Aphids are a serious threat to agriculture, despite being a rather small group of insects. The about 4,000 species worldwide engage in highly interesting and complex relationships with their microbial fauna. One of the key symbionts in arthropods is Wolbachia, an α-Proteobacterium implicated in many important biological processes and believed to be a potential tool for biological control. Aphids were thought not to harbour Wolbachia; however, current data suggest that its presence in aphids has been missed, probably due to the low titre of the infection and/or to the high divergence of the Wolbachia strains of aphids. The goal of the present study is to map the Wolbachia infection status of natural aphids populations, along with the characterization of the detected Wolbachia strains. Out of 425 samples from Spain, Portugal, Greece, Israel and Iran, 37 were found to be infected. Our results, based mainly on 16S rRNA gene sequencing, indicate the presence of two new Wolbachia supergroups prevailing in aphids, along with some strains belonging either to supergroup B or to supergroup A.
Life history traits of Patella candei were studied for the first time, including weight versus length relationship, growth, age structure, sexual maturity, recruitment pattern, mortality rates and yield and biomass-per-recruit of an exploited population in Madeira Island, north-eastern Atlantic using monthly length -frequency data from January to December 1999. The growth pattern of P. candei showed positive allometric nature of growth (b . 3, P , 0.05). The estimated growth parameters showed an asymptotic length (L 1 ) and growth coefficient (K) estimated at 80.81 mm and K at 0.32 year 21 with a growth performance index (w ′ ) calculated as 3.32 based on the collected data. This species is moderately long-lived reaching up to 9.36 years and achieving sexual maturity at 36.7 mm of shell length. The recruitment pattern was continuous, displaying a major peak event per year, occurring in January (25.12%). The estimated total mortality rate (Z) was 1.79 year 21 while natural mortality rate (M) was 0.55 year 21 and fishing mortality rate (F) was 1.24 year 21 . The probability of capture shows that the length at first capture (Lc) was 42.7 mm, the exploitation rate (E) 0.693 and the maximum allowable limit of exploitation (E max ) was 0.779 for the highest yield. The exploitation rate was less than the predicted E max values, showing that the stock of P. candei was found to be under-exploited in the investigated area; however, its slow growth and long life make it extremely vulnerable to over-exploitation.
Chromosomal inversion polymorphism affects nucleotide variation at loci associated with inversions. In Drosophila subobscura, a species with a rich chromosomal inversion polymorphism and the largest recombinational map so far reported in the Drosophila genus, extensive genetic structure of nucleotide variation was detected in the segment affected by the O(3) inversion, a moderately sized inversion at Muller's element E. Indeed, a strong genetic differentiation all over O(3) and no evidence of a higher genetic exchange in the center of the inversion than at breakpoints were detected. In order to ascertain, whether other polymorphic and differently sized inversions of D. subobscura also exhibited a strong genetic structure, nucleotide variation in 5 gene regions (P236, P275, P150, Sxl, and P125) located along the A(2) inversion was analyzed in A(st) and A(2) chromosomes of D. subobscura. A(2) is a medium-sized inversion at Muller's element A and forms a single inversion loop in heterokaryotypes. The lower level of variation in A(2) relative to A(st) and the significant excess of low-frequency variants at polymorphic sites indicate that nucleotide variation at A(2) is not at mutation-drift equilibrium. The closest region to an inversion breakpoint, P236, exhibits the highest level of genetic differentiation (F(ST)) and of linkage disequilibrium (LD) between arrangements and variants at nucleotide polymorphic sites. The remaining 4 regions show a higher level of genetic exchange between A(2) and A(st) chromosomes than P236, as revealed by F(ST) and LD estimates. However, significant genetic differentiation between the A(st) and A(2) arrangements was detected not only at P236 but also in the other 4 regions separated from the nearest breakpoint by 1.2-2.9 Mb. Therefore, the extent of genetic exchange between arrangements has not been high enough to homogenize nucleotide variation in the center of the A(2) inversion. A(2) can be considered a typical successful inversion of D. subobscura according to its relative length. Chromosomal inversion polymorphism of D. subobscura might thus cause the genome of this species to be highly structured and to harbor different gene pools that might contribute to maintain adaptations to particular environments.
Nucleotide variation at the nuclear ribosomal protein 49 (rp49) gene region has been analysed by fine restriction mapping in a sample of 47 lines from a population from Madeira. Five restriction-site (out of 37 sites scored) and 3 length polymorphisms have been detected, resulting in 14 different haplotypes. This population shows less variation than both continental and Canary Island populations. The population from Madeira shows some differentiation from mainland populations, which does not favor the idea of extensive migration between the continent and Madeira. Chromosomal and restriction-map variation of the rp49 region in D. subobscuru populations, together with data on sequence comparison of this nuclear region in D. guanche and D. mudeirensis clearly indicate that the Canary Islands underwent at least two colonization events from the nearby continent. Although the data for Madeira are compatible with a single colonization event by a continental sample polymorphic for gene arrangements 0, and 0, + 4, an alternative scenario with at least two colonization events seems more likely.
Field and laboratory studies were performed to verify whether Culex theileri Theobald functions as a natural vector of Dirofilaria immitis (Leidy) on Madeira Island, Portugal. CO2-baited light traps (EVS traps) were use to sample mosquitoes monthly basis between February 2002 and February 2003 in the area of Quebradas (Funchal). Three mosquito species were captured, including 58 Culex pipiens L., 790 Cx. theileri, and three Culiseta longiareolata (Macquart). Only C. theileri tested positive for D. immitis. The presence of this filarial worm was detected by direct observation, infectivity assay dissection technique, and polymerase chain reaction methods. Infected mosquitoes were recovered in October and December 2002 and January 2003. These data provide evidence that Cx. theileri could be the main vector of D. immitis in Funchal, Madeira.
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