Phymactis papillosa is a rocky shore sea anemone that is commonly found in the Pacific Ocean, from the Gulf of California to Tierra del Fuego, and in the Mar del Plata region, Argentina. The genus Phymactis is closely related to Bunodosoma and, due to character plasticity, a number of misidentifications have occurred. Therefore, the presence of P. papillosa in Argentina has been doubted but the matter had not been investigated in detail. Here we analyse P. papillosa specimens from Argentina and compare them, using molecular and morphological markers, to specimens from the species' type locality. In a phylogenetic analysis using 19 allozyme markers and ribosomal internal transcribed spacers sequences of different sea anemone genera, including all West Atlantic Bunodosoma species, we have found that the specimens from Argentina were genetically divergent from P. papillosa from Chile and closely related to West Atlantic Bunodosoma species. The genetic and morphological analyses indicate that those specimens belong to a new species of the genus Bunodosoma, described here as B. zamponii sp. nov.

Seven populations of Holothuria polii were sampled from the eastern and western Mediterranean coastal waters of Tunisia and screened electrophoretically for genetic variation at 11 allozyme loci. Six among the seven polymorphic loci were out of Hardy–Weinberg equilibrium (HWE) in at least one population. In the same way, the multilocus test showed deviation from HWE in all populations. These populations showed heterozygote deficiency. Genetic variability was relatively low. The number of alleles per locus ranged from 2.09 to 2.27 (average = 2.15), and the observed heterozygosity varied between 0.14 and 0.20 (average = 0.17). The observed overall differentiation among populations was slight but significant, with a mean FST value of 0.024 (P < 0.001). Pairwise FST values reflected the differentiation of the two populations, which were at the margins of the range sampled, from all the others. Our data suggest a population structure consistent with separation by Mediterranean Sea basins that might reflect different local biogeographical zones.

Genetic diversity of loggerhead turtles, Caretta caretta, nesting on the Kuriat Islands, the most important Tunisian nesting beach (central Mediterranean), was investigated using both nuclear and mitochondrial markers. Allozyme electrophoresis of 63 hatchlings from four different clutches showed a low genetic diversity. The genotypic composition of two clutches did not match Mendelian expectations suggesting the occurrence of multiple paternity. The analysis of 380 bp of the mitochondrial DNA control region revealed no genetic variability. Only one haplotype was described in our sample (N = 16), which corresponds to the sequence of the most common haplotype found on the Mediterranean nesting beaches (CC-A2). The low genetic diversity detected by both mitochondrial and allozyme markers is discussed taking into account available data about past and present situations of loggerhead nesting activity in this site. Adequate conservation measures should be urgently taken to protect the nesting population in this area.

In Guatemalan villages people commonly rear pigs, and both hosts may be infected with Ascaris. This study was designed to ask whether both humans and pigs are potential hosts in a single parasite transmission cycle in such villages, or alternatively, if there are two separate transmission cycles, one involving pigs and one involving human hosts. Parasites were collected from both host species from locations in the north and south of Guatemala. Allelic variation in the nuclear genome of Ascaris was measured using enzyme electrophoresis, while mitochondrial DNA (mtDNA) sequence variation was quantified using restriction mapping. Low levels of enzyme polymorphism were found in Ascaris, but allele frequencies at two loci, mannose phosphate isomerase and esterase, suggest that there is little gene exchange between parasite populations from humans and pigs. MtDNA haplotypes fall into two distinct clusters which differ in sequence by 3–4%; the two clusters broadly correspond to worms collected from humans and those collected from pigs. However, some parasites collected from humans have mtDNA characteristic of the ‘pig Ascaris’ haplotype cluster, while some parasites collected from pigs have mtDNA characteristic of the ’ haplotype cluster. These shared haplotypes are unlikely to represent contemporary cross-infection events. Patterns of phylogenetic similarity and geographical distribution of these haplotypes suggest, instead, that they are the result of two historical introgressions of mtDNA between the two host-associated Ascaris populations. The results clearly demonstrate that Ascaris from humans and pigs are involved in separate transmission cycles in Guatemala.

