicularis reef worms and associated mobile fauna in Loch Creran, on the west coast of Scotland, was recorded for six weeks using underwater television. The reef worms were extended almost continually, generally only retracting when stimulated by the close proximity of a predator. The length of hiding time varied according to the species triggering the reaction. The mean length of time retracted was 21 s. Fauna associated with the reefs included the corkwing wrasse Crenilabrus melops, ballan wrasse Labrus bergylta, black squat lobster Galathea squamifera and velvet swimming crabs Necora puber. There were few significant associations between tidal cycle and the presence/activity of associated fauna but a number of fish and crustacean species appeared to be influenced by the diurnal cycle. Gobies were recorded during daylight only with peak observations around midday while wrasse, butterfish Pholis gunnellus and cod Gadus morhua were crepuscular. The dawn and dusk appearance of juvenile cod in large shoals is suggestive of a nightly migration.

The efficacy of a single intraperitoneal injection of oxolinic acid in the treatment of bacterial infections in goldsinny wrasse (Ctenolabrus rupestris) and corkwing wrasse (Symphodus melops) studied under field and laboratory conditions.

The efficacy of a single intraperitoneal injection of oxolinic acid to control an outbreak of atypical Aeromonas salmonicida infection in goldsinny wrasse (Ctenolabrus rupestris) and in the treatment of systemic vibriosis in corkwing wrasse (Symphodus melops) was examined. In addition a field study was performed to examine the effect of medication on the survival rate of goldsinny wrasse in Atlantic salmon cages. Four groups of wild caught goldsinny wrasse, each of 50 fish, were treated with an intraperitoneal injection of propylene glycol:saline (50:50) (control) or 50 mg/kg oxolinic acid at a concentration of 50 mg/mL. Three days after medication the fish in all groups were treated by an intraperitoneal injection of prednisolone acetate and an increase in seawater temperature from 9.0 to 11.5 degrees C. Cumulative mortalities were 18% in the two groups treated with oxolinic acid and 94 and 100% in the unmedicated control groups, giving a 'relative percentage survival' (RPS) value of 82%. A laboratory maintained population of originally wild caught corkwing wrasse experiencing high daily mortality was treated with oxolinic acid (50 mg/kg) or propylene glycol:saline (control). Cumulative mortalities were 84% (control) and 42% (oxolinic acid medicated group) giving an RPS value of 50%. In a field investigation using goldsinny wrasse approximately 30% were medicated with oxolinic acid (50 mg/kg) prior to stocking in cages with Atlantic salmon. In two of three cages the cumulative mortality was significantly lower (P = 0.025 and P < 0.001) in the medicated groups.

Characterization of strains of Vibrio splendidus and V. tapetis isolated from corkwing wrasse Symphodus melops suffering vibriosis.

Two vibrio bacteria pathogenic to the corkwing wrasse Symphodus melops were isolated. Vibriosis-inducing strain LP1 was isolated as the dominanting bacterium in kidney samples of dead and moribund wrasse from a population suffering vibriosis and high daily mortality in 1998 on the Norwegian west coast. The other vibriosis-inducing strain, LP2, was isolated from wrasse captured the following year. Re-infection experiments have confirmed that these strains cause vibriosis in corkwing wrasse. Both strains were typical vibrios sharing the traits of fermentative Gram-negative curved rods with motility and a positive oxidase reaction. Detailed biochemical and genetic characterisation revealed a close affiliation to known species of the marine environment. The first isolate, LP1, is a form of the ubiquitous seawater organism Vibrio splendidus, while the second isolate, LP2, is closely related to V. tapetis (previously only known as the brown ring disease agent in clams). Identification of the new wrasse pathogens V. splendidus LP1 and V. tapetis LP2 is facilitated by break points observed in this study.

Variation in Plasma Steroids and Reproductive Traits in Dimorphic Males of Corkwing Wrasse (Symphodus melops L.)

Differences in plasma steroid levels may reflect behavioral, gonadal, or morphological specializations that characterize different male reproductive phenotypes in teleost fishes. In the present study, we investigated whether differences in plasma steroid levels exist between two distinct male morphs shown in the corkwing wrasse Symphodus melops. In addition, we examined differences in male reproductive traits, including gonad mass, sperm motility, and sperm concentration, between the two male types. Possible associations between plasma steroid levels and reproductive traits were also investigated. The results indicated that males with typical male secondary sexual characters (territorial males) had higher plasma levels of 11-ketotestosterone (11kT) compared to smaller males with female secondary sexual characters (female mimics). The female mimics in turn had higher plasma levels of both testosterone (T) and 17β-estradiol. In addition, female mimics had relatively larger gonads and longer-lived sperm than territorial males. Relative gonad mass covaried significantly with the plasma levels of 11kT and T among the morphs, indicating that the relative gonad size correlates positively with the plasma level of these two steroids. However, there was no significant covariance between sperm traits and plasma steroids for the two male morphs. Hence, our results do not indicate any causal link between sperm quality and hormones.

