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Shellfish growing areas in rural catchments are occasionally affected by elevated faecal contamination from diffuse sources and may be subject to frequent harvest closures/classification downgrades. We combined traditional risk management methods based on sanitary surveys and monitoring of Escherichia coli in seawater and shellfish with faecal source tracking, bacterial source apportionment, and hydrometeorological modelling to determine the causes of elevated E. coli concentrations contributing to harvest closures in Papanui Inlet (Aotearoa New Zealand). These multiple lines of evidence were used to inform a weight of evidence assessment of bacterial contamination in the inlet. Ruminant livestock was estimated to contribute 80% of the faecal coliform loading. Concentrations of E. coli in seawater were low (≤ 87 MPN 100 ml− 1) while those in tuaki (Austrovenus stutchburyi) occasionally exceeded the “Approved” classification limit (230 MPN 100 g− 1). The most frequent positive genetic markers in seawater were the seagull (Catellicoccus marimammalium) (54% of seawater samples), and in shellfish, the bovine and seagull markers (both 12.5% of shellfish samples). Solar radiation was negatively correlated with E. coli in tuaki. We found that the growing area is affected by faecal inputs from animal and, to a lesser extent, human (septic tank discharges) sources which elevate contamination to levels detectable in tuaki but not in seawater, particularly in the summer months. The innovative approach can enhance the management of shellfish growing areas affected by intermittent contamination and enables more targeted action to reduce pollution to improve shellfish water quality.

We assessed the usefulness of environmental DNA (eDNA) for monitoring the introduced pest fish Gambusia affinis by filtering water samples from streams in the Nelson and Tasman regions, South Islan...

Abiotic and biotic factors control ecosystem biodiversity, but their relative contributions remain unclear. The ultraoligotrophic ecosystem of the Antarctic Dry Valleys, a simple yet highly heterogeneous ecosystem, is a natural laboratory well-suited for resolving the abiotic and biotic controls of community structure. We undertook a multidisciplinary investigation to capture ecologically relevant biotic and abiotic attributes of more than 500 sites in the Dry Valleys, encompassing observed landscape heterogeneities across more than 200 km2. Using richness of autotrophic and heterotrophic taxa as a proxy for functional complexity, we linked measured variables in a parsimonious yet comprehensive structural equation model that explained significant variations in biological complexity and identified landscape-scale and fine-scale abiotic factors as the primary drivers of diversity. However, the inclusion of linkages among functional groups was essential for constructing the best-fitting model. Our findings support the notion that biotic interactions make crucial contributions even in an extremely simple ecosystem., Charles Lee, Daniel Laughlin et al. use structural equation modeling to analyze ecological data from more than 500 sites in the Antarctic Dry Valleys. They find that although abiotic factors are the primary drivers of biodiversity variation, biotic interactions are needed to explain the data fully and may play previously underestimated roles.

Apicomplexan-X (APX) is a significant pathogen of the flat oyster Ostrea chilensis in New Zealand. The life cycle and host range of this species are poorly known, with only the zoite stage identified. Here, we report the use of molecular approaches and histology to confirm the presence of APX in samples of green-lipped mussels Perna canaliculus, Mediterranean mussels Mytilus galloprovincialis and hairy mussels Modiolus areolatus collected from widely distributed locations in New Zealand. The prevalence of APX infection estimated by PCR was 22.2% (n = 99) and 50% (n = 30) in cultured green-lipped mussels from Nelson and Coromandel, respectively; 0.8% (n = 258), 3.3% (n = 150) and 35.3% (n = 17) in wild Mediterranean mussels from Nelson, Foveaux Strait and Golden Bay, respectively; and 46.7% (n = 30) in wild hairy mussels from Foveaux Strait. Histology detected all cases of PCR that were positive with APX and appeared to be more sensitive. The prevalence of APX estimated by histology in green-lipped mussels from Coromandel was 60% versus 50% by PCR, and 4.3%, 10.7% and 52.9% by histology versus 0.8%, 3.3% and 35.3% by PCR in wild Mediterranean mussels from Nelson, Foveaux Strait and Golden Bay, respectively. The specific identity of the parasite found in mussels was determined by sequencing PCR products for a portion (676 bp) of the 18S rRNA gene; the resulting sequences were 99-100% similar to APX found in flat oysters. Phylogenetic analyses also confirmed that all isolates from green-lipped, Mediterranean and hairy mussels grouped with APX isolates previously identified from flat oysters. This study indicates the wide geographical distribution of APX and highlights the potentially multi-host specific distribution of the parasite in commercially important bivalve shellfish.

Apicomplexa (sensu stricto) are a diverse group of obligate parasites to a variety of animal species. Gregarines have been the subject of particular interest due to their diversity, phylogenetically basal position, and more recently, their symbiotic relationships with their hosts. In the present study, four new species of marine eugregarines infecting ascidian hosts (Lankesteria kaiteriteriensis sp. nov., L. dolabra sp. nov., L. savignyii sp. nov., and L. pollywoga sp. nov.) were described using a combination of morphological and molecular data. Phylogenetic analysis using small subunit rDNA sequences suggested that gregarines that parasitize ascidians and polychaetes share a common origin as traditionally hypothesized by predecessors in the discipline. However, Lankesteria and Lecudina species did not form clades as expected, but were instead intermixed amongst each other and their respective type species in the phylogeny. These two major genera are therefore taxonomically problematic. We hypothesize that the continued addition of new species from polychaete and tunicate hosts as well as the construction of multigene phylogenies that include type-material will further dissolve the currently accepted distinction between Lankesteria and Lecudina. The species discovered and described in the current study add new phylogenetic and taxonomic data to the knowledge of marine gregarine parasitism in ascidian hosts.