New biological species and high levels of inter- and intraspecific genetic divergence were discovered in an allozyme study of some North European members of the acanthocephalan genus Echinorhynchus (sensu lato), parasites of fish and malacostracan crustaceans. (i) A strong differentiation between the marine E. gadi and the fresh- and brackish-water E. salmonis (genetic identity I ≃ 0) supports a generic distinction between these taxa; however, the subdivision would not entirely concur with the concepts of Echinorhynchus (sensu stricto) and Metechinorhynchus suggested earlier. (ii) Samples of E. gadi from the Baltic, Norwegian and North Seas included three distinct, partially sympatric biological species (spp. I–III; I ≃ 0·5). (iii) E. bothniensis, previously only known from the northern Baltic Sea, represents a complex of freshwater taxa with an intermediate host relationship to the ‘glacial relict’ Mysis spp. and with a distributional and host analogy to the North American E. leidyi. A population in a northern lake in the Barents Sea basin is closely related to E. bothniensis of the Baltic area, but is probably specifically distinct; the divergence between these populations (I ≃ 0·6) is similar to that between their Mysis host species. (iv) Considerable intraspecific differentiation (Fst = 0·25), probably reflecting post-glacial population bottlenecks, was found between Baltic and nearby lacustrine E. bothniensis, and between Atlantic and Baltic E. gadi sp. I.

Pine beauty moth, Panolis flammea (Denis & Schiffermüller), is a recent but persistent pest of lodgepole pine plantations in Scotland, but exists naturally at low levels within remnants and plantations of Scots pine. To test whether separate host races occur in lodgepole and Scots pine stands and to examine colonization dynamics, allozyme, randomly amplified polymorphic DNA (RAPD) and mitochondrial variation were screened within a range of Scottish samples. RAPD analysis indicated limited long distance dispersal (FST = 0.099), and significant isolation by distance (P < 0.05); but that colonization between more proximate populations was often variable, from extensive to limited exchange. When compared with material from Germany, Scottish samples were found to be more diverse and significantly differentiated for all markers. For mtDNA, two highly divergent groups of haplotypes were evident, one group contained both German and Scottish samples and the other was predominantly Scottish. No genetic differentiation was evident between P. flammea populations sampled from different hosts, and no diversity bottleneck was observed in the lodgepole group. Indeed, lodgepole stands appear to have been colonized on multiple occasions from Scots pine sources and neighbouring populations on different hosts are close to panmixia.

Allozyme and Random Amplified Polymorphic DNA (RAPD) variation was surveyed in the freshwater crayfish Cherax destructor Clark, an ecologically and commercially important species that is widespread throughout the freshwater systems of central Australia. At the intra-population level, allozymes revealed a similar level of variation to that found in other freshwater crayfish; RAPDs showed less diversity than allozymes, which was unexpected. At the inter-population level, both techniques revealed significant population structure, both within and between drainages. RAPD results were consistent with phylogeographic patterns previously identified using mtDNA. Although allozyme data showed little geographic pattern in relation to genetic variation based on multidimensional-scaling (MDS) plots on matrices of genetic distance, results of AMOVA and Mantel tests indicated significant population structuring. Each of the mtDNA lineages proposed in a previous study also showed significant genetic structure at similar levels as revealed by RAPDs but different levels by allozymes. These results reject hypotheses previously put forward on genetic homogenisation within the species due to wide-scale translocation. The implications of the findings for conservation and aquaculture of C. destructor are also discussed.

Allozyme variation within and among populations of 3 species of the genus Lecithochirium (Trematoda: Hemiuridae) was studied by starch gel electrophoresis. In total, 19 loci were analysed in 7 populations. The level of genetic variability was relatively high in all populations. The percentage of polymorphic loci (0·95 criterion) observed per population varied from 21·0% to 55·5%, and expected heterozygosity levels varied from 0·082 to 0·197. All populations showed significant heterozygote deficiencies. In Lecithochirium fusiforme most of the deviations from Hardy–Weinberg proportions were within the populations and this species showed moderate population structuring (FIS=0·486, FST=0·142, Nm=1·51) and accordingly low intraspecific genetic distances (D=0·003 to 0·027). A significant lack of heterozygotes for several polymorphic loci was revealed in Lecithochirium rufoviride and Lecithochirium musculus. The most probable cause of the population genetic subdivision in L. rufoviride is the presence of at least 1 cryptic species in the populations studied. Although the lowest percentage of fixed genetic differences was that between L. fusiforme and L. musculus, two different algorithms for the construction of evolutionary trees on a matrix of genetic distances confirmed that L. fusiforme and L. rufoviride are phenetically the most closely related species.

