Zika Virus Emergence in Mosquitoes in Southeastern Senegal, 2011

Background

Zika virus (ZIKV; genus Flavivirus, family Flaviviridae) is maintained in a zoonotic cycle between arboreal Aedes spp. mosquitoes and nonhuman primates in African and Asian forests. Spillover into humans has been documented in both regions and the virus is currently responsible for a large outbreak in French Polynesia. ZIKV amplifications are frequent in southeastern Senegal but little is known about their seasonal and spatial dynamics. The aim of this paper is to describe the spatio-temporal patterns of the 2011 ZIKV amplification in southeastern Senegal.

Methodology/Findings

Mosquitoes were collected monthly from April to December 2011 except during July. Each evening from 18∶00 to 21∶00 hrs landing collections were performed by teams of 3 persons working simultaneously in forest (canopy and ground), savannah, agriculture, village (indoor and outdoor) and barren land cover sites. Mosquitoes were tested for virus infection by virus isolation and RT-PCR. ZIKV was detected in 31 of the 1,700 mosquito pools (11,247 mosquitoes) tested: Ae. furcifer (5), Ae. luteocephalus (5), Ae. africanus (5), Ae. vittatus (3), Ae. taylori, Ae. dalzieli, Ae. hirsutus and Ae. metallicus (2 each) and Ae. aegypti, Ae. unilinaetus, Ma. uniformis, Cx. perfuscus and An. coustani (1 pool each) collected in June (3), September (10), October (11), November (6) and December (1). ZIKV was detected from mosquitoes collected in all land cover classes except indoor locations within villages. The virus was detected in only one of the ten villages investigated.

Conclusions/Significance

This ZIKV amplification was widespread in the Kédougou area, involved several mosquito species as probable vectors, and encompassed all investigated land cover classes except indoor locations within villages. Aedes furcifer males and Aedes vittatus were found infected within a village, thus these species are probably involved in the transmission of Zika virus to humans in this environment.     

Temporal distribution and spatial pattern of abundance of the Rift Valley fever and West Nile fever vectors in Barkedji, Senegal

The temporal distribution and spatial pattern of abundance of mosquito vectors of Rift Valley fever (RVf) and West Nile fever (WNf) were studied during the 2005 and 2006 rainy seasons at Barkedji, Senegal. Mosquitoes were collected every two weeks with CDC light traps with dry ice at 79 sites including temporary ponds, barren, shrubby savannah, wooded savannah, steppes, and villages at different distances (between 0 and 600 m) from the nearest pond. The temporal distributions of these vectors varied between 2005 and 2006 and were positively correlated with rainfall for Aedes (Aedimorphus) vexans Patton, with rainfall after a lag time of one month for Culex (Culex) poicilipes (Theobald) and Culex (Culex) neavei Theobald. All the vectors had their highest abundances and parity rates between September and November. The highest vector abundances were observed in the barren and temporary ponds. The distance of trap location to the nearest ponds was negatively correlated to the abundance of the vectors. Taking into account the linear regression equations, it was predicted that mosquitoes would not disperse and be collected by the light trap, up to 1,500 m to the nearest ponds. The implications of these findings in the epidemiology and control of RVF and WNF at Barkedji are discussed.     

Tropheryma whipplei: a common bacterium in rural Senegal

Background

Tropheryma whipplei is known as the cause of Whipple''s disease, but it is also an emerging pathogen, detected in stool, that causes various chronic localized infections without histological digestive involvement and is associated with acute infections, including gastroenteritis and bacteremia.

Methods/Principal Findings

We conducted a study in 2008 and 2009 using 497 non-diarrheic and diarrheic stool samples, 370 saliva samples, 454 sera samples and 105 samples obtained from water samples in two rural Sine-Saloum villages (Dielmo and Ndiop) in Senegal. The presence of T. whipplei was investigated by using specific quantitative PCR. Genotyping was performed on positive samples. A serological analysis by western blotting was performed to determine the seroprevalence and to detect seroconversion. Overall, T. whipplei was identified in 31.2% of the stool samples (139/446) and 3.5% of the saliva samples (13/370) obtained from healthy subjects. The carriage in the stool specimens was significantly (p<10⁻³) higher in children who were between 0 and 4 years old (60/80, 75%) compared to samples obtained from individuals who were between 5 to 10 years old (36/119, 30.2%) or between 11 and 99 years old (43/247, 17.4%). The carriage in the stool was also significantly more common (p = 0.015) in subjects with diarrhea (25/51, 49%). We identified 22 genotypes, 16 of which were new. Only one genotype (#53) was common to both villages. Among the specific genotypes, one (#52) was epidemic in Dielmo (15/28, 53.4%, p<10⁻³) and another (#49) in Ndiop (27.6%, p = 0.002). The overall seroprevalence was estimated at 72.8% (291/400). Seroconversion was detected in 66.7% (18/27) of children for whom PCR became positive in stools between 2008 and 2009.

