Testing the applicability of molecular genetic markers to population analyses of scleratinian corals

The abundance of coral reefs worldwide is in decline, and despite the ecological importance of reefs, only a limited number of DNA markers have been identified for scleractinian coral genetic studies. This paper addresses the search for new coral molecular markers and investigates the applicability of the cytochrome c oxidase subunit I (COI), the internal transcribed spacer region 1 (ITS1), and the pocilloporin gene to the question of intraspecific variation in the scleractinian coral Pocillopora verrucosa along the southeast African coastline. The COI fragment was 710 bp long and was identical for P. verrucosa (n = 10) and P. damicornis (n = 3). Only two different ITS1 sequences were found (differing by 13 bp insertion), but more importantly, 24% of the sequences were heterogenous indicating that different multiple copies of the sequence exist. Pocilloporin is an intronless gene that was absolutely conserved throughout all P. verrucosa populations (n = 50). Thus, the three DNA regions studied appear unsuitable for the population genetic analyses of P. verrucosa.

Analysis of toxin profiles in three different fish species causing ciguatera fish poisoning in Guadeloupe, French West Indies

A grey snapper (Lutjanus griseus), a grouper (Serranidae) and a blackjack (Caranx lugubris) were implicated in three different ciguatera poisonings in Guadeloupe, French West Indies. A mouse bioassay indicated toxicity for each specimens: 0.5-1, greater than or equal to 1 and > 1 M Ug g(-1), respectively. After purification by gel filtration chromatography, the samples were analysed by high-performance liquid chromatography coupled to mass spectrometry (LC-MS). The toxin profiles differ from one fish to another. C-CTX-1 was detected at 0.24, 0.90 and 13.8 ng g(-1) flesh in the snapper, grouper and jack, respectively. It contributed only to part of the whole toxicity determined by the mouse bioassay. Other toxins identified were C-CTX-2 (a C-CTX-1 epimer), three additional isomers of C-CTX-1 or -2, and five ciguatoxin congeners (C-CTX-1127, C-CTX-1143 and its isomer C-CTX-1143a, and C-CTX-1157 and its isomer C-CTX-1157b). Putative hydroxy-polyether-like compounds were also detected in the flesh of the grouper with [M+ + H](+) ions at m/z 851.51, 857.50, 875.51, 875.49 and 895.54 Da. Some of these compounds have the same mass range as some known dinoflagellate toxins. In conclusion, this study confirms the usefulness of LC-MS analysis to determine the ciguatoxins levels and the toxin profile in fish flesh hazardous to humans.

Systemic administration of interleukin-1 beta activates select populations of central amygdala afferents

The central nucleus of the amygdala (CeA) is activated robustly by an immune challenge such as the systemic administration of the proinflammatory cytokine interleukin-1beta (IL-1beta). Because IL-1beta is not believed to cross the blood-brain barrier in any significant amount, it is likely that IL-1beta elicits CeA cell recruitment by means of activation of afferents to the CeA. However, although many studies have investigated the origins of afferent inputs to the CeA, we do not know which of these also respond to IL-1beta. Therefore, to identify candidate neurons responsible for the recruitment of CeA cells by an immune challenge, we iontophoretically deposited a retrograde tracer, cholera toxin b-subunit (CTb), into the CeA of rats 7 days before systemic delivery of IL-1beta (1 mug/kg, i.a.). By using combined immunohistochemistry, we then quantified the number of Fos-positive CTb cells in six major regions known to innervate the CeA. These included the medial prefrontal cortex, paraventricular thalamus (PVT), ventral tegmental area, parabrachial nucleus (PB), nucleus tractus solitarius, and ventrolateral medulla. Our results show that after deposit of CTb into the CeA, the majority of double-labeled cells were located in the PB and the PVT, suggesting that CeA cell activation by systemic IL-1beta is likely to arise predominantly from cell bodies located in these regions. These findings may have significant implications in determining the central pathways involved in generating acute central responses to a systemic immune challenge. J. Comp. Neurol. 452:288-296, 2002. (C) 2002 Wiley-Liss, Inc.

Dispersal patterns in a regional koala population in south-east Queensland

Koala dispersal was investigated as part of a detailed ecological study of a nationally significant koala population located 20 km south-east of Brisbane, Queensland. From 1996 to 2000, 195 koalas from three sites were captured and fitted with radio-collars. A total of 40 koalas ( 23 males and 17 females) dispersed from these sites. Most (93%) dispersing individuals were 20 - 36 months of age. Three adult females ( more than 36 months old) dispersed and no adult males dispersed during the study. A significantly higher proportion of young males dispersed than females. Dispersal occurred between June and December, with most dispersal of males commencing in July and August and that of females commencing between September and November prior to, and early in, the annual breeding season. The mean straight-line distance between the natal and breeding home ranges for males and females was similar and was measured at 3.5 km ( range 1.1 - 9.7 km) and 3.4 km ( range 0.3 - 10.6 km) respectively. Dispersing males and females tended to successfully disperse south and west of their natal home ranges and were generally unable to successfully disperse to urban areas within the study area, as a high proportion of the mortality of dispersing koalas was associated with attacks by domestic dogs and with collisions with vehicles on roads. Information from other studies indicates that most young koalas disperse from their natal areas. It is likely that the social behaviour and mating systems of koala populations provide mechanisms for young koalas to disperse. The potential role of dispersal in the dynamics of regional koala populations is discussed.