Domesticated/captive stocks and wild/feral populations of common carp from Europe, Central Asia and East/South-East Asia were examined for allozyme (23 populations), microsatellite (11 populations) and mitochondrial DNA (21 populations) variation. Allozyme variability (1.06–1.81 alleles per locus, expected heterozygosity 0.006–0.136 at 16 loci) was much lower than microsatellite variability (2.5–14.0 alleles per locus, expected heterozygosity 0.426–0.887 at four loci). Differences in variability between domesticated/captive stocks and wild-caught ones were more pronounced at microsatellite loci than at allozyme loci, suggesting that microsatellites are better suited to detect population bottlenecks and loss of variation due to inbreeding. All but one European population were fixed for a single composite mtDNA haplotype, which also dominated in Central Asia but was completely missing in East/South-East Asia, indicating a single origin of European carp in Central Asia. All three classes of genetic markers clustered populations into two highly divergent groups: Europe/Central Asia and East/South-East Asia. Hierarchical partition of genetic diversity showed that for microsatellite loci most of variation was due to the within-population component while the highest proportion of mtDNA variation and substantial proportion of allozyme variation was accounted for by differences between geographical regions. Genetic data support the subspecies status of C. c. carpio assigned to the European carp and C. c. haematopterus assigned to the East/South-East Asian carp but do not justify a separate subspecies status (C. c. aralensis) for the Central Asian carp. As demonstrated for a wild/feral carp population from R. Danube, Germany, the genetic markers used in our study may be effectively applied to detect mixing and introgression of intra-species units in the presence of sufficient genetic differentiation.

Allozyme markers were used to study genetic variation in Lecithochirium fusiforme within a natural population of Conger conger. Six of 16 enzyme-coding loci studied were found to be polymorphic. These loci were surveyed in 12 infrapopulations of adult flukes. High levels of genetic variation were detected (P=0·375); Ho=0·048; He=0·085). However, the population did not conform to Hardy-Weinberg expectations, as it showed a significant deficit of heterozygotes. L. fusiforme also exhibited low differentiation between infrapopulations (FST=0·064). Despite significant linkage disequilibrium at Pgm-1 and Pgm-2 (P<0·05), mating system does not appear to be the principal reason for the deficit of heterozygotes detected, because some polymorphic loci were in Hardy-Weinberg equilibrium. Association between FIS and FST statistics suggests the existence of the Wahlund effect. However, all infrapopulations showed a strong deficit of heterozygotes for most polymorphic loci (FIS=0·409). Detection of significant genetic differentiation among temporal samples and the existence of paratenic hosts in the life-cycle suggests the Wahlund effect, caused by the mixture of genetically distinct temporal samples in the infrapopulations. Occasional temporal gene flow also might explain the high estimated genetic polymorphism.

The taxonomic relationship between two toothed South African river crabs, Potamonautes warreni and P. unispinus, is unclear. The problem stems from the widespread variation in carapace dentition patterns amongst P. warreni individuals over its biogeographic range, where single toothed individuals may appear similar in carapace morphology to P. unispinus. Ten populations of P. warreni and 18 populations of P. unispinus were collected and the morphometric and genetic differentiation between the two taxa quantified. Patterns of morphometric and genetic variation were examined using multivariate statistics and protein gel electrophoresis, respectively. Principal component analyses of carapace characters showed that the two species are morphologically indistinguishable. However, discriminate functions analyses and additional statistical results corroborate the morphological distinction between the two taxa. Allozyme electrophoresis of 17 protein coding loci, indicated a close genetic similarity between the two species (I = 0.92). A fixed allelic difference at one locus (LT-2) and extensive genetic variability at another locus (PGM-1) indicate that two gene pools are present and that the two taxa are genetically isolated. Intraspecific genetic I values for both species were >0.97 and indicated no apparent genetic structuring on a micro or macro-geographic scale. The variation in carapace dentition among P. warreni populations possesses no genetic basis and may possibly be the product of ecogenesis. The value of dentition patterns in the systematics of river crabs is discussed. Dentition patterns among river crab species appear to be conserved and reliable as species specific diagnostic markers, but should ideally be used in combination with other morphological data sets and genetic evidence.

