Morphological variation among populations of Aphanius fasciatus Nardo, 1827 (Teleostei, Cyprinodontidae) from the Mediterranean

The amount of osteological variation among 11 Italian killifish Aphanius fasciatus populations was examined by the univariate and multivariate analysis of 40 morphometric and meristic variables of the skull and vertebral column. Populations were sampled in three geographically distinct areas (the Adriatic, Sardinia and Sicily). The statistical analysis confirmed that several populations were well differentiated. In particular, discriminant analysis revealed a strong discriminating power of the morphometric variables. Morphometrics of the vertebrae, bony elements of the pharyngeal jaws, supraoccipital and parasphenoid were the most important in discriminating populations. The dendrogram obtained by UPGMA cluster analysis shows the separation of the south-eastern Sicilian populations, that of the Sardinian populations and that of the central-northern Sicilian plus Adriatic populations, as well as the isolation of the Sicilian population from Pantano Viruca and of the Sardinian populations from Pauli Figu from all the others. The significance of the observed differentiation pattern is discussed.     

Ontogenetic pattern,morphological sexual and side dimorphism in the saccular otolith of a scaleless killifish Aphanius furcatus (Teleostei: Aphaniidae)

The relationships between ontogenetic and sexual factors with otolith morphology are poorly studied in killifish genus Aphanius Nardo, 1827. We studied the ontogeny, morphological sexual and side dimorphism in the saccular otolith of a scaleless killifish Aphanius furcatus inhabiting a high‐salinity environment in Southern Iran. The results highlighted growth‐dependent variability in the otolith of A. furcatus, which might be useful to define its certain developmental stages. We underlined those characters that were consistent during fish development, which could be valuable to identify the species. Considering the ontogenetic pattern of otolith in A. furcatus, it was found that the otoliths of young individuals have largely been influenced by sexes and that it was reduced during the fish development. Morphological side dimorphism in the otolith of A. furcatus tends to be increased during fish development. The outcomes of this study are essential since they have provided new information on ontogenetic changes and the sexual and side dimorphism of Aphanius otolith.     

Genetic structure of the killifish Aphanius fasciatus, Nardo 1827 (Teleostei, Cyprinodontidae), results of mitochondrial DNA analysis

Aphanius fasciatus is a cyprinodont distributed in the salty coastal water of the central and eastern Mediterranean Sea and occasionally in internal fresh water. In this work, the authors have investigated the genetic structure of eight populations of the killifish A. fasciatus from Sardinia and Sicily. The comparison of the mtDNA control region of 237 individuals revealed a total of 49 haplotypes. Several unique haplotypes were present in each population, and no common haplotype was found among Sicilian and Sardinian populations. Almost all Sardinian populations shared a common haplotype, and indeed the four Sicilian populations examined did not share any as determined by the parsimony network analysis. The analysis of molecular variance showed that the percentage of variation among populations is much higher than within each population of A. fasciatus . The overall F _ST value is very high (0·78) and supports an extensive genetic structure of the populations. The observed genetic differentiations of A. fasciatus populations were discussed taking into account the palaeogeographic and palaeoclimatic events that interested the Mediterranean area from Miocenic to Pleistocenic age. The results provide new insight into the knowledge of the pattern of genetic structure and of evolutionary processes occurring in this species.     

Geographical differentiation of Aphanius dispar (Teleostei: Cyprinodontidae) from Southern Iran

The Arabian cyprinodontid Aphanius dispar (Rüppell, 1829) is known to show considerable morphological variation. It has remained unknown, however, whether this variation is a result of environmental differences or allopatric divergence owing to geographical isolation. In this study, 11 populations of A. dispar from three geographically separated basins were analysed, that is, the Makran Basin (I, one river system), the Hormuzgan Basin (II, five rivers and three hot springs) and the Helleh Basin (III, two hot springs) in southern Iran. Statistical analyses do not indicate significant differences between the fishes from river and hot spring habitats (T‐test, p < 0.05), which is also supported by the Canonical Discriminant Analysis (CDA). Nevertheless, morphometric and meristic characters of the fishes, as well as otolith morphology and morphometry, demonstrate that six phenotypic characters discriminate the A. dispar populations of the three basins, that is, (1) predorsal distance (Prdd.SL), (2) head length (HL.SL), (3) pelvic fin length (Lplf.SL), (4) number of pelvic fin rays, as well as relative length of both the (5) medial part and (6) rostrum of the otolith. In addition, these characters display a consistent pattern of variation, thus providing support for the assumption that the phenotypically different A. dispar populations are a result of geographical isolation and not related to environmental differences. It is likely that the geological history of the drainage systems caused isolation event(s) that may date back to the Pleistocene (1.8 million years before present). The high phenotypic differences might suggest that the A. dispar populations from the three studied basins represent separate subspecies or even species.     

