Complete genomic sequence of the Australian south-west genotype of Sindbis virus: Comparisons with other Sindbis strains and identification of a unique deletion in the 3 '-untranslated region

Our previous studies have shown that two distinct genotypes of Sindbis (SIN) virus occur in Australia. One of these, the Oriental/Australian type, circulates throughout most of the Australian continent, whereas the recently identified south-west (SW) genetic type appears to be restricted to a distinct geographic region located in the temperate south-west of Australia. We have now determined the complete nucleotide and translated amino acid sequences of a SW isolate of SIN virus (SW6562) and performed comparative analyses with other SIN viruses at the genomic level. The genome of SW6562 is 11,569 nucleotides in length, excluding the cap nucleotide and poly (A) tail. Overall this virus differs from the prototype SIN virus (strain AR339) by 23% in nucleotide sequence and 12.5% in amino acid sequence. Partial sequences of four regions of the genome of four SW isolates were determined and compared with the corresponding sequences from a number of SIN isolates from different regions of the World. These regions are the non-structural protein (nsP3), the E2 gene, the capsid gene, and the repeated sequence elements (RSE) of the 3'UTR. These comparisons revealed that the SW SIN viruses were more closely related to South African and European strains than to other Australian isolates of SIN virus. Thus the SW genotype of SIN virus may have been introduced into this region of Australia by viremic humans or migratory birds and subsequently evolved independently in the region. The sequence data also revealed that the SW genotype contains a unique deletion in the RSE of the 3'UTR region of the genome. Previous studies have shown that deletions in this region of the SIN genome can have significant effects on virus replication in mosquito and avian cells, which may explain the restricted distribution of this genotype of SIN virus.

Flavivirus isolations from mosquitoes collected from western Cape York Peninsula, Australia, 1999-2000

After the 1st appearance of Japanese encephalitis virus (JE) on mainland Australia in 1998, a study was undertaken to investigate whether JE had become established in enzootic transmission cycles on western Cape York Peninsula. Adult mosquitoes were collected during the late wet season from Kowanyama and Pormpuraaw in April 1999, and Pormpuraaw and Barr's Yard in April 2000. Despite processing 269,270 mosquitoes for virus isolation, no isolates of JE were obtained. However, other flaviviruses comprising Murray Valley encephalitis virus, Kunjin virus, Alfuy virus, and Kokobera virus (KOK) were isolated. Isolates of the alphaviruses Ross River virus, Barmah Forest virus (BF), and Sindbis virus (SIN) also were obtained. The majority (88%) of isolates were from members of the Culex sitiens subgroup. Single isolates of KOK, BF and SIN were obtained from Ochlerotatus vigilax, Oc. normanensis, and Anopheles bancroftii, respectively. The isolations of flaviviruses during the late wet season indicate that conditions were suitable for flavivirus activity in the area. No evidence was found to suggest that JE has become established in enzootic transmission cycles on western Cape York, although study sites and field trips were limited.

Host- rather than virus-related factors reduce health-related quality of life in hepatitis C virus infection.

BACKGROUND: Hepatitis C virus (HCV) infection is associated with decreased health-related quality of life (HRQOL). Although HCV has been suggested to directly impair neuropsychiatric functions, other factors may also play a role. PATIENTS AND METHODS: In this cross-sectional study, we assessed the impact of various host-, disease- and virus-related factors on HRQOL in a large, unselected population of anti-HCV-positive subjects. All individuals (n = 1736) enrolled in the Swiss Hepatitis C Cohort Study (SCCS) were asked to complete the Short Form 36 (SF-36) and the Hospital Anxiety Depression Scale (HADS). RESULTS: 833 patients (48%) returned the questionnaires. Survey participants had significantly worse scores in both assessment instruments when compared to a general population. By multivariable analysis, reduced HRQOL (mental and physical summary scores of SF-36) was independently associated with income. In addition, a low physical summary score was associated with age and diabetes, whereas a low mental summary score was associated with intravenous drug use. HADS anxiety and depression scores were independently associated with income and intravenous drug use. In addition, HADS depression score was associated with diabetes. None of the SF-36 or HADS scores correlated with either the presence or the level of serum HCV RNA. In particular, SF-36 and HADS scores were comparable in 555 HCV RNA-positive and 262 HCV RNA-negative individuals. CONCLUSIONS: Anti-HCV-positive subjects have decreased HRQOL compared to controls. The magnitude of this decrease was clinically important for the SF-36 vitality score. Host and environmental, rather than viral factors, seem to impact on HRQOL level.

Differential Expression of Immune Response Genes in Steller Sea Lions (Eumetopias jubatus): An Indicator of Ecosystem Health?

