Direct and indirect effects of egg parasitism by Neopolyscystus Girault sp. (Hymenoptera: Pteromalidae) on Paropsis atomaria Olivier (Coleoptera: Chrysomelidae)

Abstract Neopolycystus sp. is the only primary egg parasitoid associated with the pest beetle Paropsis atomaria in subtropical eucalypt plantations, but its impact on its host populations is unknown. The simplified ecosystem represented by the plantation habitat, lack of interspecific competition for host and parasitoid, and the multivoltinism of the host population makes this an ideal system for quantifying the direct and indirect effects of egg parasitism, and hence, effects on host population dynamics. Within-, between- and overall-egg-batch parasitism rates were determined at three field sites over two field seasons, and up to seven host generations. The effect of exposure time (egg batch age), host density proximity to native forest and water sources on egg parasitism rates was also tested. Neopolycystus sp. exerts a significant influence on P. atomaria populations in Eucalyptus cloeziana. plantations in south-eastern Queensland, causing the direct (13%) and indirect (15%) mortality of almost one-third of all eggs in the field. Across seasons and generations, 45% of egg batches were parasitised, with a within-batch parasitism rate of around 30%. Between-batch parasitism increased up to 5–6 days after oviposition in the field, although within-batch parasitism rates generally did not. However, there were few apparent patterns to egg parasitism, with rates often varying significantly between sites and seasons.

An egg apparatus-specific enhancer of Arabidopsis, identified by enhancer detection

Despite a central role in angiosperm reproduction, few gametophyte-specific genes and promoters have been isolated, particularly for the inaccessible female gametophyte (embryo sac). Using the Ds-based enhancer-detector line ET253, we have cloned an egg apparatus-specific enhancer (EASE) from Arabidopsis (Arabidopsis thaliana). The genomic region flanking the Ds insertion site was further analyzed by examining its capability to control gusA and GFP reporter gene expression in the embryo sac in a transgenic context. Through analysis of a 5' and 3' deletion series in transgenic Arabidopsis, the sequence responsible for egg apparatus-specific expression was delineated to 77 bp. Our data showed that this enhancer is unique in the Arabidopsis genome, is conserved among different accessions, and shows an unusual pattern of sequence variation. This EASE works independently of position and orientation in Arabidopsis but is probably not associated with any nearby gene, suggesting either that it acts over a large distance or that a cryptic element was detected. Embryo-specific ablation in Arabidopsis was achieved by transactivation of a diphtheria toxin gene under the control of the EASE. The potential application of the EASE element and similar control elements as part of an open-source biotechnology toolkit for apomixis is discussed.

History behind the egg: Women's comprehensive histories and IVF outcomes, Australia, 2008-2009 [abstract]

The individual history of infertile women, as well as their age, may influence their response to in vitro fertilisation (IVF) cycles. This study examined the associations between women’s histories and two IVF outcomes: eggs aspirated (EA) and proportion with normal, two-pronuclei (2PN), fertilisation. This is a cross-sectional survey of infertile women (n=141, 27-46 years) from a multi-centre clinical sample. Participants completed a survey of socio-demographic, relationship, lifestyle, reproductive and fertility factors, medical conditions and recurrent symptoms. Among participants with heterosexual partners (n=122), associations between women’s histories and EA or 2PN fertilisation were analysed using linear and logistic modelling, respectively, adjusted for age at EA and accounting for multiple IVF cycles (n=313 cycles). Participants aged 35+ years had reproductive histories of miscarriage only (16.9%), termination only (9.9%) or birth+termination (5.6%) that were 2-, 3- and 4-fold higher, respectively, than those aged <35 years (7.1%, 2.9%, 1.4%). More years of oral contraceptive use were associated with a lower mean EA: never used, 14.6 EA; 0-2 years, 11.7 EA; 3-5 years, 8.6 EA; 6þ years, 8.2 EA (p=.04). Participants with polycystic ovary syndrome had a higher mean EA (11.5) than those without the condition (8.3 EA, p<.01). Participants in trade or service occupations had lower proportions of 2PN fertilisation (51.7%) than participants in other occupations (professional, 58.6%; manual/other, 63.6%, p<.02). Increasing women’s age and prolonged used of oral contraceptives were associated with lower EA from IVF cycles; PCOS was associated with higher EA. Occupational exposures may have a detrimental effect on normal fertilisation rates.

