Validation of a Competitive Enzyme-Linked Immunosorbent Assay for Detection of Babesia bigemina Antibodies in Cattle

A competitive enzyme-linked immunosorbent assay (cELISA) based on a broadly conserved, species-specific, B-cell epitope within the C terminus of Babesia bigemina rhoptry-associated protein 1a was validated for international use. Receiver operating characteristic analysis revealed 16% inhibition as the threshold for a negative result, with an associated specificity of 98.3% and sensitivity of 94.7%. Increasing the threshold to 21% increased the specificity to 100% but modestly decreased the sensitivity to 87.2%. By using 21% inhibition, the positive predictive values ranged from 90.7% (10% prevalence) to 100% (95% prevalence) and the negative predictive values ranged from 97.0% (10% prevalence) to 48.2% (95% prevalence). The assay was able to detect serum antibody as early as 7 days after intravenous inoculation. The cELISA was distributed to five different laboratories along with a reference set of 100 defined bovine serum samples, including known positive, known negative, and field samples. The pairwise concordance among the five laboratories ranged from 100% to 97%, and all kappa values were above 0.8, indicating a high degree of reliability. Overall, the cELISA appears to have the attributes necessary for international application.

Diel variation in mangrove fish abundances and trophic guilds of northeastern Australian estuaries with a proposed trophodynamic model

Diel activity patterns of tropical fish assemblages in turbid, mangrove-dominated estuaries remain largely undocumented, leading to uncertainty about ecological processes in these systems. To capture active fishes by day and night, gill nets were set perpendicular to mangrove shorelines, in six northeastern Australian estuaries during 13 bimonthly trips. Fish were sampled with eight large mesh (102-151 mm) nets, set for 6 hrs (1500-2100), and checked hourly (1146 day, 635 dusk, 872 night checks). Four smaller mesh (19-51 mm) nets were also set for 1 hr before and after sunset (77 day, 78 night checks). Of 157 total species, 22 were netted exclusively before sunset and 47 exclusively after sunset. All of the top 26 species were present both day and night, but of these, 46% were primarily nocturnal (diel index > 0.65). An average of 77.2 fish hr−1 were netted by day vs 171.4 by night. Within the 400 km coastal region, assemblages differed between two northern wave-dominated (WD) estuaries and four southern tide-dominated ('I'D) estuaries. In all six estuaries Lates calcarifer (Bloch, 1790) dominated night assemblages. In 'I'D estuaries, night assemblages were also dominated by Thryssa hamiltoni Gray, 1835 and Eleutheronema tetradactylum (Shaw, 1804); while in WD estuaries Herklotsichthys castelnaui (Ogilby, 1897), Leiognathus equulus (Forsskål, 1775), and Megalops cyprinoids (Broussonet, 1782) were dominant at night. Nocturnal species included planktivores and carnivores, while daytime assemblages were dominated by detritivores (Mugillidae). Higher night catch rates are attributed to increased activity by mobile fishes moving from mangrove to adjacent habitats to forage, especially immediately post-sunset. Although day-night diets and forage resources have yet to be compared in mangrove systems, previously unrecognized trophic relationships involving variation in diel activity among important fishery species (Centropomidae, polynemidae, Carangidae) and their prey may be key ecological processes in these tropical mangrove estuaries. A proposed hypothesis explaining diel variation in mangrove fish assemblages of tropical estuaries is presented through a conceptual model.

Rickettsia-like-organisms and phytoplasmas associated with diseases in Australian strawberries

Strawberry lethal yellows (SLY) disease in Australia is associated with the phytoplasmas Candidatus Phytoplasma australiense and tomato big bud, and a rickettsia-like-organism (RLO). Ca. P. australiense is also associated with strawberry green petal (SGP) disease. This study investigated the strength of the association of the different agents with SLY disease. We also documented the location of SLY or SGP plants, and measured whether they were RLO or phytoplasma positive. Symptomatic strawberry plants collected from south-east Queensland (Australia) between January 2000 and October 2002 were screened by PCR for both phytoplasmas and the RLO. Two previously unreported disease symptoms termed severe fruit distortion (SFD) and strawberry leaves from fruit (SLF) were observed during this study but there was no clear association between these symptoms and phytoplasmas or the RLO. Only two SGP diseased plants were observed and collected, compared with 363 plants with SLY disease symptoms. Of the 363 SLY samples, 117 tested positive for the RLO, 67 tested positive for Ca. P. australiense AGY strain and 11 plants tested positive for Ca. P. australiense PYL variant strain. On runner production farms at Stanthorpe, Queensland the RLO was detected in SLY diseased plants more frequently than for the phytoplasmas. On fruit production farms on the Sunshine Coast, Queensland, Ca. P. australiense was detected in SLY disease plants more frequently than the RLO.