Responses of the killifish (Aphanius dispar) to long-term exposure to elevated temperatures: growth,survival and microstructure of gill and heart tissues

Some fish species, such as killifish, that normally inhabit temperate water environments are also found in extreme thermal environments such as thermal springs. The extent of the adaptations involved is not known. In the present laboratory study, we exposed killifish (Aphanius dispar) acclimated to a normal thermal environment to elevated temperatures (37–40 °C) in which related killifish species live permanently. Our objective was to determine whether there is evidence that killifish have heat-shock characteristics that make permanent adaptation likely. The fish was exposed to this temperature for a period of 44 days and then compared with control fish kept at their normal temperature (24 °C) with respect to growth, survival and histopathology of gill and heart tissues. At the end of the experimental time, the percentage of body weight gain and specific growth rate were significantly lower in fish kept in thermal stress compared with the control group. Feed conversion ratio (FCR) was also significantly affected by water temperature, so that during thermal stress the values of FCR were negative. Fish condition (Condition factor: CF) did not differ significantly between both groups at the end of the experiment. On days 11 and 33, however, CF was significantly lower in the thermal stress group. The gill showed blood congestion in primary lamellae and shortened secondary lamellae in fish kept at 37–40 °C. No specific alterations were found in the cardiac tissue of fish kept in thermal stress conditions. Under thermal stress, 40% of fish survived until the end of the experiment. A preliminary conclusion drawn from this work is that A. dispar, which lives at normal temperatures, shows evidence of adaptability to elevated temperatures that could be a factor in the ability of killifish to adapt permanently over time to thermal environment.     

Geographic variation in Austrolebias luteoflamulatus Vaz‐Ferreira, Sierra & Scaglia (Cyprinodontiformes, Rivulidae)

The morphological variation in the annual killifish Austrolebias luteoflamulatus was studied using meristic and pigmentation characters analysed with univariate and multivariate analysis, and morphometric characters analysed with a geometric morphometry approach (thin plate spline). The results showed that morphological variation in this species was associated with a geographical pattern. In the case of meristic and pigmentation characters, the strongest distinction occured in males, while females were more similar among different drainages. Males from Rocha (eastern Uruguay) exhibited the greatest differences from the others. The number of dorsal fin rays and bars on the flanks were the characters that contributed most to this segregation. When morphometrics were considered, the three populations analysed could be distinguished well from each other in both sexes. Some of the characters that contributed to this pattern were dorsal fin origin, position of eyes and length of snout. Since pigmentation characteristics and some characters of the dorsal fin in males were involved in the differences observed, sexual selection could have played an important role in the differentiation of populations of this species. Since Austrolebias luteoflamulatus has a restricted geographical distribution, conservation of this species should include consideration of these populations individually since they might represent different evolutionary units.     

The scale characteristics of two Aphanius species from southern Iran (Teleostei: Aphaniidae)

Fish scales are morphologically diverse among species, within species, and even among individuals. The present study investigated macro- and micromorphology of the scales between two closely related Aphanius species in southern Iran including Aphanius hormuzensis, collected from Gotab/Mehran River and A. stoliczkanus, from Howba hot sulphuric spring. The results indicated clear interspecific differences in scale macromorphology and microstructures. The scales of A. hormuzensis are characterised by a rounded shape, 7–9 (mean 8.0±0.83) primary radii, and a relatively large focus with oblong shape that positioned centrally on the scale, while the scales of A. stoliczkanus are characterised by a polygonal shape with few spines in posterior region, 8–11 (mean 10.0±0.78) primary radii and a small rounded focus that positioned posterocentraly on the scale. These species are morphologically close but clearly distinguished by their scale morphology and habitat requirements (e.g. water depth and food availability). Therefore, it can be assumed that clear differences in their scale morphology have been resulted by the combination of both genetic and environmental factors.     

Isolation and characterization of eight polymorphic microsatellite markers in Aphanius fasciatus (Teleostei: Cyprinodontidae)

The first set of eight polymorphic microsatellites markers was successfully optimized from a partial genomic library enriched for an AC motif of the killifish Aphanius fasciatus Nardo, 1827 (Teleostei, Cyprinodontidae), a brackish‐water fish widely distributed along the coasts of the Mediterranean Sea. The developed loci proved to be polymorphic, displaying from two to six alleles per locus with expected heterozygosity ranging from 0.277 to 0.752. All loci were under Hardy–Weinberg equilibrium, except Af8 that showed significant heterozygotes deficiency, and there was no evidence of linkage associations between any pair of loci. Markers reported here will be potentially useful in monitoring of genetic population structure in this species.     