Bacteria are ubiquitous in all marine habitats and can be beneficial or detrimental to the survival and growth of shellfish raised in aquaculture. To reduce the exposure of cultured shellfish to potential pathogens, antibiotics can be used. However, risks associated with this practice have led to the development of alternative techniques such as ultraviolet (UV) light exposure for seawater disinfection. In this study, we used 16S rRNA gene metabarcoding to measure the effect of low (50 mJoules/cm2) and high (200 mJoules/cm2) UV treatment of seawater on the bacterial communities present in hatchery rearing water and in Pacific oyster Crassostrea gigas larvae. 81 samples were collected between 13 and 29 March 2017 for high-throughput DNA sequencing of bacterial communities, including differentially-treated seawater, the microalgae fed to the oysters and oyster larvae (fertilized eggs, early veliger larval stage [D-larvae], and pre-settlement stage) samples. Differences in larval mortality between low and high UV treatments were also assessed. We found that the two UV treatments influenced the overall bacterial community diversity and affected its composition in both seawater and oyster larval samples. While alpha-diversity was mostly driven by the UV treatment, we found that the microbiome composition was primarily affected by temporal changes in the water source. Compared to the bacterial microbiome associated with seawater, the oyster larvae microbiome changed more significantly in response to both UV treatments and sampling dates with marked shifts in the dominant bacteria associated with fertilized eggs (class Gammaproteobacteria, families Rhodospirillaceae and Pelagibacteraceae), the D-larvae (family Alteromonadaceae), and the pre-settlement larvae (families Flavobacteriaceae and Rhodobacteraceae). As larval development progressed, an increasingly complex bacterial community structure was observed. These bacterial community changes were likely driven by multiple factors, including the microbiome associated with microalgal cultures, which increased in complexity over time. Despite the clear response of bacterial communities to both low and high UV treatments, no significant effect was observed in relation to the oyster larval mortality rate. These results suggest that increasing the UV treatment to 200 mJoules/cm2, if required for more efficacious disinfection and biosecurity, would not be detrimental to Pacific oyster larval survival. Further research is required to increase understanding of the dynamics of functionally critical bacterial taxa and their successive roles in post-larval development of Pacific oysters.

Distinguishing morphologically cryptic taxa, by definition, requires genetic data such as DNA sequences. However, DNA sequences may not be obtained easily for taxa from remote sites. Here we provide the details of a high-resolution melt-curve-based method using taxon-specific primers that can distinguish two taxa of Adelie penguins, and that will be usable in Antarctica when combined with some of the newly developed field-deployable thermal cyclers. We suggest that the wider adoption of field-deployable polymerase-chain-reaction-based techniques will enable faster assignation of haplotype to individuals in situ, and so allow the targeting of observations and sample collection to specimens relevant to the research question. Targeting individuals will also reduce the need to repeatedly handle animals and reduce the time and travel required to complete field work.

Until recently, only one native and three apparently introduced Daphnia species were known from New Zealand. We demonstrate that (1) Daphnia in subalpine habitats in southern New Zealand differ morphologically and genetically from the native taxon previously labelled Daphnia carinata to merit species nova status and (2) the name of the latter should revert to D. thomsoni, used by Sars (1894) for Daphnia described from New Zealand mud. We compare some key characteristics and cytochrome c oxidase subunit 1 (CO1) sequences of the New Zealand native and other morphologically similar species. Distinctive characteristics of subalpine populations, described as Daphnia tewaipounamu sp. nov., are a wide cephalic shield with lateral flanges curving dorsally via rounded fornices, dorsal cervical depression variably expressed as a ‘step’ in the cephalic shield exuviae and retention of ephippia within shed carapace exoskeletons long after ecdysis. CO1 sequences revealed that D. tewaipounamu sp. nov. belongs to the D. carinata complex but is highly divergent (>14%) from other known members of this complex. New Zealand D. thomsoni is divergent (>15%) from D. carinata s.s. However, it is not endemic to New Zealand, as we confirmed its presence in Tasmania, and some Australian populations are closely related to it.

Described here is a polymerase chain reaction (PCR) test to detect the apicomplexan-X (APX) parasite of a flat oyster species, Ostrea chilensis, endemic to New Zealand. The test primers target sequences in the in situ hybridisation probes identified to bind specifically to APX 18S rRNA and amplify a 723 bp DNA product. The test did not amplify 18S rRNA gene sequences of other apicomplexan species, including Toxoplasma gondii, Neospora caninum, Selenidium spp., Cephaloidophorida spp., Lecudina spp. and Thiriotia sp. Of 73 flat oysters identified by histology to be infected with APX at different severities, 69 (95%) tested PCR-positive. Failure to amplify an internal control indicated the presence of PCR inhibitors in the 4 PCR-negative samples. The high analytical sensitivity, specificity and speed of the PCR test should make it a useful tool for detecting APX.