Conclusions/Significance

T. whipplei is a common bacterium in the Sine-Saloum area of rural Senegal that is contracted early in childhood. Epidemic genotypes suggest a human transmission of the bacterium.     

Spermiogenesis and the spermatozoon ultrastructure of Robphildollfusium fractum (Digenea: Gyliauchenidae), an intestinal parasite of Sarpa salpa (Pisces: Teleostei)

Spermiogenesis in Robphildollfusium fractum begins with the formation of a differentiation zone containing: two centrioles, each bearing striated rootlets, nucleus, several mitochondria and an intercentriolar body constituted by seven electron-dense layers. The two centrioles originate two free flagella growing orthogonally to the median cytoplasmic process. Later, the free flagella rotate and undergo proximodistal fusion with the median cytoplasmic process. Nuclear and mitochondrial migrations occur before this proximodistal fusion. Finally, the young spermatozoon detaches from the residual cytoplasm after the constriction of the ring of arched membranes. The spermatozoon of R. fractum exhibits two axonemes of different length of the 9 + ‘1’ trepaxonematan pattern, nucleus, two mitochondria, two bundles of parallel cortical microtubules, external ornamentation of the plasma membrane, spine-like bodies and granules of glycogen. Additionally, a shorter axoneme, which does not reach the nuclear region, the presence of an electron-dense material in the anterior spermatozoon extremity and the morphologies of both spermatozoon extremities characterize the mature sperm of R. fractum.     

Population diversity and antibody selective pressure to Plasmodium falciparum MSP1 block2 locus in an African malaria-endemic setting

Background

Genetic evidence for diversifying selection identified the Merozoite Surface Protein1 block2 (PfMSP1 block2) as a putative target of protective immunity against Plasmodium falciparum. The locus displays three family types and one recombinant type, each with multiple allelic forms differing by single nucleotide polymorphism as well as sequence, copy number and arrangement variation of three amino acid repeats. The family-specific antibody responses observed in endemic settings support immune selection operating at the family level. However, the factors contributing to the large intra-family allelic diversity remain unclear. To address this question, population allelic polymorphism and sequence variant-specific antibody responses were studied in a single Senegalese rural community where malaria transmission is intense and perennial.

Results

Family distribution showed no significant temporal fluctuation over the 10 y period surveyed. Sequencing of 358 PCR fragments identified 126 distinct alleles, including numerous novel alleles in each family and multiple novel alleles of recombinant types. The parasite population consisted in a large number of low frequency alleles, alongside one high-frequency and three intermediate frequency alleles. Population diversity tests supported positive selection at the family level, but showed no significant departure from neutrality when considering intra-family allelic sequence diversity and all families combined. Seroprevalence, analysed using biotinylated peptides displaying numerous sequence variants, was moderate and increased with age. Reactivity profiles were individual-specific, mapped to the family-specific flanking regions and to repeat sequences shared by numerous allelic forms within a family type. Seroreactivity to K1-, Mad20- and R033 families correlated with the relative family genotype distribution within the village. Antibody specificity remained unchanged with cumulated exposure to an increasingly large number of alleles.

Conclusion

The Pfmsp1 block2 locus presents a very large population sequence diversity. The lack of stable acquisition of novel antibody specificities despite exposure to novel allelic forms is reminiscent of clonal imprinting. The locus appears under antibody-mediated diversifying selection in a variable environment that maintains a balance between the various family types without selecting for sequence variant allelic forms. There is no evidence of positive selection for intra-family sequence diversity, consistent with the observed characteristics of the antibody response.     

Increase of malaria attacks among children presenting concomitant infection by Schistosoma mansoni in Senegal

Background

Parasites incur periodic mutations which must ultimately be eliminated to maintain their genetic integrity.

Methods

It is hypothesised that these mutations are eliminated not by the conventional mechanisms of competition between parasites in different hosts but primarily by competition between parasites within the same infection.

Results

This process is enhanced by the production of a large number of parasites within individual infections, and this may significantly contribute to parasitic virulence.

Conclusions

Several features of the most virulent human malaria parasite Plasmodium falciparum can usefully be re-interpreted in this light and lend support to this interpretation. More generally, it constitutes a novel explanation for the evolution of virulence in a wider range of microparasites.     