History behind the IVF egg: Are women's comprehensive histories associated with the number of eggs collected, or fertilised normally, from repeated IVF cycles? [abstract]

Most studies of in vitro fertilisation (IVF) outcomes use cycle-based data and fail to account for women who use repeated IVF cycles. The objective of this study was to examine the association between the number of eggs collected (EC) and the percentage fertilised normally, and women’s self-reported medical, personal and social histories. This study involved a crosssectional survey of infertile women (aged 27-46 years) recruited from four privately-owned fertility clinics located in major cities of Australia. Regression modeling was used to estimate the mean EC and mean percentage of eggs fertilised normally: adjusted for age at EC. Appropriate statistical methods were used to take account of repeated IVF cycles by the same women. Among 121 participants who returned the survey and completed 286 IVF cycles, the mean age at EC was 35.2 years (SD 4.5). Women’s age at EC was strongly associated with the number of EC: <30 years, 11.7 EC; 30.0-< 35 years, 10.6 EC; 35.0-<40.0 years, 7.3 EC; 40.0+ years, 8.1 EC; p<.0001. Prolonged use of oral contraceptives was associated with lower numbers of EC: never used, 14.6 EC; 0-2 years, 11.7 EC; 3-5 years, 8.5 EC; 6þ years, 8.2 EC; p=.04. Polycystic ovary syndrome (PCOS) was associated with more EC: have PCOS, 11.5 EC; no, 8.3 EC; p=.01. Occupational exposures may be detrimental to normal fertilisation: professional roles, 58.8%; trade and service roles, 51.8%; manual and other roles, 63.3%; p=.02. In conclusion, women’s age remains the most significant characteristic associated with EC but not the percentage of eggs fertilised normally.

A comparison of lungworm faecal larval counts and trichostrongyloid faecal egg counts between Red Deer (Cervus elaphus) and Red DeerXWapiti F1 hybrids.

To determine if breed differences in susceptibility to trichostrongyloid and lungworm infection exist, two groups of weaner deer containing seven red deer and red deer X wapiti F1 hybrids were compared using faecal egg counts and faecal larval counts. All animals were run on the same pasture at the same time and treated with the same anthelmintics at the same time. Results indicated that there were significant differences between red deer and red deer X wapiti hybrids, with red deer having higher faecal lungworm counts and red deer X wapiti hybrids having significantly higher faecal egg counts. It is likely that these differences were due to breed. Differences in the efficacy of anthelmintic treatments were also noted between the two groups, with oral oxfendazole being less effective at reducing faecal lungworm counts in red deer X wapiti hybrids than red deer.

"The last egg in the basket?" Elderly primiparity: A review of findings

The recent trend of later childbearing among primiparous women is examined with attention to its possible motivations and how they might relate to maternal and infant outcomes. The influence of psychological factors is examined in the context of the inconsistencies in the obstetric literature regarding the risk factors associated with advancing maternal age. An interplay of psychological and biologic factors in determining the outcome of pregnancy and adaptation to motherhood in elderly primiparous women is proposed in terms of the compensatory as opposed to cumulative effect of psychological factors. The adoption of a broad perspective in the study of the experience of later motherhood is recommended as the direction for future research.

