Species-specific genetic markers for identification of early life-history stages of Cape hakes, Merluccius capensis and Merluccius paradoxus in the southern Benguela Current

The correct identification of fish egg and larval stages is crucial for inferring spawning areas and subsequent dispersal routes for marine fishes. The authors use species-specific mtDNA polymerase chain reaction to estimate proportions of Merluccius capensis and Merluccius paradoxus eggs and larvae and to elucidate early life stage distribution patterns in the southern Benguela system.     

Evaluating the resolution power of new microsatellites for species identification and stock delimitation in the Cape hakes Merluccius paradoxus and Merluccius capensis (Teleostei: Merlucciidae)

The utility of 15 new and 17 previously published microsatellite markers was evaluated for species identification and stock delimitation in the deep‐water hake Merluccius paradoxus and the shallow‐water hake Merluccius capensis. A total of 14 microsatellites were polymorphic in M. paradoxus and 10 in M. capensis. Two markers could individually discriminate the species using Bayesian clustering methods and a statistical power analysis showed that the set of markers for each species is likely to detect subtle genetic differentiation (F_ST < 0·006) that will be valuable to delimit and characterize genetic stocks.     

Mitochondrial DNA analyses of the Cape hakes reveal an expanding, panmictic population for Merluccius capensis and population structuring for mature fish in Merluccius paradoxus

The Cape hake species, Merluccius capensis and Merluccius paradoxus are the most important resource of the South African and Namibian demersal fishery, but it is unclear whether there is a single population of each shared by both countries. We analysed the population structure and evolutionary history of these two species using the variable 5' region of the mtDNA control region for 311 specimens of M. capensis and 333 specimens of M. paradoxus sampled between Lüderitz (southern Namibia) to south of Cape Point (South Africa). 107 haplotypes for M. capensis and eight haplotypes for M. paradoxus were recovered. AMOVA and pairwise Phi(st) analyses revealed no structure in M. capensis, however significant genetic differentiation between Namibian and South African 'populations' was detected for M. paradoxus. This was only restricted to mature fish older than 3 and 4 years and not for juvenile fish younger than 3 years. Analyses reveal that M. capensis has undergone population expansion (Fu's Fs=-26.65, P<0.001), possibly within the last 4500-23,000 years, whereas M. paradoxus has not. Our study highlights the utility of genetic markers to unravel the evolutionary history of sympatric species, as well as addressing management issues within regions where commercially valuable fish stocks are shared between nations.     

Spatial,ontogenetic and interspecific variability in stable isotope ratios of nitrogen and carbon of Merluccius capensis and Merluccius paradoxus off South Africa

General linear models (GLMs) were used to determine the relative importance of interspecific, ontogenetic and spatial effects in explaining variability in stable isotope ratios of nitrogen (δ¹⁵N) and carbon (δ¹³C) of the co‐occurring Cape hakes Merluccius capensis and Merluccius paradoxus off South Africa. Significant GLMs were derived for both isotopes, explaining 74 and 56% of observed variance in Merluccius spp. δ¹⁵N and δ¹³C, respectively. Spatial effects (west or south coast) contributed most towards explaining variability in the δ¹⁵N model, with Merluccius spp. off the west coast having higher (by c. 1·4‰) δ¹⁵N levels than Merluccius spp. off the south coast. Fish size and species were also significant in explaining variability in δ¹⁵N, with both species showing significant linear increases in δ¹⁵N with size and M. capensis having higher (by c. 0·7‰) δ¹⁵N values than M. paradoxus. Species and coast explained most and similar amounts of variability in the δ¹³C model, with M. capensis having higher (by c. 0·8‰) δ¹³C values than M. paradoxus, and values being lower (by c. 0·7‰) for fishes off the west coast compared with the south coast. These results not only corroborate the knowledge of Merluccius spp. feeding ecology gained from dietary studies, in particular the ontogenetic change in trophic level corresponding to a changing diet, but also that M. capensis feeds at a slightly higher trophic level than M. paradoxus. The spatial difference in Merluccius spp. δ¹⁵N appears due to a difference in isotopic baseline, and not as a result of Merluccius spp. feeding higher in the food web off the west than the south coast, and provides new evidence that corroborates previous observations of biogeographic differences in isotopic baselines around the South African coast. This study also provides quantitative data on the relative trophic level and trophic width of Cape hakes over a large size range that can be used in ecosystem models of the southern Benguela.     

