Biology of slugs (Agriolimacidae and Arionidae: Mollusca) in New Zealand hill country pastures

Summary Life cycles of the slugs Deroceras reticulatum and Arion intermedius were studied over a 2.5-year period in a sheep-grazed pasture in the Kaimai Range, New Zealand. D. reticulatum approximated a bivoltine phenology, with intervals between consecutive generations ranging from 4 to 7 months and maximum life span from 8 to 12 months. Egg laying occurred in autumn and spring-early summer. Periods of egg hatching were followed by a phase of juvenile growth characterised by linear increase in the logarithm of population mean body weights. This was followed by a phase of reproductive activity. A. intermedius had an annual life cycle. Eggs laid during late summer and autumn hatched during autumn-winter. Immature slugs were characterised by a period of low growth rate during winter followed by a period of rapid growth culminating in reproductive maturity. In both species, the phase of rapid growth rate was associated with enlargement of the hermaphrodite gland. The size of the hermaphrodite gland relative to body weight reached a maximum during the spermatozoon stage, generally coincident with maximum body weight. During the reproductive period of D. reticulatum and A. intermedius the body weight remained relatively stable but the hermaphrodite gland became progressively smaller as the slugs approached the post-reproductive stage. The albumen gland attained maximum weight at the oocyte stage of the hermaphrodite gland, at the onset of reproductive activity. The pasture, strongly dominated by grasses, was heterogenous in slope, vegetation and treading/grazing influences from sheep due to stratification of the habitat into contour tracks, associated with sheep movement around the slope contours, and intertrack areas. High population densities of D. reticulatum were associated with the track edges (kerbs) and the upper intertrack slopes, while densities of A. intermedius were highest for the intertrack slopes. Low densities of both species occurred on the tracks and at sheep campsites. Analysis of cohort life tables indicated that these dispersion patterns were important in the dynamics of the populations. The stage mortality contributing most to the variance in reproductive populations of both species was that between hatching and onset of reproduction; most of the mortality occurred shortly after hatching on south-facing slopes, but tended to occur later in the life cycle on north slopes. For D. reticulatum this mortality was inversely related to initial density and associated, at least in part, with predation by carabids and birds and with treading by sheep. Density relationships, both at whole plot and plot stratum level, indicated that variations in natality had a stabilizing influence on D. reticulatum populations.     

Characterization and Elucidation of Coordination Requirements of Adenine Nucleotides Complexes with Fe(II) Ions

Abstract

In spite of the significant role of iron ions-nucleotide complexes in living cells, these complexes have been studied only to a limited extent. Therefore, we fully characterized the ATP:Fe(II) complex including stoichiometry, geometry, stability constants, and dependence of Fe(II)-coordination on pH. A 1:1 stoichiometry was established for the ATP:Fe(II) complex based on volumetric titrations, UV and SEM/EDX measurements. The coordination sites of ferrous ions in the complex with ATP, established by ¹H-, ³¹P-, and ¹⁵N-NMR, involve the adenine N7 as well as Pα, Pβ, and Pγ. Coordination sites remain the same within the pH range of 3.1–8.3. By applying fluorescence monitored Fe(II)-titration, we established a log K value of 5.13 for the Fe(ATP)^2? complex, and 2.31 for the Fe(HATP)^? complex. Ferrous complexes of ADP^3? and AMP^2? were less stable (log K 4.43 and 1.68, respectively). The proposed major structure for the Fe(ATP)^2? complex is the ‘open’ structure. In the minor ‘closed’ structure N7 nitrogen is probably coordinated with Fe(II) through a bridging water molecule. The electronic and stereochemical requirements for Fe(II)-coordination with ATP^4? were probed using a series of modified-phosphate or modified-adenine ATP analogues. We concluded that: Fe(II) coordinates solely with the phosphate-oxygen atom, and not with sulfur, amine, or borane in the cases of phosphate-modified analogues of ATP; a high electron density on N7 and an anti conformation of the adenine-nucleotide are required for enhanced stability of ATP analogues:Fe(II) complexes as compared to ATP complexes (up to more than 100-fold); there are no stereochemical preferences for Fe(II)-coordination with either Rp or Sp isomers of ATP-α-S or ATP-α-BH₃ analogues.     