Supporting ICT diffusion in Australian construction organisations

Construction organisations comprise geographically dispersed virtually-linked suborganisations that work together to realise projects. They increasingly do so using information and communication technology (ICT) to communicate, coordinate their activities and to solve complex problems. One salient problem they face is how to effectively use requisite ICT tools. One important tool at their disposal is the self-help group, a body of people that organically spring up to solve shared problems. The more recognised term for this organisational form is a community of practice (COP). COPs generate knowledge networks that enhance and sustain competitive advantage and they are also used to help COP members actually use ICT tools. Etienne Wenger defines communities of practice as “groups of people informally bound together by shared expertise and passion for a joint enterprise” (Wenger and Snyder 2000, p139). This ‘chicken-or-egg’ issue about needing a COP to use the tools that are needed to effective broaden COPs (beyond co-located these groups) led us to explore how best to improve the process of ICT diffusion through construction organisations— primarily using people supported by technology that improves knowledge sharing. We present insights gained from recent PhD research results in this area. A semistructured interview approach was used to collect data from ICT strategists and users in the three large Australian construction organisations that are among the 10 or so first tier companies by annual dollar turnover in Australia. The interviewees were categorised into five organisational levels: IT strategist, implementer, project or engineering manager, site engineer and foreman. The focus of the study was on the organisation and the way that it implements ICT diffusion of a groupware ICT diffusion initiative. Several types of COP networks from the three Australian cases are identified: withinorganisation COP; institutional, implementer or technical support; project manager/engineer focussed; and collegial support. Also, there are cross-organisational COPs that organically emerge as a result of people sharing an interest or experience in something significant. Firstly, an institutional network is defined as a strategic group, interested in development of technology innovation within an organisation. This COP principally links business process domain experts with an ICT strategist.

Factors affecting reproductive performance of the prawn, Penaeus monodon

The growth of the Penaeus monodon prawn aquaculture industry in Australia is hampered by a reliance on wild-caught broodstock. This species has proven difficult to breed from if broodstock are reared in captivity. Studies were therefore carried out to investigate factors controlling reproduction and influencing egg quality. Results of the studies revealed that patterns of nutrient accumulation during early ovary development are altered by captive conditions, possibly contributing to reduce larval quality. The sinus gland hormones were shown, together with the environment, to regulate two stages of ovary development. In a separate study it was further revealed that the hormone methyl farnesoate (MF) could negatively regulate the final stages of ovary development. Lastly it was shown that broodstock reared in captivity are less likely to mate and that this is due to inherent problems in both the male and the female prawns.

Natural enemies of Paropsis atomaria Olivier (Coleoptera: Chrysomelidae) in south-eastern Queensland eucalypt plantations

Abstract A field survey for natural enemies of Paropsis atomaria was conducted at two south-eastern Queensland Eucalyptus cloeziana plantation sites during 2004–2005. Primary egg and larval parasitoids and associated hyperparasitoids were identified to genus or species, and parasitism rates were determined throughout the season. Predators were identified to family level but their impact was not quantified. P. atomaria adults were also examined as potential hosts for parasitic mites and nematodes. An undescribed species of Neopolycystus (Pteromalidae) was the major primary egg parasitoid species reared from egg batches, parasitising half of all egg batches collected. Three hyperparasitoid species (Baeoanusia albifunicle (Encyrtidae), Neblatticida sp. (Encyrtidae) and Aphaneromella sp. (Platygasteridae) were present, representing around one-quarter to one-third of all emergent wasps; this is the first host association record for Neopolycystus–B. albifunicle. In contrast to populations of P. atomaria from the Australian Capital Territory, primary larval parasitism was very low, around 1%, and attributable only to the tachinid flies Anagonia sp. and Paropsivora sp. However, the presence of the sit-and-wait larval hyperparasitoid, Perilampus sp. (Perilampidae) was high, emerging from around 17% of tachinid pupae, with planidia infesting a further 40% of unparasitised hosts. Three species of podapolipid mites parasitised sexually mature P. atomaria adults, while no nematodes were found in this study. Spiders were the most common predators and their abundance was positively correlated with P. atomaria adult and egg numbers. Although natural enemy species composition was identical between our two study sites, significant differences in abundance and frequency were found between sites

Conceptions of school-based youth health nursing

The aim of this research is to explore the meaning of the experience of school-based youth health nursing in Queensland, Australia. The research follows a qualitative approach and is based on indepth interviews. The dominant experience is negative because participants feel they have to battle to gain respect and survive in the school environment. The small, positive experience of school-based youth health nursing is related to student consultations. Student consultations are a ‘golden egg’ because participants gain a sense of reward from making a difference to student wellbeing. This paper proposes operational recommendations including those related to health promotion and professional development and strategic recommendations regarding this model of school nursing. The authors conclude, first, that this ‘golden egg’ should be promoted to ensure all school nurses reap the rewards, second, that this model of school nursing is not the most effective model.

