Evolutionary relationships between bat coronaviruses and their hosts

Recent studies have suggested that bats are the natural reservoir of a range of coronaviruses (CoVs), and that rhinolophid bats harbor viruses closely related to the severe acute respiratory syndrome (SARS) CoV, which caused an outbreak of respiratory illness in humans during 2002-2003. We examined the evolutionary relationships between bat CoVs and their hosts by using sequence data of the virus RNA-dependent RNA polymerase gene and the bat cytochrome b gene. Phylogenetic analyses showed multiple incongruent associations between the phylogenies of rhinolophid bats and their CoVs, which suggested that host shifts have occurred in the recent evolutionary history of this group. These shifts may be due to either virus biologic traits or host behavioral traits. This finding has implications for the emergence of SARS and for the potential future emergence of SARS-CoVs or related viruses.

Emerging Viruses: Coming in on a Wrinkled Wing and a Prayer

The role that bats have played in the emergence of several new infectious diseases has been under review. Bats have been identified as the reservoir hosts of newly emergent viruses such as Nipah virus, Hendra virus, and severe acute respiratory syndrome–like coronaviruses. This article expands on recent findings about bats and viruses and their relevance to human infections. It briefly reviews the history of chiropteran viruses and discusses their emergence in the context of geography, phylogeny, and ecology. The public health and trade impacts of several outbreaks are also discussed. Finally, we attempt to predict where, when, and why we may see the emergence of new chiropteran viruses.

Henipavirus in Pteropus vampyrus Bats, Indonesia

The emergence of Nipah virus (NiV) in Malaysia in 1999 resulted in 265 known human infections (105 fatal), widespread infection in pigs (with >1 million culled to control the outbreak), and the collapse of the Malaysian pig export market. As with the closely related Hendra virus (HeV) that emerged in Australia in 1994 and caused fatal disease in horses and humans, bats of the genus Pteropus (commonly known as flying foxes) were identified as the major reservoir of Nipah virus in Malaysia. This report describes a serologic survey of Pteropus vampyrus in neighboring Indonesia.

Lumometsän syli : Anni Swanin satusymbolismi 1896-1923

Lumometsän syli, Anni Swanin satusymbolismi 1896-1923 on suomenkielisen satukirjallisuuden poetiikkaa ja 1900-luvun alun modernia naiseutta selvittävä feministiseen tutkimustraditioon liittyvä tutkimus. Sen kohteena ovat lasten- ja nuortenkirjailija Anni Swanin (1875-1958) satukokoelmat vuosilta 1901-1923 ja Uusi Suometar -lehden sadunomaiset novellit vuosilta 1896-1904. Tutkimus tuo uutta tietoa lastenkirjallisuuden osalta 1900-luvun alun modernin ihmisen problematiikasta. Se sisältää naissubjektin kehityskaaren ja sisäisen kasvun kohti naistaiteilijuutta. Yksityiskohtaisen tarkastelun kohteina ovat sadut Veli ja sisar (1917), Ihmekukka (1905), Marjaanan helmikruunu (1912), Aaltojen salaisuus (1901), Jääkukka (1905), Tyttö ja kuolema (1917), Merenkuningatar ja hänen poikansa (1905), Lumolinna (1905) ja Tarina Kultasirkasta (1901). Tutkimuksessa tarkastellaan Swanin satujen poeettista kieltä ja naiseuden tematiikkaa ranskalaisen postmodernin ajan feministisen viitekehyksen valossa. Siinä keskeisiä ovat Julia Kristevan psykoanalyyttispohjaiset näkemykset ja Hélène Cixous´n sekä Luce Irigarayn ajatukset feminiinisestä kirjoituksesta. Sadut kontekstualisoidaan ajankohdan symbolistiseen taidevirtaukseen ja Suomen taiteen kultakauteen. Satuja tulkitaan naiskirjailijan lajina ja erityisenä naisen metaforisen ilmaisun muotona. Satujen feministinen lukutapa purkaa perinteisiä lukemiskonventioita ja merkitsee satutekstin lukemista "toisin". Se avaa varhaista modernia naiseutta ja sille ominaista naisen ilmaisukielen erityisyyttä sekä mykkää ei-kielellistä, melankolian ilmaisua. Tutkimus tuo esiin uudenlaisen naiskirjailijan aistimusvoimaisen kielen. Swanin satusymbolismi on luonnon kauneuden synesteettista ja aistimusvoimaista kerrontaa, jolle on luonteenomaista aistiestetiikka, metaforisuus, metonymisyys ja metamorfoosit. Swan vahvistaa osaltaan naisen sankaruutta, omaa ilmaisukieltä ja ääntä. Tuloksena paljastuu satuperinteeseen verrattuna uudenlaisia tyttöyden, äitiyden, naistaiteilijuuden ja perheen malleja ja niiden representaatioita. Satumallit osoittautuvat aikanaan moderneiksi tyttösankareiksi, osin ambivalenteiksi uudenlaista naiseutta ja suhteessa oloa heijastaviksi ja ovat siten varhaisia feministisen sadun tunnusmerkkejä. Tutkimus selvittää, miten Swan rakentaa omaperäisen satusymboliikan. Satumetsä on luonnonkauniin suomalaismetsän symbolinen mielenmaisema ja samanaikaisesti sadun myyttis-symbolinen topos. Swanin luontokäsitys sisältää luonnonsuojelun ja varhaisen ekokriittisen näkemyksen. Tutkimus osoittaa Swanin satujen kytkeytyvän 1900-luvun alun modernismiin ja Suomen taiteen kultakauteen. Swan on suomenkielisen symbolistisen taidesadun kehittäjä ja feministisen sadun aloittaja.