International audience; Advances in the sequencing of DNA extracted from media such as soil and water offer huge opportunities for biodiversity monitoring and assessment, particularly where the collection or identification of whole organisms is impractical. However, there are myriad methods for the extraction, storage, amplification and sequencing of DNA from environmental samples. To help overcome potential biases that may impede the effective comparison of biodiversity data collected by different researchers, we propose a standardised set of procedures for use on different taxa and sample media, largely based on recent trends in their use. Our recommendations describe important steps for sample pre-processing and include the use of (a) Qiagen DNeasy PowerSoil ® and PowerMax ® kits for extraction of DNA from soil, sediment, faeces and leaf litter; (b) DNeasy PowerSoil ® for extraction of DNA from plant tissue; (c) DNeasy Blood and Tissue kits for extraction of DNA from animal tissue; (d) DNeasy Blood and Tissue kits for extraction of DNA from macroorganisms in water and ice; and (e) DNeasy PowerWater ® kits for extraction of DNA from microorganisms in water and ice. Based on key parameters, including the specificity and inclusivity of the primers for the target sequence, we recommend the use of the following primer pairs to amplify DNA for analysis by Illumina MiSeq DNA sequencing: (a) 515f and 806RB to target bacterial 16S rRNA genes (including regions V3 and V4); (b) #3 and #5RC to target eukaryote 18S rRNA genes (including regions V7 and V8); (c) #3 and #5RC are also recommended for the routine analysis of protist community DNA; (d) ITS6F and ITS7R to target the chromistan ITS1 internal transcribed spacer region; (e) S2F and S3R to target the ITS2 internal transcribed spacer in terrestrial plants; (f) fITS7 or gITS7, and ITS4 to target the fungal ITS2 region; (g) NS31 and AML2 to target glomeromycota 18S rRNA genes; and (h) mICOIintF and jgHCO2198 to target cytochrome c oxidase subunit I (COI) genes in animals. More research is currently required to confirm primers suitable for the selective amplification of DNA from specific vertebrate taxa such as fish. Combined, these recommendations represent a framework for efficient, comprehensive and robust DNA-based investigations of biodiversity, applicable to most taxa and ecosystems. The adoption of standardised protocols for biodiversity assessment and monitoring using DNA extracted from environmental samples will enable more informative comparisons among datasets, generating significant benefits for ecological science and biosecurity applications.

We review recent advances in the use of molecular techniques as they apply to monitoring restoration efforts in lakes. Using DNA sequence data, biodiversity can now be assessed to levels previously unattainable using traditional, morphological assessments. In particular, DNA barcoding, the use of small standardised fragments of DNA, has become an increasingly widespread and common approach to identify species. Global initiatives such as the International Barcode of Life (iBOL) have coordinated these efforts and facilitated publically accessible reference databases such as the Barcode of Life Datasystems (BOLD). Such databases can be used for routine identification of specimens as well as for the assessment of community composition and monitoring of changes over time. Through the application of Next Generation Sequencing techniques, multiple samples can be run simultaneously (metabarcoding), greatly automating and streamlining the monitoring process. Reference databases can also be applied to environmental DNA (DNA that is shed into the environment by plants and animals). Here, species can be identified “sight unseen” through analyses of environmental samples (e.g. water, sediment). This latter method has proven useful for the monitoring of exotic fish species, particularly following eradication efforts. Ongoing developments in sequencing technology are likely to further enhance the utility of molecular techniques for assessing and monitoring restoration efforts in New Zealand.

Recent studies have advocated biomonitoring using DNA techniques. In this study, two high-throughput sequencing (HTS)-based methods were evaluated: amplicon metabarcoding of the cytochrome C oxidase subunit I (COI) mitochondrial gene and gene enrichment using MYbaits (targeting nine different genes including COI). The gene-enrichment method does not require PCR amplification and thus avoids biases associated with universal primers. Macroinvertebrate samples were collected from 12 New Zealand rivers. Macroinvertebrates were morphologically identified and enumerated, and their biomass determined. DNA was extracted from all macroinvertebrate samples and HTS undertaken using the illumina miseq platform. Macroinvertebrate communities were characterized from sequence data using either six genes (three of the original nine were not used) or just the COI gene in isolation. The gene-enrichment method (all genes) detected the highest number of taxa and obtained the strongest Spearman rank correlations between the number of sequence reads, abundance and biomass in 67% of the samples. Median detection rates across rare (1% of the total abundance or biomass), moderately abundant (1-5%) and highly abundant (5%) taxa were highest using the gene-enrichment method (all genes). Our data indicated primer biases occurred during amplicon metabarcoding with greater than 80% of sequence reads originating from one taxon in several samples. The accuracy and sensitivity of both HTS methods would be improved with more comprehensive reference sequence databases. The data from this study illustrate the challenges of using PCR amplification-based methods for biomonitoring and highlight the potential benefits of using approaches, such as gene enrichment, which circumvent the need for an initial PCR step.

Marine biofilms are precursors for colonization by larger fouling organisms, including non-indigenous species (NIS). In this study, high-throughput sequencing (HTS) of 18S rRNA metabarcodes was used to investigate four sampling methods (modified syringe, sterilized sponge, underwater tape and sterilized swab) for characterizing eukaryotic communities in marine biofilms. PerspexTM plates were sampled in and out of water. DNA collected with tape did not amplify. Otherwise, there were no statistical differences in communities among the remaining three sampling devices or between the two environments. Sterilized sponges are recommended for ease of use underwater. In-depth HTS analysis identified diverse eukaryotic communities, dominated by Metazoa and Chromoalveolata. Among the latter, diatoms (Bacillariophyceae) were particularly abundant (33% of reads assigned to Chromalveolata). The NIS Ciona savignyi was detected in all samples. The application of HTS in marine biofilm surveillance could facilitate early detection of NIS, improving the probability of successful eradication.

