An impact assessment of the alien lizard Plestiodon japonicus (Scincidae, Reptilia) on a threatened island population of the native lizard P. latiscutatus at an early phase of the biological invasion

On the island of Hachijojima, central Japan, a formerly abundant population of native lizard, Plestiodon latiscutatus has become endangered by predation from an alien weasel (Mustela itatsi). Recently, an alien lizard, P. japonicus, became established on the island. The impact of the alien lizard and current distribution of the native lizard were assessed by field surveys and DNA analyses. The native lizard was sparsely distributed in the western part of the island. The alien lizard occurred in the northeastern region of the island. Between these regions, several hybrid populations were identified, including some with non-F1 genotypes, suggesting that introgression is occurring. The distribution of the weasel was examined based on field observations, allowing us to estimate predator impacts on both lizard species. We found evidence that the weasel was present throughout the entire island, suggesting that the alien lizard was able to become established despite the presence of the weasel. This indicates that the alien lizard is likely to expand its habitat range and that introgression may spread. We conclude that P. latiscutatus populations will become increasingly endangered by the synergism of genetic introgression from P. japonicus and predation pressure from the weasel.     

Pesticide Residues and Bees – A Risk Assessment

Bees are essential pollinators of many plants in natural ecosystems and agricultural crops alike. In recent years the decline and disappearance of bee species in the wild and the collapse of honey bee colonies have concerned ecologists and apiculturalists, who search for causes and solutions to this problem. Whilst biological factors such as viral diseases, mite and parasite infections are undoubtedly involved, it is also evident that pesticides applied to agricultural crops have a negative impact on bees. Most risk assessments have focused on direct acute exposure of bees to agrochemicals from spray drift. However, the large number of pesticide residues found in pollen and honey demand a thorough evaluation of all residual compounds so as to identify those of highest risk to bees. Using data from recent residue surveys and toxicity of pesticides to honey and bumble bees, a comprehensive evaluation of risks under current exposure conditions is presented here. Standard risk assessments are complemented with new approaches that take into account time-cumulative effects over time, especially with dietary exposures. Whilst overall risks appear to be low, our analysis indicates that residues of pyrethroid and neonicotinoid insecticides pose the highest risk by contact exposure of bees with contaminated pollen. However, the synergism of ergosterol inhibiting fungicides with those two classes of insecticides results in much higher risks in spite of the low prevalence of their combined residues. Risks by ingestion of contaminated pollen and honey are of some concern for systemic insecticides, particularly imidacloprid and thiamethoxam, chlorpyrifos and the mixtures of cyhalothrin and ergosterol inhibiting fungicides. More attention should be paid to specific residue mixtures that may result in synergistic toxicity to bees.     

Electrocardiographic study in Ghanaian children with uncomplicated malaria,treated with artesunate-amodiaquine or artemether-lumefantrine

Background

In order to prepare the field site for future interventions, the prevalence of asymptomatic Plasmodium falciparum infection was evaluated in a cohort of children living in Brazzaville. Plasmodium falciparum merozoite surface protein 2 gene (msp2) was used to characterize the genetic diversity and the multiplicity of infection. The prevalence of mutant P. falciparum chloroquine resistance transporter (pfcrt) allele in isolates was also determined.

Methods

Between April and June 2010, 313 children below 10 years of age enrolled in the cohort for malaria surveillance were screened for P. falciparum infection using microscopy and polymerase chain reaction (PCR). The children were selected on the basis of being asymptomatic. Plasmodium falciparum msp2 gene was genotyped by allele-specific nested PCR and the pfcrt K76T mutation was detected using nested PCR followed by restriction endonuclease digestion.

Results

The prevalence of asymptomatic P. falciparum infections was 8.6% and 16% by microscopy and by PCR respectively. Allele typing of the msp2 gene detected 55% and 45% of 3D7 and FC27 allelic families respectively. The overall multiplicity of infections (MOI) was 1.3. A positive correlation between parasite density and multiplicity of infection was found. The prevalence of the mutant pfcrt allele (T76) in the isolates was 92%.

Conclusion

This is the first molecular characterization of P. falciparum field isolates in Congolese children, four years after changing the malaria treatment policy from chloroquine (CQ) to artemisinin-based combination therapy (ACT). The low prevalence of asymptomatic infections and MOI is discussed in the light of similar studies conducted in Central Africa.     

