Habitat specific growth rates and condition indices for the sympatric soles Solea solea (Linnaeus, 1758) and Solea senegalensis Kaup 1858, in the Tagus estuary, Portugal, based on otolith daily increments and RNA-DNA ratio

Habitat specific growth rates and condition indices were estimated for Solea solea and Solea senegalensis, in two nursery areas within the Tagus estuary, at the end of the estuarine colonization process, in 2005. While in the uppermost nursery area the two species of sole live in sympatry, in the lower nursery only S. senegalensis is present. Daily increments of left lapillar otoliths were used to estimate age (in days) and determine growth rates (mm per day). Condition indices were assessed through RNA‐DNA ratio in muscle samples. Growth rates were higher for S. senegalensis (0.970and 1.180 mm per day in nursery A and B, respectively) than for S. solea (0.767 mm per day in nursery A). Growth rates of S. senegalensis from the uppermost nursery area were lower when compared to those obtained for the other nursery. The RNA/DNA condition index followed the general trend given by the growth rate estimates, i.e. values were higher for S. senegalensis than for S. solea. However, no significant differences were detected in S. senegalensis from the two nurseries. Larger variations in salinity (10‰ amplitude in the uppermost nursery vs 0.2‰ in the lower nursery) and highest pollution loads may be important factors lowering the habitat quality of the uppermost nursery in comparison to the lower nursery. The use of growth rate estimates based on otolith readings and the RNA/DNA index as tools for habitat quality assessment was discussed.     

Genetic differences between wild and hatchery‐bred brown trout (Salmo trutta L.) in single nucleotide polymorphisms linked to selective traits

To study effects from natural selection acting on brown trout in a natural stream habitat compared with a hatchery environment, 3,781 single nucleotide polymorphism (SNP) markers were analyzed in three closely related groups of brown trout (Salmo trutta L.). Autumn (W/0+, n = 48) and consecutive spring (W/1+, n = 47) samples of brown trout individuals belonging to the same cohort and stream were retrieved using electrofishing. A third group (H/1+, n = 48) comprised hatchery‐reared individuals, bred from a mixture of wild parents of the strain of the two former groups and from a neighboring stream. Pairwise analysis of F_ST outliers and analysis under a hierarchical model by means of ARLEQUIN software detected 421 (10.8%) candidates of selection, before multitest correction. BAYESCAN software detected 10 candidate loci, all of which were included among the ARLEQUIN candidate loci. Body length was significantly different across genotypes at 10 candidate loci in the W/0+, at 34 candidate loci in the W/1+ and at 21 candidate loci in the H/1+ group. The W/1+ sample was tested for genotype‐specific body length at all loci, and significant differences were found in 10.6% of all loci, and of these, 14.2% had higher frequency of the largest genotype in the W/1+ sample than in W/0+. The corresponding proportion among the candidate loci of W/1+ was 22.7% with genotype‐specific body length, and 88.2% of these had increased frequency of the largest genotype from W/0+ to W/1+, indicating a linkage between these loci and traits affecting growth and survival under this stream's environmental conditions. Bayesian structuring of all loci, and of the noncandidate loci suggested two (K = 2), alternatively four clusters (K = 4). This differed from the candidate SNPs, which suggested only two clusters. In both cases, the hatchery fish dominated one cluster, and body length of W/1+ fish was positively correlated with membership of one cluster both from the K = 2 and the K = 4 structure. Our analysis demonstrates profound genetic differentiation that can be linked to differential selection on a fitness‐related trait (individual growth) in brown trout living under natural vs. hatchery conditions. Candidate SNP loci linked to genes affecting individual growth were identified and provide important inputs into future mapping of the genetic basis of brown trout body size selection.     