Apicomplexa is a large phylum of parasitic protists renowned for significant negative health impacts on humans and livestock worldwide. Despite the prevalence and negative impacts of apicomplexans across many animal groups, relatively little attention has been given to apicomplexan parasites of invertebrates, especially marine invertebrates. Previous work has reported an apicomplexan parasite ‘X’ (APX), a parasite that has been histologically and ultrastructurally identified from the commercially important flat oyster Ostrea chilensis in New Zealand. This apicomplexan may exacerbate host vulnerability to the infectious disease bonamiosis. In this study, we report 18S rRNA sequences amplified from APX-infected O. chilensis tissues. Phylogenetic analyses clearly established that the 18S sequences were of apicomplexan origin; however, their detailed relationship to known apicomplexan groups is less resolved. Two specific probes, designed from the putative APX 18S rRNA sequence, co-localised with APX cells in in situ hybridisations, further supporting our hypothesis that the 18S sequences were from APX. These sequences will facilitate the future development of inexpensive and sensitive molecular diagnostic tests for APX, thereby assisting research focussed on the biology and ecology of this organism and its role in morbidity and mortality of O. chilensis.

The microbial diversity of faecal communities co-existing with mega fauna is not well understood even though these faecal communities are critical for health and development. Additionally, the transfer of microbial taxa among host animals is little studied. Here, we used 16S sequences obtained from clone libraries to characterise the faecal microbiota of Weddell seals breeding in McMurdo Sound and at White Island, Antarctica. Faecal bacterial communities were dominated by four phyla; Actinobacteria (20 %), Bacteroidetes (13 %), Firmicutes (23 %), and Proteobacteria (13 %). We also used automated ribosomal intergenic spacer analysis to examine the dispersal of bacteria between populations of Weddell seals breeding at White Island and in McMurdo Sound. The Weddell seals at White Island are isolated by the Ross Ice Shelf from the larger population of Weddell seals breeding in McMurdo Sound. We found that the faecal bacteria communities of the seals at White Island had lower diversity and that the community composition was significantly different compared with the seals in the McMurdo Sound area.

The assessment of biological samples is critical for measuring the ‘health’ of New Zealand aquatic environments. Analysis of these samples commonly requires species identification and enumeration, which usually involves microscopy or microbiological methods. These techniques can be time-consuming, laborious, and are dependent on taxonomic expertise. Recent advances in molecular methods provide promising tools for assessing environmental samples. A range of molecular techniques are now used in New Zealand including: fluorescent in situ hybridisation; automated ribosomal intergenic spacer analysis; quantitative polymerase chain reaction; and, most recently, next-generation sequencing. The organisms (or targets) and environments monitored are equally diverse, ranging from cyanobacteria, rotifers and invasive fish in lakes, to macroinvertebrates, and biofilm communities in rivers, to bacteria, micro- and macro-algae and invertebrates in marine ecosystems. Despite research and validation demonstrating their po...

We used a DNA barcoding approach to identify specimens of the Daphnia pulex complex occurring in New Zealand lakes, documenting the establishment of non-indigenous North American Daphnia ‘pulex’. Morphological delineation of species in this complex is problematic due to a lack of good morphological traits to distinguish the species, as there is a relatively high degree of morphological stasis within the group through evolutionary time. Accordingly, genetic analyses were used to determine the specific identity and likely geographic origin of this species. Morphologically, individuals most closely resembled Daphnia pulicaria or Daphnia pulex sensu lato, which cannot be separated morphologically. Furthermore, each of these taxa comprises separate species in North America and Europe, despite carrying the same names. We identified individuals using a 658 bp nucleotide portion of the mitochondrial cytochrome c oxidase subunit 1 gene (COI) as North American Daphnia ‘pulex’, being distinct from European Daphnia pulex sensu stricto and D. pulicaria from Europe or North America. Cellulose allozyme electrophoresis was used to confirm that individuals were not hybrids with D. pulicaria. North American Daphnia ‘pulex’ in New Zealand were first recorded in New Zealand from South Island lakes that are popular for overseas recreational fishers, indicating a possible source of introduction for this species (e.g. on/in fishing gear). Our study provides an additional example of how genetic techniques can be used for the accurate identification of non-indigenous taxa, particularly when morphological species determination is not possible. The growth of global databases such as GenBank and Barcode of Life Datasystems (BOLD) will further enhance this identification capacity.

We examined mitochondrial DNA (COI) variability and distribution of Stereotydeus spp. in Victoria Land and the Transantarctic Mountains, and constructed Neighbour Joining (NJ) and Maximum Likelihood (ML) phylogenetic trees using all publicly available COI sequences for the three Stereotydeus species present (S. belli, S. mollis and S. shoupi). We also included new COI sequences from Miers, Marshall and Garwood valleys in southern Victoria Land (78°S), as well as from the Darwin (79°S) and Beardmore Glacier (83°S) regions. Both NJ and ML methods produced trees which were similar in topology differing only in the placement of the single available S. belli sequence from Cape Hallett (72°S) and a S. mollis haplotype from Miers Valley. Pairwise sequence divergences among species ranged from 9.5–18.1%. NJ and ML grouped S. shoupi from the Beardmore Glacier region as sister to those from the Darwin with pairwise divergences of 8%. These individuals formed a monophyletic clade with high bootstrap support basal to S. mollis and S. belli. Based on these new data, we suggest that the distributional range of S. shoupi extends northward to Darwin Glacier and that a barrier to dispersal for Stereotydeus, and possibly other arthropods, exists immediately to the north of this area.