Diversity and structure of fragmented populations of a threatened endemic cyprinodontid (Aphanius sophiae) inferred from genetics and otolith morphology: Implications for conservation and management

The assessment of population structure and genetic diversity is crucial for the management and conservation of threatened species. Natural and artificial barriers to dispersal (i.e., gene flow) increase populations’ differentiation and isolation by reducing genetic exchange and diversity. Freshwater ecosystems are highly fragmented because of human activities. Threatened species with small population sizes are more sensitive to habitat fragmentation effects. Here, we investigate the genetic population structure and gene flow among seven populations of Aphanius sophiae in the Kor Basin by using sequences of the complete Cyt b gene and otolith morphometry. The Cyt b gene showed low level of genetic variation, only 4.12% of the identified sites were variable, and 2.42% were parsimony informative. Overall, haplotype diversity was low to moderate and nucleotide diversity was low to extremely low. Fish populations exhibited high levels of genetic differentiation, suggesting limited gene flow among them. These differences were obtained not only among geographically distant populations, but also among neighboring localities. Genetic population structure was supported by the AMOVA analysis and by the haplotype network (only one of 21 haplotypes were shared by two localities). Otolith morphometric analysis was in agreement with genetic results, the two most distant and isolated populations were clearly separated, and genetically close populations showed less differences in morphometry. A significant pattern of isolation by distance was also detected among A. sophiae populations, with genetic distance more correlated with hydrological distance than with geographic distance. Results suggested that limited gene flow due to habitat fragmentation is an important factor contributing to genetic structuring and to the loss of genetic variation of A. sophiae populations. Aphanius sophiae population structure seems to be the result of habitat fragmentation and water pollution, but other factors such as introduced species should be considered. Given the high degree of genetic structuring, the definition of conservation groups is of particular importance for A. sophiae, which should be considered endangered according to the IUCN criteria. Conservation plans must recognize the genetic independence of populations and manage separately preventing the loss of locally adapted genotypes.     

Papyrus swamps, hypoxia, and faunal diversification: variation among populations of Barbus neumayeri

To test whether patches of papyrus swamp contribute to diversification of populations of non-air-breathing fishes, the gill morphology of Barbus neumayeri was compared between a papyrus swamp and several tributaries which differed in oxygen regime. Total gill filament length differed among sites and was negatively related to dissolved oxygen availability, supporting strong selection pressure for low-oxygen tolerance in the swamp interior. Among recaptures of marked B. neumayeri over a 4·5-year period among the focal swamp and connected stream and river sites, 93% of fish were recovered at the site of capture. Some of the individuals that moved crossed physicochemical gradients and traversed long distances within the swamp/stream system. This movement rate would theoretically be sufficient to homogenize gene frequencies among populations. However, randomly amplified polymorphic DNA (RAPD) markers indicated significant genetic differentiation among sites and no relationship between genetic differences and geographical distances among sites suggesting habitat-specific selection pressures on dispersers, rather than insufficient dispersal.     

Effects of pesticide temephos on the gills of Aphanius dispar (Pisces: Cyprinodontidae)

The changes in the behavior of Aphanius dispar, a cyprinodont fish when exposed to different concentrations of temephos, an organophosphorus pesticide widely used to control mosquito larvae in aquatic habitats are noted. The histopathology of gills exposed to temephos is studied in detail using light microscopy (LM), transmission electron microscopy (TEM) and scanning electron microscopy (SEM). The gill lesions were seen in all exposed concentrations of 1, 5, 10 and 25 ppm and the magnitude of the damage increased with the increase in concentrations. The most adverse effects of temephos are the destruction of cartilage and the large-scale cell death. Other effects included the degradation of chloride cells (CCs), desquamation, epithelial lifting, hypertrophy and lamellar fusion. The histological changes of microridges did not follow a consistent pattern in exposed concentrations. However, lamellar curling and collapsing was common in all exposed fish.     