Salmonella and on-farm risk factors in healthy slaughter-age cattle and sheep in eastern Australia

To examine healthy slaughter-age cattle and sheep on-farm for the excretion of Salmonella serovars in faeces and to identify possible risk factors using a questionnaire. The study involved 215 herds and flocks in the four eastern states of Australia, 56 with prior history of salmonellosis. Production systems examined included pasture beef cattle, feedlot beef cattle, dairy cattle, prime lambs and mutton sheep and animals were all at slaughter age. From each herd or flock, 25 animals were sampled and the samples pooled for Salmonella culture. All Salmonella isolated were serotyped and any Salmonella Typhimurium isolates were phage typed. Questionnaires on each production system, prepared in Epi Info 6.04, were designed to identify risk factors associated with Salmonella spp excretion, with separate questionnaires designed for each production system. Salmonellae were identified in all production systems and were more commonly isolated from dairies and beef feedlots than other systems. Statistical analysis revealed that dairy cattle were significantly more likely to shed Salmonella in faeces than pasture beef cattle, mutton sheep and prime lambs (P < 0.05). A wide diversity of Salmonella serovars, all of which have been isolated from humans in Australia, was identified in both cattle and sheep. Analysis of the questionnaires showed access to new arrivals was a significant risk factor for Salmonella excretion on dairy properties. For beef feedlots, the presence of large numbers of flies in the feedlot pens or around stored manure were significant risk factors for Salmonella excretion. Dairy cattle pose the highest risk of all the slaughter-age animals tested. Some of the identified risk factors can be overcome by improved management practices, especially in relation to hygiene.

Global trade in ornamental fish from an Australian perspective: The case for revised import risk analysis and management strategies

Over 1 billion ornamental fish comprising more than 4000 freshwater and 1400 marine species are traded internationally each year, with 8-10 million imported into Australia alone. Compared to other commodities, the pathogens and disease translocation risks associated with this pattern of trade have been poorly documented. The aim of this study was to conduct an appraisal of the effectiveness of risk analysis and quarantine controls as they are applied according to the Sanitary and Phytosanitary (SPS) agreement in Australia. Ornamental fish originate from about 100 countries and hazards are mostly unknown; since 2000 there have been 16-fold fewer scientific publications on ornamental fish disease compared to farmed fish disease, and 470 fewer compared to disease in terrestrial species (cattle). The import quarantine policies of a range of countries were reviewed and classified as stringent or non-stringent based on the levels of pre-border and border controls. Australia has a stringent policy which includes pre-border health certification and a mandatory quarantine period at border of 1-3 weeks in registered quarantine premises supervised by government quarantine staff. Despite these measures there have been many disease incursions as well as establishment of significant exotic viral, bacterial, fungal, protozoal and metazoan pathogens from ornamental fish in farmed native Australian fish and free-living introduced species. Recent examples include Megalocytivirus and Aeromonas salmonicida atypical strain. In 2006, there were 22 species of alien ornamental fish with established breeding populations in waterways in Australia and freshwater plants and molluscs have also been introduced, proving a direct transmission pathway for establishment of pathogens in native fish species. Australia's stringent quarantine policies for imported ornamental fish are based on import risk analysis under the SPS agreement but have not provided an acceptable level of protection (ALOP) consistent with government objectives to prevent introduction of pests and diseases, promote development of future aquaculture industries or maintain biodiversity. It is concluded that the risk analysis process described by the Office International des Epizooties under the SPS agreement cannot be used in a meaningful way for current patterns of ornamental fish trade. Transboundary disease incursions will continue and exotic pathogens will become established in new regions as a result of the ornamental fish trade, and this will be an international phenomenon. Ornamental fish represent a special case in live animal trade where OIE guidelines for risk analysis need to be revised. Alternatively, for countries such as Australia with implied very high ALOP, the number of species traded and the number of sources permitted need to be dramatically reduced to facilitate hazard identification, risk assessment and import quarantine controls. Lead papers of the eleventh symposium of the International Society for Veterinary Epidemiology and Economics (ISVEE), Cairns, Australia

