Identification of diverse full-length endogenous betaretroviruses in megabats and microbats

Background: Betaretroviruses infect a wide range of species including primates, rodents, ruminants, and marsupials. They exist in both endogenous and exogenous forms and are implicated in animal diseases such as lung cancer in sheep, and in human disease, with members of the human endogenous retrovirus-K (HERV-K) group of endogenous betaretroviruses (βERVs) associated with human cancers and autoimmune diseases. To improve our understanding of betaretroviruses in an evolutionarily distinct host species, we characterized βERVs present in the genomes and transcriptomes of mega- and microbats, which are an important reservoir of emerging viruses.Results: A diverse range of full-length βERVs were discovered in mega- and microbat genomes and transcriptomes including the first identified intact endogenous retrovirus in a bat. Our analysis revealed that the genus Betaretrovirus can be divided into eight distinct sub-groups with evidence of cross-species transmission. Betaretroviruses are revealed to be a complex retrovirus group, within which one sub-group has evolved from complex to simple genomic organization through the acquisition of an env gene from the genus Gammaretrovirus. Molecular dating suggests that bats have contended with betaretroviral infections for over 30 million years.Conclusions: Our study reveals that a diverse range of betaretroviruses have circulated in bats for most of their evolutionary history, and cluster with extant betaretroviruses of divergent mammalian lineages suggesting that their distribution may be largely unrestricted by host species barriers. The presence of βERVs with the ability to transcribe active viral elements in a major animal reservoir for viral pathogens has potential implications for public health. © 2013 Hayward et al.; licensee BioMed Central Ltd.

Influence of dingoes on sheep distribution in Australia

Objective To describe the influence of the dingo (Canis lupus dingo) on the past, present and future distributions of sheep in Australia. Design The role of the dingo in the rise and fall of sheep numbers is reviewed, revised data are provided on the present distribution and density of sheep and dingoes, and historical patterns of sheep distribution are used to explore the future of rangeland sheep grazing. Results Dingoes are a critical causal factor in the distribution of sheep at the national, regional and local levels. Dingo predation contributed substantially to the historical contraction of the sheep industry to its present-day distribution, which is almost exclusively confined to areas within fenced dingo exclusion zones. Dingo populations and/or their influence are now present and increasing in all sheep production zones of Australia, inclusive of areas that were once dingo free'. Conclusions Rangeland production of wool and sheep meat is predicted to disappear within 30-40 years if the present rate of contraction of the industry continues unabated. Understanding the influence of dingoes on sheep production may help refine disease response strategies and help predict the future distribution of sheep and their diseases.

Priority list of endemic diseases for the red meat industries

This report provides a systematic review of the most economically damaging endemic diseases and conditions for the Australian red meat industry (cattle, sheep and goats). A number of diseases for cattle, sheep and goats have been identified and were prioritised according to their prevalence, distribution, risk factors and mitigation. The economic cost of each disease as a result of production losses, preventive costs and treatment costs is estimated at the herd and flock level, then extrapolated to a national basis using herd/flock demographics from the 2010-11 Agricultural Census by the Australian Bureau of Statistics. Information shortfalls and recommendations for further research are also specified. A total of 17 cattle, 23 sheep and nine goat diseases were prioritised based on feedback received from producer, government and industry surveys, followed by discussions between the consultants and MLA. Assumptions of disease distribution, in-herd/flock prevalence, impacts on mortality/production and costs for prevention and treatment were obtained from the literature where available. Where these data were not available, the consultants used their own expertise to estimate the relevant measures for each disease. Levels of confidence in the assumptions for each disease were estimated, and gaps in knowledge identified. The assumptions were analysed using a specialised Excel model that estimated the per animal, herd/flock and national costs of each important disease. The report was peer reviewed and workshopped by the consultants and experts selected by MLA before being finalised. Consequently, this report is an important resource that will guide and prioritise future research, development and extension activities by a variety of stakeholders in the red meat industry. This report completes Phase I and Phase II of an overall four-Phase project initiative by MLA, with identified data gaps in this report potentially being addressed within the later phases. Modelling the economic costs using a consistent approach for each disease ensures that the derived estimates are transparent and can be refined if improved data on prevalence becomes available. This means that the report will be an enduring resource for developing policies and strategies for the management of endemic diseases within the Australian red meat industry.

Study of Interaction Force between Antigen and Antibody Using Flow Chamber Method

A parallel plate flow chamber was used to study the interaction force between human IgG (immobilized on a chip surface as ligand) and goat anti-human IgG (immobilized on microspheres surface as receptor). First, it was demonstrated that the binding force between the microspheres and the chip surface came from the bio-specific interaction between the antigen and the antibody. Secondly, it was obtained that the critical shear rate to detach microspheres from the chip surface increases with the ligand surface concentration. Finally, two models to estimate the antigen-antibody bond strength considering bonds' positions were proposed and analyzed.