Oceanic interchange and nonequilibrium population structure in the estuarine dependent Indo-Pacific tasselfish, Polynemus sheridani

We assayed mtDNA haplotype [300 base pairs (bp) control region] geography and genealogy in the Indo-Pacific tasselfish, Polynemus sheridani from its contiguous estuarine distribution across northern Australia (n = 169). Eight estuaries were sampled from three oceanographic regions (Timor Sea, Gulf of Carpentaria and the Coral Sea) to assess the impact of Pleistocene sea level changes on the historical connectivity among P. sheridani populations. Specifically, we investigated the genetic consequences of disruption to Indian-Pacific Ocean connectivity brought about by the closure of the Torres Strait. Overall there was significant population subdivision among estuaries (F-ST = 0.161, (Phi(ST) = 0.187). Despite a linear distribution, P. sheridani did not show isolation by distance over the entire sampled range because of genetic similarity of estuaries greater than 3000 km apart. However, significant isolation by distance was detected between estuaries separated by less than 3000 km of coastline. Unlike many genetic studies of Indo-Pacific marine species, there was no evidence for an historical division between eastern and western populations. Instead, phylogeographical patterns were dominated by a starlike intraspecific phylogeny coupled with evidence for population expansion in both the Gulf of Carpentaria and the Coral Sea but not the Timor Sea. This was interpreted as evidence for recent west to east recolonization across of northern Australia following the last postglacial marine advance. We argue that although sufficient time has elapsed postcolonization for populations to approach gene flow-drift equilibrium over smaller spatial scales (< 3000 km), the signal of historical colonization persists to obscure the expected equilibrium pattern of isolation by distance over large spatial scales (> 3000 km).

Segmenting the market of West Australian senior tourists using an artificial neural network

Measuring perceptions of customers can be a major problem for marketers of tourism and travel services. Much of the problem is to determine which attributes carry most weight in the purchasing decision. Older travellers weigh many travel features before making their travel decisions. This paper presents a descriptive analysis of neural network methodology and provides a research technique that assesses the weighting of different attributes and uses an unsupervised neural network model to describe a consumer-product relationship. The development of this rich class of models was inspired by the neural architecture of the human brain. These models mathematically emulate the neurophysical structure and decision making of the human brain, and, from a statistical perspective, are closely related to generalised linear models. Artificial neural networks or neural networks are, however, nonlinear and do not require the same restrictive assumptions about the relationship between the independent variables and dependent variables. Using neural networks is one way to determine what trade-offs older travellers make as they decide their travel plans. The sample of this study is from a syndicated data source of 200 valid cases from Western Australia. From senior groups, active learner, relaxed family body, careful participants and elementary vacation were identified and discussed. (C) 2003 Published by Elsevier Science Ltd.

Kangaroos and feral goats as economic resources for graziers: Some views from south-west Queensland

Fifteen years ago it was proposed that the conversion of kangaroos from a pest to an economically valuable resource would allow graziers to reduce the numbers of domestic stock and thereby lower total grazing pressure. Since then, little progress towards this goal has been achieved. This is believed to be due mainly to the low prices obtained for kangaroo products. A survey of graziers in south-west Queensland was carried out to discover their opinions on kangaroos as a potential economic resource. Questions on the harvesting of feral goats were also included in the survey because of the contrast this industry provides to kangaroo harvesting in terms of grazier involvement. The results of the survey are discussed in relation to resource ownership rights; kangaroo product prices and marketing; and competition within the kangaroo harvesting industry. They show that while low kangaroo product prices do act as a disincentive to graziers, other administrative, legal and institutional factors are also important impediments to their entry to the industry. It is concluded that until the focus of attention widens to include consideration of these as well as just market factors, little progress will be made towards integrating graziers into the kangaroo harvesting industry.

Movements of the ricefield rat, Rattus argentiventer, near a trap-barrier system in rice crops in West Java, Indonesia

The movements of the ricefield rats (Rattus argentiventer) near a trap-barrier system (TBS) were assessed in lowland flood-irrigated rice crops in West Java, Indonesia, to test the hypothesis that a TBS with a 'trap-crop' modifies the movements of rats within 200 m from the trap-crop. The home range use and locations of rat burrows were assessed using radiotelemetry at two sites, one with a TBS with trap-crop (Treatment site, the crop inside the fence was planted 3 weeks earlier than the surrounding crop) and the other with a TBS without trap-crop (Control site, the crop inside the fence was planted at the same time as the surrounding crop). Each TBS was a 50 x 50 m plastic fence with eight multiple-capture rat traps set at the base. More than 700 rats were caught in the TBS with trap-crop, whereas only 10 rats were caught in the TBS without trap-crop. The home range size of females was significantly smaller at the Treatment site (0.96 ha) than the Control site (2.99 ha), but there was no difference for males. Seventy-eight per cent of rats caught in the TBS and fitted with radiocollars had their daytime burrow locations within 200 m of the TBS. We could not determine if the rats caught in the TBS were residents or transients according to demographic parameters. Our results support the hypothesis that a TBS with a trap-crop protects the surrounding rice crop out to a distance of at least 200 m.

