The large-sized tapeworm Dibothriocephalus latus is known as the broad or fish-borne cestode of mammals that is capable to infect humans and cause diphyllobothriosis. Recently, molecular data on D. latus has been accumulating in the literature and a complete genome sequence has been published; however, little is known about the karyotype and chromosome architecture. In this study, an in-depth karyological analysis of 2 D. latus specimens was carried out. The plerocercoids originated from a perch caught in subalpine Lake Iseo (Italy) and the tapeworms were reared in hamsters. Both specimens contained cells with a highly variable number of chromosomes ranging from18 to 27. Nevertheless, the largest portion of mitotic figures (47%) showed a number corresponding to the triploid set, 3n = 27. Accordingly, the karyotype of the analyzed specimens consisted of 9 triplets of metacentric chromosomes. Fluorescence in situ hybridization (FISH) with the 18S rDNA probe clearly demonstrated the presence of 3 clusters of hybridization signals on the triplet of chromosome 7, thus confirming the triploid status of the specimens. FISH with a telomeric (TTAGGG)n probe confined hybridization signals exclusively to the terminal chromosomal regions, supporting the earlier findings that this repetitive motif is a conserved feature of tapeworm telomeres.
- MeSH
- chromozomy genetika MeSH
- cytogenetické vyšetření MeSH
- difylobotriáza parazitologie MeSH
- Diphyllobothrium genetika metabolismus MeSH
- hybridizace in situ fluorescenční MeSH
- karyotyp MeSH
- RNA helmintů analýza MeSH
- RNA ribozomální 18S analýza MeSH
- triploidie * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
An original cytogenetic study combining classical karyotype analysis and modern fluorescence in situ hybridization using telomeric (TTAGGG)n and ribosomal sequences (18S rDNA) was performed in Khawia abbottinae (Cestoda, Caryophyllidea), a parasite of Chinese false gudgeon (Abbottina rivularis) from China. Analyses based on conventional Giemsa staining, DAPI, YOYO-1 dye, and silver (Ag) staining were also carried out. The karyotype is composed of eight pairs of metacentric and telocentric chromosomes (2n = 16, n=5m + 3t). Constitutive heterochromatin was mainly positioned at pericentromeric regions, and telomeric sequences (TTAGGG)n were restricted to the end of all chromosomes. In mitotic preparations stained with Giemsa, both homologues of chromosome pair 4 showed a distinct secondary constriction. FISH with rDNA probe confirmed that this secondary constriction contains a nucleolar organizer region (NOR). The process of spermatocyte meiosis and the dynamics of nucleolus degradation in dividing cell were scrutinized. The present study and its results enhance the limited knowledge on basic karyotype characteristics and 18S rDNA clusters location in caryophyllidean tapeworms.
- MeSH
- Cestoda klasifikace genetika izolace a purifikace MeSH
- chromozomy genetika MeSH
- Cyprinidae genetika MeSH
- DNA helmintů genetika MeSH
- heterochromatin genetika MeSH
- hybridizace in situ fluorescenční MeSH
- karyotyp MeSH
- karyotypizace MeSH
- proteiny červů genetika MeSH
- ribozomální DNA genetika MeSH
- ribozomy genetika MeSH
- telomery genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Čína MeSH
The history and value of cytogenetic features for addressing questions of the evolution and systematics of tapeworms (Cestoda) are briefly reviewed along with instructions for collecting karyological data. As a supplement to worm morphology, chromosome number and morphology have been helpful in determining the systematic status of some genera in the Diphyllobothriidae and species in the Bothriocephallidea. In addition, many new techniques for chromosome analysis have been recently applied in morphological and molecular studies of invertebrates, including tapeworms. Methods of molecular karyology, fluorescence in situ hybridisation, and chromosomal location of satellite DNA, microsatellites or histone genes may also provide useful data to inference of taxonomic relationships and for revealing trends or general lines of chromosome evolution. However, as karyological data are available only for few tapeworms, they are seldom an integral part of evolutionary and taxonomic studies of cestodes. A primary reason for this lack of karyological data may lie in general difficulties in working with tapeworm chromosomes. To address these problems, herein we present a well-tested, step-by-step illustrated guide on the fixation of tapeworm material and preparation of their chromosomes for cytogenetic studies. The technique requires standard glassware, few reagents and simple equipment such as needles; it can also be used on other neodermatan flatworms.
- MeSH
- Cestoda klasifikace genetika MeSH
- chromozomy * MeSH
- cytogenetika metody MeSH
- parazitologie metody MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
This paper reports results of the first cytogenetic study carried out on a recently described monozoic tapeworm, Khawia saurogobii Xi et al., 2009, from the Chinese lizard gudgeon (Saurogobio dabryi). The karyotype of this species is composed of eight pairs of metacentric and telocentric chromosomes (2n = 16; n = 3m + 5t), metacentric chromosomes representing the first, sixth, and eight pairs. All chromosomes except the largest pair displayed 4',6-diamidino-2-phenylidole (DAPI) positive heterochromatin in centromeric regions. In mitotic preparations stained with Giemsa, one of the homologues of a smaller metacentric chromosome pair (No. 7) showed a distinct secondary constriction, whereas the other did not. Fluorescent in situ hybridization (FISH) with 18S ribosomal DNA (rDNA) probe revealed that the chromosomes No. 7 carry each a cluster of ribosomal genes associated with the centromeric heterochromatin and confirmed that this chromosome pair contains a nucleolar organizer region (NOR). The rDNA-FISH also confirmed heteromorphism in the size of NOR (i.e., secondary constriction) observed after Giemsa staining. The present cytogenetic analysis revealed species-specific characters of K. saurogobii and showed that FISH may represent a new valuable cytogenetic tool suitable for comparative taxonomic or phylogenetic studies within the order Caryophyllidea in the future.
- MeSH
- barvení a značení MeSH
- Cestoda genetika MeSH
- chromozomy MeSH
- cytogenetické vyšetření MeSH
- heterochromatin metabolismus MeSH
- hybridizace in situ fluorescenční MeSH
- indoly metabolismus MeSH
- karyotypizace MeSH
- mapování chromozomů MeSH
- organizátor jadérka MeSH
- ribozomální DNA genetika MeSH
- RNA ribozomální 18S genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
