ELISA absorbance cut-off method affects malaria sporozoite rate determination in wild Afrotropical Anopheles

ABSTRACT. Malaria sporozoite infection rates in a mixed species group of 244 Anopheles gambiae Giles sensu lato and 115 An. funestus Giles wild female mosquitoes were compared using three methods to determine cutoff absorbance values for positivity of a Plasmodium falciparum Welch enzyme-linked immunosorbent assay (ELISA). Positive controls were based on P. falciparum circumsporozoite protein. As negative controls, four wild male Anopheles were included on each microtitre plate; tests were repeated on four consecutive days for each plate.
Infection rates were estimated at 13.1–22.8% using the mean absorbance value of negative controls plus three standard deviations, 11.7–12.8% using double the mean and 12.5–13.6% using the fixed cut-off value of 0.20 (allowing for 20% variation in negative control absorbance values).
Observed agreement for positivity or negativity among samples tested four times was 98.6% for the 2× mean method, 97.2% for the fixed cut-off 0.20 value, but only 82.7% for the mean +3 SD method. It was concluded that the 2× mean cut-off method is most reliable for field studies. P. falciparum sporozoite rates of 12.2% in An. funestus and 11.9% in An. gambiae s. l . were thus determined on the basis of the 2× mean cut-off method.
This comparative evaluation demonstrates that vector infectivity rates can be seriously over-estimated from sporozoite ELISA tests, by as much as 87% in one case considered here, depending on the absorbance cut-off method applied for negative controls.     

Quantitation of malaria sporozoites transmitted in vitro during salivation by wild Afrotropical Anopheles

Abstract. The malaria transmission potential of wild, infective Anopheles from western Kenya was evaluated by determining the number of sporozoites transmitted in vitro by salivation when their mouthparts were inserted into capillary tubes containing either sucrose or blood. With sucrose, 86.6% of 102 infective Anopheles transmitted a geometric mean (GM) of 3.84 sporozoites (range 1–34). With blood, 23.1% of 104 infective Anopheles , tested on the day of collection, transmitted a GM of 2.30 sporozoites (range 1–117). For Anopheles held 5 days postcapture before testing with blood, 53.6% of 56 transmitted a GM of 6.04 sporozoites (range 1–420). Transmitting Anopheles contained significantly more salivary gland sporozoites than non-transmitters. No significant differences were detected between Anopheles gambiae Giles sensu lato and Anopheles funestus Giles in sporozoite transmission by individuals with sporozoites in their salivary glands.
Sporozoites were detected microscopically in the salivary duct from heads in 80.3% of 117 infective Anopheles (GM=11.2, range 1–71). Sporozoite detection in mosquito heads by ELISA was 25% less efficient than microscopic detection.
Over 98% of the infective Anopheles transmitted less than twenty-five sporozoites. Transmitted sporozoites represented only about 3% of the total sporozoites in the salivary glands suggesting that sporozoite transmission may be restricted to sporozoites in the salivary duct at the time of feeding. Results are discussed in relation to anti-sporozoite vaccine development.     

Selenium in Aquatic Birds From Central California

ABSTRACT After mostly freshwater replaced agricultural drainage water used for wetland management in 1985, Selenium (Se) concentrations in 3 wintering waterfowl species and black-necked stilts (Himantopus mexicanus) from the North and South Grasslands of central California, USA, declined in the years from 1986 to 1988 and 1989 to 1994. However, Se concentrations were still above the threshold for potential reproductive impairment and exceeded background levels for some species. Consequently, we measured Se concentrations in aquatic birds in 2005 after long-term use (20 yr) of predominately freshwater for wetland management in the Grasslands. As in 1986–1994, Se concentrations in 2005 were higher for birds from the South Grasslands, which historically received more undiluted drainage water compared with the North Grasslands. Liver Se concentrations for stilts from the South Grasslands were at levels associated with potential reproductive impairment. All species from the South Grasslands, as well as mallards (Anas platyrhynchos), northern pintails (A. acuta), and American coots (Fulica americana) from the North Grasslands, were above species-specific background levels. From 1994 to 2005, Se levels in some aquatic birds stabilized above background levels likely indicating long-term cycling within the Grasslands. We recommend Se-contaminated drainage water (≥ 2 ppb Se) not be used for management or allowed as an input into arid wetlands throughout the western United States.     

