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Screening of some British simuliids for susceptibility to experimental Onchocerca lienalis infection.

Ten species of Simulium from Britain were tested for susceptibility to Onchocerca lienalis infection by intrathoracic injection of microfilariae recovered from the skin of cattle at slaughter. All of the species examined supported development of a proportion of the microfilariae to third-stage larvae, but this was greatest at 53% in S. ornatum s.l., which is a known natural vector in Britain. Inocula of 20, 30 or 50 microfilariae resulted in similar levels of mortality in S. ornatum s.l. (42-53%), but a rise in the dose above 20 microfilariae per fly increased the proportion which developed to third-stage larvae. In a series of comparative experiments employing S. ornatum s.l. as a reference positive for blackflies of unknown susceptibility to infection, 6 other species (S. reptans, S. erythrocephalum, S. lineatum, S. equinum, S. nitidifrons and S. austeni) allowed a proportion of the microfilariae to reach third-stage larvae equivalent to 37-50% of those which developed in S. ornatum s.l. A small sample of S. angustipes supported development as well as its respective control group of S. ornatum s.l., but S. vernum and S. aureum were relatively refractory and were the only species in which third-stage larvae did not accumulate in the head. Fly survival rates varied between the species, but all were below those in S. ornatum s.l. It is concluded that some of these blackflies offer promise as surrogate vectors of O. lienalis in the laboratory and their possible role in natural transmission should receive further attention.

Animals↗

Ultrastructure study of the excretory system and the genital primordium of the infective stage of Onchocerca volvulus (Nematoda:Filarioidea).

The electron microscopic investigation of the anterior part of the infective third-stage juvenile of Onchocerca volvulus provides first insights into the structure of the excretory system of this developmental stage of the parasite. The most anterior part of this system consists of a cell process of the syncytial excretory cells. At this height the excretory cells enclose the cuticle-lined excretory channel. The channel is in the process of elongation in the anterior-posterior direction, indicated by cell division in this region. More posteriad an ampulla-like structure is forming in the cytoplasm of the excretory cells. The inner surface of this ampulla is lined with a small number of single microvilli. In this part of the system the cytoplasm of the excretory cells is rich in Golgi bodies and endocytic vesicles. The ampulla has direct access to the exterior by the excretory duct. The excretory duct is a cuticle-lined structure surrounded by supporting fibres of an additional cell. This duct cell connects the excretory duct to the body-wall cuticle at the excretory pore. Adjacent to the region of the excretory system a cell is found that resembles a gland cell. This cell is in close contact to the ventral nerve cord. The genital primordia of the third-stage juvenile consist of several dividing cells. The female genital primordium is seen at the junction of the muscular with the glandular oesophagus and the male primordium can be found at the junction of the glandular oesophagus with the gut.

Animals↗

Toxicity of microencapsulated permethrin to selected nontarget aquatic invertebrates.

Microencapsulated permethrin (penncapthrin) was evaluated under laboratory conditions for its toxicity toward several nontarget aquatic invertebrates. Average LC50 estimates for selected lotic invertebrates, based on a one hour dosing regime, were: 2.71 mg/L for Simulium vittatum, 4.59 mg/L for Hydropsyche spp., and 13.41 mg/L for Isonychia bicolor. In acute static tests with Daphnia magna, there was no significant difference (p less than or equal to 0.05) between the toxicity of penncapthrin at 96 h (LC50 range: 6.80-22.5 micrograms/L) and the EC formulation at 72 h (LC50 range: 0.6-21 micrograms/L). Comparatively, the toxicity of microencapsulated methyl parathion (penncap-m) was not significantly different from that of penncapthrin toward D. magna, the former having LC50 estimates ranging form 0.3-12.25 micrograms/L. LC50 estimates associated with Daphnia pulex ranged from 19 to 131 micrograms/L. The toxicity of penncapthrin and penncap-m toward D. pulex was difficult to determine because of frequent control mortality due to food deprivation resulting from the need to run tests for longer than 48 h. In successful tests, LC50 estimates ranged from 19 to 28 micrograms/L for penncapthrin and 0.08 to 25 micrograms/L for penncap-m after 72 h exposure. In long term toxicity tests, 95% of D. magna at 1 microgram/L, 44% at 10 micrograms/L, and 20% at 15 micrograms/L survived after 39 days exposure. Less than 15% of D. pulex survived over the same concentration range following 32 days exposure. Despite same drawbacks, long-term toxicity tests were more appropriate than short-term tests for evaluating microencapsulated pesticides because of reduced variability in LC50 estimates and lower control mortality.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Trypanosoma avium: experimental transmission from black flies to canaries.

