Biosynthesis of cyanogenic glucosides: in vitro analysis of the glucosylation step

The last step in the biosynthesis of cyanogenic glucosides, the glucosylation of the cyanohydrin intermediate, has been investigated in detail using Triglochin maritima seedlings. The glucosyltransferase activity is not associated with membranes and appears to be a "soluble" enzyme. The cyanohydrin intermediate, which is formed by hydroxylation of 4-hydroxyphenylacetonitrile by a membrane-bound enzyme, is free to equilibrate in the presence of the glucosyltransferase and UDPG, because it can be trapped very efficiently. This indicates that this intermediate is not channeled (unlike some of the other intermediates), although it is probably the most labile of all of them. The glucosyltransferase of T. maritima responsible for the glucosylation of the cyanohydrin was separated from another glucosyltransferase, which used 4-hydroxybenzylalcohol as a substrate, and purified over 200-fold. It catalyzed the glucose transfer from UDPG to only 4-hydroxymandelonitrile and 3,4-dihydroxymandelonitrile, giving rise to the respective cyanogenic glucosides. Although the activities with these two substrates behaved differently in certain respects (e.g., extent of inactivation during purification and difference in activation by higher salt concentrations), most of the data acquired favor the view that only one enzyme in T. maritima is responsible for the glucosylation of both substrates.     

Artificial Spawning Habitats Improve Egg Production of a Declining Diadromous Fish,Galaxias maculatus (Jenyns, 1842)

Anthropogenic impacts from urbanization, deforestation, and agriculture have degraded the riparian margins of waterways worldwide. In New Zealand, such impacts have caused changes in native vegetation, enhanced invasion by exotic grasses, and altered river bank morphology. One consequence has been a great reduction in obligate spawning habitat of a diadromous fish, Galaxias maculatus. Juvenile G. maculatus comprise a culturally important fishery that has been considerably reduced over recent decades. Rehabilitation of riparian vegetation needed for spawning is relatively straightforward, but slow. We hypothesized that artificial spawning habitats could accelerate restoration of fish egg production by creating an environment that would support at least the same density and survival of eggs as non‐impacted vegetation. We tested three artificial devices (straw bales, straw tubes, and moss tubes) in degraded and intact sites. Eggs were laid in all of these with numbers and survival usually exceeding that in riparian grasses. Where habitat was degraded, artificial spawning habitats yielded up to 10,000 eggs compared to none in nearby natural spawning habitat. The ground‐level environment of artificial habitat was similar to that of intact vegetation in buffering ambient temperature and humidity fluctuations. Crucial properties of the artificial habitats were (1) shelter to provide shade and hold moisture; (2) accessibility to allow adult fish to deposit and fertilize eggs; and (3) robustness to provide reliable surfaces and protection for the eggs during their development. We show that artificial spawning habitats are a viable short‐term alternative to rehabilitating spawning habitat while legacy effects abate and riparian vegetation recovers.     

Population structure of Placostylus hongii (Gastropoda: Paryphantidae) on the Poor Knights Islands

Abstract

Populations of the native land snail Placostylus hongii (Lesson, 1830) on the Poor Knights Islands were investigated during visits in 1976, 1977, and 1978. Marking and recapture of individual snails showed that linear growth of the shell ceased when a lip was formed around the outer margin of the aperture. Measurements of snails suggested that increase in outer lip width was a good indication of age, but that the rate of lip thickening varied between populations. Large Placostylus showed a restricted range of movement. Both tagging studies and random sampling revealed that mortality was size specific. Most small snails (<60 mm shell length) located were dead, whereas most larger snails were alive. Population densities of living snails averaged 2.2 individuals per 0.25 m² in the study area. The local distribution was patchy owing to the tendency of snails to be associated with specific features of the vegetation.     

Experimental gill-netting of reef fish: Species-specific responses modify capture probability across mesh sizes

Gill-nets are highly selective in terms of the sizes of fish they catch, but often unselective in terms of the suite of fish species they capture. We investigated gill-net selectivity from the point of view of behavioural interactions between the fish and the gear. We observed interactions between fish and gill-nets of three mesh sizes (65 mm, 88 mm & 110 mm) set over rocky reefs in southern New Zealand. There were significant differences among eight species of mobile reef fish in their response to gill-nets and in their capture rates. Some species were more vulnerable because of their use of habitat, swimming motion or morphology. Species that occupied low visibility habitats (e.g., the herbivorous Odax pullus, which mostly swims beneath the algal canopy) were more susceptible to being caught because they had little time to detect and avoid the gill-nets. Species with carangiform or sub-carangiform swimming motion (e.g., Latridopsis ciliaris or O. pullus) were more susceptible to being caught because once in the gill-net, they could only attempt to force their way forwards becoming wedged further into the mesh. Species whose morphology makes tangling in the mesh more likely (e.g., large or protruding spines (Aplodactylus arctidens), fins (L. ciliaris) or opercula) are also more susceptible to being caught. Some species, particularly the common labrid Notolabrus celidotus, were less susceptible than other species to being caught. Fewer than 1% of 538 N. celidotus observed within one metre of the gill-nets were caught. Most N. celidotus altered their swimming direction near the gill-nets and did not hit the mesh. N. celidotus that swam through the nets were smaller than those that swam over the gill-nets or turned away. The fact that different size classes had different responses suggests that interactions with the gill-net are actively controlled. To divers, it appeared that this species could readily detect the gill-nets and treated them as part of the seascape. Furthermore, their labriform swimming motion allowed them to swim backwards out of gill-nets to avoid becoming caught. The species-specific responses of reef fish near the gill-nets and behavioural differences may explain the low numbers of some common reef fish that are caught in gill-nets and the disproportionately high numbers of others. The potentially great ancillary effects from by-catch of important species of untargeted reef fish, birds and marine mammals make gill-nets a somewhat blunderbuss method of catching fish on coastal reefs.     

