The Human-Baited Double Net Trap: An Alternative to Human Landing Catches for Collecting Outdoor Biting Mosquitoes in Lao PDR

Estimating the exposure of individuals to mosquito-borne diseases is a key measure used to evaluate the success of vector control operations. The gold standard is to use human landing catches where mosquitoes are collected off the exposed limbs of human collectors. This is however an unsatisfactory method since it potentially exposes individuals to a range of mosquito-borne diseases. In this study several sampling methods were compared to find a method that is representative of the human-biting rate outdoors, but which does not expose collectors to mosquito-borne infections. The sampling efficiency of four odour-baited traps were compared outdoors in rural Lao PDR; the human-baited double net (HDN) trap, CDC light trap, BG sentinel trap and Suna trap. Subsequently the HDN, the best performing trap, was compared directly with human landing catches (HLC), the ‘gold standard’, for estimating human-biting rates. HDNs collected 11–44 times more mosquitoes than the other traps, with the exception of the HLC. The HDN collected similar numbers of Anopheles (Rate Ratio, RR = 1.16, 95% Confidence Intervals, 95% CI = 0.61–2.20) and Culex mosquitoes (RR = 1.26, 95% CI = 0.74–2.17) as HLC, but under-estimated the numbers of Aedes albopictus (RR = 0.45, 95% CI = 0.27–0.77). Simpson’s index of diversity was 0.845 (95% CI 0.836–0.854) for the HDN trap and 0.778 (95% CI 0.769–0.787) for HLC, indicating that the HDN collected a greater diversity of mosquito species than HLC. Both HLC and HDN can distinguish between low and high biting rates and are crude ways to measure human-biting rate. The HDN is a simple and cheap method to estimate the human-biting rate outdoors without exposing collectors to mosquito bites.     

Genetic differentiation of Macrognathus siamensis within the Mekong River between Laos and Cambodia

This study investigated the genetic variation in the Mastacembelidae fish Macrognathus siamensis by sequencing 1047 bp of the mitochondrial ND2 gene of 48 fish samples collected in the Mekong River. They were divided into 15 mtDNA haplotypes. The haplotype network and Bayesian tree revealed striking genetic differences between the Laos and Cambodian samples. The high pairwise F statistic values (0.93–0.97) also confirmed this genetic differentiation. Analysis of the molecular clock indicated that these two samples separated approximately four million years ago. Most likely, the genetic variation was influenced by immigration and isolation events as well as by environmental factors. Furthermore, the present study results could be valuable in identifying the conservation and resource management units of this species.     

A high variability of mixed infections and recent recombinations of hepatitis B virus in Laos

In Lao PDR, where more than 8% of the population are chronic carriers of HBsAg, multiple genotypes and subgenotypes co-circulate and are prone to generate recombinant viruses. Phylogenetic analyses of multiple clones per donor revealed mixed infections of subgenotypes B1, B2, B4, C1, C5, I1 and I2 in almost 6% of HBsAg positive rejected blood donors. Recombination analyses and distance calculations furthermore showed that about 65% (17/26) of the mixed infected donors showed recombinations in the S-gene alone, involving the predominant genotypes B and C. These results suggest that, at least in Laos, hepatitis B virus (HBV) mixed infections lead to frequent recombinations. In many donors with recombinant strains, the recombinant fragment and a non-recombinant strain of the same genotype co-existed (127/185 analysed recombinant fragments). For a large proportion of these (60/127), the most closely related known virus was found, although not always exclusively, in the same donor. Recombinant virus strains are largely distinct. This is reflected in an unexpected diversity in recombination breakpoints and the relatively rare recombinations with identical recombination patterns of the same genotypes in different donors. Recent recombination events would explain the limited spread of each of the recombinants. Using a published mutation rate of 4.2 × 10(-5) mutations per site and year, the observed minimum genetic distances of 0-0.60% between parent strain and recombinant fragment would correspond to 0-71 years of evolution from a most recent common ancestor (MRCA). Thus several lines of evidence are suggestive of recent independent recombination events, a proportion of these even occurring within the same donors. In conclusion, our analyses revealed a high variability of mixed infections as a very probable breeding ground of multiple variable recombination events in Laos that so far have not led to new dominant strains.