First Colombian multicentric newborn screening for congenital toxoplasmosis

Aims

To determine the incidence of congenital toxoplasmosis in Colombian newborns from 19 hospital or maternal child health services from seven different cities of five natural geographic regions (Caribbean, Central, Andean, Amazonia and Eastern).

Materials and Methods

We collected 15,333 samples from umbilical cord blood between the period of March 2009 to May 2010 in 19 different hospitals and maternal-child health services from seven different cities. We applied an IgM ELISA assay (Vircell, Spain) to determine the frequency of IgM anti Toxoplasma. The results in blood cord samples were confirmed either by western blot and repeated ELISA IgM assay. In a sub-sample of 1,613 children that were negative by the anti-Toxoplasma IgM assay, the frequency of specific anti-Toxoplasma IgA by the ISAGA assay was determined. All children with positive samples by IgM, IgA, clinical diagnosis or treatment during pregnancy were recalled for confirmatory tests after day 10 of life.

Results

61 positive samples for specific IgM (0.39%) and 9 positives for IgA (0.5%) were found. 143 questionnaires were positive for a clinical diagnosis or treatment for toxoplasmosis during pregnancy. 109 out of the 218 children that had some of the criteria for postnatal confirmatory tests were followed. Congenital toxoplasmosis infection was confirmed in 15 children: 7 were symptomatic, and three of them died before the first month of life (20% of lethality). A significant correlation was found between a high incidence of markers for congenital toxoplasmosis and higher mean annual rainfall for the city.

Conclusions

Incidence for congenital toxoplasmosis is significantly different between hospitals or maternal child health services from different cities in Colombia. Mean annual rainfall was correlated with incidence of congenital toxoplasmosis.     

Contrasting Effect of Prepulse Signals on Performance of Toxoplasma-Infected and Toxoplasma-Free Subjects in an Acoustic Reaction Times Test

Background

About 30% of people on Earth have latent toxoplasmosis. Infected subjects do not express any clinical symptoms, however, they carry dormant stages of parasite Toxoplasma for the rest of their life. This form of toxoplasmosis is mostly considered harmless, however, recent studies showed its specific effects on physiology, behaviour and its associations with various diseases, including psychiatric disorders such as schizophrenia. Individuals who suffer from schizophrenia have about 2.7 times higher prevalence of Toxoplasma-seropositivity than controls, which suggests that some traits characteristic of schizophrenic patients, including the sex difference in schizophrenia onset, decrease of grey matter density in specific brain areas and modification of prepulse inhibition of startle reaction could in fact be caused by toxoplasmosis for those patients who are Toxoplasma-seropositive.

Methodology/Principal Findings

We measured the effect of prepulse inhibition/facilitation of the startle reaction on reaction times. The students, 170 women and 66 men, were asked to react as quickly as possible to a startling acoustic signal by pressing a computer mouse button. Some of the startling signals were without the prepulse, some were 20 msec. preceded by a short (20 msec.) prepulse signal of lower intensity. Toxoplasma-seropositive subjects had longer reaction times than the controls. Acoustic prepulse shorted the reaction times in all subjects. This effect of prepulse on reaction times was stronger in male subjects and increased with the duration of infection, suggesting that it represented a cumulative effect of latent toxoplasmosis, rather than a fading out after effect of past acute toxoplasmosis.

Conclusions

Different sensitivity of Toxoplasma-seropositive and Toxoplasma-seronegative subjects on effect of prepulses on reaction times (the toxoplasmosis-prepulse interaction) suggested, but of course did not prove, that the alternations of prepulse inhibition of startle reaction observed in schizophrenia patients probably joined the list of schizophrenia symptoms that are in fact caused by latent toxoplasmosis.     

Toxoplasmosis-Associated Difference in Intelligence and Personality in Men Depends on Their Rhesus Blood Group but Not ABO Blood Group

Background

The parasite Toxoplasma gondii influences the behaviour of infected animals and probably also personality of infected humans. Subjects with a Rhesus-positive blood group are protected against certain behavioural effects associated with Toxoplasma infection, including the deterioration of reaction times and personality factor shift.