基于形态分析探究鳞头犬牙南极鱼各生活史阶段耳石形态变化

鳞头犬牙南极鱼为南极最为重要的底层鱼类,具有重要的生态作用和极高的商业价值,鱼类耳石形态随着生活史过程的推移有所差异,故耳石形态分析可用于推断鳞头犬牙南极鱼的生活史过程。本研究利用采集自南极罗斯海、阿蒙森海、威德尔海及拉扎列夫海120尾分属4个生活史阶段的鳞头犬牙南极鱼耳石,结合传统测量分析与椭圆傅里叶分析两种形态学研究方法,对其各生活史阶段耳石形态的差异进行研究。结果表明: 鳞头犬牙南极鱼各生活史阶段耳石形态存在差异,随着生活史过程的推移,耳石的变化趋势为整体上由轮廓平滑、复杂度较低向轮廓曲折、高度复杂化转变,且纵轴方向增长速度低于横轴方向,翼叶等特征部位变化显著。针对耳石形态的线性判别分析结果显示,椭圆傅里叶分析判别值较高,为85.4%,而传统测量分析判别值较低,为71.9%,表明相较于传统测量分析,椭圆傅里叶分析更具优势。     

Morphological variation in the polyps of the scleractinian coral Favia speciosa (Dana) around Singapore

A photographic technique was used to examine morphological differences in the living polyps of Favia speciosa sampled from three sites around Singapore. Eight characters were measured, seven of which differed significantly between the three study sites. Sedimentation rates and character size were much higher at the site closest to the mainland than at the two sites further from shore. Land reclamation and dredging contribute to high sediment rates in Singapore waters; these rates decrease with increasing distance from shore. Large polyps close to the main island of Singapore are possibly a plastic, or selected for, response to high levels of sediment.     

Morphological variation in the palaeonemertean Tubulanus annulatus (Montagu, 1804)

Previous studies on the palaeonemertean Tubulanus annulatus have hinted on possible geographic variation, or even a species complex. This study describes the variation in morphology, focusing on the characters that have been supposed to vary geographically. Specimens from the Swedish west coast (Tjärnö and Kristineberg), Scottish west coast (Millport), and the Mediterranean (Naples and Split) were included in the study. It is concluded that the pattern of variation in the studied characters do not correspond to geography and most of the character states previously thought to vary geographically are in fact found within one and the same population.     

Intraspecific variation in the direction and degree of sex-biased dispersal among sea-snake populations

Higher rates of dispersal in one sex than the other are widespread, and often attributed to the genetic advantages of reduced inbreeding. The direction of sex-biased dispersal shows strong phylogenetic conservatism (e.g. males disperse more than females in most mammals, but the reverse is true in most birds). By contrast, our genetic data reveal strong inter-population variation in the relative dispersal rates of two species of sea snakes (Laticauda saintgironsi and L. laticaudata) in the Noumea Lagoon of New Caledonia. Assignment methods using microsatellite data identified parallel variation in sex-specific dispersal in both species: dispersal was female-biased in the north-west of the sampling area (in islands far from the main island), but male-biased in the south-east (in islands closer to the main island). This flexibility may reflect sex differences in diets, with spatial variation in sex-specific resources generating spatial variation in sex-specific dispersal distances.     

Incidence of spinal deformities in natural populations of Aphanius fasciatus Nardo, 1827 from the Gulf of Gabes, Tunisia

Spinal deformities in Aphanius fasciatus were observed in fish collected from polluted (S1) and non-polluted (S2) areas in the Gulf of Gabes in Tunisia. Kyphosis, scoliosis and lordosis were detected and these basic deformities frequently co-occur in varying degrees of severity. A total of 612 specimens of A. fasciatus were caught and 54 were found to be deformed, which represents 8.82%. Deformed fish were eight times more frequent in S1 than in S2, possibly indicating a relationship between pollution levels and skeletal malformation. In both sexes, variation of deformities incidence with length was very similar. The highest occurrence of deformities was observed in fish with total length <25 mm and decreasing thereafter with fish length. The possible causes for such deformities as well as the suitability of this kind of study for environmental monitoring are discussed.     

A preliminary investigation of allozyme genetic variation and population geographical structure in Aphanius fasciatus from Italian brackish-water habitats

A total of 150 individuals from Aphanius fasciatus from coastal brackish-water habitats was analysed by allozyme electrophoresis to collect data on its genetic variation. From 22 enzymes, 43 putative enzyme-coding loci were resolved, 12 of which were polymorphic at P_0·99 level. Only one of the 31 probability tests showed a significant departure from the Hardy-Weinberg equilibrium. Aphanius fasciatus showed low levels of genetic polymorphism, with expected heterozygosity values ranging from 0·027 (S.E.=0·013) to 0·064 ( s.e. =0·023). Nei's genetic distances between populations ranged from 0·002 to 0·042. Weir & Cockerham F -statistics showed high levels of genetic heterogeneity among populations (jackknifed θ=0·302, s.e. =0·045) and estimates of N _m were < 1, indicating restricted gene flow. Significant positive correlation between genetic distance and geographical distance matrices, detected by Mantel's test ( g =1·941; P 0·001), is consistent with the prediction that the species is genetically structured by isolation-by-distance.