DNA vaccine coding for the full-length infectious Kunjin virus RNA protects mice against the New York strain of West Nile virus

A plasmid DNA directing transcription of the infectious full-length RNA genome of Kunjin (KUN) virus in vivo from a mammalian expression promoter was used to vaccinate mice intramuscularly. The KUN viral cDNA encoded in the plasmid contained the mutation in the NS1 protein (Pro-250 to Leu) previously shown to attenuate KUN virus in weanling mice. KUN virus was isolated from the blood of immunized mice 3-4 days after DNA inoculation, demonstrating that infectious RNA was being transcribed in vivo; however, no symptoms of virus-induced disease were observed. By 19 days postimmunization, neutralizing antibody was detected in the serum of immunized animals. On challenge with lethal doses of the virulent New York strain of West Nile (WN) or wild-type KUN virus intracerebrally or intraperitoneally, mice immunized with as little as 0.1-1 mug of KUN plasmid DNA were solidly protected against disease. This finding correlated with neutralization data in vitro showing that serum from KUN DNA-immunized mice neutralized KUN and WN,viruses with similar efficiencies. The results demonstrate that delivery of an attenuated but replicating KUN virus via a plasmid DNA vector may provide an effective vaccination strategy against virulent strains of WN virus.

Ex vivo analysis of T-cell responses to Epstein-Barr virus-encoded oncogene latent membrane protein 1 reveals highly conserved epitope sequences in virus isolates from diverse geographic regions

Epstein-Barr virus (EBV)-encoded oncogene latent membrane protein (LMP) 1, which is consistently expressed in multiple EBV-associated malignancies, has been proposed as a potential target antigen for any future vaccine designed to control these malignancies. However, the high degree of genetic variation in the LMP1 sequence has been considered a major impediment for its use as a potential immunotherapeutic target for the treatment of EBV-associated malignancies. In the present study, we have employed a highly efficient strategy, based on ex vivo functional assays, to conduct an extensive sequence-wide analysis of LMP1-specific T-cell responses in a large panel of healthy virus carriers of diverse ethnic origin and nasopharyngeal carcinoma patients. By comparing the frequencies of T cells specific for overlapping peptides spanning LMP1, we mapped a number of novel HLA class I- and class II-restricted LMP1 T-cell epitopes, including an epitope with dual HLA class I restriction. More importantly, extensive sequence analysis of LMP1 revealed that the majority of the T-cell epitopes were highly conserved in EBV isolates from Caucasian, Papua New Guinean, African, and Southeast Asian populations, while unique geographically constrained genetic variation was observed within one HLA A2 supertype-restricted epitope. These findings indicate that conserved LMP1 epitopes should be considered in designing epitope-based immunotherapeutic strategies against EBV-associated malignancies in different ethnic populations.

Epitope-blocking enzyme-linked immunosorbent assays for the detection of serum antibodies to West Nile virus in multiple avian species

We report the development of epitope-blocking enzyme-linked immunosorbent assays (ELISAs) for the rapid detection of serum antibodies to West Nile virus (WNV) in taxonomically diverse North American avian species. A panel of flavivirus-specific monoclonal antibodies (MAbs) was tested in blocking assays with serum samples from WNV-infected chickens and crows. Selected MAbs were further tested against serum samples from birds that represented 16 species and 10 families. Serum samples were collected from birds infected with WW or Saint Louis encephalitis virus (SLEV) and from noninfected control birds. Serum samples from SLEV-infected birds were included in these experiments because WNV and SLEV are closely related antigenically, are maintained in similar transmission cycles, and have overlapping geographic distributions. The ELISA that utilized MAb 3.11126 potentially discriminated between WW and SLEV infections, as all serum samples from WNV-infected birds and none from SLEV-infected birds were positive in this assay. Assays with MAbs 2132 and 6B6C-1 readily detected serum antibodies in all birds infected with WNV and SLEV, respectively, and in most birds infected with the other virus. Two other MAbs partially discriminated between infections with these two viruses. Serum samples from most WNV-infected birds but no SLEV-infected birds were positive with MAb 3.676, while almost all serum samples from SLEV-infected birds but few from WNV-infected birds were positive with MAb 6B5A-5. The blocking assays reported here provide a rapid, reliable, and inexpensive diagnostic and surveillance technique to monitor WNV activity in multiple avian species.

Epitope-blocking enzyme-linked immunosorbent assays for detection of West Nile virus antibodies in domestic mammals

We evaluated the ability of epitope-blocking enzyme-linked immunosorbent assays (ELISAs) to detect West Nile virus (WNV) antibodies in domestic mammals. Sera were collected from experimentally infected horses, cats, and pigs at regular intervals and screened in ELISAs and plaque reduction neutralization tests. The diagnostic efficacies of these techniques were similar.