Methodology/Principal Findings

Here, we searched for differences in the toxoplasmosis-associated effects between RhD-positive and RhD-negative subjects by testing 502 soldiers with two personality tests and two intelligence tests. The infected subjects expressed lower levels of all potentially pathognomic factors measured with the N-70 questionnaire and in neurasthenia measured with NEO-PI-R. The RhD-positive, Toxoplasma-infected subjects expressed lower while RhD-negative, Toxoplasma-infected subjects expressed higher intelligence than their Toxoplasma-free peers. The observed Toxoplasma-associated differences were always larger in RhD-negative than in RhD-positive subjects.

Conclusions

RhD phenotype plays an important role in the strength and direction of association between latent toxoplasmosis and not only psychomotor performance, but also personality and intelligence.     

Detection of Echinococcus multilocularis in carnivores in Razavi Khorasan province, Iran using mitochondrial DNA

Background

Echinococcus multilocularis is the source of alveolar echinococcosis, a potentially fatal zoonotic disease. This investigation assessed the presence of E. multilocularis infection in definitive hosts in the Chenaran region of Razavi Khorasan Province, northeastern Iran.

Methodology/Principal Findings

Fecal samples from 77 domestic and stray dogs and 14 wild carnivores were examined using the flotation/sieving method followed by multiplex PCR of mitochondrial genes. The intestinal scraping technique (IST) and the sedimentation and counting technique (SCT) revealed adult Echinococcus in the intestines of five of 10 jackals and of the single wolf examined. Three jackals were infected only with E. multilocularis but two, and the wolf, were infected with both E. multilocularis and E. granulosus. Multiplex PCR revealed E. multilocularis, E. granulosus, and Taenia spp. in 19, 24, and 28 fecal samples, respectively. Echinococcus multilocularis infection was detected in the feces of all wild carnivores sampled including nine jackals, three foxes, one wolf, one hyena, and five dogs (6.5%). Echinococcus granulosus was found in the fecal samples of 16.9% of dogs, 66.7% of jackals, and all of the foxes, the wolf, and the hyena. The feces of 16 (21.8%) dogs, 7 of 9 (77.8%) jackals, and all three foxes, one wolf and one hyena were infected with Taenia spp.

Conclusions/Significance

The prevalence of E. multilocularis in wild carnivores of rural areas of the Chenaran region is high, indicating that the life cycle is being maintained in northeastern Iran with the red fox, jackal, wolf, hyena, and dog as definitive hosts.     

Physical Factors Influencing Pleasant Touch during Passive Fingertip Stimulation

Objective

Tactile explorations with the fingertips provide information regarding the physical properties of surfaces and their relative pleasantness. Previously, we performed an investigation in the active touch domain and linked several surface properties (i.e. frictional force fluctuations and net friction) with their pleasantness levels. The aim of the present study was to investigate physical factors being important for pleasantness perception during passive fingertip stimulation. Specifically we were interested to see whether factors, such as surfaces'' topographies or their frictional characteristics could influence pleasantness. Furthermore, we ascertained how the stimulus pleasantness level was impacted by (i) the normal force of stimulus application (F_N) and (ii) the stimulus temperature (T_S).

Methods and Results

The right index fingertips of 22 blindfolded participants were stimulated using 27 different stimuli, which varied in average roughness (Ra) and T_S. A 4-axis robot moved the stimuli horizontally under participants'' fingertips with three levels of F_N. The robot was equipped with force sensors, which recorded the F_N and friction force (F_T) during stimulation. Participants rated each stimulus according to a three-level pleasantness scale, as very pleasant (scored 0), pleasant (scored 1), or unpleasant (scored 2). These ordinal pleasantness ratings were logarithmically transformed into linear and unidimensional pleasantness measures with the Rasch model. Statistical analyses were conducted to investigate a possible link between the stimulus properties (i.e. Ra, F_N, F_T, and T_S) and their respective pleasantness levels. Only the mean Ra and F_T values were negatively correlated with pleasantness. No significant correlation was detected between F_N or T_S and pleasantness.