Monthly survival of African Sylvia warblers in a seasonally arid tropical environment

Latitudinal gradients of life-history traits in animals are thought to be shaped by environmental variables. For example, it has been suggested that the increase in avian clutch size from the tropics towards the northern temperate regions is caused by a reduced survival of adult birds in the north due to increasing environmental seasonality. However, the tropical savannahs of East Africa show pronounced seasonality in resources caused by distinct rainy and dry seasons. This raises the question of whether survival and other life-history traits of birds living in these tropical savannahs are influenced by this seasonality, making them more similar to northern temperate species. We used 2-year monthly resighting data, a multistate modelling approach and the program MARK to test whether survival, transition probabilities between breeding states and other life-history traits of two resident Kenyan Sylvia species (Aves: Passeriformes: Sylviidae) are shaped by seasonality of rainfall in their environment. Contradicting our hypotheses, the two species showed only very slight influence of seasonality of rainfall on their survival. Survival in the dry months was hardly lower than in the rainy months. The species in the more seasonal environment ( S. boehmi , annual survival 71%) survived as well as the one in the more constant environment ( S. lugens , 56%). The observed survival rates correspond well to other life-history traits of the two species and are of similar magnitude to survival rates of other tropical passerines. This implies that either seasonality is not the driving force behind the life-history traits of the two species or the birds do not experience their environment as seasonal, as might be suggested by fluctuations in rainfall.     

Phylogenetic relationships of colubroid snakes based on mitochondrial DNA sequences

We used a 694 bp length of the mitochondrial ND4 gene from 40 genera to infer phylogenetic relationships among colubroid snakes. The goals of this study were to identify conserved subsets of ND4 sequence data that could be used to address (1) which nominal higher-level colubroid taxa are monophyletic, and (2) the relationships among the monophyletic lineages identified. Use of transversions only proved the most reliable and efficient means of retrieving colubroid relationships. Transversion parsimony and neighbour-joining analyses identify similar monophyletic higher-level taxa, but relationships among these lineages differ considerably between the two analyses. These differences were affected by the inclusion/exclusion of (1) transitions, (2) autapomorphies, and (3) the boid outgroups. Saturation effects among the transitions, uninformativeness of autapomorphies for clustering taxa, and long-branch and base-compositional problems among the boids lead us to regard the tree resulting from transversion parsimony analysis rooted with Acrochordus as the best current estimate of colubroid phylogenetic relationships. However, several aspects of this proposed phylogeny need further testing (e.g. the apparent diphyly of Natricinae is especially controversial). Relationships retrieved using all colubroid taxa are not obtained when sparsely or unevenly sampled experimental subsets of taxa are used instead, suggesting that long-branch problems can severely compromise elucidation of colubroid relationships if limited taxonomic sampling strategies are followed. We discuss the importance of this finding for previous molecular attempts to assess colubroid relationships. Our analyses confirm the historical validity of several nominal colubroid families and subfamilies, establish polyphyly of a few, but generally fail to resolve relationships among the monophyletic taxa we identify. More conservative character information will be required to confidently resolve the last issue.     

MICROCLIMATE VERSUS PREDATION RISK IN ROOST AND COVERT SELECTION BY BOBWHITES

Abstract: Knowledge of factors that influence habitat selection by wildlife leads to better understanding of habitat ecology and management. Therefore, we compared microclimate and predation risk as factors influencing the selection of stopping points (mid-day coverts, nocturnal roosts) by northern bobwhites (Colinus virginianus). Stopping points were located using radiomarked bobwhites in the Texas Panhandle, USA, during 2002–2003. We obtained blackbody temperatures of microclimates and assessed predation risk (angles of obstruction for aerial predators, vegetation profiles for terrestrial predators) at stopping points and paired random points. Summer coverts showed fewer degree-minutes of hyperthermic exposure (blackbody temperatures >39°C; x̄ = 655.0, SE = 4.1 for coverts, x̄ = 2,255.5, SE = 4.9 for random; 1200–1600 hr) and a lower risk to predators (e.g., 95% confidence intervals [CIs] of angles of obstruction = 87.8–90.8° for coverts, 55.9–70.6° for random). Summer roost temperatures were similar to paired random sites (x̄ = −13.9°C, SE = 0.6 for roost, x̄ = 13.9°C, SE = 0.7 for random) as were winter roost temperatures (x̄ = −1.3°C, SE = 0.7 for roosts, x̄ = −1.4°C, SE = 0.8 for random). There were minor issues of habitat selection of winter or summer roosts based on predation risk (e.g., 95% CIs of vegetation profiles of summer roosts and random sites did not overlap at lower strata). We concluded other selection factors likely exist for winter roosts because microclimate and predation risk assessments between winter roosts and random sites showed no difference. Similarly, other selection factors may exist for summer roosts, as they showed only a weak difference in terrestrial predation risk and no difference in microclimate in comparison to random sites. We concluded microclimate was the primary selection factor for coverts because prevention of hyperthermia necessitated that bobwhites select cooler microclimates within the study area.     