Infection with Menangle virus in flying foxes (Pteropus spp.) in Australia

Objective: To examine flying foxes (Pteropus spp.) for evidence of infection with Menangle virus. Design: Clustered non-random sampling for serology, virus isolation and electron microscopy (EM). Procedure: Serum samples were collected from 306 Pteropus spp. in northern and eastern Australia and tested for antibodies against Menangle virus (MenV) using a virus neutralisation test (VNT). Virus isolation was attempted from tissues and faeces collected from 215 Pteropus spp. in New South Wales. Faecal samples from 68 individual Pteropus spp. and four pools of faeces were examined by transmission EM following routine negative staining and immunogold labelling. Results: Neutralising antibodies (VNT titres ≥ 8) against MenV were detected in 46% of black flying foxes (P. alecto), 41% of grey-headed flying foxes (P. poliocephalus), 25% of spectacled flying foxes (P. conspicillatus) and 1% of little red flying foxes (P. scapulatus) in Australia. Positive sera included samples collected from P. poliocephalus in a colony adjacent to a piggery that had experienced reproductive disease caused by MenV. Virus-like particles were observed by EM in faeces from Pteropus spp. and reactivity was detected in pooled faeces and urine by immunogold EM using sera from sows that had been exposed to MenV. Attempts to isolate the virus from the faeces and tissues from Pteropus spp. were unsuccessful. Conclusion: Serological evidence of infection with MenV was detected in Pteropus spp. in Australia. Although virus-like particles were detected in faeces, no viruses were isolated from faeces, urine or tissues of Pteropus spp.

Letter to the editor - Name that plant scientifically, please!

The report of Brunfelsia poisoning of dogs in Sydney by Drs Singh, Cowan and Child is very welcome, but it raises one important scientific point that has been further highlighted by the letter from Dr Seavers. Neither in the case report nor in Dr Seavers' letter is the species or cultivar of the plants identified. There are about 40 known Brunfelsia species, a number of which are cultivated in tropical to temperate climate gardens in Australia. It is noteworthy that Dr Seavers speculates about the Brunfelsia plants involved in dog poisonings in Australia, suggesting that 'dwarf' plants may be more poisonous ...

Semantic Fluency in Mild and Moderate Alzheimer's Disease

Alzheimer's disease (AD) is characterized by an impairment of the semantic memory responsible for processing meaning-related knowledge. This study was aimed at examining how Finnish-speaking healthy elderly subjects (n = 30) and mildly (n=20) and moderately (n = 20) demented AD patients utilize semantic knowledge to performa semantic fluency task, a method of studying semantic memory. In this task subjects are typically given 60 seconds to generate words belonging to the semantic category of animals. Successful task performance requires fast retrieval of subcategory exemplars in clusters (e.g., farm animals: 'cow', 'horse', 'sheep') and switching between subcategories (e.g., pets, water animals, birds, rodents). In this study, thescope of the task was extended to cover various noun and verb categories. The results indicated that, compared with normal controls, both mildly and moderately demented AD patients showed reduced word production, limited clustering and switching, narrowed semantic space, and an increase in errors, particularly perseverations. However, the size of the clusters, the proportion of clustered words, and the frequency and prototypicality of words remained relatively similar across the subject groups. Although the moderately demented patients showed a poor eroverall performance than the mildly demented patients in the individual categories, the error analysis appeared unaffected by the severity of AD. The results indicate a semantically rather coherent performance but less specific, effective, and flexible functioning of the semantic memory in mild and moderate AD patients. The findings are discussed in relation to recent theories of word production and semantic representation. Keywords: semantic fluency, clustering, switching, semantic category, nouns, verbs, Alzheimer's disease