Conclusion

Pleasantness perception, resulting from passive fingertip stimulation, seems to be influenced by the surfaces'' average roughness levels and average F_T occurring during fingertip stimulation.     

Effects of Latent Toxoplasmosis on Autoimmune Thyroid Diseases in Pregnancy

Background

Toxoplasmosis, one of the most common zoonotic diseases worldwide, can induce various hormonal and behavioural alterations in infected hosts, and its most common form, latent toxoplasmosis, influences the course of pregnancy. Autoimmune thyroid diseases (AITD) belong to the well-defined risk factors for adverse pregnancy outcomes. The aim of this study was to investigate whether there is a link between latent toxoplasmosis and maternal AITD in pregnancy.

Methods

Cross-sectional study in 1248 consecutive pregnant women in the 9–12^th gestational weeks. Serum thyroid-stimulating hormone (TSH), thyroperoxidase antibodies (TPOAb), and free thyroxine (FT4) were assessed by chemiluminescence; the Toxoplasma status was detected by the complement fixation test (CFT) and anti-Toxoplasma IgG enzyme-linked immunosorbent assay (ELISA).

Results

Overall, 22.5% of the women were positive for latent toxoplasmosis and 14.7% were screened positive for AITD. Women with latent toxoplasmosis had more often highly elevated TPOAb than the Toxoplasma-negative ones (p = 0.004), and latent toxoplasmosis was associated with decrease in serum TSH levels (p = 0.049). Moreover, we found a positive correlation between FT4 and the index of positivity for anti-Toxoplasma IgG antibodies (p = 0.033), which was even stronger in the TPOAb-positive Toxoplasma-positive women, (p = 0.014), as well as a positive correlation between FT4 and log₂ CFT (p = 0.009).

Conclusions

Latent toxoplasmosis was associated with a mild increase in thyroid hormone production in pregnancy. The observed Toxoplasma-associated changes in the parameters of AITD are mild and do not seem to be clinically relevant; however, they could provide new clues to the complex pathogenesis of autoimmune thyroid diseases.     

Physical Factors Influencing Pleasant Touch during Tactile Exploration

Background

When scanning surfaces, humans perceive some of their physical attributes. These percepts are frequently accompanied by a sensation of (un)pleasantness. We therefore hypothesized that aspects of the mechanical activity induced by scanning surfaces with fingertips could be objectively associated with a pleasantness sensation. Previously, we developed a unidimensional measure of pleasantness, the Pleasant Touch Scale, quantifying the pleasantness level of 37 different materials. Findings of this study suggested that the sensation of pleasantness was influenced by the average magnitude of the frictional forces brought about by sliding the finger on the surface, and by the surface topography. In the present study, we correlated (i) characteristics of the fluctuations of frictional forces resulting from the interaction between the finger and the surface asperities as well as (ii) the average friction with the sensation of pleasantness.

Results

Eight blindfolded participants tactually explored twelve materials of the Pleasant Touch Scale through lateral sliding movements of their index fingertip. During exploration, the normal and tangential interaction force components, f_N and f_T, as well as the fingertip trajectory were measured. The effect of the frictional force on pleasantness sensation was investigated through the analysis of the ratio f_T to f_N, i.e. the net coefficient of kinetic friction, μ. The influence of the surface topographies was investigated through analysis of rapid f_T fluctuations in the spatial frequency domain. Results showed that high values of μ were anticorrelated with pleasantness. Furthermore, surfaces associated with fluctuations of f_T having higher amplitudes in the low frequency range than in the high one were judged to be less pleasant than the surfaces yielding evenly distributed amplitudes throughout the whole spatial frequency domain.

Conclusion

Characteristics of the frictional force fluctuations and of the net friction taking place during scanning can reliably be correlated with the pleasantness sensation of surfaces.     

The pollination of a self-incompatible, food-mimic orchid, Coelogyne fimbriata (Orchidaceae), by female Vespula wasps

Background and Aims

The study of specialized interactions between species is crucial to our understanding of processes in evolutionary ecology due to their profound effect on life cycles and diversification. Obligate pollination by a single wasp species is rare in Orchidaceae except in species with sexually deceptive flowers that are pollinated exclusively by male insects. The object of this study was to document pollination of the food-deceptive flowers of Coelogyne fimbriata, a species pollinated exclusively by female wasps.

