Limited hybridisation and introgression despite stocking among endemic Interior Highlands black basses (Centrarchidae: Micropterus)

Aim

Smallmouth Bass (Micropterus dolomieu; SMB) are globally popular among anglers and have been widely introduced (i.e. stocked) for population management and sportfishing. Importantly, stocking was prevalent before cryptic diversity within the SMB complex was known, which now includes three newly elevated species: Neosho Bass (M. velox; NB), Little River Bass (M. sp. cf. dolomieu Little River; LRB) and Ouachita Bass (M. sp. cf. dolomieu Ouachita River; OB). We sought to quantify population structure and hybridisation and introgression in these three recently described species.

Location

Species-level diversity, particularly in the basin-restricted LRB and OB in the Ouachita Mountains within the Central Interior Highlands (CIH), North America, suggests the presence of distinct genetic variation that could be eroded by introgression.

Methods

We estimated interspecific introgression and intraspecific population differentiation in the Smallmouth Bass species complex (SMB-C) using 472 specimens comprising SMB, NB, LRB and OB, including the naturally sympatric Spotted Bass (M. punctulatus; SPB). Genomic samples were genotyped on a SNP panel of 192 loci designed to detect allele-sharing on multiple hierarchical levels.

Results

We found low range-wide hybridisation between species in the SMB-C and SPB (mostly SMB-C backcrosses), and interspecific heterozygosity varied, indicating differential introgression. Range-wide hybridisation between species in the CIH and SMB was similar overall (but mostly F₂ and CIH backcrosses) and was observed in streams with known SMB stocking in connected reservoirs. Interspecific heterozygosity in SMB hybrids was also generally lower, indicating later-generation backcrosses. We found strong population structure in the Ouachita Mountains (LRB and OB).

Main Conclusions

Despite isolated incidences of natural (SPB) and human-mediated (SMB) introgression, genomic identity appears intact in endemic LRB and OB, suggesting potential ecological or behavioural isolating mechanisms preventing cross-species reproduction. Our findings reveal that genetic variation remains in cryptic, basin-restricted species in the Ouachita Mountains ecoregion that may be managed for long-term conservation.     

Hydrogen production by immobilized cells in the nozzle loop bioreactor

Summary The continuous production of hydrogen in a Nozzle Loop Bioreactor was investigated using immobilized Rhodospirillum rubum KS-301 with glucose as the growth-limiting substrate. The maximum hydrogen production rate in the experimental range was 91mL/h at dilution rate 0.4h^-1, initial glucose concentration 5.4g/L, and circulation rate 70h^-1 .     

Sequence and evolution of cattle MHC class I cDNAs: concerted evolution has not taken place in cattle

To explore genetic mechanisms responsible for major histocompatibility complex (MHC) class I evolution in the artiodactyls, we cloned and sequenced MHC class I cDNAs from a Bos taurus bull heterozygous for cattle MHC (BoLA) class I serological specificities w2 and w30. Four unique cDNAs were found, indicating the presence of at least two MHC class I loci. Analysis of these four cDNAs and all previously published BoLA cDNA sequences suggested that there may be three cattle MHC class I loci. Additionally, comparison of all of the BoLA class I cDNAs to MHC class I cDNAs of other artiodactyls showed that some of the BoLA class I cDNAs were more similar to certain sheep cDNAs than they were to other cattle cDNAs. These data indicate that each BoLA class I locus has evolved independently after an ancestral gene duplication event and that inter-locus segmental exchange o or concerted evolution has not occurred rapidly enough to cause extensive divergence between the orthologous MHC class I loci of sheep and cattle.The nucleotide sequence data reported in this paper have been submitted to the GenBank nucleotide sequence database and have been assigned the accession numbers L02832–L02835. Correspondence to: T. L. Garber at the present address.     

Isolation and characterization of a putative multidrug resistance pump from Vibrio cholerae

Multidrug-resistant strains of Vibrio cholerae (the causative agent of the diarrhoeal disease cholera) have recently been described. In an attempt to identify a homologue of the Escherichia coli TolC in V . cholerae , we isolated a DNA fragment (pVC) that enabled an E . coli tolC mutant to grow in the presence of 0.05% deoxycholate (DOC). However, other TolC defects were not complemented. Nucleotide sequence analysis of this fragment revealed the presence of two open reading frames (ORF1 and ORF2) separated by 9 bp and encoding 42.4 and 55.8 kDa proteins respectively. The translational products of these two ORFs correlated closely with the molecular weights of the predicted proteins. The deduced amino acid sequences of ORF1 and ORF2 showed a high degree of similarity with conserved regions of the E . coli efflux pump proteins, EmrA and EmrB. The presence of pVC2 within the E . coli efflux pump mutants defective in either the emrAB or the acrAB genes provided the mutants with resistance against several antibiotics. A V . cholerae isogenic mutant defective in ORF2 was constructed by gene replacement. Characterization of this mutant has shown it to be more sensitive to CCCP, PMA, PCP, nalidixic acid and DOC than the parent strain. These results suggest that ORF1 and ORF2 constitute an operon encoding two components of a putative multidrug resistance pump in V . cholerae . In addition, the presence of both structural and functional similarities between VceAB and EmrAB suggests that VceAB is a homologue of EmrAB.     