Identifying footprints of selection in stocked brown trout populations: a spatio‐temporal approach

Studies of interactions between farmed and wild salmonid fishes have suggested reduced fitness of farmed strains in the wild, but evidence for selection at the genic level is lacking. We studied three brown trout populations in Denmark which have been significantly admixed with stocked hatchery trout (19–64%), along with two hatchery strains used for stocking. The wild populations were represented by contemporary samples (2000–2006) and two of them by historical samples (1943–1956). We analysed 61 microsatellite loci, nine of which showed putative functional relationships [expressed sequence tag (EST)‐linked or quantitative trait loci]. F_ST‐based outlier tests provided support for diversifying selection at chromosome regions marked by three loci, two anonymous and one EST‐linked. Patterns of differentiation suggested that the loci were candidates for being under diversifying hitch‐hiking selection in hatchery vs. wild environments. Analysis of hatchery strain admixture proportions showed that in one wild population, two of the loci showed significantly lower admixture proportions than the putatively neutral loci, implying contemporary selection against alleles introduced by hatchery strain trout. In the most strongly admixed population, however, there was no evidence for selection, possibly because of immigration by stocked trout overcoming selection against hatchery‐derived alleles or supportive breeding practices allowing hatchery strain trout to escape natural selection. To our knowledge, this is the first study demonstrating footprints of selection in wild salmonid populations subject to spawning intrusion by farmed fish.     

Immunogenetic studies of segregation of protein specificities in backcross hybrids in columbidae

Antisera recognizing five specificities of plasma proteins inStreptopelia senegalensis were produced inS. risoria. Immunizations were made with serum ofS. senegalensis or with plasma from selected backcross hybrids obtained from mating toS. risoria hybrids from the cross ofS. senegalensis ×S. risoria. The genetic segregation in the first backcross generation of the presence or absence of reactivity with each antiserum could be explained by the assumption that single genes inS. senegalensis effected each protein specificity.     

Do stocked hatchery‐reared juveniles ecologically suppress wild juveniles in Salvelinus leucomaenis?

The dominancy of semi‐wild and hatchery‐reared white‐spotted charr Salvelinus leucomaenis juveniles was evaluated using pair‐wise enclosure tests and field stocking tests. The semi‐wild S. leucomaenis originated in a hatchery, being stocked into the test stream as eyed‐eggs. In the pair‐wise enclosure test, the semi‐wild S. leucomaenis dominated the hatchery S. leucomaenis that were of a similar standard length (L_S). The semi‐wild S. leucomaenis were subordinate to hatchery S. leucomaenis that were > 11% larger in L_S. In the field stocking test, the abundance and growth of semi‐wild S. leucomaenis was decreased in the presence of larger hatchery S. leucomaenis (14% larger L_S). Taken together, these results suggest that larger hatchery S. leucomaenis ecologically suppress the smaller semi‐wild S. leucomaenis. Salvelinus leucomaenis juveniles that are stocked with the intention of supplementing natural populations should be < 10% larger than their wild counterparts at the time of stocking to minimize their competitive advantage. The semi‐wild and hatchery S. leucomaenis used in both tests were genetically similar individuals, suggesting that the differences are due to the early rearing environment of either a natural stream or hatchery. The hatchery S. leucomaenis have lower levels of aggression as a result of selection in the hatchery rearing environment. Rearing in a natural stream from the eyed‐egg stage is likely to increase their lowered aggression.     

Stratification by CARD15 variant genotype in a genome-wide search for inflammatory bowel disease susceptibility loci

Previously we have conducted a genome-wide search for inflammatory bowel disease susceptibility loci in a large European cohort. Results from this study demonstrated suggestive evidence of linkage to loci at chromosomes 1q, 6p, and 10p and replicated linkages on chromosomes 12 and 16. Recently, NOD2/CARD15 on chromosome 16q12 has been found to be strongly associated with Crohn's disease. In order to determine if there are other loci in the genome that interact with the three associated functional variants in CARD15 (R702W, G908R, 1007fs), we have stratified our large inflammatory bowel disease genome scan cohort by dividing pedigrees into two groups stratified by CARD15 variant genotype. The two pedigree groups were analysed using non-parametric allele sharing methods. The group of pedigrees that contained one of the three CARD15 variants had two suggestive linkage results occurring in 6p (lod = 3.06 at D6S197, IBD phenotype) and 10p (lod=2.29 at D10S197, CD phenotype). In addition, at 16q12 where CARD15 is located, the original genome scan had a peak lod score of 2.18 at D16S415 (CD phenotype). The stratified pedigree cohort containing one of three CARD15 variants had a peak lod score of 0.90 at D16S415 (CD phenotype), accounting for approximately less than half of the genetic evidence for linkage at this locus. This result is in agreement with the existence of a substantial number of private variants at the NOD2/CARD15 locus. Interaction with NOD2/CARD15 needs to be considered in future gene identification efforts on chromosomes 6 and 10.     