Characterization of morphospecies and strains of the genus Microcystis (Cyanobacteria) for a reconsideration of species classification

For characterization of Microcystis species and strains, cell size, growth temperature optimum, salinity tolerance, dark chemoheterotrophy, photoheterotrophy, guanine + cytosine content in DNA, total fatty acid composition and restriction fragment length polymorphism of a polymerase chain reaction product (PCR-RFLP) of the cpcBA intergenic spacer and flanking region were examined using 24 strains of Microcystis isolated from various lakes and ponds in Japan. From the results obtained it was observed that Microcystis spp. displayed low phenotypic diversity. Cell diameters of these strains were overlapping and there was no clear correlation with morphospecies. Slight differences in growth temperature optimum and salinity tolerance were observed among all strains. No strains showed either chemoheterotrophy or photoheterotrophy. The fatty acids present were the same in different strains although the amounts were different. All the strains had a similar G + C content ranging from 39 to 43 mol%. The phonogram constructed from the PCR-RFLP analysis showed that the species assignment for Microcystis species by morphology did not correspond with the genetic background.     

The colonization of Europe by the freshwater crustacean Asellus aquaticus (Crustacea: Isopoda) proceeded from ancient refugia and was directed by habitat connectivity

Recent continental-scale phylogeographic studies have demonstrated that not all freshwater fauna colonized Europe from the classic Mediterranean peninsular refugia, and that northern or central parts of the continent were occupied before, and remained inhabited throughout the Pleistocene. The colonization history of the ubiquitous aquatic isopod crustacean Asellus aquaticus was assessed using mitochondrial COI and a variable part of nuclear 28S rDNA sequences. Phylogeographic analysis of the former suggested that dispersion proceeded possibly during late Miocene from the western part of the Pannonian basin. Several areas colonized from here have served as secondary refugia and/or origins of dispersion, well before the beginning of the Pleistocene. Postglacial large-scale range expansion was coupled with numerous separate local dispersions from different refugial areas. Connectivity of the freshwater habitat has played an important role in shaping the current distribution of genetic diversity, which was highest in large rivers. The importance of hydrographic connections for the maintenance of genetic contact was underscored by a discordant pattern of mtDNA and nuclear rDNA differentiation. Individuals from all over Europe, differing in their mtDNA to a level normally found between species or even genera (maximal within population nucleotide divergence reached 0.16 +/- 0.018), shared the same 28S rRNA gene sequence. Only populations from hydrographically isolated karst water systems in the northwestern Dinaric Karst had distinct 28S sequences. Here isolation seemed to be strong enough to prevent homogenization of the rRNA gene family, whereas across the rest of Europe genetic contact was sufficient for concerted evolution to act.     

Genotypic diversity of the Killer Cell Immunoglobulin-like Receptors (KIR) and their HLA class I Ligands in a Saudi population

Killer Cell Immunoglobulin-like Receptors (KIR) have been used as good markers for the study of genetic predisposition in many diseases and in human genetic population dynamics. In this context, we have investigated the genetic diversity of KIR genes and their main HLA class I ligands in Saudi population and compared the data with other studies of neighboring populations. One hundred and fourteen randomly selected healthy Saudi subjects were genotyped for the presence or absence of 16 KIR genes and their HLA-C1, -C2, -Bw4^Thr80 and Bw4^Ile80 groups, using a PCR-SSP technique. The results show the occurrence of the framework genes (3DL2, 3DL3 and 2DL4) and the pseudogenes (2DP1 and 3DP1) at highest frequencies. All inhibitory KIR (iKIR) genes appeared at higher frequencies than activating genes (aKIR), except for 2DS4 with a frequency of 90.35%. A total of 55 different genotypes were observed appearing at different frequencies, where 12 are considered novel. Two haplotypes were characterized, AA and Bx (BB and AB), which were observed in 24.5% and 75.5% respectively of the studied group. The frequencies of iKIR + HLA associations were found to be much higher than aKIR + HLA. KIR genes frequencies in the Saudi population are comparable with other Middle Eastern and North African populations.     