Genetic diversity and differentiation at MHC genes in island populations of tuatara (Sphenodon spp.)

Neutral genetic markers are commonly used to understand the effects of fragmentation and population bottlenecks on genetic variation in threatened species. Although neutral markers are useful for inferring population history, the analysis of functional genes is required to determine the significance of any observed geographical differences in variation. The genes of the major histocompatibility complex (MHC) are well‐known examples of genes of adaptive significance and are particularly relevant to conservation because of their role in pathogen resistance. In this study, we survey diversity at MHC class I loci across a range of tuatara populations. We compare the levels of MHC variation with that observed at neutral microsatellite markers to determine the relative roles of balancing selection, diversifying selection and genetic drift in shaping patterns of MHC variation in isolated populations. In general, levels of MHC variation within tuatara populations are concordant with microsatellite variation. Tuatara populations are highly differentiated at MHC genes, particularly between the northern and Cook Strait regions, and a trend towards diversifying selection across populations was observed. However, overall our results indicate that population bottlenecks and isolation have a larger influence on patterns of MHC variation in tuatara populations than selection.     

Robust dynamics of Amazon dieback to climate change with perturbed ecosystem model parameters

Climate change science is increasingly concerned with methods for managing and integrating sources of uncertainty from emission storylines, climate model projections, and ecosystem model parameterizations. In tropical ecosystems, regional climate projections and modeled ecosystem responses vary greatly, leading to a significant source of uncertainty in global biogeochemical accounting and possible future climate feedbacks. Here, we combine an ensemble of IPCC‐AR4 climate change projections for the Amazon Basin (eight general circulation models) with alternative ecosystem parameter sets for the dynamic global vegetation model, LPJmL. We evaluate LPJmL simulations of carbon stocks and fluxes against flux tower and aboveground biomass datasets for individual sites and the entire basin. Variability in LPJmL model sensitivity to future climate change is primarily related to light and water limitations through biochemical and water‐balance‐related parameters. Temperature‐dependent parameters related to plant respiration and photosynthesis appear to be less important than vegetation dynamics (and their parameters) for determining the magnitude of ecosystem response to climate change. Variance partitioning approaches reveal that relationships between uncertainty from ecosystem dynamics and climate projections are dependent on geographic location and the targeted ecosystem process. Parameter uncertainty from the LPJmL model does not affect the trajectory of ecosystem response for a given climate change scenario and the primary source of uncertainty for Amazon ‘dieback’ results from the uncertainty among climate projections. Our approach for describing uncertainty is applicable for informing and prioritizing policy options related to mitigation and adaptation where long‐term investments are required.     

Effect of Heliothis subflexa herbivory on fruit abscission by Physalis species: the roles of mechanical damage and chemical factors

1. Insect oral secretions are important for the induction of a number of plant responses, but the relative role of mechanical damage in the induction of these responses is often not well understood. Damage from the frugivore Heliothis subflexa, a specialist on Physalis species, causes herbivore‐induced fruit abscission. In this field study, we examined the separate and combined effects of mechanical damage and H. subflexa oral secretions on Physalis fruit abscission. 2. To determine the relative role of mechanical and chemical factors, the following treatments were administered to fruit: (1) three levels of mechanical damage, (2) natural herbivore damage by control larvae and by larvae surgically treated to inhibit saliva secretion, and (3) injection of H.subflexa oral secretions and a water control. Abscission of mechanically damaged fruit with and without the addition of oral secretions was also compared. 3. Mechanical damage was sufficient to cause fruit abscission, and the addition of oral secretions to mechanically damaged fruit did not cause an increase in fruit abscission. Normal caterpillars and those treated to inhibit saliva secretion caused similar abscission rates. 4. Though most studies examining the effects of insect oral secretions on induced plant responses find these chemical stimuli to be important or essential, the results of the present study showed that oral secretions are not necessary for fruit abscission. Future work is needed to determine the relative importance of mechanical damage in herbivore‐induced plant responses in other systems.