Biochemical properties and immunohistochemical localization of carbonic anhydrase in the sacculus of the inner ear in the salmon Oncorhynchus masou

Carbonic anhydrase (CA) in the inner ear sacculus was examined by activity assay, Western blotting and immunohistochemistry to determine its role in otolith calcification. An immunoreactive protein with a molecular mass of approximately 28 kDa was detected by Western blotting. The CO2 hydration activity in the cytosol fraction of the sacculus was 5.4 units/mg protein, while little or no activity was detected in the nuclear and mitochondrial fractions. The enzyme activity was highly inhibited by acetazolamide. The concentration of 50% inhibition was 8.16 nM and the inhibition constant of the activity was 8.25 nM. Transitional and squamous epithelial cells of the sacculus were immunopositive with an anti-CA II antibody, but sensory epithelial cells and mitochondria-rich cells in the transitional epithelium were not. These results suggest that transitional epithelial cells other than mitochondria-rich cells and squamous epithelial cells play an important role in otolith calcification by supplying bicarbonate to otoliths and/or by eliminating protons from endolymph.     

Morphogenesis of the inner ear of Rana temporaria (Amphibia,Anura)

Summary Differentiation of the inner ear of Rana temporaria temporaria Linné, 1758 begins with invagination of the epidermis to form the otocyst. The first sensory epithelium to form is the macula communis. Not until this is complete are the semicircular canals produced as protrusions from the otocyst; at the same time the ampullar cristae develop as structures for the detection of rotation, separate from the macula communis. The formation of utricle and saccule, organs for the gravitational sense, occurs by concentric constriction between the superior and inferior parts of the otocyst, dividing the macula communis into two parts. At a time when the utricle and the semicircular canals have completely differentiated, the saccule region of the inner ear is still undergoing morphological development, with the formation first of the two auditory papillae (the basilar and amphibian papillae) and then of the lagena, an evagination of the saccule.Abbreviations aa Anterior ampulla - al Lateral ampulla - c Cartilage - ca Ampullar crista - cc Crus commune - csa Anterior semicircular canal - csl Lateral semicircular canal - csp Posterior semicircular canal - cu Cupula - ed Epidermis - fus Utriculo-saccular foramen - g Ganglion - kc Kinocilium - l Lagena - m Medulla oblongata - mc Macula communis - mes Mesenchyme - ms Macula of the saccule - mu Macula of the utricle - on Organic network - pa Amphibian papilla - pb Basilar papilla - pi Inferior part of otocyst - pm Presumptive medulla oblongata - po Preotolith - ps Superior part of otocyst - raa Anterior acoustic ramus - rl Recess of labyrinth - s Saccule - tm Tectorial membrane - u Utricle - * Perilymphatic space     

Diet of two large sympatric teleosts, the ling (Genypterus blacodes) and hake (Merluccius australis)

Ling and hake are tertiary consumers, and as a result both may have an important structuring role in marine communities. The diets of 2064 ling and 913 hake from Chatham Rise, New Zealand, were determined from examination of stomach contents. Ling was a benthic generalist, and hake a demersal piscivore. The diet of ling was characterised by benthic crustaceans, mainly Munida gracilis and Metanephrops challengeri, and demersal fishes, mainly Macrourids and scavenged offal from fishing vessels. The diet of hake was characterised by teleost fishes, mainly macrourids and merlucciids. Multivariate analyses using distance-based linear models found the most important predictors of diet variability were depth, fish length, and vessel type (whether the sample was collected from a commercial or research vessel) for ling, and fish length and vessel type for hake. There was no interspecific predation between ling and hake, and resource competition was largely restricted to macrourid prey, although the dominant macrourid species predated by ling and hake were different. Cluster analysis of average diet of intraspecific groups of ling and hake confirmed the persistent diet separation. Although size is a central factor in determining ecological processes, similar sized ling and hake had distinctly different foraging ecology, and therefore could influence the ecosystem in different ways, and be unequally affected by ecosystem fluctuations.     