Observation of brown-throated three-toed sloths: mating behaviour and the simultaneous nurturing of two young

Bradypus variegatus is a member of the Order Pilosa, Family Bradypodidae, and is distributed in many subtropical and tropical countries in South and Central America. However, studies on this species in the wild are relatively limited and many aspects of its reproductive behaviour are unknown or unclear. The current report presents new observations of the reproductive behaviour of B. variegatus in its natural environment. These include details of both a male–female copulation and the simultaneous nurturing of two young sloths.     

The role of mass spectrometry in the characterization of biologic protein products

Introduction: Mass spectrometry (MS) is widely used in the characterization of biomolecules including peptide and protein therapeutics. These biotechnology products have seen rapid growth over the past few decades and continue to dominate the global pharmaceutical market. Advances in MS instrumentation and techniques have enhanced protein characterization capabilities and supported an increased development of biopharmaceutical products.

Areas covered: This review describes recent developments in MS-based biotherapeutic analysis including sequence determination, post-translational modifications (PTMs) and higher order structure (HOS) analysis along with improvements in ionization and dissociation methods. An outlook of emerging applications of MS in the lifecycle of product development such as comparability, biosimilarity and quality control practices is also presented.

Expert commentary: MS-based methods have established their utility in the analysis of new biotechnology products and their lifecycle appropriate implementation. In the future, MS will likely continue to grow as one of the leading protein identification and characterization techniques in the biopharmaceutical industry landscape.     

Comparative phylogeography of coastal limpets across a marine disjunction in New Zealand

Cook Strait, which separates the North and South Island of New Zealand, has been a transient, but re-occurring feature of the New Zealand land mass throughout the Pleistocene, maintaining its current width and depth for the past 5000 years. Historic land fragmentation coupled with the complex hydrography of the Greater Cook Strait region has created both biogeographic and phylogeographic disjunctions between the North and South Island in several marine species. Here we use mitochondrial cytochrome b DNA sequences of three endemic intertidal limpets, Cellana ornata, Cellana radians and Cellana flava to assess intraspecific phylogeographic patterns across Cook Strait and to look for interspecific concordance of ecological and evolutionary processes among closely related taxa. We sequenced 328-359 bp in 85-321 individuals from 8-31 populations spanning the biogeographic range of the three species. Intraspecific phylogeographic analyses show moderate to strong genetic discontinuity among North and South Island populations due to allopatric fragmentation. This pattern was broadly concordant across the three species and the observed divergence among this group of intertidal limpets (0.3-2.0%) is similar to that of previously studied subtidal organisms. For each species, divergence time calculations suggest contemporary North and South Island lineages diverged from their respective most recent common ancestor approximately 200 000 to 300 000 years before present (bp), significantly earlier than previous estimates in other coastal marine taxa that arose from a miscalculation of divergence time.     

Population sinks resulting from degraded habitats of an obligate life-history pathway

In networks of plant–animal mutualisms, different animal groups interact preferentially with different plants, thus forming distinct modules responsible for different parts of the service. However, what we currently know about seed dispersal networks is based only on birds. Therefore, we wished to fill this gap by studying bat–fruit networks and testing how they differ from bird–fruit networks. As dietary overlap of Neotropical bats and birds is low, they should form distinct mutualistic modules within local networks. Furthermore, since frugivory evolved only once among Neotropical bats, but several times independently among Neotropical birds, greater dietary overlap is expected among bats, and thus connectance and nestedness should be higher in bat–fruit networks. If bat–fruit networks have higher nestedness and connectance, they should be more robust to extinctions. We analyzed 1 mixed network of both bats and birds and 20 networks that consisted exclusively of either bats (11) or birds (9). As expected, the structure of the mixed network was both modular (M = 0.45) and nested (NODF = 0.31); one module contained only birds and two only bats. In 20 datasets with only one disperser group, bat–fruit networks (NODF = 0.53 ± 0.09, C = 0.30 ± 0.11) were more nested and had a higher connectance than bird–fruit networks (NODF = 0.42 ± 0.07, C = 0.22 ± 0.09). Unexpectedly, robustness to extinction of animal species was higher in bird–fruit networks (R = 0.60 ± 0.13) than in bat–fruit networks (R = 0.54 ± 0.09), and differences were explained mainly by species richness. These findings suggest that a modular structure also occurs in seed dispersal networks, similar to pollination networks. The higher nestedness and connectance observed in bat–fruit networks compared with bird–fruit networks may be explained by the monophyletic evolution of frugivory in Neotropical bats, among which the diets of specialists seem to have evolved from the pool of fruits consumed by generalists.