Molecular Evidence for Introgression and Loss of Genetic Variability in Salmo (trutta) macrostigma as a Result of Massive Restocking of Apennine Populations (Northern and Central Italy)

We assessed structural gene variation (allozymes and mtDNA) of brown trout to evaluate the genetic variability of Apennine stream populations (Northern and Central Italy) and the possibility of introgression by alien genomes after massive restocking with hatchery strains (Atlantic stocks). Genetic variability within and between Apennine populations was extremely low in our samples. Only two allozyme loci were polymorphic and mean hetero-zygosity was also reduced compared to other brown trout populations. Allelic frequencies determined for both loci were similar to the ones detected in the corresponding hatchery spawners. The reduction or total absence of the Mediterranean nuclear (LDH-5) and mitochondrial (16S rDNA) diagnostic markers suggests the domestic origin of most populations, and the introgression effects carried out by non-native genomes. From a taxonomic point of view, a clear differentiation emerges among basins placed on opposite sides of the Apennine chain (Tyrrhenian and Adriatic regions). In particular, the presence of Mediterranean genotypes and haplotypes characterizing Salmo (trutta) macrostigma is sporadic along the eastern Apennine side, adding additional doubts on the original presence and wide distribution of this salmonid along the Adriatic side of the mountain chain. In spite of conservation programs devoted to preservation of local genetic characteristics of S. t. macrostigma, massive restocking practices with hatchery strains obtained by a few spawners is the major cause of significant `founder effect' and `inbreeding depression' even in Apennine regions.     

New polymorphic microsatellite markers in Pacific abalone Haliotis discus hannai and their application to genetic characterization of wild and aquaculture populations

Seven new microsatellite markers were developed for the Pacific abalone (Haliotis discus hannai, Haliotidae), and allelic variability was compared between a wild population and a hatchery population in Yeosu, Korea. All loci amplified readily and demonstrated allelic variability, with the number of alleles ranging from 6 to 15 in the wild population and from 3 to 12 in farmed populations. Average observed and expected heterozygosities were estimated at 0.65 and 0.77 in the hatchery samples, and 0.79 and 0.87 in the wild samples. These results indicated lower genetic variability in the hatchery population, as compared with the wild population and significant genetic differentiation between the wild population and the hatchery samples (F _ST=0.055, p<0.001). These microsatellite loci may be valuable for future population genetic studies and for tracking hatchery samples used in stock enhancement programs.     

A review of the culture potential of <Emphasis Type="Italic">Solea solea</Emphasis> and <Emphasis Type="Italic">S. senegalensis</Emphasis>

A number of scientific studies have investigated aspects of soles(Solea soleaandS. senegalensis) ecology, population genetics and biology in their natural environment, and the species have been extensively studied in captivity during the last decade. Studies on the genetic population structure of sole indicate that several distinct breeding populations exist within its distributional range in European waters. Recent studies suggest a phylogenetic relatedness ofS. soleaand S. senegalensis, being found as closest sister lineages in most reconstructions. However, studies on molecular genetics and morphological traits give diagnostic differences that consistently lead to their taxonomic separation at the specific rank. Studies show that sole spawn readily in captivity, and the buoyant, fertilized eggs are easily collected. Stocking density during maturation should be 1–1.5kg/m², and temperature should be kept above 16°C (S. senegalensis) or between 8 and 12°C (S. solea). In nature, the onset of spawning is related to a rise in temperature occurring during spring (March–June). Salinity should be kept constant around 33–35 and the fish reared under simulated natural photoperiod (LDN). In other cultured flatfish species, a change in the photoperiod is the key environmental signal used to manipulate and control maturation, but at present time there are no published work that verifies or contradicts this for either S. senegalensisor S. solea. Studies indicate that a mixture of inert and live food may increase the weaning success of sole fry, and this can be further enhanced by using attractants in the dry feed. Future experiments are needed to determine the ideal time to commence weaning and determine the minimum duration of this period. Studies on alternative feeding strategies are also required. The effect of temperature and photoperiod on juvenile growth has not been studied systematically in neither of the two species and the relative importance of a direct photoperiod effect on growth in sole therefore remains to be defined.     

