Last-come, best served? Mosquito biting order and Plasmodium transmission.

A pervasive characteristic of parasite infections is their tendency to be overdispersed. Understanding the mechanisms underlying this overdispersed distribution is of key importance as it may impact the transmission dynamics of the pathogen. Although multiple factors ranging from environmental stochasticity to inter-individual heterogeneity may explain parasite overdispersion, parasite infection is also overdispersed in an inbred host population maintained under laboratory conditions, suggesting that other mechanisms are at play. Here, we show that the aggregated distribution of malaria parasites within mosquito vectors is partially explained by a temporal heterogeneity in parasite infectivity triggered by the bites of mosquitoes. Parasite transmission tripled between the mosquito's first and last blood feed in a period of only 3 h. Surprisingly, the increase in transmission is not associated with an increase in parasite investment in production of the transmissible stage. Overall, we highlight that Plasmodium is capable of responding to the bites of mosquitoes to increase its own transmission at a much faster pace than initially thought and that this is partly responsible for overdispersed distribution of infection. We discuss the underlying mechanisms as well as the broader implications of this plastic response for the epidemiology of malaria.

Effects of copper on gill structure and transport function in the rainbow trout, Oncorhynchus mykiss.

Effects of copper were studied in freshwater adapted rainbow trout using the perfused head preparation. In its monovalent chemical form, copper at millimolar concentrations had no significant effects on Na+ and water transport. By contrast, the divalent form produced an increase in gill perfusion pressure, a significant reduction in Na+ influx and water fluxes and reversed Na+ net flux. Observations by light microscopy showed important cell damage (oedema, mucus production, cellular desquamation). By electron microscopy there was smoothing of apical membranes, swelling of the tubular system and destruction of mitochondria. The Na, K-ATPase activity was totally suppressed and residual ATPase activity largely inhibited by 1 mM Cu2+. There was inhibition of the Na,K-ATPase activity with an IC50 of approximately 10 microM of total copper (free and bound cupric fractions). As active sodium transport is located on the secondary lamellae, our results show that its entry mechanism is inhibited at that level by cupric ions only. Results are discussed in relation to hydromineral balance of the trout.

Effects of lead loads on branchial osmoregulatory mechanisms in the rainbow trout Oncorhynchus mykiss.

Exposure of rainbow trout to lead chloride (PbCl2, 1 ppm) in fresh water killed all animals within 16 days. Exposure to this lethal dose for 6 days only showed a significant increase in the haematocrit. Calcium, sodium and chloride concentrations in plasma were not notably modified. Both the influx and the net flux of sodium fluctuated much less than the diffusional water fluxes through secondary lamellae in gills. Branchial Na,K-ATPase, Ca-ATPase and HCO3-ATPase activities were not sensitive to lead toxicity. Lead caused a cellular 'wave-shaped' degeneration and renewal with modification in the number and morphology of chloride cells. Results are discussed in relation to the hydromineral balance of the trout.

Measurement of branchial vascular space of trout, Oncorhynchus mykiss: effects of adrenaline.

Using the isolated-perfused head preparation at a constant flow rate, hemodynamic effects of adrenaline were studied in trout gills. The calculation of the vascular spaces was performed with the isotopic pulse technique allowing measurement of the distribution space of the tracer. The results show that the branchial arterial circuit was cleared more quickly than the branchial venous and cephalic circuits. Adrenaline addition significantly increased the volume of the branchial arterial circuit at the expense of the venous circuit, illustrating the closing of arterio-venous sphincters under catecholamine control. The increase of the arterial volume could be explained by a vasodilation of the arterial circuit, rather than resulting from lamellar recruitment. Furthermore, the flow rate of the cephalic circuit represented 5% of the total branchial flow rate.

Kinetics of potassium transport across trout gills.

