Non-Invasive Sampling of Schistosomes from Humans Requires Correcting for Family Structure

For ethical and logistical reasons, population-genetic studies of parasites often rely on the non-invasive sampling of offspring shed from their definitive hosts. However, if the sampled offspring are naturally derived from a small number of parents, then the strong family structure can result in biased population-level estimates of genetic parameters, particularly if reproductive output is skewed. Here, we document and correct for the strong family structure present within schistosome offspring (miracidia) that were collected non-invasively from humans in western Kenya. By genotyping 2,424 miracidia from 12 patients at 12 microsatellite loci and using a sibship clustering program, we found that the samples contained large numbers of siblings. Furthermore, reproductive success of the breeding schistosomes was skewed, creating differential representation of each family in the offspring pool. After removing the family structure with an iterative jacknifing procedure, we demonstrated that the presence of relatives led to inflated estimates of genetic differentiation and linkage disequilibrium, and downwardly-biased estimates of inbreeding coefficients (F[subscript IS]). For example, correcting for family structure yielded estimates of F[[subscript ST] among patients that were 27 times lower than estimates from the uncorrected samples. These biased estimates would cause one to draw false conclusions regarding these parameters in the adult population. We also found from our analyses that estimates of the number of full sibling families and other genetic parameters of samples of miracidia were highly intercorrelated but are not correlated with estimates of worm burden obtained via egg counting (Kato-Katz). Whether genetic methods or the traditional Kato-Katz estimator provide a better estimate of actual number of adult worms remains to be seen. This study illustrates that family structure must be explicitly accounted for when using offspring samples to estimate the genetic parameters of adult parasite populations

Interactions between Natural Populations of Human and Rodent Schistosomes in the Lake Victoria Region of Kenya: A Molecular Epidemiological Approach

One of the world's most prevalent neglected diseases is schistosomiasis, which infects approximately 200 million people worldwide. Schistosoma mansoni is transmitted to humans by skin penetration by free-living larvae that develop in freshwater snails. The origin of this species is East Africa, where it coexists with its sister species, S. rodhaini. Interactions between these species potentially influence their epidemiology, ecology, and evolutionary biology, because they infect the same species of hosts and can hybridize. Over two years, we examined their distribution in Kenya to determine their degree of overlap geographically, within snail hosts, and in the water column as infective stages. Both species were spatially and temporally patchy, although S. mansoni was eight times more common than S. rodhaini. Both species overlap in the time of day they were present in the water column, which increases the potential for the species to coinfect the same host and interbreed. Peak infective time for S. mansoni was midday and dawn and dusk for S. rodhaini. Three snails were coinfected, which was more common than expected by chance. These findings indicate a lack of obvious isolating mechanisms to prevent hybridization, raising the intriguing question of how the two species retain separate identities

Relationship between HIV stage and psychomotor speed neurocognitive score at a Kenyan sub-county hospital

Background: Human immunodeficiency virus (HIV) and acquired immunodeficiency syndrome (AIDS) is associated with cognitive impairment which affects psychomotor speed. Psychomotor slowing is a predictor of dementia and death in people living with HIV and AIDS. The purpose of this study was to assess the relationship between HIV disease stage and psychomotor speed neurocognitive score which will add to the body of knowledge required to manage patients with HIV and AIDS. Objective: To determine the relationship between psychomotor speed neurocognitive score and the HIV disease stage in adults at initiation of care. Setting: This study was conducted at Kangundo Sub-county hospital comprehensive care centre. Methods: This was a cross-sectional study. All HIV seropositive patients aged 18 to 50 years recently initiated into care were studied. A pretested questionnaire was used to collect data. The World Health Organization (WHO) stage was used during data collection to classify study participants into asymptomatic and symptomatic groups. The grooved pegboard test was used to obtain psychomotor speed neurocognitive scores. Descriptive statistics were used to summarise data. Mann–Whitney U test, Spearman’s rho and multiple linear regression were employed in the analysis; p-value of 0.05 was considered significant. Results: The WHO stage did not have a significant effect on the psychomotor speed neurocognitive score (p ≥ 0.05). The CD4 count had a significant effect on psychomotor speed neurocognitive score (p = 0.001). Conclusions: There was a significant correlation between CD4 counts and psychomotor speed neurocognitive score. Efforts should be made to ensure that the CD4 counts of people living with HIV and AIDS do not continue to fall after initiation into care in order to preserve psychomotor function

Development of a Loop Mediated Isothermal Amplification for Diagnosis of Ascaris lumbricoides in Fecal Samples

Ascaris lumbricoides is a nematode parasite that causes the common tropical infection ascariasis in humans. It is also considered among the neglected tropical diseases. Diagnosis relies mainly on microscopy-based methods which are laborious, are limited by low sensitivity, and require high expertise. We have developed a loop mediated isothermal amplification (LAMP) for diagnosis of ascariasis in fecal samples, based on the first internal transcribed (ITS-1) spacer region of the ribosomal DNA. We used Primer Explorer V4 software to design primers. Ascaris adult and ova were obtained from naturally infected school children, whose parents/guardians gave consent for their participation in the study. Genomic DNA was extracted using alkaline lysis method and amplified by LAMP at 63°C for 45 minutes. LAMP products were visualized by naked eyes after adding SYBR Green dye and also on agarose gel. LAMP successfully and reliably detected Ascaris DNA from a single egg and in fecal samples. The assay specifically detected Ascaris DNA without amplifying DNA from ova of other parasites which commonly coexist with A. lumbricoides in feces. The developed LAMP assay has great potential for use in ascariasis diagnosis at the point of care and in low infection intensity situation that characterize control and elimination campaigns

Minimum evolution tree of several specimens of <i>Schistosoma mansoni</i> and <i>S. rodhaini</i> based on 512 bp of mitochondrial DNA including part of the 16S ribosomal RNA gene, all of thetRNA-Cys gene, and part of the 12S ribosomal RNA gene.

<p>Uncorrected p-distance is given for scale.</p

Collection sites for snails of the genus <i>Biomphalaria</i> and their schistosome parasites in Western Kenya.

<p>Global Positioning System coordinates are projected in North American Datum 1983 decimal degrees.</p

Results of the Fisher's Exact tests to determine if the proportions of genotypes of <i>S. mansoni</i> released from snails are consistent over timed replicates.

<p>P indicates the probability of significance, n indicates the number of worms sampled, R is the number of replicates tested, and G is the number of genotypes present in a snail. Mode indicates whether the proportions remained constant over replicates, followed a replacement pattern, or was variable.</p