Effect of temperature and salinity on the gastric evacuation of juvenile sole Solea solea and Solea senegalensis

The juveniles of Senegal sole, Solea senegalensis, Kaup 1858, and common sole, Solea solea (Linnaeus 1758) concentrate in estuarine and coastal nurseries of widely differing temperatures and salinities. Yet, little is known about the effect of these physiologically important variables on the gastric evacuation rates of these species. Gastric evacuation experiments were performed on juveniles of S. senegalensis and S. solea. Three temperatures were tested, 26, 20 and 14°C at a salinity of 35‰. A low salinity experiment was also carried out at 15‰, at 26°C. Experimental conditions intended to reflect conditions in estuarine and coastal nurseries where juveniles of these species spend their first years of life. The relation between stomach contents and time was best described by exponential regression models for both species. An analysis of covariance (ancova ) was performed in order to test differences in evacuation rate due to temperature and salinity (slope of evacuation time against stomach contents) for each species. While increasing temperature increased evacuation rates in both species (although not at 26°C in S. solea), the effect of low salinity differed among species, leading to a decrease in gastric evacuation rate in that of S. senegalensis and an increase in S. solea. Differences in gastric evacuation rate between species were related to its metabolic optimums and to its distribution in the nursery area where fish were captured. Implications for the habitat use of estuarine and coastal nurseries are discussed.     

Migration barriers protect indigenous brown trout (<Emphasis Type="Italic">Salmo trutta</Emphasis>) populations from introgression with stocked hatchery fish

Brown trout populations in the Belgian rivers Scheldt and Meuse have been intensively stocked in the past decades, often with material of uncertain origin. Moreover, the species habitat has become increasingly fragmented, preventing gene flow between neighboring populations. We assessed how this impacted genetic diversity and population structure by analyzing 12 wild populations (total n=309) and seven hatchery stocks (n=200) at the mitochondrial control region with SSCP and at 27 RAPD loci. Historical records indicate that brown trout from distant locations have been used to supplement hatchery stocks; nevertheless we detected non-Atlantic mitochondrial genomes in only one population of the Scheldt basin and in one hatchery. In general, the hatchery samples displayed a higher genetic diversity and differentiated less among each other (global F_ST(mtDNA)=0.311/F_ST(RAPD)=0.029) compared to the wild populations (global F_ST(mtDNA)=0.477/F_ST(RAPD)=0.204). This is due to frequent exchanges between hatcheries and regular supplementation from several indigenous populations. Gene pools present in most downstream sections from tributaries of the Meuse were similar to each other and to the hatchery samples, despite the presence of migration barriers. Assignment analyses indicated that the contribution of hatchery material to the upstream parts was limited or even completely absent in populations separated by a physical barrier. Intensive stocking and exchange between hatcheries has homogenized the downstream sections of the Meuse River, whereas the migration barriers preserved the indigenous upstream populations. As such, uncontrolled removal of barriers might result in an irreversible loss of the remnant indigenous gene pools.     