Space allowances for confined livestock and their determination from allometric principles

The amount of space provided to animals governs important elements of their behaviour and, hence, is critical for their health and welfare. We review the use of allometric principles and equations to estimate the static space requirements of animals when standing and lying, and the space required for animals to feed, drink, stand-up and lie-down. We use the research literature relating to transportation and intensive housing of sheep and cattle to assess the validity of allometric equations for estimating space allowances. We investigated these areas because transportation and intensive housing provide points along a continuum in terms of the duration of confinement, (from hours to months) and spatial requirements are likely to increase with increasing duration of confinement, as animals will need to perform a greater behavioural repertoire for long-term survival, health and welfare. We find that, although there are theoretical reasons why allometric relationships to space allowances may vary slightly for different classes of stock, space allowances that have been demonstrated to have adverse effects on animal welfare during transportation correlated well with an inability to accommodate standing animals, as estimated from allometry. For intensive housing, we were able to detect a space allowance below which there were adverse effects on welfare. For short duration transportation during which animals remain standing, a space allowance per animal described by the allometric equation: area (m^2) = 0.020W^0.66, where W = liveweight (kg), would appear to be appropriate. Where it is desirable for all animals to lie simultaneously, then a minimum space allowance per animal described by the allometric equation: area (m^2) = 0.027W^0.66 appears to permit this, given that animals in a group time-share space. However, there are insufficient data to determine whether this allowance onboard a vehicle/vessel would enable animals to move and access food and water with ease. In intensive housing systems, a minimum space allowance per animal described by the allometric equation: area (m^2) = 0.033W^0.66 appears to be the threshold below which there are adverse effects on welfare. These suggested space allowances require verification with a range of species under different thermal conditions and, for transportation, under different conditions of vehicular/vessel stability. The minimum length of trough per animal (L in m) required for feeding and drinking can be determined from L = 0.064W^0.33, with the number of animals required to feed/drink simultaneously taken into account, together with any requirement to minimise competition. This also requires verification with a range of species. We conclude that allometric relationships are an appropriate basis for the formulation of space allowances for livestock.

Herpesviral haematopoietic necrosis virus (CyHV-2) infection: case studies from commercial goldfish farms

Herpesviral haematopoietic necrosis is a disease of goldfish, Carassius auratus, caused by Cyprinid herpesvirus-2 (CyHV-2) infection. Quantitative PCR was carried out on tissue homogenates from healthy goldfish fingerlings, broodfish, eggs and fry directly sampled from commercial farms, from moribund fish submitted to our laboratory for disease diagnosis, and on naturally-infected CyHV-2 carriers subjected to experimental stress treatments. Healthy fish from 14 of 18 farms were positive with copy numbers ranging from tens to 10(7) copies mu g(-1) DNA extracted from infected fish. Of 118 pools of broodfish tested, 42 were positive. The CyHV-2 was detected in one lot of fry produced from disinfected eggs. Testing of moribund goldfish, in which we could not detect any other pathogens, produced 12 of 30 cases with 10(6)-10(8) copies of CyHV-2 mu g(-1) DNA extracted. Subjecting healthy CyHV-2 carriers to cold shock (22-10 degrees C) but not heat, ammonia or high pH, increased viral copy numbers from mean copy number (+/- SE) of 7.3 +/- 11 to 394 +/- 55 mu g(-1) DNA extracted after 24 h. CyHV-2 is widespread on commercial goldfish farms and outbreaks apparently occur when healthy carriers are subjected to a sharp temperature drop followed by holding at the permissive temperature for the disease.