Methods

Field observations and experiments were conducted in two populations of C. fimbriata. Floral phenology was recorded, and functional floral architecture was measured. Insect visitors to flowers were observed from 2005 to 2007. Bioassay experiments were conducted to check whether the floral odour attracted pollinators. Natural (insect-mediated) rates of pollinarium removal, pollinium deposition on stigmas, and fruit set were recorded. To determine the importance of cross-pollination, the breeding system was assessed via controlled, hand-pollination experiments.

Key Results

Two populations of C. fimbriata with fragrant, nectarless flowers are pollinated by females of the same Vespula species (Vespidae, Hymenoptera). Experiments on wasps show that they crawl towards the source of the odour. The flowering period appears to coincide with an annual peak in Vespula colony expansion when additional workers forage for carbohydrates. Rates of pollinarium removal (0·069–0·918) and pollinium deposition on stigmas (0·025–0·695) are extremely variable. However, fruit set in C. fimbriata is always low (0·014–0·069) and appears to be based on self-incompatibility coupled with intraclonal (geitonogamous) deposition of pollinia.

Conclusions

Coelogyne fimbriata and Steveniella satyrioides are now the only orchid species known to have food-deceptive flowers that are pollinated exclusively by eusocial, worker wasps. In C. fimbriata, floral odour appears to be the major attractant. Sub-populations may go through flowering seasons when pollinators are abundant or infrequent, but fruit set always remains low because the obligate pollinator does not often appear to transfer pollinaria between intercompatible genets.Key words: Coelogyne fimbriata, Vespula wasps, food deception, floral odour, pollinarium removal, pollinium deposition, self-incompatibility     

Effect of pitcher age on trapping efficiency and natural prey capture in carnivorous Nepenthes rafflesiana plants

Background and Aims

Nepenthes pitchers are sophisticated traps that employ a variety of mechanisms to attract, capture and retain prey. The underlying morphological structures and physiological processes are subject to change over the lifetime of a pitcher. Here an investigation was carried out on how pitcher properties and capture efficiency change over the first 2 weeks after pitcher opening.

Methods

Prey capture, trapping efficiency, extrafloral nectar secretion, pitcher odour, as well as pH and viscoelasticity of the digestive fluid in N. rafflesiana pitchers were monitored in the natural habitat from pitcher opening up to an age of 2 weeks.

Key Results

Pitchers not only increased their attractiveness over this period by becoming more fragrant and secreting more nectar, but also gained mechanical trapping efficiency via an enhanced wettability of the upper pitcher rim (peristome). Consistently, natural prey capture was initially low and increased 3–6 d after opening. It was, however, highly variable within and among pitchers. At the same time, the pH and viscoelasticity of the digestive fluid decreased, suggesting that the latter is not essential for effective prey capture.

Conclusions

Prey capture and attraction by Nepenthes are dynamic processes strongly influenced by the changing properties of the pitcher. The results confirm insect aquaplaning on the peristome as the main capture mechanism in N. rafflesiana.Key words: Carnivorous plants, pitcher development, prey attraction, prey capture, insect aquaplaning, extrafloral nectar, Nepenthes rafflesiana     

A comparison of urinary mercury between children with autism spectrum disorders and control children

Background

Urinary mercury concentrations are used in research exploring mercury exposure. Some theorists have proposed that autism is caused by mercury toxicity. We set out to test whether mercury concentrations in the urine of children with autism were significantly increased or decreased compared to controls or siblings.

Methods

Blinded cohort analyses were carried out on the urine of 56 children with autism spectrum disorders (ASD) compared to their siblings (n = 42) and a control sample of children without ASD in mainstream (n = 121) and special schools (n = 34).

Results

There were no statistically significant differences in creatinine levels, in uncorrected urinary mercury levels or in levels of mercury corrected for creatinine, whether or not the analysis is controlled for age, gender and amalgam fillings.