Mass culture of Undaria gametophyte clones and their use in sporeling culture

Undaria pinnatifida (Harv.) Sur. is one of the three main seaweed species under commercial cultivation in China. In the mid-1990s the annual production was about 20000 tons dry. The supply of healthy sporelings is key to the success of commercial cultivation of Undaria. Previous studies demonstrated that instead of the zoospore collection method, sporelings can be cultured through the use of gametophyte clones. This paper reports the experimental results on mass culture of clones and sporeling raising in commercial scale. Light had an obvious effect on growth of gametophyte clones. Under an irradiance of 80 mumol m(-2) s(-1) and favorable temperature of 22-25degreesC, mean daily growth rate may reach as high as 37%. Several celled gametophyte fragments were sprayed onto the palm rope frame. Gametogenesis occurred after 4-6 days. Juvenile sporeling growth experiments showed that nitrate and phosphate concentrations of 2.9 10(-4) mol 1(-1) and 1.7 10(-5) mol 1(-1) were sufficient to enable the sporelings to maintain a high daily growth rate. Sporelings can reach a length of 1 cm in a month. Since 1997, extension of the clone technique has been carried out in Shandong Province. Large-scale production of sporelings for commercial cultivation of 14 and 31 hectares in 1997 and 1998 had been conducted successfully.

Spatial distribution of upland beetles in relation to landform, vegetation and grazing management

Dennis, P., Aspinall, R. J., Gordon, I. J. (2002). Spatial distribution of upland beetles in relation to landform vegetation and grazing management. Basic and Applied Ecology, 3 (2), 183?193. Sponsorship: SEERAD RAE2008

Grandparents and parenting: towards a theory of grandparenthood

This paper explores the changing role of contemporary grandparents with many demonstrating a willingness and ability to take on parental responsibilities for their grandchildren, where they may face challenges and opportunities in difficult times. Three main forms of grand parenting are identified in the literature, those who have primary responsibility and are raising their grand children as their main carers perhaps in response to crisis situations, those who live in extended families and participate in care, and those who provide day care while the child’s parents work. The latter has increased because of the increasing frequency of divorce, single parenting and the lack of available or subsidised child care in the United Kingdom. When grandparents step into a troubled situation and attempt to offer stability and security for their grandchildren they may have to manage the combined responsibilities of family caregivers and parental figures. Grandparenthood is a tenuous role, lacking clear agreement on behaviour norms. In the culture of advice and parenting support, while care must be taken not to undermine parenting skills or make judgements about the ability to cope with the demands of childcare, an exploration of the impact on grandparents and children must be undertaken. Due to the complex web of interrelated factors the process and outcomes of care giving by grandparents is not well known in the literature. It is proposed therefore that it is timely for research to be undertaken to explore and develop a theory of Grandparenthood.

Strategies for water pollution control in Dhaka, Bangladesh: an institutional and economic analysis

This paper presents primary data based on research carried out as part of a large World Bank project. Results from our survey show that water pollution in Dhaka watershed has reached alarming levels and is posing significant threats to health and economic activity, particularly among the poor and vulnerable. Rice productivity in the watershed area, for example, has declined by 40% in recent years and vegetable cultivation in the riverbeds has been severely damaged. We also found significant correlation between water pollution and diseases such as jaundice, diarrhoea and skin problems. It was reported that the cost of treatment of skin diseases for one episode could be as high as 29% of the weekly earnings of poor households. Given the magnitude of the contamination problem, a multi-agent stakeholder approach was necessary to analyse the institutional and economic constraints that would need to be addressed in order to improve environmental management. This approach, in turn, enabled core strategies to be developed. The strategies were better understood around three types of actors in industrial pollution, i.e. (1) principal actors, who contribute directly to industrial pollution; (2) stakeholders, who exacerbate the situation by inaction; and (3) the potential actors in mitigation of water contamination. Within a carrot-and-stick framework, nine strategies leading to the strengthening of environmental management were explored. They aim at improving governance and transparency within public agencies and private industry through the setting up of incentive structures to advance compliance and enforcement of environmental standards. Civil society and the population at large are, on the other hand, encouraged to contribute actively to the mitigation of water pollution by improving the management of environmental information and by raising public awareness.