Dopamine depletion in nucleus accumbens reduces ACTH1–24-induced excessive grooming

Animals were pretreated with 6-OHDA or ascorbate vehicle injected into the nucleus accumbens and tested 10 days later for excessive grooming induced by intracerebroventricular injection of ACTH_1–24. The animals pretreated with 6-OHDA showed a significant decrease in excessive grooming produced by the neuropeptide and this reduction was seen only in the last 30 minutes of a 60-min test session. The results suggest an interaction of ACTH with dopamine systems on the onset and maintenance of excessive grooming.     

Differential Effects of Fluoronorepinephrines on Phosphatidylinositol Turnover in Rat Pinealocytes

(+/-)-Norepinephrines (NAs), substituted with fluorine at positions 2, 5, and 6 of the ring, were compared with the unsubstituted compound with respect to their capacity for eliciting increased incorporation of [32P]orthophosphate into phosphatidylinositol of pinealocytes. 5F-NA and 6F-NA were approximately equipotent with NA, whereas 2F-NA required a higher concentration and gave lower maximum stimulation. Inhibition of these effects by prazosin confirmed the participation of alpha 1-adrenergic receptors. The results are comparable with those reported for alpha 1-adrenergic receptor-mediated events in other systems and different from the beta-adrenergic receptor-mediated elevation of cyclic AMP in pinealocytes. These and earlier results emphasize the importance of the hydroxyl group at position 3 of the ring and at the beta-position of the side chain in catecholamine activation of the pineal alpha-adrenergic receptors, which are involved in alterations of phospholipid metabolism.     

Labeling of the centromeric region on human chromosome 8 by in situ hybridization

Summary Probe DNA that binds preferentially to the centromeric region of human chromosomes 8 was synthesized. Alpha satellite probe DNA molecules were selectively amplified from sorter-purified human chromosomes 8 by in vitro DNA amplification using the polymerase chain reaction (PCR). Probe labeling was performed during PCR by incorporation of biotinylated deoxyuridine. In situ hybridization of unpurified probe DNA comprised of alpha satellite monomer and higher molecular weight DNA fragments with metaphase chromosome spreads showed binding to the centromeric regions of numerous chromosomes. However, blocking with unlabeled total human alphoid DNA dramatically reduced crosshybridization to chromosomes other than 8. Under these conditions, the degenerate probe DNA allowed unambiguous visualization of domains occupied by centromeric DNA of chromosome 8 in metaphase spreads and interphase cell nuclei, thus greatly facilitating the detection of numerical chromosome aberrations in tumor cells. In situ hybridization of size-fractionated alpha satellite DNA identified the monomeric fraction as the major cause of crosshybridization. Alpha satellite dimers and higher molecular weight DNA fragments showed relatively high specificity for human chromosomes 8.     

Polymers of Cu/Zn Superoxide Dismutase Produced by Cross-Linking with Glutaraldehyde

Soluble polymers of bovine Cu/Zn superoxide dismutase (EC 1.15.1.1) have been prepared using the homobifunctional cross-linking reagent, glutaraldehyde. A form of the enzyme, a tetramer. with a molecular weight of 64, 000 has been purified by gel filtration. The functional properties of the tctrarner have been investigated. Reconstitution with copper and zinc was required for full activity. After metal reconstitution, the specific activity of the tetramer was shown to be close to 90% that of the native dimerism enzyme.

The serum half-life of the tetramer in rats was found to be increased by a factor of six when compared with native superoxide dismutase. The tissue distribution of the two forms was also found to be direrent with the tetrarner accumulating predominantly in the liver.     

The rate of fusion of phospholipid vesicles and the role of bilayer curvature

Kinetics of Ca²⁺-induced fusion of phosphatidylserine vesicles is studiied for lipid concentrations varying from 1 μM to 100 μM. Fusion is monitored by mixing of aqueous vesicle contents and by explicitly accounting for leakage. The analysis provides separately rates of aggregation and fusion. The rate of fusion per se decreases steeply with vesicle size.