Nest Building and Mating in Relation to Male Size in Corkwing Wrasse, Symphodus Melops

This study examines nest building and mating in relation to size in male corkwing wrasse, Symphodus melops, in the absence of male–male competition. We kept males of varying size together with mature females in tanks containing adequate materials and sites for nest construction. Our results indicate that the size of territorial males does not influence the ability to construct a nest. However, larger males initiated nest building earlier than smaller males and nest size correlated positively with male size. Furthermore, male size did not affect whether or not a nesting male succeeded in mating, indicating that male size was not essential for female acceptance (i.e. whether or not a female is willing to mate) of a male in the current situation.

Differential processing of yolk proteins during oocyte hydration in marine fishes (Labridae) that spawn benthic and pelagic eggs

The yolk proteins and free amino acids (FAA) were investigated during oocyte matura- tion in 4 species of Labridae with either benthic or pelagic eggs. Benthic eggs and oocytes of both types had a small FAA pool dominated by taurine, while pelagic eggs had a large and characteristic FAA pool with about equal amounts of indispensable and dispensable amino acids. Electrophoresis (SDS-PAGE) of the yolk proteins revealed the presence of high molecular weight yolk proteins in the oocytes of all 4 species. In 2 species with benthic eggs (Labrus bergylta, L. mixtus), no buildup of an FAA pool occurred (nmol µg -1 dry mass) during final maturation, and the yolk protein profile remained essentially unchanged. In a third species (Crenilabrus melops) with benthic eggs, partial degradation of a 112 kDa protein coincided with the buildup of a small FAA pool. In the species with pelagic eggs (Ctenolabrus rupestris), the buildup of a large FAA pool coincided with the disappear- ance of the 112 kDa yolk protein. The calculated yield of FAA by hydrolysis of the disappearing fraction of the 112 kDa yolk protein for both species matched well with the measured increase in the FAA pool. It is suggested that differential processing of the high molecular yolk proteins causes differences in teleost egg behaviour, benthic or pelagic.

Male dimorphism, sperm traits and immunology in the corkwing wrasse ( Symphodus melops L.)

The evolution of alternative male mating phenotypes inevitably involves variation in risk or intensity of sperm competition. Males that typically mate in disfavoured roles (i.e. sneakers/parasitic males) are predicted to experience higher magnitudes of sperm competition than males mating in favoured roles (preferred by females/exhibiting mate monopolisation). Here, we investigated whether two distinct male phenotypes in corkwing wrasse (Symphodus melops) differ with respect to adaptations for sperm competition in terms of sperm quantity, sperm motility and fertilising capacity. We found that males exhibiting female mimicry (disfavoured role) had both absolutely and relatively larger gonads than males with typical secondary sexual characters (favoured role). Moreover, a higher proportion of sperm from female mimics was motile 5 min after activation compared to sperm of territorial males. There were, however, no differences in sperm concentration or in in vitro fertilisation ability between the two male morphs. We also examined whether the male phenotypes differed in some immune parameters and if any such variation is associated with sperm traits. Current theories predict that ejaculate quality should be negatively correlated with the level of immunocompetence. Territorial males had relatively heavier spleens than female mimics, but whether this reflects differing immunocompetence is uncertain, since the spleen is also an important organ for erythrocyte storage. Furthermore, there were no differences in the densities of circulating lymphocytes or granulocytes, and the immune parameters did not covary with the sperm traits. Differences in immunocompetence are therefore unlikely to provide a major proximate explanation for variations in sperm traits in corkwing wrasse.

Evidence of uptake, biotransformation and DNA binding of polyaromatic hydrocarbons in Atlantic cod and corkwing wrasse caught in the vicinity of an aluminium works

Feral Atlantic cod ( Gadus morhua) and corkwing wrasse ( Symphodus melops) were investigated for polyaromatic hydrocarbon (PAH) contamination in the Karmsund strait, western Norway. This strait is highly contaminated with PAHs, and a main source is the chronic release of gas-scrubbing effluents from a local aluminium works. In both species, the level of biliary PAH metabolites and hepatic DNA adducts were higher in fish collected near the aluminium works. Interestingly, a significantly higher level of both biliary PAH metabolites and hepatic DNA adducts was found in corkwing wrasse as compared to cod, indicating a higher potential for genotoxic effects in this species. Hepatic cytochrome P4501A (CYP1A) in cod estimated by ethoxyresorufin- O-deethylase and an immunoassay technique (ELISA), seemed to be weakly induced at the contaminated sites. At the most contaminated site, skin ulcers and fin erosion were detected in about 70 and 45% of the cods, respectively. The data demonstrated that both cod and corkwing wrasse may be suitable target species for PAH pollution monitoring.