w e tested the hypothesis that cytochrome c oxidase subunit 1 (COI) sequences would successfully discriminate recognised species of New Zealand caddisflies. w e further examined whether phylogenetic analyses, based on the COI locus, could recover currently recognised superfamilies and suborders. COI sequences were obtained from 105 individuals representing 61 species and all 16 families of Trichoptera known from New Zealand. No sequence sharing was observed between members of different species, and congeneric species showed from 2.3 to 19.5% divergence. Sequence divergence among members of a species was typically low (mean = 0.7%; range 0.0-8.5%), but two species showed intraspecific divergences in excess of 2%. Phylogenetic reconstructions based on COI were largely congruent with previous conclusions based on morphology, although the sequence data did not support placement of the purse-cased caddisflies (Hydroptilidae) within the uncased caddisflies, and, in particular, the Rhyacophiloidea. w e conclude that sequence variation in the COI gene locus is an effective tool for the identification of New Zealand caddisfly species, and can provide preliminary phylogenetic inferences. Further research is needed to ascertain the significance of the few instances of high intra-specific divergence and to determine if any instances of sequence sharing will be detected with larger sample sizes.

The gut of animals changes quickly from a totally sterile environment before birth to a numerous and highly diverse microbial community shortly after birth. However, few studies have examined the source of the bacteria colonizing the gut. We used a genetic based approach to investigate the distribution and acquisition of faecal microbial communities by Adelie penguins, Pygoscelis adeliae, breeding in the Ross Sea region of Antarctica by cloning a portion of the 16S rRNA gene and by automated ribosomal intergenic spacer analysis (ARISA). We hypothesized that bacteria were either acquired from the penguins' neighbours or inherited from their ancestors. Samples were collected from Adelie penguin breeding colonies at the north-western edge of the Ross Sea coast through to the southernmost Adelie penguin colonies on Ross Island. Most of the bacterial sequences we obtained were only distantly homologous with previously published sequences. Bacterial taxa appear to have a restricted distribution as the majority of 16S rRNA clones were isolated from only one or two hosts. Faecal bacterial community similarity was strongly negatively correlated with the genetic distance between hosts suggesting that bacterial communities are inherited. There was little support for a correlation between distance between collection sites and community similarity.

Blue penguins, Eudyptula minor, breeding on Penguin Island, Western Australia are considerably larger than other blue penguins in Australia. If genetic isolation is the cause, it may have implications for the conservation status of some blue penguin populations. We compared the sequences of two mitochondrial gene regions (cytochrome‐b and the control region) from Western Australian blue penguins with other populations of blue penguins from Australia and New Zealand. We found few differences between sequences from Western Australia, Phillip Island, Victoria and Otago, New Zealand, although all three differed considerably from other New Zealand blue penguins. Sequences for the control region from the Western Australian blue penguins and 30 more birds breeding at various Australasian sites provided further support for two major clades within Eudyptula; an Australian clade (including Otago) and a New Zealand clade.

Spiders have been advocated as valuable bio-indicators of forest ecosystem ''health.'' However, the numbers and types of spiders that are recorded at a site will usually be highly dependent on the sampling method employed. The use of lethal, indiscriminate invertebrate sampling techniques is undesirable when investigating rare species, or sampling within areas of high conservation status. There- fore we used non-lethal artificial tree-mounted shelters to monitor arboreal spiders in nature reserves near Christchurch, New Zealand. After three months, over 60% of the shelters had been used by spiders, increasing to 91% after twelve months. There were significant differences in the numbers of spiders found in the shelters at the different sites. However, factors such as the species of tree the shelter was attached to, ground vegetation, and levels of incident light did not affect the likelihood of a shelter being occupied. The species composition of the spider faunas in those sites regarded as high quality forest remnants was dissimilar to the faunas found in the low quality reserves. However, although spiders were more abundant in the high quality sites compared with the poorest stands of woodland, they were not more species rich. The shelters are inexpensive and easy to manufacture and are useful for long-term non-lethal monitoring of spider communities. They also have good potential as a tool for studying spider phenology, population dynamics, behavior, and as a collection/carriage device for live specimens used in conservation translo- cations.

The taxonomy of New Zealand oyster‐catchers is controversial. Some authorities assign full species status to all three oystercatcher taxa breeding in New Zealand, whereas others classify the variable oystercatcher as a full species and the Chatham island oystercatcher and South island pied oystercatcher as distinct only at the subspecies level. The debate is not just of academic interest, as the IUCN lists the Chatham island oystercatcher as endangered and the New Zealand department of Conservation has carried an intensive management programme to conserve it. We obtained genetic data from four regions of the mitochondrial genome of all three taxa, and found support for classifying the Chatham island and South island pied oystercatchers as full species, rather than subspecies.