Characterization of the polygenic and polymorphic features of the Steller sea lion major histocompatibility complex (MHC) provides an ideal window for evaluating immunologic vigor of the population and identifying emergence of new genotypes that reflect ecosystem pressures. MHC genotyping can be used to measure the potential immunologic vigor of a population. However, since ecosystem-induced changes to MHC genotype can be slow to emerge, measurement of differential expression of these genes can potentially provide real-time evidence of immunologic perturbations. MHC DRB genes were cloned and sequenced using peripheral blood mononuclear leukocytes derived from 10 Steller sea lions from Southeast Alaska, Prince William Sound, and the Aleutian Islands. Nine unique DRB gene sequences were represented in each of 10 animals. MHC DRB gene expression was measured in a subset of six sea lions. Although DRB in genomic DNA was identical in all individuals, relative levels of expressed DRB mRNA was highly variable. Selective suppression of MHC DRB genes could be indicative of geographically disparate environmental pressures, thereby serving as an immediate and sensitive indicator of population and ecosystem health.

Host- rather than virus-related factors reduce health-related quality of life in hepatitis C virus infection

BACKGROUND: Hepatitis C virus (HCV) infection is associated with decreased health-related quality of life (HRQOL). Although HCV has been suggested to directly impair neuropsychiatric functions, other factors may also play a role. PATIENTS AND METHODS: In this cross-sectional study, we assessed the impact of various host-, disease- and virus-related factors on HRQOL in a large, unselected population of anti-HCV-positive subjects. All individuals (n = 1736) enrolled in the Swiss Hepatitis C Cohort Study (SCCS) were asked to complete the Short Form 36 (SF-36) and the Hospital Anxiety Depression Scale (HADS). RESULTS: 833 patients (48%) returned the questionnaires. Survey participants had significantly worse scores in both assessment instruments when compared to a general population. By multivariable analysis, reduced HRQOL (mental and physical summary scores of SF-36) was independently associated with income. In addition, a low physical summary score was associated with age and diabetes, whereas a low mental summary score was associated with intravenous drug use. HADS anxiety and depression scores were independently associated with income and intravenous drug use. In addition, HADS depression score was associated with diabetes. None of the SF-36 or HADS scores correlated with either the presence or the level of serum HCV RNA. In particular, SF-36 and HADS scores were comparable in 555 HCV RNA-positive and 262 HCV RNA-negative individuals. CONCLUSIONS: Anti-HCV-positive subjects have decreased HRQOL compared to controls. The magnitude of this decrease was clinically important for the SF-36 vitality score. Host and environmental, rather than viral factors, seem to impact on HRQOL level.

Spatial and temporal definition of Ochlerotatus camptorhynchus (Thomson) (Diptera : Culicidae) in the Gippsland Lakes system of eastern Victoria

The confirmed vector of Ross River virus, Ochlerotatus camptorhynchus (Thomson), is the dominant mosquito species inhabiting saline marshes in coastal Victoria. This paper re-examines previously published data on Oc. camptorhynchus, plus additional data collected since that time, and provides greater spatial and temporal definition of Oc. camptorhynchus numbers at seven sites across the Gippsland Lakes system of eastern Victoria. A total of 357 672 Oc. camptorhynchus was captured from 1188 trap-nights across the seven trap sites during trapping seasons from 1990 to 2001. The dominance of Oc. camptorhynchus across the seven sites averaged 75%, with significant differences in mean abundance of Oc. camptorhynchus found between all trap sites. Significant differences in monthly abundance of Oc. camptorhynchus were observed for Wellington Shire. Increase in populations of Oc. camptorhynchus was associated with increases in rainfall at all trap sites, higher minimum temperatures at two of the seven trap sites, and wind speed at one trap site. Prioritisation of mosquito control may be applied based on spatial and temporal factors according to the findings of this study.

Development time and survival of Verrallina funerea (Theobald) (Diptera : Culicidae) immatures and other brackish water mosquito species in southeast Queensland, Australia

Verrallina funerea (Theobald) is a brackish water mosquito that is recognised as an important pest and vector in southeast Queensland, Australia. Immature development time and survival of Ve. funerea was defined in the laboratory in response to a range of temperatures (17-34 degrees C) and salinities (0-35 parts per thousand (p.p.t)). The expression of autogeny in this species was also assessed. Salinity only had a slight effect on mean development time from hatching to adult emergence (7.0-7.4 d at salinities of 0, 17.5 and 31.5 p.p.t) and survival was uniformly high (97.5-99.0%). Mean development times were shorter at 26, 29 and 32 degrees C (7.0, 6.8 and 6.8 d, respectively) and longest at 17 degrees C (12.2 d). The threshold temperature (t) was 5.8 degrees C and the thermal constant (K) was 142.9 degree-days above t. Survival to adulthood decreased from > 95% (at 17-29 degrees C) to 78% (at 32 degrees C) and 0% (at 34 degrees C). No expression of autogeny was observed. Immature development times of Ve. funerea, Ochlerotatus vigilax (Skuse) and Oc. procax (Skuse) were then determined under field conditions at Maroochy Shire. Following tide and rain inundation, cohorts of newly hatched larvae were monitored daily by dipping, and time until pupation was noted. Tidal inundation triggered hatching of Ve. funerea and Oc. vigilax larvae whereas Oc. procax larvae were found only after rain inundation. Estimates of Ve. funerea and Oc. vigilax field development times were similar (8-9 d) while Oc. procax development time was slightly longer (9-10 d). Based on these survey results, control activities targeting Ve. funerea must be initiated 4 d (if using Bacillus thuringiensis var. israelensis de Barjac) or 5 d (if using s-methoprene) after inundation. However, Casuarina glauca Sieber canopy and branchlets covering breeding habitats may present a problem for the penetration of such treatments.