Conclusions

This study lends no support for the hypothesis of differences in urinary mercury excretion in children with autism compared to other groups. Some of the results, however, do suggest further research in the area may be warranted to replicate this in a larger group and with clear measurement of potential confounding factors.     

Masterpiece of epigenetic engineering – how Toxoplasma gondii reprogrammes host brains to change fear to sexual attraction

The protozoan parasite Toxoplasma gondii is known to induce specific behavioural changes in its intermediate hosts, including humans, that are believed to increase the chance of its successful transmission to the definitive host, the cat. The most conspicuous change is the so‐called fatal attraction phenomenon, the switch from the mice's and rats’ natural fear of the smell of cats toward an attraction to this smell. The mechanism of this manipulation activity is unknown; however, many indices suggest that changes in the concentrations of dopamine and testosterone are involved. In this issue of Molecular Ecology, Hari Dass & Vyas ( 2014 ) present results of a study showing that, by hypomethylation of certain regulatory elements of key gene, Toxoplasma is able to reprogramme the brain's genetic machinery in such a way that cat odour activates and changes the wiring of the medial amygdala circuits responsible for sexual behaviour. This study delivers the first clear evidence of a parasite's ability to use sophisticated epigenetic engineering techniques for the manipulation of the phenotype of its infected host.     

Genetic and environmental influences on female sexual orientation, childhood gender typicality and adult gender identity

Background

Human sexual orientation is influenced by genetic and non-shared environmental factors as are two important psychological correlates – childhood gender typicality (CGT) and adult gender identity (AGI). However, researchers have been unable to resolve the genetic and non-genetic components that contribute to the covariation between these traits, particularly in women.

Methodology/Principal Findings

Here we performed a multivariate genetic analysis in a large sample of British female twins (N = 4,426) who completed a questionnaire assessing sexual attraction, CGT and AGI. Univariate genetic models indicated modest genetic influences on sexual attraction (25%), AGI (11%) and CGT (31%). For the multivariate analyses, a common pathway model best fitted the data.

Conclusions/Significance

This indicated that a single latent variable influenced by a genetic component and common non-shared environmental component explained the association between the three traits but there was substantial measurement error. These findings highlight common developmental factors affecting differences in sexual orientation.     

Ocular sequelae of congenital toxoplasmosis in Brazil compared with Europe

Background

Toxoplasmic retinochoroiditis appears to be more severe in Brazil, where it is a leading cause of blindness, than in Europe, but direct comparisons are lacking. Evidence is accumulating that more virulent genotypes of Toxoplasma gondii predominate in South America.

Methods

We compared prospective cohorts of children with congenital toxoplasmosis identified by universal neonatal screening in Brazil and neonatal or prenatal screening in Europe between 1992 and 2003, using the same protocol in both continents.

Results

Three hundred and eleven (311) children had congenital toxoplasmosis: 30 in Brazil and 281 in Europe, where 71 were identified by neonatal screening. Median follow up was 4.1 years in Europe and 3.7 years in Brazil. Relatively more children had retinochoroiditis during the first year in Brazil than in Europe (15/30; 50% versus 29/281; 10%) and the risk of lesions by 4 years of age was much higher: the hazard ratio for Brazil versus Europe was 5.36 (95%CI: 3.17, 9.08). Children in Brazil had larger lesions, which were more likely to be multiple and to affect the posterior pole (p<0.0001). In Brazil, visual impairment (<6/12 Snellen) was predicted for most affected eyes (87%, 27/31), but not in Europe (29%; 20/69, p<0.0001). The size of newly detected lesions decreased with age (p = 0.0007).

Conclusions

T. gondii causes more severe ocular disease in congenitally infected children in Brazil compared with Europe. The marked differences in the frequency, size and multiplicity of retinochoroidal lesions may be due to infection with more virulent genotypes of the parasite that predominate in Brazil but are rarely found in Europe.     

The Impact of Psychological Stress on Men's Judgements of Female Body Size

Background

Previous work has suggested that the experience of psychological stress may influence physical attractiveness ideals, but most evidence in favour of this hypothesis remains archival. The objective of this study was to experimentally investigate the impact of stress on men''s judgements of female body size.