Previous estimates of a generic level phylogeny for the ubiquitous parasitoid wasp subfamily Microgastrinae (Hymenoptera) have been problematic due to short internal branches deep in the phylogeny. These short branches might be attributed to a rapid radiation among the taxa, the use of genes that are unsuitable for the levels of divergence being examined, or insufficient quantity of data. We added over 1200 nucleotides from four nuclear genes to a dataset derived from three genes to produce a dataset of over 3000 nucleotides per taxon. While the number of well-supported short branches in the phylogeny increased, we still did not obtain strong bootstrap support for every node. Parametric and nonparametric bootstrap simulations projected that an enormous, and likely unobtainable, amount of data would be required to get bootstrap support greater than 50% for every node. However, a marked increase in the number of well-supported nodes was seen when we conducted a Bayesian analysis of a combined dataset generated from morphological characters added to the seven gene dataset. Our results suggest that, in some cases, combining morphological and genetic characters may be the most practical way to increase support for short branches deep in a phylogeny.

When monitoring rare insect species, or when surveying faunas within nature reserves, it is desirable not to use indiscriminate lethal sampling techniques. In this investigation we assessed the usefulness of simple tree-mounted wooden shelters to monitor endemic weta (Orthoptera) in nature reserves in Canterbury, New Zealand. Fifty shelters were placed out at six sites and examined at three-monthly intervals for a year. A wide variety of invertebrates were found utilizing the shelters, with Arachnida, Blattodea and Collembola being the most common occupants. After three months over 80% of the shelters exhibited signs of use by invertebrates, increasing to 96% after 12 months. Only seven tree weta (Anostostomatidae) and one (dead) ground weta (Hemiandrus sp.) were observed in the shelters over the full 12 month period. There were 52 observations of cave weta (Rhaphidophoridae) in the shelters, 36 of which occurred at one site, Orton Bradley Park. Occupation of the shelters by cave weta was not affected by soil conditions, light intensity or aspect of the shelter. However, cave weta exhibited a preference for shelters less than 50 cm above the ground and for shelters attached to kanuka and vines. Although weta were found in only a small proportion (9%) of the shelters, this method proved useful in confirming the presence of weta without risk of harming vulnerable populations. These shelters are inexpensive and easy to manufacture and have potential for long-term non-lethal monitoring of weta and as a collection/carriage device for live specimens used in conservation translocations.

International audience; The taxonomy of rockhopper penguins, Eudyptes chrysocome (Forster 1781), is contentious. Some authorities recognise three subspecies based on morphological differences and geographical separation of breeding populations while others suggest that morphological differences support classifying rockhopper penguins as two distinct species. The taxonomy of rockhopper penguins is of more than academic interest as breeding colonies worldwide have declined markedly in size since the 1930s and rockhopper penguins are currently listed as vulnerable by the IUCN. We compared the genetic distances between three mitochondrial gene regions from the three putative rockhopper penguin subspecies with the distances between various penguin sister species to clarify the taxonomy and systematics of rockhopper penguins. Genetic distances between the rockhopper penguin taxa, relative to other closely related penguin species, support reclassifying the three rockhopper penguin subspecies as species. Reclassification of rockhopper penguins as three species could result in their conservation status being upgraded from vulnerable to endangered.

We describe and illustrate Austrogoniodes vanalphenae, a new species of chewing louse from the yellow‐eyed penguin (Megadyptes antipodes) in New Zealand. We amend the key to the species of the genus Austrogoniodes published in 1967 by Clay to incorporate this new species. We also report the following new host records: Austrogoniodes bifasciatus (Piaget, 1885) from Spheniscus humboldti, and A. demersus Keler, 1952 from Spheniscus mendiculus.

Studies of cophylogenetic associations between hosts and parasites have become increasingly common. Historically, congruence between host and parasite phylogenies has been seen as evidence for cospeciation. Analyses of such coevolutionary relationships, however, are made extremely difficult by the complex interplay of cospeciation, host switching, sorting (extinction), duplication (intrahost speciation) and inertia (lack of parasite speciation) events, all of which may produce incongruence between host and parasite phylogenies. Here we review several methods of analysing cospeciation. We illustrate these methods with an example from a Procellariiformes (seabird) and chewing louse (Halipeurus) association.

Integrated management guidelines for postcontract land use Conservation Reserve Program lands in semiarid regions are generally lacking. We determined the relative efficacy of four systems of transitional conservation practices for producing Old World' bluestem (OWB) (Bothriochlora ischaemum L.) and dryland wheat (Triticum aestivum L.) and cotton (Gossypium hirsutum L.) on former CRP fields. The sites were located on Dalhart fine sandy loam (Aridic Paleustalf) and La Casa-Aspermont clay loam (Typic Paleustoll) near Forgan and Duke, OK, respectively. Removing old growth increased cumulative OWB yields between 1994 and 1997. Applications of 67 kg N and 16.5 kg P ha -1 increased yields by 0, 70, and 180% at Forgan and 290, 70, and 280% at Duke in 1995 to 1997, respectively. Removing the old dry matter and regrowth vigor also enhanced chemical suppression and killing of the grass, the performance of conservation tillage, and achieving a uniform crop stand. Early OWB suppression conserved stored water that was vital to cool-season crop production in the year the contract expired. First-year wheat yields averaged 970, 490, and 1002 kg ha -1 at Forgan and 1590, 600, and 830 kg ha ' at Duke under unfavorable weather conditions (i.e., drought, late freeze ) of 1995 through 1997, respectively. No-till generally produced higher yields, averaging 10 and 35% greater than conservation systems at Forgan and Duke, respectively. In variable semiarid environment, the chance of success for agronomic production decreased in the order of grass production, NT wheat, tilled wheat, and dryland cotton on former CRP lands.