A novel mechanism for protein delivery: granzyme B undergoes electrostatic exchange from serglycin to target cells

The molecular interaction of secreted granzyme B-serglycin complexes with target cells remains undefined. Targets exposed to double-labeled granzyme B-serglycin complexes show solely the uptake of granzyme B. An in vitro model demonstrates the exchange of the granzyme from serglycin to immobilized, sulfated glycosaminoglycans. Using a combination of cell binding and internalization assays, granzyme B was found to exchange to sulfated glycosaminoglycans and, depending on the cell type, to higher affinity sites. Apoptosis induced by purified granzyme B and cytotoxic T-cells was diminished in targets with reduced cell surface glycosaminoglycan content. A mechanism of delivery is proposed entailing electrostatic transfer of granzyme B from serglycin to cell surface proteins.     

Worms can worsen malaria: towards a new means to roll back malaria?

Recent studies in Africa and Asia indicate that different helminthic infections adversely affect the clinical outcome of malaria infections. This suggests that helminths can influence the acquisition of immunity against Plasmodium. Worms could constitute a confounding factor in the assessment of efficacy of malaria-control intervention, including vaccine prototypes in clinical trials. These observations have fundamental and practical consequences; if the deleterious effect of worms on malaria is confirmed, treatment of helminths would offer an affordable, strongly effective and novel means to roll back malaria. With this article, we hope to induce others to conduct similar studies in different regions.     

P2X7 nucleotide receptors mediate caspase-8/9/3-dependent apoptosis in rat primary cortical neurons

Apoptosis is a major cause of cell death in the nervous system. It plays a role in embryonic and early postnatal brain development and contributes to the pathology of neurodegenerative diseases. Here, we report that activation of the P2X₇ nucleotide receptor (P2X₇R) in rat primary cortical neurons (rPCNs) causes biochemical (i.e., caspase activation) and morphological (i.e., nuclear condensation and DNA fragmentation) changes characteristic of apoptotic cell death. Caspase-3 activation and DNA fragmentation in rPCNs induced by the P2X₇R agonist BzATP were inhibited by the P2X₇R antagonist oxidized ATP (oATP) or by pre-treatment of cells with P2X₇R antisense oligonucleotide indicating a direct involvement of the P2X₇R in nucleotide-induced neuronal cell death. Moreover, Z-DEVD-FMK, a specific and irreversible cell permeable inhibitor of caspase-3, prevented BzATP-induced apoptosis in rPCNs. In addition, a specific caspase-8 inhibitor, Ac-IETD-CHO, significantly attenuated BzATP-induced caspase-9 and caspase-3 activation, suggesting that P2X₇R-mediated apoptosis in rPCNs occurs primarily through an intrinsic caspase-8/9/3 activation pathway. BzATP also induced the activation of C-jun N-terminal kinase 1 (JNK1) and extracellular signal-regulated kinases (ERK1/2) in rPCNs, and pharmacological inhibition of either JNK1 or ERK1/2 significantly reduced caspase activation by BzATP. Taken together, these data indicate that extracellular nucleotides mediate neuronal apoptosis through activation of P2X₇Rs and their downstream signaling pathways involving JNK1, ERK and caspases 8/9/3.     

Disruption of Maize Kernel Growth and Development by Heat Stress (Role of Cytokinin/Abscisic Acid Balance)

Temperature stress during kernel development affects maize (Zea mays L.) grain growth and yield stability. Maize kernels (hybrid A619 x W64A) were cultured in vitro at 3 d after pollination and either maintained at 25[deg]C or transferred to 35[deg]C for 4 or 8 d, then returned to 25[deg]C until physiological maturity. Kernel fresh and dry matter accumulation was severely disrupted by the long-term heat stress (8 d at 35[deg]C) and did not recover when transferred back to 25[deg]C, resulting in abortion of 97% of the kernels. Kernels exposed to 35[deg]C for 4 d (short-term heat stress) exhibited a recovery in kernel growth and water content at about 18 d after pollination and kernel abortion was reduced to about 23%. During the cell division phase, abscisic acid (ABA) levels showed a steady decline in the control but maintained a moderate level in the heat-stressed kernels. However, later in development heat-stressed kernels had significantly higher levels of ABA than the control. Cytokinin analysis confirmed a peak in zeatin riboside and zeatin levels in control kernels at 10 to 12d after pollination. In contrast, kernels subjected to 4 d of heat stress had no detectable levels of zeatin and the zeatin riboside peak was reduced by 70% and delayed until 18 d after pollination. The long-term heat-stressed kernels showed low to nondetectable levels of either zeatin riboside or zeatin. Regression analysis of ABA level against cytokinin level during the endosperm cell division phase revealed a highly significant negative correlation in nonstressed kernels but no correlation in kernels exposed to short-term or long-term heat stress. Application of benzyladenine to heat-stressed, growth-chamber-grown plants increased thermotolerance in part by reducing kernel abortion at the tip and middle positions on the ear. These results confirm that shift in hormone balance of kernels is one mechanism by which heat stress disrupts maize kernel development. The maintenance of high levels of cytokinins in the kernels during heat stress appears to be important in increasing thermotolerance and providing yield stability of maize.