Methods

Men were randomly assigned to either an experimental group, in which they took part in a task that heightened stress (experimental group, n = 41) or in which they did not take part in such a task (control group, n = 40). Both groups rated the attractiveness of female bodies varying in size from emaciated to obese, completed a measure of appetite sensation, and had their body mass indices (BMIs) measured.

Results

Between-groups analyses showed that the experimental group was matched with the control group in terms of mean age, BMI, and appetite sensation. Further analyses showed that men in the experimental group rated a significantly heavier female body size as maximally attractive than the control group. Men in the experimental group also rated heavier female bodies as more attractive and idealised a wider range of female figures than did the control group.

Conclusion

This study found that the experience of stress was associated with a preference among men for heavier female body sizes. These results indicate that human attractiveness judgements are sensitive to variations in local ecologies and reflect adaptive strategies for dealing with changing environmental conditions.     

Accuracy of Urine Circulating Cathodic Antigen (CCA) Test for Schistosoma mansoni Diagnosis in Different Settings of C?te d'Ivoire

Background

Promising results have been reported for a urine circulating cathodic antigen (CCA) test for the diagnosis of Schistosoma mansoni. We assessed the accuracy of a commercially available CCA cassette test (designated CCA-A) and an experimental formulation (CCA-B) for S. mansoni diagnosis.

Methodology

We conducted a cross-sectional survey in three settings of Côte d''Ivoire: settings A and B are endemic for S. mansoni, whereas S. haematobium co-exists in setting C. Overall, 446 children, aged 8–12 years, submitted multiple stool and urine samples. For S. mansoni diagnosis, stool samples were examined with triplicate Kato-Katz, whereas urine samples were tested with CCA-A. The first stool and urine samples were additionally subjected to an ether-concentration technique and CCA-B, respectively. Urine samples were examined for S. haematobium using a filtration method, and for microhematuria using Hemastix dipsticks.

Principal Findings

Considering nine Kato-Katz as diagnostic ‘gold’ standard, the prevalence of S. mansoni in setting A, B and C was 32.9%, 53.1% and 91.8%, respectively. The sensitivity of triplicate Kato-Katz from the first stool and a single CCA-A test was 47.9% and 56.3% (setting A), 73.9% and 69.6% (setting B), and 94.2% and 89.6% (setting C). The respective sensitivity of a single CCA-B was 10.4%, 29.9% and 75.0%. The ether-concentration technique showed a low sensitivity for S. mansoni diagnosis (8.3–41.0%). The specificity of CCA-A was moderate (76.9–84.2%); CCA-B was high (96.7–100%). The likelihood of a CCA-A color reaction increased with higher S. mansoni fecal egg counts (odds ratio: 1.07, p<0.001). A concurrent S. haematobium infection or the presence of microhematuria did not influence the CCA-A test results for S. mansoni diagnosis.

Conclusion/Significance

CCA-A showed similar sensitivity than triplicate Kato-Katz for S. mansoni diagnosis with no cross-reactivity to S. haematobium and microhematuria. The low sensitivity of CCA-B in our study area precludes its use for S. mansoni diagnosis.     

Flower morphology, pollination biology and mating system of the complex flower of Vigna caracalla (Fabaceae: Papilionoideae)

Background and Aims

Vigna caracalla has the most complex flower among asymmetrical Papilionoideae. The objective of this study was to understand the relationships among floral characteristics, specialization, mating system and the role of floral visitors under different ecological contexts.

Methods

Five populations were studied in north-western Argentina, from 700 to 1570 m a.s.l. Anthesis, colour and odour patterns, stigmatic receptivity, visitors and pollination mechanism were examined and mating-system experiments were performed.

Key Results

The petals are highly modified and the keel shows 3·75–5·25 revolutions. The sense of asymmetry was always left-handed. Hand-crosses showed that V. caracalla is self-compatible, but depends on pollinators to set seeds. Hand-crossed fruits were more successful than hand-selfed ones, with the exception of the site at the highest elevation. Bombus morio (queens and workers), Centris bicolor, Eufriesea mariana and Xylocopa eximia trigger the pollination mechanism (a ‘brush type’). The greatest level of self-compatibility and autonomous self-pollination were found at the highest elevation, together with the lowest reproductive success and number of pollinators (B. morio workers only).