To understand how the social and physical environment influences behaviour, reproduction and survival, studies of underlying hormonal processes are crucial; in particular, interactions between stress and reproductive responses may have critical influences on breeding schedules. Several authors have examined the timing of breeding in relation to environmental stimuli, while others have independently described endocrine profiles. However, few studies have simultaneously measured endocrine profiles, breeding behaviour, and offspring survival across seasons. We measured sex and stress hormone concentrations (oestrogens, testosterone, and corticosterone), timing of breeding, and chick survival, in Adelie penguins (Pygoscelis adeliae) at two colonies in two different years. Clutch initiation at Cape Bird South (CBS; year 1, ∼14,000 pairs) occurred later than at Cape Crozier East (CCE; year 2, ∼25,000 pairs); however, breeding was more synchronous at CBS. This pattern was probably generated by the persistence of extensive sea ice at CBS (year 1). Higher corticosterone metabolite and lower sex hormone concentrations at CBS correlated with later breeding and lower chick survival compared to at CCE - again, a likely consequence of sea ice conditions. Within colonies, sub-colony size (S, 50-100; M, 200-300; L, 500-600; XL, >1000 pairs) did not influence the onset or synchrony of breeding, chick survival, or hormone concentrations. We showed that the endocrine profiles of breeding Adelie penguins can differ markedly between years and/or colonies, and that combining measures of endocrinology, behaviour, and offspring survival can reveal the mechanisms and consequences that different environmental conditions can have on breeding ecology. Refereed/Peer-reviewed

The Montreal Protocol has been very effective in reducing the consumption of the major ozone depleting chemical, methyl bromide (MB), and represents an excellent model for future phaseout of other environmentally damaging products, such as those involved with climate change. Over a ten year period, 85% of MB (c. 45,000 tonnes) used for preplant soil fumigation has been phased out and a wide range of chemical and non chemical technologies adopted for disease and weed control in agriculture. Restrictions on the use of MB have stimulated new research and knowledge on: (1) soil health and relationships between soil microbial diversity and crop growth, and (2) new crop protection agents and production systems that moderate the need for harsh pesticides in agriculture. This has also led to increased use of substrate systems, grafting and plant resistance for disease control which, in most cases, avoid the need for soil disinfestation. Also other fumigants, such as 1,3-dichloropropene (1,3D)/chloropicrin (Pic), metham sodium, iodomethane (MI)/Pic, drip applied fumigants and barrier films have been adopted by growers as alternatives strategies for soil disinfestation. Implementation of a wide range of alternatives has led to a 50% fall in anthropogenic bromine in the troposphere and 30% reduction in effective chlorine load to date in the stratosphere. This has been hugely significant to the start of ozone layer recovery which should be observed within the next few years. Internationally, pressure is mounting to restrict use of all fumigants worldwide (EC Reg 2037/US Cluster Analysis) and this will further stimulate new technologies for plant disease control. There are also moves to restrict MB, approx. 10,300 tonnes a year, used for quarantine and pre-shipment (QPS). The pressure on fumigant use globally is stimulating development of more sustainable crop protection, biosecurity and integrated pest management (IPM) strategies.

Physiological measurements of both stress and sex hormones are often used to estimate the consequences of natural or human-induced change in ecological studies of various animals. Different methods of hormone measurement exist, potentially explaining variation in results across studies; methods should be cross-validated to ensure that they correlate. We directly compared faecal and plasma hormone measurements for the first time in a wild free-living species, the Adelie penguin (Pygoscelis adeliae). Blood and faecal samples were simultaneously collected from individual penguins for comparison and assayed for testosterone and corticosterone (or their metabolites). Sex differences and variability within each measure, and correlation of values across measures were compared. For both hormones, plasma samples showed greater variation than faecal samples. Males had higher mean corticosterone concentrations than females, but the difference was only statistically significant in faecal samples. Plasma testosterone, but not faecal testosterone, was significantly higher in males than females. Correlation between sample types was poor overall, and weaker in females than in males, perhaps because measures from plasma represent hormones that are both free and bound to globulins, whereas measures from faeces represent only the free portion. Faecal samples also represent a cumulative measure of hormones over time, as opposed to a plasma ‘snapshot’ concentration. Our data indicate that faecal sampling appears more suitable for assessing baseline hormone concentrations, whilst plasma sampling may best define immediate responses to environmental events. Consequently, future studies should ensure that they select the most appropriate matrix and method of hormone measurement to answer their research questions. Refereed/Peer-reviewed