Mycobacterium avium subsp. paratuberculosis in foods: current evidence and potential consequences

Mycobacterium avium ssp. paratuberculosis (MAP), the cause of Johne's disease in cattle, sheep and goats, may have a role in Crohn's disease in humans. Animals with Johne's disease shed viable MAP in their milk and faeces. The organism is also widely disseminated in the blood and tissues of infected animals. Consequently, transmission to humans via consumption of animal-derived foods is a distinct possibility. Milk, other dairy products, beef and water have been identified as possible food vehicles of transmission. To date, viable MAP has been cultured from raw cows', sheep and goats' milk, retail pasteurized cows' milk, and some retail cheeses in several countries during recent studies. MAP has not been isolated from retail beef to date, although limited testing has been carried out. The public health consequences, if any, of low numbers of viable MAP being periodically consumed by susceptible individuals are uncertain. An association between MAP and Crohn's disease is not proven, but neither can it be discounted on the basis of current evidence. A precautionary approach is therefore warranted in relation to the existence of MAP in food, and action is needed to reduce the prevalence of Johne's disease in the cattle population worldwide, in order to minimize public exposure to this potential human pathogen.

Production and characterisation of polyclonal antibodies to a range of nitroimidazoles.

The nitroimidazoles dimetridazole and ronidazole are metabolised to hydroxydimetridazole, while metronidazole is metabolised to hydroxymetronidazole. To screen for a large number of samples by immunoassay for the presence of this family of drugs and metabolites, it was necessary to produce an antibody with broad-spectrum recognition. Metronidazole and hydroxydimetridazole were selected as antigens as they could be coupled to large (immunogenic) carrier proteins at two different positions of the general nitroimidazole structure. The resulting conjugates were used to immunise rabbits, sheep and goats. Seventeen out of thirty-nine animals immunised produced a detectable antibody titre and these antibodies were consequently characterised as regards sensitivity and cross-reactivity.

The panel of antisera produced exhibited IC50 ranging from 1.26 to 73.76 ng ml-1 using a competitive ELISA assay. Cross-reactivity studies showed that sera from several animals were capable of significant binding of six of the seven nitroimidazole compounds tested.

Fasciola hepatica cathepsin L-like proteases: biology, function and potential in the development of first generation liver fluke vaccines.

Fasciola hepatica secretes cathepsin L proteases that facilitate the penetration of the parasite through the tissues of its host, and also participate in functions such as feeding and immune evasion. The major proteases, cathepsin L1 (FheCL1) and cathepsin L2 (FheCL2) are members of a lineage that gave rise to the human cathepsin Ls, Ks and Ss, but while they exhibit similarities in their substrate specificities to these enzymes they differ in having a wider pH range for activity and an enhanced stability at neutral pH. There are presently 13 Fasciola cathepsin L cDNAs deposited in the public databases representing a gene family of at least seven distinct members, although the temporal and spatial expression of each of these members in the developmental stage of F. hepatica remains unclear. Immunolocalisation and in situ hybridisation studies, using antibody and DNA probes, respectively, show that the vast majority of cathepsin L gene expression is carried out in the epithelial cells lining the parasite gut. Within these cells the enzyme is packaged into secretory vesicles that release their contents into the gut lumen for the purpose of degrading ingested host tissue and blood. Liver flukes also express a novel multi-domain cystatin that may be involved in the regulation of cathepsin L activity. Vaccine trials in both sheep and cattle with purified native FheCL1 and FheCL2 have shown that these enzymes can induce protection, ranging from 33 to 79%, to experimental challenge with metacercariae of F. hepatica, and very potent anti-embryonation/hatch rate effects that would block parasite transmission. In this article we review the vaccine trials carried out over the past 8 years, the role of antibody and T cell responses in mediating protection and discuss the prospects of the cathepsin Ls in the development of first generation recombinant liver fluke vaccines. Author Keywords: Helminths; Trematodes; Parasites; Cathepsins; Proteases; Vaccines; Immunology; Biochemistry

The prediction of intake potential and organic matter digestibility of grass silages by near infrared spectroscopy analysis of undried samples