Conclusions

Self-fertilization may have evolved in the peripheral population at the highest site of V. caracalla because of the benefits of reproductive assurance under reduced pollinator diversity.Key words: Fabaceae, Vigna caracalla, asymmetry, breeding system, complex flowers, elevational gradient, pollination     

Pollinator convergence and the nature of species' boundaries in sympatric Sardinian Ophrys (Orchidaceae)

Background and Aims

In the sexually deceptive Ophrys genus, species isolation is generally considered ethological and occurs via different, specific pollinators, but there are cases in which Ophrys species can share a common pollinator and differ in pollen placement on the body of the insect. In that condition, species are expected to be reproductively isolated through a pre-mating mechanical barrier. Here, the relative contribution of pre- vs. post-mating barriers to gene flow among two Ophrys species that share a common pollinator and can occur in sympatry is studied.

Methods

A natural hybrid zone on Sardinia between O. iricolor and O. incubacea, sharing Andrena morio as pollinator, was investigated by analysing floral traits involved in pollinator attraction as odour extracts both for non-active and active compounds and for labellum morphology. The genetic architecture of the hybrid zone was also estimated with amplified fragment length polymorphism (AFLP) markers, and pollination fitness and seed set of both parental species and their hybrids in the sympatric zone were estimated by controlled crosses.

Key Results

Although hybrids were intermediate between parental species in labellum morphology and non-active odour compounds, both parental species and hybrids produced a similar odour bouquet for active compounds. However, hybrids produced significantly lower fruit and seed set than parental species, and the genetic architecture of the hybrid zone suggests that they were mostly first-generation hybrids.

Conclusions

The two parental species hybridize in sympatry as a consequence of pollinator overlap and weak mechanical isolation, but post-zygotic barriers reduce hybrid frequency and fitness, and prevent extensive introgression. These results highlight a significant contribution of late post-mating barriers, such as chromosomal divergence, for maintaining reproductive isolation, in an orchid group for which pre-mating barriers are often considered predominant.Key words: AFLP markers, floral scent variation, hybrid zone, hybrid fitness, Ophrys iricolor, Ophrys incubacea, reproductive isolation, sexual deception     

Floral odour chemistry defines species boundaries and underpins strong reproductive isolation in sexually deceptive orchids

Background and Aims

The events leading to speciation are best investigated in systems where speciation is ongoing or incomplete, such as incipient species. By examining reproductive barriers among incipient sister taxa and their congeners we can gain valuable insights into the relative timing and importance of the various barriers involved in the speciation process. The aim of this study was to identify the reproductive barriers among sexually deceptive orchid taxa in the genus Chiloglottis.

Methods

The study targeted four closely related taxa with varying degrees of geographic overlap. Chemical, morphological and genetic evidence was combined to explore the basis of reproductive isolation. Of primary interest was the degree of genetic differentiation among taxa at both nuclear and chloroplast DNA markers. To objectively test whether or not species boundaries are defined by the chemistry that controls pollinator specificity, genetic analysis was restricted to samples of known odour chemistry.

Key Results

Floral odour chemical analysis was performed for 600+ flowers. The three sympatric taxa were defined by their specific chiloglottones, the semiochemicals responsible for pollinator attraction, and were found to be fully cross-compatible. Multivariate morphometric analysis could not reliably distinguish among the four taxa. Although varying from very low to moderate, significant levels of genetic differentiation were detected among all pairwise combinations of taxa at both nuclear and chloroplast loci. However, the levels of genetic differentiation were lower than expected for mature species. Critically, a lack of chloroplast DNA haplotype sharing among the morphologically indistinguishable and most closely related taxon pair confirmed that chemistry alone can define taxon boundaries.