Inermicapsifer beveridgei n. sp. (Cestoidea: Anoplocephalidae) from Procavia capensis (Hyracoidea) in Israel, with notes on two species of Hymenolepis

Inermicapsifer beveridgei n. sp. from Procavia capensis differs from all other species in the genus in having 150-174 testes per segment. The closest species is I. hyracis (Rudolphi, 1810) Janicki, 1919, with 75-120 testes. Hymenolepis vogeae Singh, 1956 and H. horrida (Linstow, 1901) Lühe, 1910, are briefly discussed.     

Mechanics of microtubule bundles in pillar cells from the inner ear.

The mechanical properties of cross-linked microtubule bundles were measured from outer pillar cells isolated from the mammalian inner ear. Measurements were made using a three-point bending test and were incorporated into a mathematical model designed to distinguish between the stiffness contributions from microtubules and their cross-linking proteins. Outer pillar cells were composed of 1000-3000 parallel bundled microtubules in a square array that was interdigitated and cross-linked with actin filaments. The average midpoint bending stiffness of intact cells was 7 x 10(-4) N/m. After removal of both the actin filaments and cross-links with detergent in the presence of DNase I, the square array was disrupted and the stiffness decreased by a factor of 4, to 1.7 x 10(-4) N/m. The bending modulus for individual microtubules was calculated to be 7 x 10(-23) Nm2, and the Young's modulus for these 15 protofilament microtubules was 2 x 10(9) Pa. The shear modulus between microtubules in intact cells was calculated to be 10(3) Pa. It was concluded that cross-linking proteins provided shear resistance between microtubules, which resulted in a fourfold increase in stiffness. The model can be used to estimate the mechanical properties of cross-linked microtubule bundles in cells from which direct measurements are not available.     

Diversity in capsule and seed morphological and phytochemical features and essential oil composition of korarima (Aframomum corrorima (braun) P. C. M. jansen) collections from Ethiopia

Korarima (Aframomum corrorima (Braun) P. C. M. Jansen) is one of the Ethiopian native perennial aromatic herbs belonging to the family Zingiberaceae. Conservation of genotypes and genetic improvement of korarima has been hampered due to lack of evidences on genetic diversity of korarima. Genetic diversity of 44 korarima collections was assessed based on joint capsules and seeds morphological and phytochemical characters. Chemical composition of essential oil in seed samples has also been investigated. Physical measurements were done manually while total organic nutrients, crude fiber and total ash in the seeds were done in terms of proximate analysis. Essential oil in the seed samples was extracted by hydro-distillation and its chemical composition was analyzed with gas chromatography and mass spectrometry (GC-MS). ANOVA revealed significant variations in all the variables considered except capsule and seed shape. Ayda collections showed the largest record for fresh capsule weight (25.3 g), dry capsule weight (6.6 g), hundred seed weight (2.8 g), seed to husk ratio (2.1), total organic matter (66.6%) and seed essential oil (7.3%). Collection from Bazet was also excellent in dry weight of seed per capsule (4.45 g) while those from Metser were best in fruit husk essential oil (1.45%). Cluster analysis of the 44 collections revealed four groups each showing large (23.82–280.13) and significant (p < 0.01) pairwise genetic distance. The pattern of grouping is weak and does not match with the region of collections indicating the presence of different genetic background within the collections as well as genetic similarity among the collections. Principal component analysis (PCA) revealed about 81.2% of the total variations in the first four PCs and the pattern of grouping is similar to cluster analysis. In Essential oil composition, Bona Kike collection was remarkable in 1, 8-Cineole (60.81%). Ayda collection showed the highest m-Mentha-1,8-diene, (+)- (10.05%) and β-farnesene (24.03%). Adele Bise collection was highest in D-Limonene (20.21%). The genetic variability in essential oil composition among the samples was revealed because of spatial separation. The result suggests the potential of Ethiopian korarima in general and the collections in particular for further improvement and commercialization. Overall, the higher extent of morphological and biochemical genetic diversity observed signpost the high opportunity for genetic improvement of the crop via hybridization and selection.     

