Production of positive controls for calcivirus-specific PCR using recombinant baculiovirus technology

Recent advances in our knowledge of the genetic structure of human caliciviruses (HuCVs) and small round-structured viruses (SRSVs) have led to the development of polymerase chain reaction (PCR)-based molecular tests specific for these viruses. These methods have been developed to detect a number of human pathogenic viruses in environmental samples including water, sewage and shellfish. HuCVs and SRSVs are not culturable, and no animal model is currently available. Therefore there is no convenient method of preparing viruses for study or for reagent production. One problem facing those attempting to use PCR-based methods for the detection of HuCVs and SRSVs is the lack of a suitable positive control substrate. This is particularly important when screening complex samples in which the levels of inhibitors present may significantly interfere with amplificiation. Regions within the RNA polymerase regions of two genetically distinct human caliciviruses have been amplified and used to produce recombinant baculoviruses which express RNA corresponding to the calicivirus polymerase. This RNA is being investigated as a positive control substrate for PCR testing, using current diagnostic primer sets. Recombinant baculovirus technology will enable efficient and cost-effective production of large quantities of positive control RNA with a specific known genotype. We consider the development of these systems as essential for successful screening and monitoring applications.

Summary report on water quantity and quality studies of Vancouver Lake, Washington

Vancouver Lake, located adjacent to the Columbia River and just north of the Vancouver-Portland metropolitan area, is a "dying" lake. Although all lakes die naturally in geologic time through the process of eutrophication,* Vancouver Lake is dying more rapidly due to man's activities and due to the resultant increased accumulation of sediment, chemicals, and wastes. Natural eutrophication takes thousands of years, whereas man-made modifications can cause the death of a lake in decades. Vancouver Lake does, however, have the potential of becoming a valuable water resource asset for the area, due particularly to its location near the Columbia River which can be used as a source of "flushing" water to improve the quality of Vancouver Lake. (Document pdf contains 59 pages) Community interest in Vancouver Lake has waxed and waned. Prior to World War II, there were relatively few plans for discussions about the Lake and its surrounding land area. A plan to drain the Lake for farming was prohibited by the city council and county commissioners. Interest increased in 1945 when the federal government considered developing the Lake as a berthing harbor for deactivated ships at which time a preliminary proposal was prepared by the City. The only surface water connection between Vancouver Lake and the Columbia River, except during floods, is Lake River. The Lake now serves as a receiving body of water for Lake River tidal flow and surface flow from creeks and nearby land areas. Seasonally, these flows are heavily laden with sediment, septic tank drainage, fertilizers and drainage from cattle yards. Construction and gravel pit operations increase the sediment loads entering the Lake from Burnt Bridge Creek and Salmon Creek (via Lake River by tidal action). The tidal flats at the north end of Vancouver Lake are evidence of this accumulation. Since 1945, the buildup of sediment and nutrients created by man's activities has accelerated the growth of the large water plants and algae which contribute to the degeneration of the Lake. Flooding from the Columbia River, as in 1968, has added to the deposition in Vancouver Lake. The combined effect of these human and natural activities has changed Vancouver Lake into a relatively useless body of shallow water supporting some wildlife, rough fish, and shallow draft boats. It is still pleasant to view from the hills to the east. Because precipitation and streamflow are the lowest during the summer and early fall, water quantity and quality conditions are at their worst when the potential of the Lake for water-based recreation is the highest. Increased pollution of the Lake has caused a larger segment of the community to become concerned. Land use and planning studies were undertaken on the Columbia River lowlands and a wide variety of ideas were proposed for improving the quality of the water-land environment in order to enhance the usefulness of the area. In 1966, the College of Engineering Research Division at Washington State University (WSU0 in Pullman, Washington, was contacted by the Port of Vancouver to determine possible alternatives for restoring Vancouver Lake. Various proposals were prepared between 1966 and 1969. During the summer and fall of 1967, a study was made by WSU on the existing water quality in the Lake. In 1969, the current studies were funded to establish a data base for considering a broad range of alternative solutions for improving the quantity and quality of Vancouver Lake. Until these studies were undertaken, practically no data on a continuous nature were available on Vancouver Lake, Lake River, or their tributaries. (Document pdf contains 59 pages)