Estrogenic modulation of CYP3A38, CYP3A40, and CYP19 in mature male medaka (Oryzias latipes)

We examined cytochrome P450 production and activity and circulating hormone concentrations in male medaka exposed to 17beta-estradiol (E2) or 17alpha-ethinylestradiol (EE2). Intraperitoneal injection of E2 at 1, 10, or 100 microg/g-fish completely suppressed CYP3A38 protein production and suppressed CYP3A40 protein levels by 89%, 52%, or 47%, respectively. CYP3A38 and CYP3A40 mRNA expression was unaltered, and CYP3A enzymatic activity initially increased and then decreased with increasing E2 dose. Males co-cultured with females were exposed to a markedly high concentration (43 ng/L) of E2 secreted by females. CYP3A protein levels in co-cultured males were suppressed. Serum testosterone (TE) and 11keto-testosterone levels in co-cultured males were downregulated to 40% of pre-exposure levels. Serum E2 levels increased in co-cultured males or males exposed to EE2. Testicular CYP19, which converts TE to E2, increased by 9.5 times in males exposed to 50 ng/L EE2 and by 21.5 times in those exposed to 100 ng/L EE2. Male medaka exposed to EE2 showed increased serum Vtg levels. Estrogenic exposure induced Vtg production, suppressed CYP3A protein production, downregulated TE metabolism, and enhanced CYP19 activity. Serum E2 endogenously induced by CYP19 could contribute to Vtg induction in male medaka.     

Electrophoretic detection of enzyme variation among Japanese red-coloured spider mites of the genus Tetranychus (Acari: Tetranychidae)

The genetic variation in phosphoglucoisomerase (PGI) and malate dehydrogenase (MDH) was studied in red-pigmented Japanese spider mites of the genus Tetranychus by means of poly-acrylamide gel electrophoresis. The analysis revealed (1) that Tetranychus kanzawai possesses five and three alleles for PGI and MDH, respectively and that PGI allele frequencies clearly differ between the Hokkaido and the Honshu populations (2) that Tetranychus urticae and Tetranychus pueraricola, two closely related species, have different alleles for PGI and (3) that two populations of Tetranychus piercei, a species which had only been found on the islands of Okinawa, were obtained from Honshu and that the PGI locus is fixed for different alleles in the two populations. The two enzyme systems are apparently useful not only for discrimination of spider mite species but also for the detection of intraspecific variation. © Rapid Science Ltd. 1998     

Significance of habitat type for the genetic population structure of Panonychus citri (Acari: Tetranychidae)

Restricted migration and habitat fragmentation promote genetic differentiation between populations: Because most of the hosts of Panonychus citri are woody plants, mainly citrus trees that are usually planted at intervals of several metres, this mite likely faces more risks (e.g., starvation) by dispersing between host plants, compared to other spider mite species that infest both herbaceous and woody plants, such as Tetranychus urticae. Such a limited gene flow between patches (host plants) can lead to differentiation of populations even within a small area. Therefore, we hypothesize that P. citri populations are genetically differentiated not only between distant populations but also within small areas, such as within a grove. To test this hypothesis, we investigated the divergence of P. citri populations in Japanese citrus groves according to a hierarchical arrangement of geographical distance, ranging from distant populations (10 groves distributed throughout different areas in two major Japanese islands; this level of analysis is referred to as ‘geographic’) to local populations (different trees in a specific grove; ‘local’). Three molecular markers were used an esterase locus, one microsatellite and a point mutation in the mitochondrial cytochrome oxidase subunit I. At a local level acaricide susceptibility tests were also performed using two acaricides: fenpyroximate (25 ppm) and etoxazole (3.33 ppm). At a broad geographic level the gene diversity decreased with decreasing area size and distance between populations. By contrast, at the local level, populations maintained a significant level of variation between trees within groves, and the divergence within groves was higher than between groves. Whereas no statistical difference of the mortalities was detected among groves for the two acaricides tested, the difference was statistically significant among trees within groves in fenpyroximate (ANOVA, p < 0.025) and=" marginal=" in=" etoxazole="><>p < 0.05). we=" concluded=" that=">P. citri populations maintain a higher level of variation between trees (or patches of trees) within groves than between groves at the local level, though the gene diversity tended to be smaller with decreasing distance between populations at the geographical level. Results are discussed in relation to the dispersal behaviour of spider mites.     

Molecular Cloning and Characterization of a Microsatellite Locus Found in an RAPD Marker of a Spider Mite, Panonychus Citri (Acari: Tetranychidae)

Genetic markers were searched using PCR with 40 kinds of decanucleotide primers to investigate DNA polymorphism in Panonychuscitri. A region consisting of a variable number of CT tandem repeats (microsatellite) was found in a fragment amplified with the OPB10 primer. The microsatellite differed in size by ca. 100bp among several P. citri populations screened and was derived from at least seven alleles. This region was characteristic of P. mori and P. osmanthi, but was lacking in P. ulmi. The flanking regions were highly conserved among these species.     