Differentiation of the inner ear sensory epithelia of Proteus anguinus (Urodela,amphibia)

The stages of differentiation of the inner ear sensory epithelia of the neotenous cave urodele, Proteus anguinus, was studied with light and electron microscopy. Comparative ultrastructural analysis among specimens of different sizes confirms that new sensory cells may be generated throughout life, particularly along the periphery of the saccular macula. The inner ear of Proteus contains at least four types of sensory cells that differ in their apical ciliary part. The lungs and air-filled buccal cavity may function as transducers of sound pressure in underwater conditions. Sound waves might be transmitted from the buccal cavity to the connected oval window. The very complex orientation of the sensory hair cells of the saccular macula and the large overlying saccular otoconial mass suggest that this macula facilitates orientation of Proteus in its underground aqueous habitat.     

Gas exchange of two CAM species of the genus Cissus (vitaceae) differing in morphological features

NADP-malate dehydrogenase extracted from darkened leaves of the C₃ plants pea, barley, wheat and spinach was activated by reduced glutathione, a monothiol, as well as by dithiothreitol (DTT). However, in the C₄ plants maize and Flaveria trinervia, only dithiothreitol could effectively activate the enzyme. There was no activation of the maize enzyme and little or no activation of the F. trinervia enzyme by glutathione. The failure of glutathione to activate NADP-MDH in leaf extracts of maize and F. trinervia may indicate there is some difference in disulfide groups of the protein compared to the C₃ plant enzyme. Both DTT and glutathione could activate NADP-malate dehydrogenase in a partially purified enzyme preparation from pea leaves with or without addition of partially purified thioredoxin. However, the required concentration of reductant was lower with addition of thioredoxin than in its absence. In extracts of C₃ species and the partially purified pea enzyme the level of activation after 40 to 60 min under aerobic conditions was higher (up to twofold) with DTT than with glutathione. Under anaerobic conditions, the initial rate of activation was about twice as high with DTT as with glutathione, but the total activation after 40 to 60 min was similar. Ascorbate was totally ineffective as a reducing agent in activating NADP-MDH from C₃ or C₄ plants, possibly due to its more positive redox potential.Abbreviations Chl Chlorophyll - DTT Dithiothreitol - GSH Reduced Glutathione - NADP-MDH NADP-malate Dehydrogenase     

Ultrastructural features,chromium content and in situ immunodetection of 5-methyl-cytosine following Cr (VI) treatment in two strains of Scenedesmus acutus M. (Chlorophyceae) with different chromium sensitivity

Two strains of the unicellular green alga Scenedesmus acutus (F.J.F. Meyen) with different sensitivity to chromium (VI) were compared to evaluate their ultrastructural morphology in chromium-free and -supplemented medium with a sub-lethal concentration of Cr(VI) for 72 hours. The ultrastructural alteration in different cytological compartments indicated that Cr(VI) induced earlier and stronger alterations in the wild type (wt) compared with the chromium-tolerant strain (Cr-t). After Cr treatments, ICP-MS (Inductively Coupled Plasma Mass Spectrometry) showed a higher Cr accumulation in the wild type than in the Cr-tolerant strain, suggesting a more efficient chromium-exclusion mechanism in the latter. The Cr treatment induced an increase in the nuclear area and a rearrangement in the eu-heterochromatic fraction, suggesting that chromatin remodelling could be at the basis of differential gene expression and metal tolerance. To gain additional information on the remodelling of the nuclear chromatin, we analysed DNA methylation by immunolocalization of 5-methyl-cytosine, before and after Cr exposure. Significant differences in the quantification of the immunolabelling of DNA cytosine-rich zones between the two strains were observed. These data suggest that an epigenetic mechanism could be at the basis of the Cr tolerance in S. acutus, as supported by previous data reporting that the acquired tolerance was inherited and maintained through the progeny.     