Australian Bat Lyssavirus

In Chapter 1, the literature relating to rabies virus and the rabies like lyssaviruses is reviewed. In Chapter 2, data are presented from 1170 diagnostic submissions for ABLV testing by fluorescent antibody test (Centocor FAT). All 27 non-bat submissions were ABLV-negative. Of 1143 bat accessions 74 (16%) were ABLV-positive, including 69 of 974 (7.1%) flying foxes (Pteropus spp.), 5 of 7 (71.4%) Saccolaimus flaviventris (Yellow-bellied sheathtail bats), none of 151 other microchiropteran bats, and none of 11 unidentified bats. Statistical analysis of data from 868 wild Black, Grey-headed, Little Red and Spectacled flying foxes (Pteropus alecto, P. poliocephalus, P. scapulatus, and P. conspicillatus) indicated that three factors; species, health status and age were associated with significant (p< 0.001) differences in the proportion of ABLV-positive bats. Other factors including sex, whether the bat bit a person or animal, region, year, and season submitted, were not associated with ABLV. Case data for 74 ABLV-positive bats, including the circumstances in which they were found and clinical signs, is presented. In Chapter 3, the aetiological diagnosis was investigated for 100 consecutive flying fox submissions with neurological signs. ABLV (32%), spinal and head injuries (29%), and neuro-angiostrongylosis (18%) accounted for most neurological syndromes in flying foxes. No evidence of lead poisoning was found in unwell (n=16) or healthy flying foxes (n=50). No diagnosis was reached for 16 cases, all of which were negative for ABLV by TaqMan PCR. The molecular diversity of ABLV was examined in Chapter 4 by sequencing 36 bases of the leader sequence, the entire N gene, and start of the P gene of 28 isolates from pteropid bats and 3 isolates from Yellow-bellied sheathtail (YBST) bats. Phylogenetic analysis indicated all ABLV isolates clustered together as a discrete group within the Lyssavirus genera closely related to rabies virus and European bat lyssavirus-2 isolates. The ABLV lineage consisted of two variants; one (ybst-ABLV) consisted of isolates only from YBST bats, the other (pteropid-ABLV) was common to Black, Grey-headed and Little Red flying foxes. No associations were found between the sequences and either the geographical location or year found, or individual flying fox species. In Chapter 5, 15 inocula prepared from the brains or salivary glands of naturally-infected bats were evaluated by intracerebral (IC) and footpad (FP) inoculation of Quackenbush mice in order to select and characterize a highly virulent inoculum for further use in bats (Inoculum 5). In Chapter 6, nine Grey-headed flying foxes were inoculated with 105.2 to 105.5 MICED50 of Inoculum 5 divided into four sites, left footpad, pectoral muscle, temporal muscle and muzzle. Another bat was inoculated with half this dose divided into the footpad and pectoral muscle only. Seven of 10 bats developed clinical disease of 1 to 4 days duration between PI-days 10 and 19 and were shown to be ABL-positive by FAT, HAM immunoperoxidase staining, virus isolation in mice, and TaqMan PCR. Five of the seven bats displayed overt aggression, one died during a seizure, and one showed intractable agitation, pacing, tremors, and ataxia. Viral antigen was demonstrated throughout the central and peripheral nervous systems and in the epithelial cells of the submandibular salivary glands (n=4). All affected bats had mild to moderate non-suppurative meningoencephalitis and severe ganglioneuritis. No ABLV was detected in three bats that remained well until the end of the experiment on day 82. One survivor developed a strong but transient antibody response. In Chapter 7, the relative virulence of inocula prepared from the brains and salivary glands of experimentally infected flying foxes was evaluated in mice by IC and FP inoculation and TaqMan assay. The effects in mice were correlated to the TaqMan CT value and indicated a crude association between virulence and CT value that has potential application in the selection of inocula. In Chapter 8, 36 Black and Grey-headed flying foxes were vaccinated with one (day 0) or two (+ day 28) doses of Nobivac rabies vaccine and co-vaccinated with keyhole limpet haemocyanin (KLH). All bats responded to the Nobivac vaccine with a rabies-RFFIT titer > 0.5 IU/mL that is nominally indicative of protective immunity. Plasma from bats with rabies titres >2 IU/mL had cross-neutralising ABLV titres >1:154. A specifically developed ELISA detected a strong but transient response to KLH.

Bats and Emerging Zoonoses: Henipaviruses and SARS.