Characterization and developmental expression of AmphiNk2-2, an NK2 class homeobox gene from amphioxus (Phylum Chordata; Subphylum Cephalochordata)

 The genome of amphioxus includes AmphiNk2-2, the first gene of the NK2 homeobox class to be demonstrated in any invertebrate deuterostome. AmphiNk2-2 encodes a protein with a TN domain, homeodomain, and NK2-specific domain; on the basis of amino acid identities in these conserved regions, AmphiNk2-2 is a homolog of Drosophila vnd and vertebrate Nkx2–2. During amphioxus development, expression of Amph- iNk2-2 is first detected ventrally in the endoderm of late gastrulae. In neurulae, endodermal expression divides into three domains (the pharynx, midgut, and hindgut), and neural expression commences in two longitudinal bands of cells in the anterior neural tube. These neural tube cells occupy a ventrolateral position on either side of the cerebral vesicle (the probable homolog of the vertebrate diencephalic forebrain). The dynamic expression patterns of AmphiNkx2-2 suggest successive roles, first in regionalizing the endoderm and nervous system and later during differentiation of specific cell types in the gut (possibly peptide endocrine cells) and brain (possibly including axon outgrowth and guidance). Received: 24 November 1997 / Accepted: 2 February 1998     

Characterization of six microsatellite loci in Myrica faya (Myricaceae) and cross amplification in the endangered endemic M. rivas-martinezii in Canary Islands, Spain

Six novel polymorphic microsatellite markers were isolated from enriched libraries in Myrica faya Ait., recently renamed Morella faya, (fayatree, firetree, or firebush) in order to examine the genetic diversity in natural populations. Also, test cross-specific amplification and genetic diversity in Myrica rivas-martinezii, which is endemic on the Canary islands. Microsatellite loci were screened in 225 individuals of both species from different islands of the Canarian archipelago. All markers were successfully amplified from both Myrica species, with an average number of 6.5 and 9.3 alleles per locus in M. rivas-martinezii and M. faya, respectively. There was no evidence for linkage disequilibrium between loci, and the probability of null alleles ranged from 0.01 to 0.17.     

Evolution of MHC-<Emphasis Type="Italic">DRB</Emphasis> class II polymorphism in the genus <Emphasis Type="Italic">Apodemus</Emphasis> and a comparison of <Emphasis Type="Italic">DRB</Emphasis> sequences within the family Muridae (Mammalia: Rodentia)

Allelic diversity at major histocompatibility complex (MHC) genes is thought to be maintained by balancing selection over long periods of time, even across multiple speciation events. Trans-species sharing of MHC alleles among genera has been supported by many studies on mammals and fish, but in rodents, the results are ambiguous. We investigated natural levels of MHC-DRB variability and evolutionary processes in the wood mouse (Apodemus sylvaticus) and the yellow-necked mouse (Apodemus flavicollis), which are common, sympatric murid rodents in European forests. Using single-strand conformation polymorphism analysis and DNA sequencing, 38 DRB exon 2 alleles were detected among 162 A. sylvaticus from nine different locations in Germany and Switzerland, and 15 DRB exon 2 alleles were detected among 60 A. flavicollis from three different locations in northern Germany. There was evidence for balancing selection in both species. Phylogenetic analysis, including additional murid taxa, showed that the DRB exon 2 sequences did not separate according to species, consistent with trans-species evolution of the MHC in these taxa.