Distinct patterns of cytokine regulation in discrete clinical forms of Plasmodium falciparum malaria

The pathogenesis of two of the most severe complications of Plasmodium falciparum malaria, cerebral malaria (CM) and severe malarial anaemia (SA) both appear to involve dysregulation of the immune system. We have measured plasma levels of TNF and its two receptors in Ghanaian children with strictly defined cerebral malaria (CM), severe malarial anaemia (SA), or uncomplicated malaria (UM) in two independent studies in an area of seasonal, hyperendemic transmission of P. falciparum. Levels of TNF, soluble TNF receptor 1 (sTNF-R1) and 2 (sTNF-R2) were found to be significantly higher in CM than in the other clinical categories of P. falciparum malaria patients. Levels of both receptors depended on clinical category, whereas only sTNF-R1 levels were significantly dependent on parasitemia. Detailed analysis of the interrelationship between these variables resolved this pattern further, and identified marked differences between the patient categories. While levels of TNF, sTNF-R1 and sTNF-R2 correlated with parasitemia in UM, this was not the case in CM and SA. Rather, there was a tendency towards high levels of TNF and its receptors in CM and low levels in SA without significant correlation to parasitemia in either category. This, and the fact that malaria-induced increases in plasma levels of IL-10 are much lower in SA compared to CM, suggest that distinct forms of dysregulation of the immune response to infection contribute to the pathogenesis of CM and SA.     

Acute toxicity of neonicotinoids and some insecticides to first instar nymphs of a non-target damselfly, <Emphasis Type="Italic">Ischnura senegalensis</Emphasis> (Odonata: Coenagrionidae), in Japanese paddy fields

Systemic insecticides such as neonicotinoids and fipronil are widely applied in rice production. These insecticides have been suspected of reducing biodiversity in paddy ecosystems and reducing wild dragonfly populations in Japan. Conventional ecotoxicological risk assessment could not confirm this, as it has not considered interspecific variation in sensitivity to insecticides. We estimated the median effect concentration (EC50) of 15 systemic insecticides to first instar nymphs of a Japanese damselfly, Ischnura senegalensis (Rambur) (Odonata: Coenagrionidae), commonly found in rice paddy fields. Damselflies were found to be highly sensitive to pyrethroid pesticides, less so to phenylpyrazole, organophosphates, and carbamates, and least sensitive to neonicotinoids, nereistoxin, and diamide. Given the acute toxicity data, the sensitivity of the damselfly to neonicotinoids was considered to be lower than that of other aquatic insects, whereas the EC50 values of the damselfly were 2–3 orders lower than that of Daphnia magna Straus (Diplostraca: Daphniidae), which is a standard test species. These results indicate that the conventional ecological risk assessment based on acute toxicity data of D. magna would underestimate the impact of neonicotinoids on Odonata diversity in paddy ecosystems. We therefore recommend using the paddy-dwelling damselfly as a new test species for insecticide bioassay.     

Worldwide migration of parasitic mites as a result of bumblebee commercialization

We investigated the status of infestation by a tracheal mite, Locustacarus buchneri, in natural populations of a Japanese native bumblebee species, Bombus hypocrita, collected on Hokkaido Island and in the Aomori prefecture between 1997 and 2001. We also investigated mite infestation in commercial colonies of the European bumblebee, Bombus terrestris, imported from the Netherlands and Belgium, and the Japanese native species, B. ignitus, imported from the Netherlands, between 1997 and 2001. We detected mites in natural populations of the two B. hypocrita subspecies and in the commercial colonies. Analysis of variations in 535 bp sequences of the mitochondrial cytochrome oxidase subunit 1 (CO1) gene showed that the mite haplotypes in the native populations and in the imported colonies did not overlap in 1997–1999, but in 2000–2001 some mites possessing European CO1 haplotypes were detected in the natural populations of Japanese native bumblebees. In addition, many mites possessing Japanese haplotypes were detected in the imported commercial colonies from Europe. Considering the fact that the Japanese native bumblebees, B. hypocrita, were once exported to Europe for commercialization, these results suggest that bumblebee commercialization has caused overseas migration and cross-infestation of parasitic mites among natural and commercial colonies. However, because the Japanese and European CO1 haplotypes were closely related, there was a possibility that the European haplotypes found in the mites in the Hokkaido Island revealed native variation. To clarify the status of mite invasion, further detailed analysis of genetic variation of the mite, using other genetic markers on additional samples, need to be performed.