Chemical contaminants, pathogen exposure and general health status of live and beach-cast Washington sea otters (Enhydra lutris kenyoni)

Analyses of blood and liver samples from live captured sea otters and liver samples from beachcast sea otter carcasses off the remote Washington coast indicate relatively low exposure to contaminants, but suggest that even at the low levels measured, exposure may be indicated by biomarker response. Evidence of pathogen exposure is noteworthy - infectious disease presents a potential risk to Washington sea otters, particularly due to their small population size and limited distribution. During 2001 and 2002, 32 sea otters were captured, of which 28 were implanted with transmitters to track their movements and liver and blood samples were collected to evaluate contaminant and pathogen exposure. In addition, liver samples from fifteen beachcast animals that washed ashore between 1991 and 2002 were analyzed to provide historical information and a basis of reference for values obtained from live otters. The results indicate low levels of metals, butyltins, and organochlorine compounds in the blood samples, with many of the organochlorines not detected except polychlorinated biphenyls (PCBs), and a few aromatic hydrocarbons detected in the liver of the live captured animals. Aliphatic hydrocarbons were measurable in the liver from the live captured animals; however, some of these are likely from biogenic sources. A significant reduction of vitamin A storage in the liver was observed in relation to PCB, dibutyltin and octacosane concentration. A significant and strong positive correlation in vitamin A storage in the liver was observed for cadmium and several of the aliphatic hydrocarbons. Peripheral blood mononuclear cell (PBMC) cytochrome P450 induction was elevated in two of 16 animals and may be potentially related to aliphatic and aromatic hydrocarbon exposure. Mean concentration of total butyltin in the liver of the Washington beach-cast otters was more than 15 times lower than the mean concentration reported by Kannan et al. (1998) for Southern sea otters in California. Organochlorine compounds were evident in the liver of beach-cast animals, despite the lack of large human population centers and development along the Washington coast. Concentrations of PCBs and chlordanes (e.g., transchlordane, cis-chlordane, trans-nonachlor, cis-nonachlor and oxychlordane) in liver of Washington beach-cast sea otters were similar to those measured in Aleutian and California sea otters, excluding those from Monterey Bay, which were higher. Mean concentrations of 1,1,1,- trichloro-2,2-bis(p-chlorophyenyl)ethanes (DDTs) were lower, and mean concentrations of cyclohexanes (HCH, e.g., alpha BHC, beta BHC, delta BHC and gamma BHC) were slightly higher in Washington beach-cast otters versus those from California and the Aleutians. Epidemiologically, blood tests revealed that 80 percent of the otters tested positive for morbillivirus and 60 percent for Toxoplasma, the latter of which has been a significant cause of mortality in Southern sea otters in California. This is the first finding of positive morbillivirus titers in sea otters from the Northeast Pacific. Individual deaths may occur from these diseases, perhaps more so when animals are otherwise immuno-compromised or infected with multiple diseases, but a population-threatening die-off from these diseases singly is unlikely while population immunity remains high. The high frequency of detection of morbillivirus and Toxoplasma in the live otters corresponds well with the cause of death of stranded Washington sea otters reported herein, which has generally been attributable to infectious disease. Washington’s sea otter population continues to grow, with over 1100 animals currently inhabiting Washington waters; however, the rate of growth has slowed over recent years. The population has a limited distribution and has not yet reached its carrying capacity and as such, is still considered at high risk to catastrophic events. (PDF contains 189 pages)

The brine and gas content of sea ice with attention to low salinities and high temperatures

Based on the well known sea ice phase diagram, equations are derived for determining the brine and gas content of sea Ice for high temperatures (range 0 to -2 °C) and low salinities. The presently widely used equations of Cox and Weeks (1982) are valid only for temperatures below -2°C. Fresh-water ice is used as a boundary condition for the equations. The relative salt concentrations in brine are_assumed to be the same as in normal (or standard) seawater. Two sets of equations are presented: 1) accurate formulae based on UNESCO standard sea water equations, and 2) approximate formulae based on general properties of weak solutions. The approximate formulae are not essentially different from the classical system which basically assumes the freezing point to be a linear function of fractional salt content. The agreement between the two approaches is excellent and the approximate system is good enough for most applications.