Analysis of gene associated tandem repeat markers in Atlantic salmon (<Emphasis Type="Italic">Salmo salar L.</Emphasis>) populations: implications for restoration and conservation in the Baltic Sea

Patterns of genetic diversity and differentiation among five wild and four hatchery populations of Atlantic salmon in the Baltic Sea were assessed based on eight assumedly neutral microsatellite loci and six gene-associated markers, including four expressed sequence tag (EST) linked and two major histocompatibility complex (MHC) linked tandem repeat markers (micro- and mini-satellites). The coalescent simulations based on the method of Beaumont and Nichols (1996, Proc. R. Soc. Lond. Ser. B – Biol. Sci., 263, 1619–1626) indicated that two loci (MHCIIα and Ssa171, with the lowest and highest overall F_ST estimates, respectively) exhibited significant departures (P<0.05) from the neutral expectations. Another coalescent-based test for selective neutrality (Vitalis et al. 2001, Genetics, 158, 1811–1823) further supported the outlier status of the Ssa171 microsatellite locus but not of the MHCIIα linked minisatellite. In addition, actin related protein linked microsatellite locus was identified with this test as an outlier in six pairwise population comparisons. All genetic diversity estimates revealed more genetic variation in hatchery stocks than in the small wild salmon populations from the Gulf of Finland. However, the wild populations possessed alleles at gene-associated markers (e.g. MHCI and IGF) not found in the hatchery stocks, which together with moderate genetic differentiation and distinctive environmental conditions justifies the special conservation measures for the last remaining native salmon populations in the Gulf of Finland.     

Microsatellite DNA markers for kinship analysis and genetic mapping in red drum,Sciaenops ocellatus (Sciaenidae,Teleostei)

Thirty‐eight nuclear‐encoded microsatellites were isolated from the marine fish Sciaenops ocellatus (red drum). The species is of economic importance in the southeastern United States, and declines in abundance have led to augmentation of the ‘wild’ fishery with hatchery‐raised fingerlings. The microsatellites will be useful for studies designed to assess larval/juvenile recruitment of hatchery‐raised individuals at varying spatial and temporal scales and for assessment of genetic components contributing to variation in performance and survival of hatchery‐produced fingerlings in the wild. The microsatellites also will prove useful as ‘anchor’ loci in constructing a genetic map.     

Species-specific primers discriminate schistosome intermediate hosts: unambiguous PCR diagnosis of Bulinus forskalii group taxa (Gastropoda: Planorbidae)

The morphological definition of taxa has proved difficult within the Bulinus forskalii group, which includes intermediate hosts of medically important Schistosoma species in West Africa. Although B. forskalii and B. senegalensis transmit different schistosome species they are conchologically similar and their distributions overlap. Randomly amplified polymorphic DNA (RAPD) allows differentiation of sibling species in the genus Bulinus, but RAPDs are difficult to standardize, impairing their value as a taxonomic tool. Hence, RAPD products diagnostic for either B. senegalensis or B. forskalii from West Africa were cloned, sequenced and a panel of species-specific primers designed. Sequencing of RAPD products identified a homology in two apparently independent RAPD loci, a problem where RAPDs are indiscriminately scored for phylogenetic analyses. Specificity of primers was confirmed by widespread sampling throughout each species' range. This approach produced a simple, robust, unambiguous PCR-based species identification strategy for this difficult group.     

Morphological and Biochemical Correlates in the Characterization of Sarcocystis spp.1

ABSTRACT. Isoenzyme electrophoretic techniques were applied to the characterization of seven Sarcocystis spp. that had been identified by conventional morphological studies. Cystozoites were harvested from macroscopic cysts from sheep, cattle, and mice and from microscopic cysts from sheep, cattle, and goats. Soluble cystozoite extracts were subjected to cellulose acetate gel electrophoresis and characterized at 15 of the 39 enzyme loci examined. Genetic relationships among isolates were examined by simple phenetic clustering. Two different morphological types of macroscopic cysts from sheep, identified as S. gigantea (syn. S. ovifelis) and S. medusiformis, consistently differed at 40% of the loci examined. Such genetic divergence confirms their separate morphotypic classification. Both differed from microscopic cyst isolates from sheep at 87% of the loci examined; however, two different morphotypes of microscopic cysts were found in the sheep sampled (thick-walled and thin-walled cysts). Until sufficient numbers of each type can be isolated and examined separately, both were regarded as belonging to the species S. tenella (syn. S. ovicanis). Macroscopic and microscopic cysts from cattle consistently differed at 80% of the loci thereby supporting their separate classification as S. hirsuta (syn. S. bovifelis) and S. cruzi (syn. S. bovicanis), respectively. Isolates from goats (microscopic cysts identified as S. capracanis) differed from S. tenella and S. cruzi at 20% and 47% of the loci, respectively. All macroscopic cyst isolates from the various host animal species (including S. muris from mice) differed from each other at nearly all loci. Isoenzyme electrophoretic techniques therefore provided genetic evidence supporting the classification of these various Sarcocystis spp. by their morphological characteristics.     