Karyotypic and allozyme analyses were carried out on 55 individuals of Microtus subterraneus and M. majori from 19 populations in Europe and Anatolia. The occurrence of M. subterraneus in Asia Minor was proven. Standard karyotypes were found in M. majori (2n=54), M. subterraneus from Poland (2n=54), and central and south-eastern Europe (2n=52). Although the karyotype of M. subterraneus populations from Anatolia (2n=54), resembled that from Poland, there were apparent differences in the X chromosomes: it was larger and submetacentric (instead of metacentric) in the Anatolian populations owing to the presence of a large block of heterochromatin. The heterochromatic segment was also found to cause the size and shape differences in the X chromosomes between M. majori and European M. subterraneus (both in the 2n=52 and 2n=54 karyotypes). However, the location of this segment differed between the latter species and M. subterraneus from Anatolia. The distribution of the 2n=52 and both the 2n=54 karyotypes of M. subterraneus suggests that the Robertsonian fusion, distinguishing these different karyotypes, originated within the range of the species (i.e. within the 54-chromosome karyotype) and has spread subsequently towards its periphery. It is also suggested that both the fusion in European populations and amplification of heterochromatin in Anatolian populations are quite recent phenomena. The allozyme data indicate that gene flow is limited between populations of M. subterraneus and they are concordant with the isolation-by-distance model. There seems to be little correspondence between patterns of chromosomal and allozyme variation in M. subterraneus. Nei's genetic distance between M. subterraneus and M. majori was low (D=0.169), suggesting their recent divergence.

Use of isozymes (including allozymes) in studies of population genetics and systematics of seaweeds has increased sufficiently in the last decade to allow some generalization. Only a single locus has been observed for about half the enzymes analysed in seaweeds, compared with 29% in vascular plants. Compared with higher plants, macroalgal species generally have low amounts of electrophoretically detectable genetic variation; the lowest levels of genetic variation found in natural populations are those reported for seaweeds. Nonetheless, seaweeds show an association between levels of genetic diversity as revealed by isozymes and species-specific attributes, such as mating system and predominance of asexual versus sexual reproduction. In systematic studies, isozymes have revealed cryptic species and identified pairs of sibling taxa. The quaternary structure of enzymes appears to be conserved at the phylum level. With the current availability of improved techniques for enzyme electrophoresis and for data interpretation, we expect future studies utilizing isozyme electrophoresis to provide further insight into population and evolutionary processes in seaweeds.

Populations of Potamonautes perlatus were sampled at various points along the Olifants Rivers (220 km), in the Western Cape to examine the degree of genetic variation. Allozyme electrophoresis revealed a near fixed allelic difference at one locus and extensive differentiation at another locus between populations from the upper and lower reaches of the Olifants River system. Limited hybridization in the region of population sympatry was observed. The narrow hybrid zone and the low occurrence of hybrids suggest selection against hybrids. These results provide evidence for two genetically distinct freshwater crab populations. We concluded that the genetic divergence between the two population groups represents a species boundary between the two populations.

Phylogenetic relationships of 31 pomacanthid species representing all genera were investigated by horizontal starch gel electrophoresis. Nei's genetic distances (D) were calculated and dendrograms constructed from allozyme variation in 23 loci. The average D value was 0.03 between conspecific colour variants, 0.40 between congeneric subgenera, and 1.27 between confamilial genera. From the dendrograms the following were revealed: (1) the generic status of Apolemichthys arcuatus and Holacanthus venustus, as well as the generic and subgeneric status of Centropyge multifasciata were confirmed; (2) the subgeneric status of Xiphypops was confirmed; (3) Pygoplites diacanthus was found to be closely related to members of the subgenus Pomacanthus; (4) Pomacanthus zonipectus was found to be biochemically distinct from its con-subgeners; (5) the genera Pomacanthus and Holacanthus are more closely related than currently thought; (6) the Genicanthus species might warrant a subfamily of their own.

Genetic variation in the pulmonate land snail Zonitoides nitidus was examined by means of allozyme electrophoresis in 17 European populations. We observed no heterozygotes, indicating that Z. nitidus consists of several homozygous multilocus genotypes (strains) probably produced by self-fertilization. Nine strains were detected. Strains appeared not to be equally distributed over the regions considered. One strain was strongly differentiated from the others, which is suggestive of a taxonomic differentiation. Anatomically, two major phally types were distinguished: euphallics, with well-developed male reproductive organs, and hemiphallics, with weakly developed male reproductive organs. Both phally types were found together in natural populations, but euphally ratios were very low (0–19%; overall mean = 7.79%) and differed between geographical regions (Belgium, Germany, and Sweden). In some populations, hemiphallic individuals were smaller compared to euphallic individuals although the difference is small and its biological significance remains to be established. We found no clear relation between phally type and allozyme data.