1. Kinetics of potassium transport across trout gills was studied, using an isolated-head preparation. 2. Potassium exchanges were shown to take place across secondary lamellae only. 3. Influx of potassium was saturable and fitted satisfactorily the lineweaver-Burk linear plot. 4. Results suggest that these exchanges occur through potassium channel. 5. Kinetics of potassium exchanges is discussed in relation to the maintenance of the osmoregulation in fish.

Estimation of sodium uptake through the gill of the rainbow trout Salmo gairdneri.

Sodium exchanges through the gill epithelia were estimated in the rainbow trout, Salmo gairdneri, using the perfused head technique. The head tissues accumulate radioactivity. In addition there exists an extrabranchial entry of sodium, which is 20 times smaller than the branchial one. The study of the evolution of venous and arterial sodium specific activities as a function of time is necessary before flux measurements are achieved. Contrary to previous studies, an uptake at the primary lamella level is demonstrated. Uptake of sodium through the secondary lamellae can be measured rapidly while at least 30 min are needed for measurement of uptake through the primary lamellae. Chloride cells of the primary lamellae contribute to about 20% of the total sodium uptake.

Glucose transport across the gill of the rainbow trout, Salmo gairdneri.

Glucose fluxes across the gills were measured in freshwater-adapted trout (Salmo gairdneri) using an in vitro, perfused-head preparation. A large asymmetry was observed for the primary lamellar pathway, glucose permeability in the serosa-to-mucosa direction being up to 24 times greater than the permeability in the mucosa-to-serosa direction. Chloride cells appeared to possess a maximal rate of transport, or TMG, of 79 mumol/hr per 100 g. Phlorizin, phloretin and, to a lesser extent, harmaline caused an increase in the rate of glucose efflux. The results suggest that the tubulo-vesicular reticulum, into which plasma is introduced under low pressure, may be regarded as a reabsorption site for glucose in a way similar to the nephron proximal tubule. Thus, essential molecules such as glucose are removed while excess or non-essential substances are excreted into the external medium.

Effects of environmental salinity on branchial permeability of rainbow trout, Salmo gairdneri.

1. The gill fluxes of various non-electrolytes were measured in fresh-water- and sea-water-adapted trout (Salmo gairdneri). The studies were performed in vitro with a 'perfused-head' preparation. 2. The results allow one to specify different transepithelial pathways according to the physico-chemical characteristics of the permeant molecules: (1) for hydrophilic the physico-chemical characteristics of the permeant molecules: (1) for hydrophilic and lipophilic molecules of small molecular radius, a transcellular pathway in the respiratory cells of the secondary lamellae, (2) for hexose, all paracellular pathways in the gill epithelium, and (3) for hexose polymers (inulin, dextran), a transcellular pathway in the chloride cells of the primary lamellae. 3. The selectivity of the respiratory cells as a function of the liposolubility of the molecules tested is low. The decrease of this selectivity in the course of salt-water adaptation taken together with the modification of lipid composition of membranes and the effect of adrenaline on the branchial permeability suggests that non-electrolytes diffuse through a lipid phase able to form hydrogen bonds. 4. The high permeability of gills to hexose polymers of high molecular weight suggests a vesicular transport, especially in fish adapted to fresh water.

Active urea transport through isolated skins of frog and toad.

Urea influxes (Ji) and effluxes (Je) were studied across the isolated skins of Rana esculenta, Bufo bufo, and B. viridis. Two symmetrical pieces of the same skin, bathed in Ringer + 2 mM urea, were used for the two fluxes. In R. esculenta the urea fluxes are passive when the animals are kept in running water but become active after dehydration in air or preadaptation in saline solutions. The ratio Ji/Je can vary between 3 and 27 and the Ji between 2 and 22 nmol . h-1 . cm-2 according to preadaptation. Only the active fluxes obey saturation kinetics. Urea absorption is always independent of sodium transport. In toads, active urea transport occurs even when hydrated. It is markedly stimulated by saline preadaptation. A correlation between the degree of active urea transport across the skin and the capacity of the species to endure dehydrating conditions would appear to exist. The physiological significance of this transport mechanism is discussed.

Active transport of urea across the skin of the euryhaline toad, Bufo viridis.