The microgastroid complex of braconid wasps is a widely recognized and biologically coherent lineage of endoparasitoids of lepidopteran larvae (caterpillars). The complex has received significant phylogenetic attention in recent years due in part to the taxons' association with mutualistic polydnaviruses, with which they compromise host immune systems. A number of previous attempts using a variety of morphological and molecular approaches have not unequivocally resolved relationships amongst the main subfamilies. This work represents a more extensive attempt to resolve the microgastroid relationships, using seven genes (16S rRNA, cytochrome oxidase I (CO1), 28S rRNA, arginine kinase (ArgK), long wavelength rhodopsin (Ops), elongation factor 1 alpha (EF1a) and wingless (Wg)) and a greater taxonomic representation. Bayesian, likelihood and parsimony phylogenetic reconstructions of this improved data set has determined that the chelonines diverged first from the remainder of the microgastroids, however the relationships amongst the other subfamilies are still unclear, suggesting a greater nucleotide sample is required to resolve them. Examination of the contribution of individual gene trees to the phylogeny demonstrates why the relationships between subfamilies are still unclear, with not all groups monophyletic for all trees. Filtered supernetworks demonstrate that monophyly of all subfamilies is only recovered when splits found in only one or two genes are excluded, but this also results in little remaining structure left in the deep nodes to resolve inter-subfamily relationships. By increasing the breadth of the study we were also able to re-evaluate previous attempts at dating the lineage and, therefore the origin of the polydnavirus association. Previous attempts used a much reduced data set and fewer fossil calibrations. Thorough literature searches have revealed a substantial increase in the fossil calibrations and these, combined with more sophisticated molecular dating analysis, have substantially increased the age of the microgastroid lineage from previous estimates of approximately 73MYA to approximately 100MYA. Examination of the resultant linearized clock tree also allows an insight into the evolution of the more species rich subfamilies. The chelonines appear to have had a steady rate of evolution, whilst the microgastrines and cardiochilines appear to have undergone a more significant "burst" of evolution. It is hypothesized that the different parasitism strategies of subfamilies (Chelonines are egg parasitoids and the remainder are larval parasitioids) may have influenced the evolutionary rates of the groups.

Many ichneumonid and braconid endoparasitoids inject a polydnavirus (PDV) into their caterpillar hosts during oviposition. The viral entities carried by wasps of these families are referred to as “ichnoviruses” (IVs) and “bracoviruses” (BVs), respectively. All IV genomes characterized to date are found in wasps of the subfamily Campopleginae; consequently, little is known about PDVs found in wasps of the subfamily Banchinae, the only other ichneumonid taxon thus far shown to carry these viruses. Here we report on the genome sequence and virion morphology of a PDV carried by the banchine parasitoid Glypta fumiferanae . With an aggregate genome size of ∼290 kb and 105 genome segments, this virus displays a degree of genome segmentation far greater than that reported for BVs or IVs. The size range of its genome segments is also lower than those in the latter two groups. As reported for other PDVs, the predicted open reading frames of this virus cluster into gene families, including the protein tyrosine phosphatase (PTP) and viral ankyrin ( ank ) families, but phylogenetic analysis indicates that ank genes of the G. fumiferanae virus are not embedded within the IV lineage, while its PTPs and those of BVs form distinct clusters. The banchine PDV genome also encodes a novel family of NTPase-like proteins displaying a pox-D5 domain. The unique genomic features of the first banchine virus examined, along with the morphological singularities of its virions (IV-like nucleocapsids, but enveloped in groups like some of the BVs), suggest that they could have an origin distinct from those of IVs and BVs.

It is generally thought that the evolution of obligate parasites should be linked intimately to the evolution of their hosts and that speciation by the hosts should cause speciation of their parasites. The penguins and their chewing lice present a rare opportunity to examine codivergence between a complete host order and its parasitic lice. We estimated a phylogeny for all 15 species of lice parasitising all 17 species of penguins from the third domain of the mitochondrial 12S ribosomal rRNA gene, a portion of the mitochondrial cytochrome oxidase subunit 1 gene and 55 morphological characters. We found no evidence of extensive cospeciation between penguins and their chewing lice using TreeMap 2.02beta. Despite the paucity of cospeciation, there is support for significant congruence between the louse and penguin phylogenies due to possible failure to speciate events (parasites not speciating in response to their hosts speciating).

Cophylogenetic studies examine the relationship between host and parasite evolution. One aspect of cophylogenetic studies that has had little modern discussion is parasites with multiple definitive hosts. Parasite species with multiple host species are anomalous as, under a codivergence paradigm, speciation by the hosts should cause speciation of their parasites. We discuss situations such as cryptic parasite species, recent host switching or failure to speciate that may generate multi-host parasites. We suggest methods to identify which of the mechanisms have led to multi-host parasitism. Applying the suggested methods may allow multi-host parasites to be integrated more fully into cophylogenetic studies.

The wide spread occurrence of mummified seal and penguincarcasses tens of kilometres from the open ocean is aninteresting phenomenon occurring in the Antarctic DryValleys. Mummified seal carcasses were first reported byScott’s expedition in 1903 (Scott 1969), and live seals andseal carcasses have since been reported many kilometresfrom the nearest ice-free ocean. For example, Stirling &Rudolph (1968) reported a live crabeater seal near MountSaunders, Marie Byrd Land, 113km from the open ocean.Seal carcasses found in the McMurdo Dry Valleys arepredominantly crabeater seals (

Penguins are parasitised by 15 species of lice in the genera Austrogoniodes and Nesiotinus and present an opportunity to analyse phylogenetic relationships of two complete genera of chewing lice parasitising a monophyletic group of hosts. Taxonomy of penguin lice has been revised several times, including the erection of the genus Cesareus to contain some of the penguin-chewing louse species. Additionally, other groups of species within Austrogoniodes have been proposed. We constructed a phylogeny for all the chewing lice parasitising penguins from 46 parsimony-informative morphological characters and found support for two groups within Austrogoniodes, but little support for the Cesareus genus. Austrogoniodes metoecus, the only Austrogoniodes species parasitising a bird other than a penguin, was basal in the phylogeny, which suggests that if A. metoecus did originate from a louse species parasitising penguins, the host-switching event was unlikely to have been recent. A�superficial comparison of louse and penguin phylogenies identified some potential instances of co-speciation. However, a full analysis of co-phylogenetic relationships between penguins and their lice awaits the publication of a better-resolved penguin phylogeny.