Species of Echinococcus (Cestoda: Taeniidae) require 2 mammalian hosts to complete their life-cycle; a carnivorous definitive host, and a herbivorous or omnivorous intermediate host. For most species of Echinococcus, the definitive host range is restricted to 1 or a few species, but the intermediate host range is very broad. Programmes to control hydatid disease attempt to break the life-cycle of the parasite and their effectiveness is therefore enhanced by an understanding of local patterns of transmission. Although it is known that the rostellar hooks of protoscoleces may be influenced by the species of intermediate host in which they develop, the application of this knowledge to infer transmission cycles has been limited, because the intermediate host effect has not been isolated from other environmental and genetic components of phenotypic variance. This study presents a method for separating these potentially confounding genetic and environmental effects, by combining quantitative genetic analyses of hook traits with data on population structure from neutral genetic markers. The method was applied to 5 hook traits (hook number, total length of large hooks, blade length of large hooks, total length of small hooks, blade length of small hooks) measured on protoscoleces from 2 intermediate host types (sheep and macropod marsupials) in Australia. Although genetic variance was similar for all traits, they differed markedly in the extent of environmental variance attributed to development in different host types. Total length of small hooks was the trait most affected, with 49–60% of phenotypic variance being explained by environmental differences between intermediate host species. Blade length of small hooks was least affected, with none of the phenotypic variance due to intermediate host origin. These data suggest that hook measurements of adult worms from naturally infected definitive hosts could be used to determine the intermediate host species from which infection was acquired, if the appropriate traits are measured.

Morphological and isozyme variations between 13 populations of the species hitherto named Patella granularis were investigated to see whether differences in shell structure between the west coast versus the south and east coasts of southern Africa are supported by other morphological features or by genetic differences. The shells showed a definite decrease in size from west to east, but this is correlated with productivity and is of no diagnostic use in distinguishing between populations. Discriminant functions analysis based on shell morphometrics failed to separate populations from the three coastal regions. Shells from the northern east coast do, however, have shell nodules with a dark pigmentation, distinctly separating them from those further south and west. No differences in radular or soft part morphology were detected between the populations, but the four northernmost populations on the east coast have a significantly shorter Z looping of the gut than the other populations along the coast. Significant microstructural differences in the sperm were also detected between these two groups of populations. Electrophoretic analysis of 16 enzyme loci failed to detect any significant differences between the west and south coast populations, but revealed a genetic identity (Nei) of 0.528 as well as four diagnostic alleles between the four northernmost populations from the east coast compared with those to the south and west. The two genetically distinct forms occurred sympatrically at one of the study sites on the east coast (Coffee Bay). It was concluded the two groupings were sufficiently different to warrant the recognition of a separate species, which is centred in KwaZulu-Natal on the east coast and extends south to Coffee Bay, from where it is replaced by P. granularis. There is, however, no evidence at all that the west coast populations are in any way separable from the remaining populations of P. granularis on the south coast.

The Quaternary dispersal of the red fox Vulpes vulpes in the Mediterranean area was evaluated through the study of allelic variation at 45 enzyme loci in 120 individuals from 10 sampling sites. A 375 bp fragment of the mitochondrial cytochrome b gene was also sequenced in a total of 41 specimens from the same sampling locations. Nine allozyme loci were polymorphic. The proportion of polymorphic loci per population (P) ranged from 0 to 15.6%, and expected average heterozygosity (H) from 0 to 4.4%. A total of 18 different Cyt b haplotypes were detected. Most of them were confined to only one population. Both allozyme and mtDNA data implied that our fox populations were genetically fairly isolated from one another, suggesting low gene flow between them. This isolation should be of comparatively recent origin according to the slight differentiation among Cyt b haplotypes. Fox populations appeared to belong to two genetically distinct groups. With a mean value of Nei's D = 0.024, genetic distance between these groups was similar to that detected at subspecies level in taxa of large mammals. This pattern may have originated from different colonization waves during Quaternary glaciations and deglaciations. Red foxes from Sardinia were more closely related to the Bulgarian foxes than to the Iberian ones. However, repeated introductions to Sardinia probably also occurred from Central Italy and Spain, as suggested by the presence of haplotype A and a typical Central Italian allele, Ck-290.