Laboratory assessment of the potential of Paranosema locustae to control immature stages of Schistocerca gregaria and Oedaleus senegalensis and vertical transmission of the pathogen in host populations

We tested the effects of Paranosema locustae spores in wheat bran formulation on the immature stages of Schistocerca gregaria and Oedaleus senegalensis under laboratory conditions. Younger instars were the most sensitive to the pathogen. While 100% infection was recorded in younger instar nymphs, older instars were less sensitive, with 16–27% of the inoculated nymphs remaining uninfected at the end of the experiment. Mortality of each instar increased with increased spore concentration. Immature survival time was significantly reduced by the pathogen and none of the nymphs inoculated as first, second, and third instar nymphs developed to adulthood (6–30% and 55–74% of nymphs inoculated as fourth and fifth instar, respectively). Sublethal effects such as delayed host growth, reduced host size, and abnormal wing and leg development (37% of emerging adults) were noted. Almost half the infected adults showed morphological abnormalities at emergence. Moreover, infection in S. gregaria and O. senegalensis by P. locustae did not affect female oviposition. However, 60% of S. gregaria and 52% of O. senegalensis progeny clearly showed infection by P. locustae with infection intensity of 1.08±0.27×10¹ and 1.19±0.32×10² spores/nymph, respectively. In view of the mortality rates, immature survival, host growth, and abnormal development in the P. locustae treatments, and the high prevalence of the pathogen in offspring from infected parents, it can be expected that the reduction in the impact of the two acridid species in the field will be considerable.     

Effects of acute handling stress on cerebral monoaminergic neurotransmitters in juvenile Senegalese sole Solea senegalensis

Juvenile Senegalese sole Solea senegalensis were subjected for short periods to two different types of handling‐related stress: air exposure stress and net handling stress. The S. senegalensis were sacrificed 2 and 24 h after the stress events and the levels of serotonin (5‐HT), noradrenaline (NA), dopamine (DA) and their respective major metabolites, 5‐hydroxyindoleacetic acid (5‐HIAA), 3‐methoxy‐4‐hydroxyphenylglycol (MHPG) and 3,4‐dihydroxyphenylacetic acid (DOPAC), were measured in three brain regions (telencephalon, hypothalamus and optic tectum) and compared with those in control, non‐stressed S. senegalensis. Neither type of stress caused any significant alteration of serotoninergic activity (5‐HIAA:5‐HT ratio) or NA levels. Dopaminergic activity (DOPAC:DA ratio) was lower in stressed fish in all of the brain regions studied. For both air exposure stress and net handling stress, DA levels were significantly higher (P < 0·05) than in the control S. senegalensis. In addition, the higher DA levels after net handling stress were always significantly higher (P < 0·05) than those observed after acute air exposure stress, except in the telencephalon after 24 h. The significantly lower DOPAC:DA ratio (P < 0·05) in all of the brain regions studied was only observed in response to net handling stress.     