Nearly 75% of all emerging infectious diseases (EIDs) that impact or threaten human health are zoonotic. The majority have spilled from wildlife reservoirs, either directly to humans or via domestic animals. The emergence of many can be attributed to predisposing factors such as global travel, trade, agricultural expansion, deforestation habitat fragmentation, and urbanization; such factors increase the interface and or the rate of contact between human, domestic animal, and wildlife populations, thereby creating increased opportunities for spillover events to occur. Infectious disease emergence can be regarded as primarily an ecological process. The epidemiological investigation of EIDs associated with wildlife requires a trans-disciplinary approach that includes an understanding of the ecology of the wildlife species, and an understanding of human behaviours that increase risk of exposure. Investigations of the emergence of Nipah virus in Malaysia in 1999 and severe acute respiratory syndrome (SARS) in China in 2003 provide useful case studies. The emergence of Nipah virus was associated with the increased size and density of commercial pig farms and their encroachment into forested areas. The movement of pigs for sale and slaughter in turn led to the rapid spread of infection to southern peninsular Malaysia, where the high-density, largely urban pig populations facilitated transmission to humans. Identifying the factors associated with the emergence of SARS in southern China requires an understanding of the ecology of infection both in the natural reservoir and in secondary market reservoir species. A necessary extension of understanding the ecology of the reservoir is an understanding of the trade, and of the social and cultural context of wildlife consumption. Emerging infectious diseases originating from wildlife populations will continue to threaten public health. Mitigating and managing the risk requires an appreciation of the connectedness between human, livestock and wildlife health, and of the factors and processes that disrupt the balance.

High-level coordination and strategy in the 2007 equine influenza outbreak response.

The equine influenza (EI) outbreak presented many challenges that required high-level coordination and decision making, as well as the development of new approaches for satisfactory and consistent resolution. This paper outlines the elements of the national coordination arrangements, preparatory arrangements in place prior to the outbreak that facilitated national coordination, and some of the issues faced and resolved in the response.

Automatic Lameness Detection in a Milking Robot : Instrumentation, measurement software, algorithms for data analysis and a neural network model

The aim of this thesis is to develop a fully automatic lameness detection system that operates in a milking robot. The instrumentation, measurement software, algorithms for data analysis and a neural network model for lameness detection were developed. Automatic milking has become a common practice in dairy husbandry, and in the year 2006 about 4000 farms worldwide used over 6000 milking robots. There is a worldwide movement with the objective of fully automating every process from feeding to milking. Increase in automation is a consequence of increasing farm sizes, the demand for more efficient production and the growth of labour costs. As the level of automation increases, the time that the cattle keeper uses for monitoring animals often decreases. This has created a need for systems for automatically monitoring the health of farm animals. The popularity of milking robots also offers a new and unique possibility to monitor animals in a single confined space up to four times daily. Lameness is a crucial welfare issue in the modern dairy industry. Limb disorders cause serious welfare, health and economic problems especially in loose housing of cattle. Lameness causes losses in milk production and leads to early culling of animals. These costs could be reduced with early identification and treatment. At present, only a few methods for automatically detecting lameness have been developed, and the most common methods used for lameness detection and assessment are various visual locomotion scoring systems. The problem with locomotion scoring is that it needs experience to be conducted properly, it is labour intensive as an on-farm method and the results are subjective. A four balance system for measuring the leg load distribution of dairy cows during milking in order to detect lameness was developed and set up in the University of Helsinki Research farm Suitia. The leg weights of 73 cows were successfully recorded during almost 10,000 robotic milkings over a period of 5 months. The cows were locomotion scored weekly, and the lame cows were inspected clinically for hoof lesions. Unsuccessful measurements, caused by cows standing outside the balances, were removed from the data with a special algorithm, and the mean leg loads and the number of kicks during milking was calculated. In order to develop an expert system to automatically detect lameness cases, a model was needed. A probabilistic neural network (PNN) classifier model was chosen for the task. The data was divided in two parts and 5,074 measurements from 37 cows were used to train the model. The operation of the model was evaluated for its ability to detect lameness in the validating dataset, which had 4,868 measurements from 36 cows. The model was able to classify 96% of the measurements correctly as sound or lame cows, and 100% of the lameness cases in the validation data were identified. The number of measurements causing false alarms was 1.1%. The developed model has the potential to be used for on-farm decision support and can be used in a real-time lameness monitoring system.