Absorption, tissue distribution and excretion of flumequine and oxolinic acid in corkwing wrasse (Symphodus melops) following a single intraperitoneal injection or bath treatment.

The pharmacokinetic properties of the antibacterial agents oxolinic acid and flumequine were studied in corkwing wrasse (Symphodus melops) after either intraperitoneal injection or bath treatment. Following intraperitoneal administration the peak plasma concentrations (Cmax) and the time to peak plasma concentrations (Tmax) were estimated to be 2.0 microg/mL and 12 h, respectively, for oxolinic acid and 2.6 microg/mL and 12 h, respectively, for flumequine. In muscle, Cmax and Tmax were estimated to 6.7 microg/g and 12 h, respectively, for oxolinic acid with corresponding values of 8.5 microg/g and 13 h, respectively, for flumequine. In liver, Cmax and Tmax were calculated to 7.0 microg/g and 12 h, respectively, for oxolinic and 12.2 microg/g and 11 h, respectively, for flumequine. Elimination half-lives (t1/2 beta) of 26, 24 and 29 h, respectively, for plasma, muscle and liver were calculated for flumequine. For oxolinic acid two distinct elimination phases were found and calculated to be 16 h (t1/2 beta) and 57 h (t1/2 gamma) in plasma, 15 and 59 h, respectively, in muscle and 20 and 72 h, respectively, in liver. Bath treatment using 150 mg/L of flumequine or 200 mg/L of oxolinic acid for 72 h resulted in flumequine concentrations of 1.0 microg/mL in plasma, 5.0 microg/g in muscle and 12.4 microg/g in liver. Corresponding values for oxolinic acid were 1.0 microg/g in plasma, 2.5 microg/g in muscle and 4.9 microg/g in liver.

The Efficacy of a Single Intraperitoneal Injection of Flumequine in the Treatment of Systemic Vibriosis in Corkwing Wrasse Symphodus melops

In this investigation, the efficacy of a single intraperitoneal injection of flumequine in the treatment of systemic vibriosis in corkwing wrasse Symphodus melops was examined. In a population of corkwing wrasse that was experiencing daily mortality, the dominant bacterium cultivated from kidney samples of dead fish was tentatively classified as a species of Vibrio resembling V. splendidus. The infected fish were randomly divided into two groups, each of 60 individuals. Group 1 was treated with flumequine (25 mg/kg of fish) administered by a single intraperitoneal injection, whereas the fish from group 2 were injected with saline. The mortality was observed daily for 21 d. The cumulative mortality was significantly (P < 0.005) higher in the control group (58%) compared with the medicated group (11%). Bacterial examination of kidneys from dead and surviving fish of both groups indicated the Vibrio to be quickly and effectively eliminated by the antibacterial agent.

Phenotypic variation between dimorphic males in corkwing wrasse

The age‐frequency distributions for sneaker males (individuals with female secondary sexual characters) and territorial males (individuals with typical male sexual characters) of corkwing wrasse Symphodus melops in a Swedish population did not differ. Otolith analysis revealed that sneaker males were smaller than their territorial male counterparts as juveniles, but there was no evidence of a clear‐cut threshold to determine the subsequent reproductive patterns. Thus, the expression of male morph in corkwing wrasse most likely is related to early growth pattern and the different morphs could be fixed for life. Adult sneaker males in general were smaller than territorial males of the same age. The relative gonad size and the abdomen in sneaker males were substantially enlarged compared with territorial males, but there was no difference in gonad water content between the two types of males. Sneaker males weighed less in relation to their length than territorial males. These results support theoretical sperm competition models which predict that the male morph with reduced attractiveness to females (i.e. sneakers) should invest more into spermatogenesis in order to achieve fertilization than the attractive male morph (i.e. territorial males).

Phenotypic variation between dimorphic males in corkwing wrasse

The age-frequency distributions for sneaker males (individuals with female secondary sexual characters) and territorial males (individuals with typical male sexual characters) of corkwing wrasse Symphodus melops in a Swedish population did not differ. Otolith analysis revealed that sneaker males were smaller than their territorial male counterparts as juveniles, but there was no evidence of a clear-cut threshold to determine the subsequent reproductive patterns. Thus, the expression of male morph in corkwing wrasse most likely is related to early growth pattern and the different morphs could be fixed for life. Adult sneaker males in general were smaller than territorial males of the same age. The relative gonad size and the abdomen in sneaker males were substantially enlarged compared with territorial males, but there was no difference in gonad water content between the two types of males. Sneaker males weighed less in relation to their length than territorial males. These results support theoretical sperm competition models which predict that the male morph with reduced attractiveness to females (i.e. sneakers) should invest more into spermatogenesis in order to achieve fertilization than the attractive male morph (i.e. territorial males).