Conclusions

The results confirmed that pollinator isolation, mediated by specific pollinator attraction, underpins strong reproductive isolation in these taxa. A combination of large effective population sizes, initial neutral mutations in the genes controlling floral scent, and a pool of available pollinators likely drives diversity in this system.     

Arbovirus detection in insect vectors by rapid, high-throughput pyrosequencing

Background

Despite the global threat caused by arthropod-borne viruses, there is not an efficient method for screening vector populations to detect novel viral sequences. Current viral detection and surveillance methods based on culture can be costly and time consuming and are predicated on prior knowledge of the etiologic agent, as they rely on specific oligonucleotide primers or antibodies. Therefore, these techniques may be unsuitable for situations when the causative agent of an outbreak is unknown.

Methodology/Principal Findings

In this study we explored the use of high-throughput pyrosequencing for surveillance of arthropod-borne RNA viruses. Dengue virus, a member of the positive strand RNA Flavivirus family that is transmitted by several members of the Aedes genus of mosquitoes, was used as a model. Aedes aegypti mosquitoes experimentally infected with dengue virus type 1 (DENV-1) were pooled with noninfected mosquitoes to simulate samples derived from ongoing arbovirus surveillance programs. Using random-primed methods, total RNA was reverse-transcribed and resulting cDNA subjected to 454 pyrosequencing.

Conclusions/Significance

In two types of samples, one with 5 adult mosquitoes infected with DENV-1- and the other with 1 DENV-1 infected mosquito and 4 noninfected mosquitoes, we identified DENV-1 DNA sequences. DENV-1 sequences were not detected in an uninfected control pool of 5 adult mosquitoes. We calculated the proportion of the Ae. aegypti metagenome contributed by each infecting Dengue virus genome (p^IP), which ranged from 2.75×10⁻⁸ to 1.08×10⁻⁷. DENV-1 RNA was sufficiently concentrated in the mosquito that its detection was feasible using current high-throughput sequencing instrumentation. We also identified some of the components of the mosquito microflora on the basis of the sequence of expressed RNA. This included members of the bacterial genera Pirellula and Asaia, various fungi, and a potentially uncharacterized mycovirus.     

Evaluation of Circulating Cathodic Antigen (CCA) Urine-Tests for Diagnosis of Schistosoma mansoni Infection in Cameroon

Background

The Kato-Katz is the most common diagnostic method for Schistosoma mansoni infection. However, the day-to-day variability in host egg-excretion and its low detection sensitivity are major limits for its use in low transmission zones and after widespread chemotherapy. We evaluated the accuracy of circulating cathodic antigen (CCA) urine-assay as a diagnostic tool of S. mansoni. In comparison, a low sensitive CCA test (CCA-L) was assessed.

Methodology

The study was conducted in three settings: two foci with single S. mansoni infections (settings A and B), and one mixed S. mansoni – S. haematobium focus (setting C). Stool and urine samples were collected from school-children on three consecutive days. Triplicate Kato-Katz readings were performed per stool sample. Each urine sample was tested with one CCA and only the first urine sample was subjected to CCA-L. Urine samples were also examined for S. haematobium eggs using the filtration method and for microhaematuria using urine reagent strips. Overall, 625 children provided three stool and three urine samples.

Principal Findings

Considering nine Kato-Katz thick smears as ‘reference’ diagnostic test, the prevalence of S. mansoni was 36.2%, 71.8% and 64.0% in settings A, B and C, respectively. The prevalence of S. haematobium in setting C was 12.0%. The sensitivities of single Kato-Katz, CCA and CCA-L from the first stool or urine samples were 58%, 82% and 46% in setting A, 56.8%, 82.4% and 68.8% in setting B, and 49.0%, 87.7% and 55.5% in setting C. The respective specificities were 100%, 64.7% and 100%; 100%, 62.3% and 91.3%; and 100%, 42.5% and 92.0%. Mixed infection with S. haematobium did not influence the CCA test results for S. mansoni diagnosis.

Conclusions/Significance

Urine CCA revealed higher sensitivity than CCA-L and triplicate Kato-Katz, and produced similar prevalence as nine Kato-Katz. It seems an attractive method for S. mansoni diagnosis.