BALD EAGLES (HALIAEETUS LEUCOCEPHALUS) AS INDICATORS OF GREAT LAKES ECOSYSTEM HEALTH

Environmental indicators have been proposed as a means to assess ecological integrity, monitoring both chemical and biological stressors. In this study, we used nestling bald eagles as indicators to quantify direct or indirect tertiary-level contaminant exposure. The spatial and temporal trends of polychlorinated biphenyl (PCB) congeners were evaluated in nestling plasma from 1999–2014. Two hexa-chlorinated congeners, PCB-138 and 153, were detected with the highest frequency and greatest concentrations throughout Michigan. Less-chlorinated congeners such as PCB-52 and 66 however, comprised a greater percentage of total PCB concentrations in nestlings proximate to urbanized areas, such as along the shorelines of Lake Erie. Toxic equivalents were greatest in the samples collected from nestlings located on Lake Erie, followed by the other Great Lakes spatial regions. Nestling plasma samples were also used to measure concentrations of the most heavily-used group of flame retardants, brominated diphenyl ethers (BDEs), and three groups of alternative flame retardants, non-BDE Brominated Flame Retardants (NBFRS), Dechloranes, and organophosphate esters (OPs). BDE-47, 99 and 100 contributed the greatest to total BDE concentrations. Concentrations of structurally similar NBFRs found in this study and recent atmospheric studies indicate that they are largely used as replacements to previously used BDE mixtures. A variety of Dechloranes, or derivatives of Mirex and Dechlorane Plus, were measured. Although, measured at lesser concentrations, environmental behavior of these compounds may be similar to mirex and warrant future research in aquatic species. Concentrations of OPs in nestling plasma were two to three orders of magnitude greater than all other groups of flame retardants. In addition to chemical indicators, bald eagles have also been proposed as indicators to identify ecological stressors using population measures that are tied to the fitness of individuals and populations. Using mortality as a population vitality rate, vehicle collisions were found to be the main source of mortality with a greater incidence for females during white-tailed deer (Odocoileus virginianus) hunting months and spring snow-melt. Lead poisoning was the second greatest source of mortality, with sources likely due to unretrieved hunter-killed, white-tailed deer carcasses, and possibly exacerbated by density-dependent effects due to the growing population in Michigan.

Evaluation of liquid Bacillus thuringiensis var. israelensis products for control of Australian Aedes arbovirus vectors

Laboratory bioassay studies were conducted in southeast Queensland, Australia,: on the efficacy of Teknar (R), VectoBac (R) 12AS, and Cybate (R) (active ingredient: 1,200 international toxic units Bacillus thuringiensis var, israelensis [Bti]) against 3rd instars of the arbovirus vectors Aedes aegypti. Ae. notoscriptus, Ae. vigilax, and Ae. camptorhynchus. Probit analyses were then used to determine LD,, (median lethal dose), LD95, and lethal dose ratios (LDR). Aedes aegypti and Ae. notoscriptus, both container-habitat species, tolerated the highest Bti concentrations compared with saltmarsh Ae. vigilax and Ae. camptorhynchus. For example, the LDR for Ae. vigilax versus Ae. notoscriptus exposed to Cybate was 0.14 (95% confidence limit [CL] 0.03-0.61). Similarly, the Cybate LDR for Ae. camptorhynchus versus Ae. notoscriptus was 0.22 (95% CL 0.07-0.70). Teknar produced similar results with an LDR of 0.21 (95% CL 0.04-1.10) for Aedes vigilax versus Aedes notoscriptus. Differences in product efficacy were found when tested against the 2 container-breeding species. Cybate was less effective than Teknar with LDRs of 1.55 (95% CL 0.65-3.67) and 1.87 (95% CL 0.68-5.15) for Aedes aegypti and Ae. notoscriptus, respectively. The significant differences in susceptibility between mosquito species and varying efficacy between products highlight the importance of evaluating concentration-response data prior to contracting with distributors of mosquito control products. This information is crucial to resistance management strategies.

Putative receptor binding sites on alphaviruses as visualized by cryoelectron microscopy.

The structures of Sindbis virus and Ross River virus complexed with Fab fragments from monoclonal antibodies have been determined from cryoelectron micrographs. Both antibodies chosen for this study bind to regions of the virions that have been implicated in cell-receptor recognition and recognize epitopes on the E2 glycoprotein. The two structures show that the Fab fragments bind to the outermost tip of the trimeric envelope spike protein. Hence, the same region of both the Sindbis virus and Ross River virus envelope spike is composed of E2 and is involved in recognition of the cellular receptor.