Trypanosomes identified as Trypanosoma avium were found in the ornithophilic black flies ( Eusimulium latipes) attacking buzzard nestlings ( Buteo buteo). Parasites formed plugs and rosettes in the hindgut of the vector and were attached on the cuticular lining of the black fly anterior intestine (ileum) by hemidesmosome-like plaques. Hindgut stages from infected black flies were experimentally transmitted into canaries ( Serinus canaria) by ingestion of vectors and by contamination of host conjunctiva. This is the first clear evidence of such transmission in avian trypanosomes. Parasites survived in peripheral blood of birds at the least 10 months. In contrast to the direct inoculation of insect stages, parasites from culture failed to produce infection in experimental birds; this has consequences for laboratory studies of host susceptibility and transmission.

Animals↗

Local modification of benthic flow environments by suspension-feeding stream insects.

Larval black flies often exhibit spatially aggregated distributions, and individuals within patches can potentially reduce the supply of suspended food particles to downstream neighbors by modifying local flow characteristics. We used hot-film anemometry to quantify the magnitude and spatial extent of flow modifications downstream from feeding Simulium vittatum larvae in a laboratory flume, and to determine whether temporal patterns of flow variation are related to movements of the larval feeding appendages. Mean velocity 1 mm downstream from feeding larvae was reduced by 75%, and the percent reduction in velocity diminished asymptotically with downstream distance. Reduced velocities were evident as much as 60 mm downstream from, and 3 mm to either side of, larvae. Turbulence intensity (i.e., the SD of the velocity time series) was generally higher in this region relative to control flow conditions. Three results demonstrate the major contribution of the larval feeding appendages (i.e., labral fans) to such flow modification. First, there was a minimal reduction in mean velocity 5 mm downstream from non-feeding larvae (i.e., with closed labral fans), whereas mean velocity at the same location was reduced markedly when larvae were feeding. Second, the power spectrum of the velocity time series exhibited greatest power at frequencies that corresponded to the frequency of labral fan motions. Third, fan flick times accounted for most of the variance in the velocity power spectrum. The large local flow modifications that we documented have potentially important consequences for the feeding performance and growth of individuals located within larval aggregations, and are likely to influence behavioral interactions and spacing patterns.

Animals↗

Balancing food availability and hydrodynamic constraint: phenotypic plasticity and growth in Simulium noelleri blackfly larvae.

Organisms through phenotypic plasticity can cope with multiple changed environmental conditions. Theory predicts that animals in streams and rivers should be able to balance demands of the needs to obtain food efficiently and to adjust response to hydrodynamic variability. This study examined effects of variations in food availability and current velocity on the feeding structure and growth rate of Simulium noelleri blackfly larvae. The larvae developed larger labral fans and more rays under slow current and low food regimes than in fast current and high food conditions. In both fast and slow current regimes, growth rates were higher and development periods to the final-instar stage were shorter in high food treatments. The estimated flux rates of food particles through labral fans under high food treatments for both fast and slow current regimes were higher than those under low food treatments. Although both food and current velocity appeared to have selected for flexibility of feeding structure and growth rate, food availability was a more important factor for phenotypic and developmental plasticity than current velocity. The results indicate a strong link between environmental changes in food availability and current velocity, phenotypic plasticity, and growth rate of S. noelleri. This study suggests that plasticity of ecomorphs with macroevolutionary significance may play a role in the early evolutionary stages of blackfly larvae.

Adaptation, Physiological↗

Silk filaments enhance the settlement of stream insect larvae.

Many aquatic organisms need to settle in suitable benthic habitats while being transported via water currents. Such settlement is especially challenging for organisms that encounter complex benthic topography and lack the ability to move easily from the water column to the bed (e.g., via swimming). We conducted flume studies to examine whether the settlement of drifting stream insects is facilitated by adhesive filaments that extend from their bodies. Using a new tripwire visualization technique, we found that neonatal black flies (Simulium tribulatum) drifted with silk threads averaging six times their body length. These threads allowed larvae to contact or snag the bed from a greater height than would be possible through direct body-to-bed contact alone, and instantly arrested their downstream movement. Thus, silk increased their probability of settlement. We then performed an experiment to examine how settlement varied with bed topography and velocity. We tested whether settlement rate differed between a flat bed and an irregular bed that mimicked key aspects of their natural cobble-bed habitat. Velocities were similar for both bed treatments. Settlement on the irregular bed was 40 times greater than on the flat bed due to silk use. Settlement rate also exhibited a marginally significant decline with increasingly velocity on the flat bed, but not on the irregular bed. Silk threads should greatly increase the settlement rate of these nonswimming larvae on coarse-grained stream beds. Thus, silk snagging can potentially reduce the downstream distance that individuals are transported during a drift event, although the effects of silk on other phases of larval dispersal may differ.