Molecules, morphology, and ecology indicate a recent, amphibious ancestry for echidnas

The semiaquatic platypus and terrestrial echidnas (spiny anteaters) are the only living egg-laying mammals (monotremes). The fossil record has provided few clues as to their origins and the evolution of their ecological specializations; however, recent reassignment of the Early Cretaceous Teinolophos and Steropodon to the platypus lineage implies that platypuses and echidnas diverged >112.5 million years ago, reinforcing the notion of monotremes as living fossils. This placement is based primarily on characters related to a single feature, the enlarged mandibular canal, which supplies blood vessels and dense electrosensory receptors to the platypus bill. Our reevaluation of the morphological data instead groups platypus and echidnas to the exclusion of Teinolophos and Steropodon and suggests that an enlarged mandibular canal is ancestral for monotremes (partly reversed in echidnas, in association with general mandibular reduction). A multigene evaluation of the echidna–platypus divergence using both a relaxed molecular clock and direct fossil calibrations reveals a recent split of 19–48 million years ago. Platypus-like monotremes (Monotrematum) predate this divergence, indicating that echidnas had aquatically foraging ancestors that reinvaded terrestrial ecosystems. This ecological shift and the associated radiation of echidnas represent a recent expansion of niche space despite potential competition from marsupials. Monotremes might have survived the invasion of marsupials into Australasia by exploiting ecological niches in which marsupials are restricted by their reproductive mode. Morphology, ecology, and molecular biology together indicate that Teinolophos and Steropodon are basal monotremes rather than platypus relatives, and that living monotremes are a relatively recent radiation.

Studies of assisted reproduction in the spotted grass frog Limnodynastes tasmaniensis: ovulation, early development and microinjection (ICSI)

Assisted Reproductive Technologies (ART) offer a wide range of techniques that have the potential to augment efforts to conserve and manage endangered amphibians and improve wild and captive population numbers. Gametes and tissues of species nearing endangered or extinct status can be cryopreserved and stored in gene banks, to provide material that can be utilised in the future as ART methods are refined. The Spotted Grass Frog, Limnodynastes tasmaniensis, is an abundant amphibian species in South-Eastern Australia of the family Myobatrachidae, that is suitable for the development of ART systems that can be applied to the threatened and endangered myobatrachid and other amphibian species native to Australia. The aim of this study was to advance the understanding of ovulation, fertilisation and embryo nic development of Lim. tasmaniensis and in vitro manipulations of reproduction and development for use in the development of advanced ART procedures such as intracytoplasmic spermatozoon injection (ICSI), androgenesis and nuclear transfer. Ovulation in amphibians can be induced by protocols utilising natural or synthetic hormones. All protocols tested on Lim. tasmaniensis in this study required two injections and the most effective protocols continued to require a first injection of pituitary extracts to induce ovulation. The second injection was, however, successfully replaced by synthetic chorionic gonadotrophin at a threshold dosage of 100 iu and halved the number of cane toads required to source the pituitaries. A combination of LHRH and Pimozide offered a less effective protocol, that did not require the use of pituitary extracts, and avoided the risk of pathogen transfer associated with unsterilised pituitary extracts. Unfertilised eggs of Lim. tasmaniensis were exposed to media of various osmolalities to determine media effects on eggs and their surrounding jelly layers that might impact on egg viability and fertilisability. Osmolality had no effect upon the egg diameter, however, rapid swelling of the jelly layers occurred within 15 minutes of exposure to various media treatments and plateaued from 30-90 minutes without further expansion. Swelling of the jelly layers was increased in hypotonic media (2.5% SAR, H2O) and minimised in the isotonic media (100% SAR). The optimal conditions for the culture of Lim. tasmaniensis eggs were identified as a holding media of 100% SAR, followed by a medium change to 2.5% SAR at insemination. This sequence of media minimised the rate of swelling of the jelly layers prior to contact with the spermatozoa, and maximised the activation of spermatozoa and eggs throughout fertilisation and embryonic development. Embryos of Lim. tasmaniensis were cultured at four temperatures (13 C, 17 C, 23 C and 29 C), to determine the effect of temperature on cleavage and embryonic development rates. Embryonic development progressed through a sequence of stages that were not altered by changes in temperature. However cleavage rates were affected by changes in temperature as compared with normal embryonic growth at 23 C. Embryonic development was suspended at the lowest temperature (13 C) while embryonic viability was maintained. A moderate decrease in temperature (17 C) slowed cleavage, while the highest temperature (29 C) increased the cleavage rate, but decreased the embryo survival. Rates of embryonic development can be manipulated by changes in temperature and this method can be used to source blastomeres of a specific size/stage at a predetermined age or halt cleavage at specific stages for embryos or embryo derived cells to be included in ART procedures. This study produced the first report of the application of Intracytoplasmic Spermatozoon Injection (ICSI) in an Australian amphibian. Eggs that were activated by microinjection with a single spermatozoon (n=50) formed more deep, but abnormal, cleavage furrows post-injection (18/50, 36%), than surface changes (12/50, 24%). This result is in contrast to eggs injected without a spermatozoon (n=42), where the majority of eggs displayed limited surface changes (36/42, 86%), and few deep, abnormal furrows (3/42, 7%). Three advanced embryos (3/50, 6%) were produced by ICSI that developed to various stages within the culture system. Technical difficulties were encountered that prevented the generation of any metamorphs from ICSI tadpoles. Nevertheless, when these blocks to ICSI are overcome, the ICSI procedure will be both directly useful as an ART procedure in its own right, and the associated refinement of micromanipulation procedures will assist in the development of other ART procedures in Lim. tasmaniensis. A greater understanding of basic reproductive and developmental biology in Lim. tasmaniensis would greatly facilitate refinement of fertilisation by ICSI. Assisted Reproductive Technologies, in conjunction with gene banks may in the future regenerate extinct amphibian species, and assist in the recovery of declining amphibian populations nationally and worldwide.

