Histological Aspects of the Early Development of the Digestive System of Burbot Lota lota L. (Lotidae, Gadiformes).

The ontogeny of the digestive tract was studied histologically in burbot, Lota lota L., from hatching to 42 days post-hatch (dph). At hatching, the digestive tract consisted of a straight tube with discernible digestive accessory glands (the liver and the pancreas) dorsally attached to the yolk sac. Most of the yolk sac reserves were consumed during the first 12 days and were completely depleted by 17 dph. The first PAS-positive goblet cells appeared at 6 dph, dispersed within the epithelium of the oesophagus and increasing substantially in number and distribution as development progressed. At 12 dph, the first vacuoles (neutral lipids) appeared in the intestine, indicating the functional absorption of nutrients from food. Differentiation of gastric glands was first noticed at 17 dph and was extensive by 27 dph. L. lota larvae have a morphologically complete digestive tract by 32 dph. These findings on the development of the digestive system in L. lota may contribute to a better understanding of its ontogeny and can be useful for improvement of the larval rearing techniques of this promising species for freshwater aquaculture diversification.

Chromosomal characteristics and distribution of rDNA sequences in the brook trout Salvelinus fontinalis (Mitchill, 1814).

Brook trout Salvelinus fontinalis (Mitchill, 1814) chromosomes have been analyzed using conventional and molecular cytogenetic techniques enabling characteristics and chromosomal location of heterochromatin, nucleolus organizer regions (NORs), ribosomal RNA-encoding genes and telomeric DNA sequences. The C-banding and chromosome digestion with the restriction endonucleases demonstrated distribution and heterogeneity of the heterochromatin in the brook trout genome. DNA sequences of the ribosomal RNA genes, namely the nucleolus-forming 28S (major) and non-nucleolus-forming 5S (minor) rDNAs, were physically mapped using fluorescence in situ hybridization (FISH) and primed in situ labelling. The minor rDNA locus was located on the subtelo-acrocentric chromosome pair No. 9, whereas the major rDNA loci were dispersed on 14 chromosome pairs, showing a considerable inter-individual variation in the number and location. The major and minor rDNA loci were located at different chromosomes. Multichromosomal location (3-6 sites) of the NORs was demonstrated by silver nitrate (AgNO3) impregnation. All Ag-positive i.e. active NORs corresponded to the GC-rich blocks of heterochromatin. FISH with telomeric probe showed the presence of the interstitial telomeric site (ITS) adjacent to the NOR/28S rDNA site on the chromosome 11. This ITS was presumably remnant of the chromosome rearrangement(s) leading to the genomic redistribution of the rDNA sequences. Comparative analysis of the cytogenetic data among several related salmonid species confirmed huge variation in the number and the chromosomal location of rRNA gene clusters in the Salvelinus genome.

Influence of heavy metals and 4-nonylphenol on reproductive function in fish.

Many industrial and agricultural chemicals (including heavy metals and alkylphenols) present in the environment have adverse effects on the reproductive function in fish. Three studies were conducted to assess toxicity of these chemicals towards reproduction of freshwater fish. It was shown that heavy metals added to the diets accumulate in brain tissue of carp, and this accumulation results in inhibition of the secretion of noradrenaline and stimulation of the secretion of dopamine in the hypothalamus. These processes results in a disturbance of hormonal equilibrium of the hypothalamo-pituitary system, which can unfavorably influence the efficiency of artificial spawning in fish. Quality of salmonid and sturgeon sperm was impaired after in vitro exposure to heavy metals. The degree of this toxic effect was species-specific. It was demonstrated that sperm motility parameters appeared to be good indicators of adverse effects of heavy metals fish sperm. The protection role of seminal plasma against toxic effects of heavy metals was suggested for salmonid fish. Oral application of 4-nonylphenol (NP) disrupted reproduction in pikeperch. In juvenile fish a decrease in the percentage of males and an increase of intersex fish was observed in relation to dose of NP and time of exposure to this alkylphenol. Exposure of adult males to the NP led to the reduction in fecundity, milt quality and fertility.

Chromosome banding studies by replication and restriction enzyme treatment in vendace (Coregonus albula) (Salmonidae, Salmoniformes).

Chromosome banding studies were performed in vendace, Coregonus albula. Original data on distribution of early and late replication regions, restriction sites (AluI, DdeI, HinfI and HaeIII) on chromosomes in this coregonid fish have been used to analyse karyotype heterochromatin differentiation. Heterochromatic bands (C-positive and not digested by restriction enzymes) have been identified as late replicating regions. Extra bands produced by the applied methods have permitted the identification of several homologous pairs. The centromeres were differentially digested by the restriction enzymes. The studied population seems to be homogenic regarding karyotype characteristics.

Chromosome study of peled (Coregonus peled, Salmoniformes).

Chromosomes of Coregonus peled were examined by Giemsa, CMA3, Ag-NOR and C-banding. The karyotype of peled had a diploid number 2n=76, arm number NF=96 and consisted of twenty meta-submeta chromosomes and 56 subtelo-acrocentric chromosomes. C-positive blocks of heterochromatin were observed on the telomeric regions of meta- and submetacentric chromosomes. Pairs no. 1 and 11 had short arms, completely heterochromatic. The NOR was observed at one acrocentric pair, no. 11. Arm length polymorphism was observed on the NOR-bearing pair.

Chromosome study of Anodonta anatina (L., 1758) (Bivalvia, Unionidae).

The chromosome complement of the freshwater mussel Anodonta anatina was studied using Giemsa, Ag-NOR and chromomycin A3 staining. The diploid chromosome number of this species is 2n=38 and the arm number (NF) = 76. The nucleolar organizer region (NOR) was found on one chromosome pair and it was connected to GC rich chromatin as visualized by CMA3 staining.

Homologous chromosomes characteristics by sequential banding procedures in rainbow trout Oncorhynchus mykiss.

The development of high resolution methods of chromosome banding helped the finding of homologous chromosomes, detecting chromosomal abnormalities, and assigning the gene loci to particular chromosomes in mammals. Unfortunately, small and numerous fish chromosomes do not show GC rich and GC poor compartments, this preventing the establishment of G banding pattern. The combination of techniques enabling the identification of constitutive heterochromatin (C-banding), heterochromatin resistant to restriction endonucleas, NOR bearing chromosomes (AgNO3 banding), or AT rich regions on chromosomes (DAPI banding) in sequential staining provides a better characteristic of fish chromosomes. In this work sequentially DAPI, DdeI, AgNO3 stained chromosomes of rainbow trout resulted in the characteristic banding pattern of some homologous chromosomes. Procedure of FISH with telomere probe and DAPI as a counterstaining fluorochrome visualized simultaneous hybridization signals and DAPI banding. Possibility of detection both FISH and DAPI signals can help in procedures of gene mapping on chromosomes.