Variation in yellowfin tuna (Thunnus albacares) catches related to El Niño-Southern Oscillation events at the entrance to the Gulf of California

Fishery catch data on yellowfin tuna (Thunnus albacares) were examined to study the effects of El Niño events between 1990 and 1999 for an area in the northeastern tropical Pacific (18−24°N, 112−104°W). The data were extracted from a database of logbook records from the Mexican tuna purse-seine f leet. Latitudinal distribution of the catches increased from south to north for the 10-year period. Highest catches and effort were concentrated between 22°N and 23°N. This area accumulated 48% of the total catch over the 10year period. It was strongly correlated with El Niño-Southern Oscillation (ENSO) events. At least two periods of exceptionally high catches occurred following El Niño events in 1991 and 1997. Peaks of catches were triggered by the arrival of positive anomalies of sea surface temperature (SST) to the area. A delay of two to four months was observed between the occurrence of maximum SST anomalies at the equator and peaks of catch. Prior to these two events, negative SST anomalies were the dominant feature in the study area and catch was extremely low. This trend of negative SST anomalies with low catches followed by positive SST anomalies and high catches may be attributed to northward yellowfin tuna migration patterns driven by El Niño forcing, a result that contrasts with the known behavior of decreasing relative abundance of these tuna after El Niño events in the eastern Pacific. However, this decrease in relative abundance may be the result of a local or subregional effect.

Individual growth rates and movement of juvenile white shrimp (Litopenaeus setiferus) in a tidal marsh nursery

We measured growth and movements of individually marked free-ranging juvenile white shrimp (Litopenaeus setiferus) in tidal creek subsystems of the Duplin River, Sapelo Island, Georgia. Over a period of two years, 15,974 juvenile shrimp (40−80 mm TL) were marked internally with uniquely coded microwire tags and released in the shallow upper reaches of four salt marsh tidal creeks. Subsequent samples were taken every 3−6 days from channel segments arranged at 200-m intervals along transects extending from the upper to lower reach of each tidal creek. These collections included 201,384 juvenile shrimp, of which 184 were marked recaptures. Recaptured shrimp were at large an average of 3−4 weeks (range: 2−99 days) and were recovered a mean distance of <0.4 km from where they were initially marked. Mean residence times in the creek subsystems ranged from 15.2 to 25.5 days and were estimated from exponential decay functions describing the proportions of marked individuals recaptured with increasing days at large. Residence time was not significantly correlated with creek length (Pearson=−0.316, P=0.684 ), but there was suggestive evidence of positive associations with either intertidal (Pearson r=0.867, P=0.133) or subtidal (Pearson r=0.946, P=0.054) drainage area. Daily mean specific growth rates averaged 0.009 to 0.013 among creeks; mean absolute growth rates ranged from 0.56−0.84 mm/d, and were lower than those previously reported for juvenile penaeids in estuaries of the southeastern United States. Mean individual growth rates were not significantly different between years (t-test, P>0.30) but varied significantly during the season, tending to be greater in July than November. Growth rates were size-dependent, and temporal changes in size distributions rather than temporal variation in physical environmental factors may have accounted for seasonal differences in growth. Growth rates differed between creeks in 1999 (t-test, P<0.015), but not in 1998 (t-test, P>0.5). We suggest that spatial variation in landscape structure associated with access to intertidal resources may have accounted for this apparent interannual difference in growth response.