Setting up a platform for plant-based influenza virus vaccine production in South Africa

Background During a global influenza pandemic, the vaccine requirements of developing countries can surpass their supply capabilities, if these exist at all, compelling them to rely on developed countries for stocks that may not be available in time. There is thus a need for developing countries in general to produce their own pandemic and possibly seasonal influenza vaccines. Here we describe the development of a plant-based platform for producing influenza vaccines locally, in South Africa. Plant-produced influenza vaccine candidates are quicker to develop and potentially cheaper than egg-produced influenza vaccines, and their production can be rapidly upscaled. In this study, we investigated the feasibility of producing a vaccine to the highly pathogenic avian influenza A subtype H5N1 virus, the most generally virulent influenza virus identified to date. Two variants of the haemagglutinin (HA) surface glycoprotein gene were synthesised for optimum expression in plants: these were the full-length HA gene (H5) and a truncated form lacking the transmembrane domain (H5tr). The genes were cloned into a panel of Agrobacterium tumefaciens binary plant expression vectors in order to test HA accumulation in different cell compartments. The constructs were transiently expressed in tobacco by means of agroinfiltration. Stable transgenic tobacco plants were also generated to provide seed for stable storage of the material as a pre-pandemic strategy. Results For both transient and transgenic expression systems the highest accumulation of full-length H5 protein occurred in the apoplastic spaces, while the highest accumulation of H5tr was in the endoplasmic reticulum. The H5 proteins were produced at relatively high concentrations in both systems. Following partial purification, haemagglutination and haemagglutination inhibition tests indicated that the conformation of the plant-produced HA variants was correct and the proteins were functional. The immunisation of chickens and mice with the candidate vaccines elicited HA-specific antibody responses. Conclusions We managed, after synthesis of two versions of a single gene, to produce by transient and transgenic expression in plants, two variants of a highly pathogenic avian influenza virus HA protein which could have vaccine potential. This is a proof of principle of the potential of plant-produced influenza vaccines as a feasible pandemic response strategy for South Africa and other developing countries.