Toxic effect of heavy metals on ovarian deformities,apoptotic changes,oxidative stress,and steroid hormones in rainbow trout

BackgroundAs is well known, the pollution in the aquatic environment in which fish grow has a direct impact on aquaculture practices. Pollution in aquatic systems because of multiple adverse effects on fish metabolic processes, especially the reproductive systems.AimThe goal of this study was to assess the severity of pollution impact in two different hatcheries, Verinag hatchery, Site 1 (S1) and Panzath hatchery, Site 2 (S2) in Anantnag region, using histopathological, ultrastructural, oxidative stress, genotoxic, and hormonal analysis in rainbow trout gonad (ovary).M&M: Fish were collected between May 2018 and April 2019 from two locations, Verinag hatchery (S1) and Panzath hatchery (S2), which were affected by heavy metals.ResultsThe histological and ultrastructural examination of rainbow trout ovaries from the Verinag hatchery (S1) revealed normal structure in growing oocytes in rainbow trout at various stages based on morphological features while the fish ovaries in the Panzath hatchery (S2) showed various deformities and irregularly shaped oocytes. The surfaces of some of these oocytes were wrinkled, rough, or distorted. Apoptotic studies revealed that the frequency of apoptotic cells collected from S2 water was significantly increased in ovarian cells (P < 0.05). The activity of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were found to be increased in fish collected from S1 but decreased in fish collected from S2. In S2 caught fish, malondialdehyde (MDA) levels were found to increase gradually, and the degree of heavy metal stress was positively correlated (p < 0.05). The comet assay was used to determine the induction of DNA damage in ovarian cells. The induction of DNA damage was found to be significantly higher (p < 0.05) in S2 fish specimens compared to fish from S1. On comparing the DNA damage of the rainbow trout from the two sampling sites, it was revealed that the fish is much more sensitive to aquatic contaminants. Regarding steroid hormones, higher levels of progesterone and estrogen were reported in the fish samples collected from S1 as compared to S2 captured fish.ConclusionIn conclusion, the comparative study of fish from two different sites viz. Verinag hatchery (S1) and Panzath hatchery (S2) revealed that S2 sampled fish suffered more heavy metal damage, including cellular deformities, apoptosis, oxidative damage, and altered steroid hormones.     

Isolation and characterization of polymorphic microsatellite DNA markers in two shrew species,Sorex unguiculatus and S. caecutiens

We isolated six microsatellite markers from the partial genomic libraries of two Sorex shrews, S. unguiculatus and S. caecutiens, and examined their allelic variation. All loci showed high allelic variation ranging from 15 to 19 alleles and all but one locus conformed to Hardy–Weinberg expectations in the species where the loci were isolated. Cross-species amplifications showed that all primers derived from S. unguiculatus were useful for S. caecutiens, while among primer sets derived from S. caecutiens only one was useful for S. unguiculatus. Accordingly, at least five microsatellite markers were useful in S. caecutiens and three in S. unguiculatus.     

Heterochromatin and rDNA patterns in Solanum species of the Morelloid and Dulcamaroid clades (Solanaceae)

The heterochromatin distribution and the position of 18-5.8-26S, and 5S rDNA loci were determined in 13 species of Solanum of the Morelloid and Dulcamaroid clades. The CMA/DAPI staining and FISH were employed. Two types of constitutive heterochromatin were determined: CMA⁺/DAPI^? associated to NOR and CMA⁺/DAPI^? distributed as terminal bands. In the Morelloid clade, CMA⁺/DAPI^? bands were found in five species while in the Dulcamaroid clade, only S. angustifidum presented this feature. In the Morelloid clade, two to four 18-5.8-26S rDNA loci occupied terminal positions and two rDNA 5S loci were found with variable positions (terminal, intercalary, and centromeric). In the Dulcamaroid clade, two terminal 18-5.8-26S rDNA loci were detected with the exception of S. salicifolium which possessed four such loci and two to four 5S rDNA loci. Solanum crispum is the only species possessing the 5S in synteny with 18-5.8-26S rDNA loci. Karyotype features chromosome banding pattern as well as the location of ribosomal genes which varied among the species, reflecting the chromosome differentiation and evolutionary divergence. The findings obtained contributed to the development of tools that can be used for establishing chromosomic homeologies among species and hence to clarify their taxonomic relationships.