Sobolevicanthus transvaalensis n.sp. (Cestoda: Hymenolepididae) from the Cape Teal (Anas capensis Gmelin, 1789)

Summary Sobolevicanthus transvaalensis n.sp. is described from the Cape Teal, Anas capensis Gmelin, 1789, collected in the Republic of South Africa. The new species possesses 8 skrjabinoid hooks 78–88 m long (mean 85 m) and a short claviform cirrus-sac 79–143 m long and resembles S. javanensis (Davis, 1945) and S. terraereginae (Johnston, 1913). It can be distinguished from S. javanensis by its shorter cirrus-sac and smaller cirrus diameter, and by differences in the morphology of the accessory sac and vagina and in their position relative to the cirrus-sac. It can be separated from S. terraereginae on the basis of cirrus length and diameter. The basal diameter of the cirrus in S. terraereginae is three times that in S. transvaalensis. ac]19830414     

Fatty acid composition and dynamics of phospholipids from hake (Merluccius hubbsi) spinal cord and brain and sea bass (Acanthustius brasilianus) brain.

This work studies the phospholipid and fatty acid composition in hake brain and spinal cord and in sea bass brain. Fluorescence anisotropy of phospholipid vesicles labeled with 1,6-diphenyl hexatriene was measured to investigate the associated dynamic properties. In all tissues studied, phosphatidylcholine and phosphatidylethanolamine were the major constituents with minor contributions of phosphatidylserine, phosphatidylinositol and sphingomyelin. Fatty acids belong to the n-9 and n-3 series exclusively. Phosphatidylinositol from hake spinal cord and phosphatidylethanolamine and phosphatidylserine from hake brain contain the greatest percentages of eicosa-5,8,11,14,17-pentaenoic (20:5) and docosa-4,7,10,13,16,19-hexaenoic (22:6), respectively. For all fractions studied the total content of saturated fatty acids increases in the order of hake spinal cord, hake brain, sea bass brain together with a decrease in the sum of monounsaturated fatty acids. The comparison between fluorescence anisotropy values and fatty acid composition clearly demonstrates that saturated acids and 20:5 and 22:6 exert a rigidizing effect.     

Tectorial structures on the inner ear sensory epithelia of Proteus anguinus (Amphibia,caudata)

The tectorial structures of the inner ear of the proteid salamander Proteus anguinus were studied with transmission and scanning electron microscopy in order to analyze the ultrastructure of the otoconial membranes and otoconial masses of the maculae and the tectorial membrane of the papilla amphibiorum. Both otoconial and tectorial membranes consist of two parts: (1) a compact part and (2) a fibrillar part that joins the membrane with the sensory epithelium. Masses of otoconia occupy the lumina above these membranes. There are two types of calcium carbonate crystals in the otoconial masses within the inner ear of Proteus anguinus. The relatively small otoconial mass of the utricular macula occupies an area no greater than the diameter of the sensory epithelium, and it is composed of calcite crystals. On the other hand, the enormous otoconial masses of the saccular macula and the lagenar macula are composed of aragonite crystals. In the sacculus and lagena, globular structures 2–9 m?m in diameter were discovered on the lower surfaces of the otoconial masses above the sensory epithelia. These globules show a progression from smooth-surfaced, small globules to large globules with spongelike, rough surfaces. It is hypothesized that these globules are precursors of the aragonite crystals and that calcite crystals develop similarly in the utriculus. The presence of globular precursors in adult animals suggests that the formation of new crystals in the otoconial membranes of the sacculus and lagena of Proteus is a continuous, ongoing process.     

Sparc (Osteonectin) functions in morphogenesis of the pharyngeal skeleton and inner ear

Sparc (Osteonectin), a matricellular glycoprotein expressed by many differentiated cells, is a major non-collagenous constituent of vertebrate bones. Recent studies indicate that Sparc expression appears early in development, although its function and regulation during embryogenesis are largely unknown. We cloned zebrafish sparc and investigated its role during development, using a mo rpholino antisense oligonucleotide-based knockdown approach. Consistent with its strong expression in the otic vesicle and developing pharyngeal cartilages, knockdown of Sparc function resulted in specific inner ear and cartilage defects that are highlighted by changes in gene expression, morphology and behavior. We rescued the knockdown phenotypes by co-injecting sparc mRNA, providing evidence that the knockdown phenotype is due specifically to impairment of Sparc function. A comparison of the phenotypes of Sparc knockdown and known zebrafish mutants with similar defects places Sparc downstream of sox9 in the genetic network that regulates development of the pharyngeal skeleton and inner ear of vertebrates.