A sudden collapse in distribution of Pacific sardine (Sardinops sagax) off southwestern Australia enables an objective re-assessment of biomass estimates

Fishery-independent estimates of spawning biomass (BSP) of the Pacific sardine (Sardinops sagax) on the south and lower west coasts of Western Australia (WA) were obtained periodically between 1991 and 1999 by using the daily egg production method (DEPM). Ichthyoplankton data collected during these surveys, specifically the presence or absence of S. sagax eggs, were used to investigate trends in the spawning area of S. sagax within each of four regions. The expectation was that trends in BSP and spawning area were positively related. With the DEPM model, estimates of BSP will change proportionally with spawning area if all other variables remain constant. The proportion of positive stations (PPS), i.e., stations with nonzero egg counts — an objective estimator of spawning area — was high for all south coast regions during the early 1990s (a period when the estimated BSP was also high) and then decreased after the mid-1990s. There was a decrease in PPS from the mid-1990s to 1999. The particularly low estimates in 1999 followed a severe epidemic mass mortality of S. sagax throughout their range across southern Australia. Deviations from the expected relationship between BSP and PPS were used to identify uncertainty around estimates of BSP. Because estimation of spawning area is subject to less sampling bias than estimation of BSP, the deviation in the relation between the two provides an objective basis for adjusting some estimates of the latter. Such an approach is particularly useful for fisheries management purposes when sampling problems are suspected to be present. The analysis of PPS undertaken from the same set of samples from which the DEPM estimate is derived will help provide information for stock assessments and for the management of purse-seine fisheries.

Biology and population dynamics of cowcod (Sebastes levis) in the southern California Bight

Cowcod (Sebastes levis) is a large (100-cm-FL), long-lived (maximum observed age 55 yr) demersal rockfish taken in multispecies commercial and recreational fisheries off southern and central California. It lives at 20–500 m depth: adults (>44 cm TL) inhabit rocky areas at 90–300 m and juveniles inhabit fine sand and clay at 40–100 m. Both sexes have similar growth and maturity. Both sexes recruit to the fishery before reaching full maturity. Based on age and growth data, the natural mortality rate is about M =0.055/yr, but the estimate is uncertain. Biomass, recruitment, and mortality during 1951–98 were estimated in a delay-difference model with catch data and abundance indices. The same model gave less precise estimates for 1916–50 based on catch data and assumptions about virgin biomass and recruitment such as used in stock reduction analysis. Abundance indices, based on rare event data, included a habitat-area–weighted index of recreational catch per unit of fishing effort (CPUE index values were 0.003–0.07 fish per angler hour), a standardized index of proportion of positive tows in CalCOFI ichthyoplankton survey data (binomial errors, 0–13% positive tows/yr), and proportion of positive tows for juveniles in bottom trawl surveys (binomial errors, 0–30% positive tows/yr). Cowcod are overfished in the southern California Bight; biomass during the 1998 season was about 7% of the virgin level and recent catches have been near 20 metric tons (t)/yr. Projections based on recent recruitment levels indicate that biomass will decline at catch levels > 5 t/yr. Trend data indicate that recruitment will be poor in the near future. Recreational fishing effort in deep water has increased and has become more effective for catching cowcod. Areas with relatively high catch rates for cowcod are fewer and are farther offshore. Cowcod die after capture and cannot be released alive. Two areas recently closed to bottom fishing will help rebuild the cowcod stock.

Environmental assessment of shrimp farming in relation to livelihoods in the south-west coastal Bangladesh

The study was designed to assess the environmental impact of shrimp farming and implications on local livelihoods at the south-west coastal area of Bangladesh. All the stakeholders reported that shrimp farming negatively affected on the environment at the coastal area. The soil and water, fish habitation, agricultural cropland, grazing land, indigenous fish, household vegetations, trees and plants, land fertility and mangroves are affected negatively by the shrimp farming in the coastal area. About 44% of stakeholders agreed that mangroves were destroyed by the extension of shrimp farming in the study area. In the case of positive impact of shrimp farming on environment about 16% of stakeholders agreed that the household vegetations increased due to alternate rice and shrimp-prawn farming.