Japanese encephalitis on Badu Island, Australia: the first isolation of Japanese encephalitis virus from Culex gelidus in the Australasian region and the role of mosquito host-feeding patterns in virus transmission cycles

During investigation of an outbreak of Japanese encephalitis (JE) in the Torres Strait, Australia, in 2000, mosquitoes were collected in Badu Island community and at a newly established communal piggery about 3 km from the community. A total of 94285 mosquitoes, comprising 91240 (96.8%) unengorged females, 1630 (1.7%) blood-engorged females and 1415 (1.5%) males, were processed for virus isolation. One isolate of JE virus was obtained from Culex gelidus, with a minimum infection rate of 12.4:1000. This is the first isolate of JE virus from Cx. gelidus in the Australasian region. No isolates were obtained from Cx. annulirostris, the primary implicated Australian JE vector. Analysis of mosquito host-feeding patterns, using gel diffusion, demonstrated that Cx. annulirostris and 5 other species fed predominately on mammals, Analysis of blood-fed mosquitoes collected within the community demonstrated that the proportion of Cx. annulirostris feeding on pigs in 2000 (2.3%) was significantly lower than that for the 1995-97 period (31.3%). The removal of the pigs from Badu Island community has limited the contact between potential amplifying hosts and mosquitoes, thus potentially reducing the risk of transmission of JE virus to the human population.

The appearance of a second genotype of Japanese encephalitis virus in the Australasian region

In mid-January 2000, the reappearance of Japanese encephalitis (JE) virus activity in the Australasian region was first demonstrated by the isolation of JE virus from 3 sentinel pigs on Badu Island in the Torres Strait. Further evidence of JE virus activity was revealed through the isolation of JE virus from Cidex gelidus mosquitoes collected on Badu Island and the detection of specific JE virus neutralizing antibodies in 3 pigs from Saint Pauls community on Moa Island. Nucleotide sequencing and phylogenetic analyses of the premembrane and envelope genes were performed which showed that both the pig and mosquito JE virus isolates (TSOO and TS4152, respectively) clustered in genotype I, along with northern Thai, Cambodian, and Korean isolates. All previous Australasian JE virus isolates belong to genotype II, along with Malaysian and Indonesian isolates. Therefore, for the first time, the appearance and transmission of a second genotype of JE virus in the Australasian region has been demonstrated.

Flaviviruses isolated from mosquitoes collected during the first recorded outbreak of Japanese encephalitis virus on Cape York Peninsula, Australia

In response to an outbreak of Japanese encephalitis (JE) virus on Cape York Peninsula, Australia, in 1998, mosquitoes were collected using CO2 and octenol-baited Centers for Disease Control and Prevention light traps. A total of 35,235 adult mosquitoes, comprising 31 species, were processed for virus isolation. No isolates of JE virus were recovered from these mosquitoes. However, 18 isolates of Kokobera virus, another flavivirus were obtained from Culex annulirostris. Twelve isolates were from western Cape York (minimum infection rate (MIR) of 0.61: 1,000 mosquitoes) and 6 were from the Northern Peninsula Area (MIR of 1.0:1,000). Potential explanations for the failure to detect JE virus in mosquitoes collected from Cape York Peninsula include the timing of collections, the presence of alternative bloodmeal hosts, differences in pig husbandry, asynchronous porcine seroconversion, and the presence of other flaviviruses.

Experimental infections of pigs with Japanese encephalitis virus and closely related Australian flaviviruses

The flavivirus Japanese encephalitis (JE) virus has recently emerged in the Australasian region. To investigate the involvement of infections with related enzootic flaviviruses, namely Murray Valley encephalitis (MVE) virus and Kunjin (KUN) virus, on immunity of pigs to JE virus and to provide a basis for interpretation of serologic data, experimental infections were conducted with combinations of these viruses. Antibody responses to primary and secondary infections were evaluated using panels of monoclonal antibody-based blocking enzyme-link-ed immuno-sorbent assays and microtiter scrum neutralization tests (mSNTs). Identification of the primary infecting virus was possible only using the mSNTs. Following challenge, unequivocal diagnosis was impossible due to variation in immune responses between animals and broadened and/or anamnestic responses. Viremia for JE virus was readily detected in pigs following primary infection, but was not detected following prior exposure to MVE or KUN viruses. Boosted levels of existing cross-neutralizing antibodies to JE virus suggested a role for this response in suppressing JE viremia.

Development of the pulmonary surfactant system in the green sea turtle, Chelonia mydas

This study describes the developmental changes in pulmonary surfactant (PS) lipids throughout incubation in the sea turtle, Chelonia mydas. Total phospholipid (PL), disaturated phospholipid (DSP) and cholesterol (Chol) harvested from lung washings increased with advancing incubation, where secretion was maximal at pipping, coincident with the onset of pulmonary ventilation. The DSP/PL ratio increased, whereas the Chol/PL and the Chol/DSP ratio declined throughout development. The phospholipids, therefore, are independently regulated from Chol and their development matches that of mammals. To explore whether hypoxia could elicit an effect on the development of the PS system, embryos were exposed to a chronic dose of 17% O-2 for the final similar to 40% of incubation. Hypoxia did not affect incubation time, absolute, nor relative abundance of the surfactant lipids, demonstrating that the development of the system is robust and that embryonic development continues unabated under mild hypoxia. Hypoxia-incubated hatchlings had lighter wet lung weights than those from normoxia, inferring that mild hypoxia facilitates lung clearance in this species. (C) 2001 Elsevier Science B.V. All rights reserved.

Temperature variation within and between nests of the green sea turtle, Chelonia mydas (Chelonia : Cheloniidae) on Heron Island, Great Barrier Reef

Four temperature data-loggers were placed in each of five green sea turtle nests on Heron Island in the 1998-99 nesting season. Temperatures in all nests increased as incubation progressed due to general sand heating and increased metabolic heat production of the developing embryos. Even at the top of nests no daily diurnal fluctuation in temperature was evident. The temperature of eggs in the middle of the nest increased above those in the nest periphery during the last third of incubation. However, this metabolic nest heating would have little effect on hatchling sex ratio because it occurred after the sex-determining period. Small differences in temperature between regions of a nest persisted throughout incubation and may be important in ensuring the production of at least some individuals of the opposite sex in nests that have temperatures close to either the all-male or all-female determining temperatures. Location and degree of shading of nests had little effect on mean nest temperature, but deeper nests were generally cooler and therefore were predicted to produce a higher proportion of males than were shallower nests. Nest temperature profile data indicated that the 1998-99 nesting season on Heron Island would have produced a strongly female-biased sex ratio amongst hatchlings.

Incubation temperature, energy expenditure and hatchling size in the green turtle (Chelonia mydas), a species with temperature-sensitive sex determination

Eggs from the Heron Island, Great Barrier Reef, nesting population of green turtles (Chelonia mydas) were incubated at all-male-determining (26 degreesC) and all-female-determining (30 degreesC) temperatures. Oxygen consumption and embryonic growth were monitored throughout incubation, and hatchling masses and body dimensions were measured from both temperatures. Eggs hatched after 79 and 53 days incubation at 26 degreesC and 30 degreesC respectively. Oxygen consumption at both temperatures increased to a peak several days before hatching, a pattern typical of turtle embryos, and the rate of oxygen was higher at 30 degreesC than 26 degreesC. The total amount of energy consumed during incubation, and hatchling dimensions, were similar at both temperatures, but hatchlings from 26 degreesC had larger mass, larger yolk-free mass and smaller residual yolks than hatchlings from 30 degreesC. Because of the difference in mass of hatchlings, hatchlings from 30 degreesC had a higher production cost.