A study was undertaken to examine a range of sample preparation and near infrared reflectance spectroscopy (NIPS) methodologies, using undried samples, for predicting organic matter digestibility (OMD g kg(-1)) and ad libitum intake (g kg(-1) W-0.75) of grass silages. A total of eight sample preparation/NIRS scanning methods were examined involving three extents of silage comminution, two liquid extracts and scanning via either external probe (1100-2200 nm) or internal cell (1100-2500 nm). The spectral data (log 1/R) for each of the eight methods were examined by three regression techniques each with a range of data transformations. The 136 silages used in the study were obtained from farms across Northern Ireland, over a two year period, and had in vivo OMD (sheep) and ad libitum intake (cattle) determined under uniform conditions. In the comparisons of the eight sample preparation/scanning methods, and the differing mathematical treatments of the spectral data, the sample population was divided into calibration (n = 91) and validation (n = 45) sets. The standard error of performance (SEP) on the validation set was used in comparisons of prediction accuracy. Across all 8 sample preparation/scanning methods, the modified partial least squares (MPLS) technique, generally minimized SEP's for both OMD and intake. The accuracy of prediction also increased with degree of comminution of the forage and with scanning by internal cell rather than external probe. The system providing the lowest SEP used the MPLS regression technique on spectra from the finely milled material scanned through the internal cell. This resulted in SEP and R-2 (variance accounted for in validation set) values of 24 (g/kg OM) and 0.88 (OMD) and 5.37 (g/kg W-0.75) and 0.77 (intake) respectively. These data indicate that with appropriate techniques NIRS scanning of undried samples of grass silage can produce predictions of intake and digestibility with accuracies similar to those achieved previously using NIRS with dried samples. (C) 1998 Elsevier Science B.V.

Preparation of highly stable oligo (ethylene glycol) derivatives-functionalized gold nanoparticles and their application in LSPR-Based detection of PSA/ACT complex

A sandwich immunoassay for PSA/ACT complex detection based on gold nanoparticle aggregation using two probes was developed. The functionalized colloidal gold nanoparticles (AuNPs) showed highly stable not only in the presence of high ionic strength but also in a wide pH range. The functionalized AuNPs were tagged with PSA/ACT complex monoclonal antibody and goat PSA polyclonal antibody and served as the probes to induce aggregation of the colloidal particles. As a result, PSA/ACT complex was detected at concentrations as low as 1 ng/ml. This is the first time that a new aggregation sandwich-immunoassay technique using two gold probes has been used, and the results are generally applicable to other LSPR-based immunoassays.

The sympathetic release test: a test used to assess thermoregulation and autonomic control of blood flow

When a subject is heated, the stimulation of temperature-sensitive nerve endings in the skin, and the raising of the central body temperature, results in the reflex release of sympathetic vasoconstrictor tone in the skin of the extremities, causing a measurable temperature increase at the site of release. In the sympathetic release test, the subject is gently heated by placing the feet and calves in a commercially available foot warming pouch or immersing the feet and calves in warm water and wrapping the subject in blankets. Skin blood flow is estimated from measurements of skin temperature in the fingers. Normally skin temperature of the fingers is 65-75 degrees F in cool conditions (environmental temperature: 59-68 degrees F) and rises to 85-95 degrees F during body heating. Deviations in this pattern may mean that there is abnormal sympathetic vasoconstrictor control of skin blood flow. Abnormal skin blood flow can substantially impair an individual's ability to thermoregulate and has important clinical implications. During whole body heating, the skin temperature from three different skin sites is monitored and oral temperature is monitored as an index of core temperature. Students determine the fingertip temperature at which the reflex release of sympathetic activity occurs and its maximal attainment, which reflects the vasodilating capacity of this cutaneous vascular bed. Students should interpret typical sample data for certain clinical conditions (Raynaud's disease, peripheral vascular disease, and postsympathectomy) and explain why there may be altered skin blood flow in these disorders.

Fasciola hepatica:development of monoclonal antibodies and their use to characterize a glycocalyx antigen in migrating flukes

Using mice harbouring early Fasciola hepatica infections, six monoclonal antibodies were prepared against a tegumental antigen present in T1 granules and glycocalyx of flukes. Blocking tests indicated that all monoclonals bound the same T1 epitope (or epitopes in close proximity on the antigen molecule), but this was not the determinant recognized by sheep and cattle. Localization of antibody binding at light and electron microscope levels showed that T1-type antigen also occurred in metacercarial tegument and in glycocalyx of gut cells and excretory ducts in juvenile and adult flukes. This indicates that the natural host-antibody response to F. hepatica may be to one antigen early in the infection. Protein A-gold labelling of monoclonal treated fluke sections revealed that the epitope was probably a polypeptide, unmodified by glycosylation in Golgi bodies. When isolated by immunoadsorption and separated electrophoretically under reducing conditions T1-type antigen was found to consist of a polypeptide mol. wt. 50 000, possibly linked to smaller entities mol. wt. 25-40 000. Tissue-specific variations in the antigen molecule might be conferred by linkage of different polypeptides or carbohydrate side-chains to an antigenic core polypeptide. A component of T1-type antigen was found to have mol. wt. of 25 000, possibly resembling a polypeptide of mol. wt. 24 000 from Schistosoma mansoni tegument.