Animals↗

Isolation by distance and a chromosomal cline in the Cayapa cytospecies of Simulium exiguum, the vector of human onchocerciasis in Ecuador.

The population genetic structure of the Cayapa cytospecies of Simulium exiguum, the vector of onchocerciasis, was analysed using allozyme frequency and chromosomal inversion polymorphism data from 6 and 15 populations respectively, collected in Ecuador. Eight allozyme loci were scored. No unique allozyme markers were found enabling us to identify biting adults of the vector from the non-vector Bucay cytotype. Mannose-phosphate isomerase (Mpi) contributed largely to the significant heterogeneity in gene frequency among populations of the Cayapa cytospecies and also to the overall population structuring (F(ST) = 0.015 +/- 0.014) which fitted the isolation by distance model. However, heterozygote deficits were recorded for Mpi in four of the six populations, which could indicate that selection is acting at this locus but this hypothesis will require further convincing evidence. Furthermore the significant population structuring of allozymes was not evident when Mpi was omitted from the analysis. All inversion polymorphisms (IIS-B, IIS-F, IIL-A and IIL-B) were in Hardy-Weinberg equilibrium, showed significant heterogeneity between populations and revealed the occurrence of an altitudinal cline in inversion IIS-B frequency. The inversion polymorphisms revealed a significant degree of population structuring (F(ST) = 0.083 +/- 0.027), which can be explained by the isolation by distance model. A UPGMA cluster analysis revealed the relatively remote, high altitude Rio Mira populations to be the most genetically distinct.

Animals↗

Onchocerca volvulus: application of the polymerase chain reaction to identification and strain differentiation of the parasite.

Previous studies have demonstrated that the genome of Onchocerca volvulus contains a variable tandemly repeated DNA sequence family with a unit length of 150 bp. The variability of the 150-bp family has been exploited to develop O. volvulus strain and species specific DNA probes. Application of these DNA probes to the study of the epidemiologically most significant life cycle stages of the parasite has been confounded by several obstacles. These include the relative insensitivity of some of the DNA probes and the difficulty in releasing genomic DNA from infective larvae and skin microfilariae in a form that may be directly detected by hybridization to the probes. DNA sequence comparison of 18 known examples of the 150-bp repeat has been used to develop two populations of degenerate oligonucleotides. These oligonucleotides have been shown to support the amplification of the 150-bp repeat family from Onchocerca DNA, using the polymerase chain reaction. Both strain and species specific members of the repeat family are faithfully amplified, allowing characterization of a parasite on the basis of hybridization of the PCR amplification products to the previously developed DNA probes. This method is shown to be applicable to all diagnostically important forms of the parasite, including adults, infective larvae, and skin microfilariae. In addition, the method is capable of detecting O. volvulus infective larvae directly in extracts of blackfly vectors.

Animals↗

Mitochondrial alleles of Simulium damnosum sensu lato infected with Onchocerca volvulus.

Onchocerca volvulus infected Simulium damnosum s.l. were analysed by directed heteroduplex analysis. Of 73 infected flies, 68 produced heteroduplex products identical to those previously identified. All 6 major sibling species, except S. leonense, were present in this group. In the 5 remaining flies, 2 new heteroduplex patterns were noted. Molecular phylogenetic analysis of these samples suggested that they belonged to the S. squamosum/S. yahense subcomplex. The ability to reliably genotype adult flies will permit studies of the vectorial capacity of the sibling species of S. damnosum s.l. for the blinding and non-blinding strains of O. volvulus.

Alleles↗

The distribution of the microfilariae of Onchocerca volvulus in the different body regions in relation to the attacking behaviour of Simulium damnosum s.l. in the Sudan savanna of northern Cameroon.

Densities of Onchocerca volvulus microfilariae in four volunteers with low to moderate infections were estimated at five body sites by paired skin snips. The landing of Simulium damnosum s.1. females on the body of these volunteers was recorded during 12 hours for six days. Most flies fed at the ankles (53% and 51%) and calves (28% and 27% respectively) in both the standing and sitting positions. The density of microfilariae in the skin was highest in the pelvic region (24.1 mf/mg) and relatively low in the calf (14.8 mf/mg) and ankle (1.0 mf/mg) regions. From the biting rate (females/body part) and the microfilarial density (mf/mg) a transmission index was calculated for the different body regions. This was highest for the calves showing that this part of the leg, if unclothed, accounts for the highest rate of contact (50 to 60% of total) between vector and parasite.

Animals↗