Urea is accumulated in considerable amounts (greater than 100 mM) in the blood of the euryhaline toad Bufo viridis, under conditions of adaptation to high salinities. Salt adaptation increases active transport of urea (inward direction) in the skin, which was measured in vitro. The active transport of urea is insensitive to ADH, and was inhibited nearly 50% by 0.5 mM phloretin. This transport system is different from the facilitated diffusion of urea which has been studied extensively in the toad urinary bladder, and may offer a simple model system for the study of active urea transport.

Non-electrolyte permeability of trout gills: effect of temperature and adrenaline.

1. The gill permeability to various non-electrolytes (P(s)) was measured in fresh-water and sea-water adapted trout (Salmo gairdneri). This study was performed in vitro using a ;head-perfused' preparation. The influence of temperature and adrenaline (10(-6)M) on permeability to non-electrolytes was also investigated.2. During salt adaptation P(butanol) and P(water) decrease, P(mannitol) rises and P(dextran) stays constant. In view of recently acquired morphological data these results back up the hypothesis of different pathways across the gill epithelium (transcellular, vesicular and paracellular) according to the physico-chemical characteristics of the molecules. The low selectivity of the gill epithelium as a function of the liposolubility of the molecules used testifies to the hydrophilic nature of diffusion across this epithelium, a feature becoming more pronounced during salt adaptation.3. The activation energies are about 4 kcal/mol, an energy comparable to diffusion in water for most of the substances tested, exceptions being butanol for fresh-water adapted gills and water for fresh-water and sea-water adapted gills. Arrhenius plots for butanol in fresh water gills show a transition temperature at 15 degrees C, suggesting an increased membrane lipid fluidity above this temperature.4. Adrenaline has no effect on P(mannitol) and P(dextran), but increases P(butanol) and P(water) selectively according to the adaptation medium (+ 160% and + 100% in fresh water and + 25% and + 20% in sea water respectively). These results point to an effect of this catecholamine on the membrane lipid fluidity.

Effects of epinephrine on branchial non-electrolyte permeability in rainbow trout.

Using isolated heads perfused at constant pressure, at rates close to those occurring in vivo, the permeability of the gills of the trout Salmo gairdneri to a range of solutes was measured. Under epinephrine-free conditions, butanol and water showed similar high branchial permeability coefficients. Urea, inulin and dextrans (mol. wt 3000 and 20 000) were 7-12 times less permeant, and mannitol 60-70 times less permeant than water or butanol. Epinephrine, at 10(-6) M, greatly increased the permeability of the gills to the small hydrophilic molecules, water and urea, and to the lipophilic substance, butanol, but did not affect the penetration of the large hydrophilic solutes, mannitol, inulin and dextrans. In the presence of 10(-6) M propanolol, a beta-blocker, epinephrine had no effect on the permeation of any of the test substances except that the permeability to urea decreased somewhat. The results suggest that epinephrine increases the permeability of the membranes of the branchial cells but does not affect the permeation of substances that cross the gill walls by paracellular routes or via an intracellular 'bulk-transport' mechanism. Such an action would be expected to increase the branchial transfer of oxygen.

Epinephrine effects on branchial water and urea flux in rainbow trout.

In isolated trout heads, perfused at constant pressure, epinephrine (10(-6) M) was found to double water and urea efflux but increased Ringer perfusion rate by only 33%. Drastic changes in perfusion rate (by clamping) produced smaller changes of both efflux rates. Epinephrine-stimulated increase in water and urea efflux, and perfusion rate, was blocked by propranolol (beta-blocker) but not by phentolamine (alpha-blocker). Both blockers together canceled out all epinephrine effects. Epinephrine increased water influx across isolated unperfused gill arches, the effect again being blocked by propranolol but not by phentolamine. Both blockers together canceled any epinephrine effect. We conclude that epinephrine alters branchial vascular flow and functional surface area via alpha- and beta-adrenergic receptors, but also increases branchial permeability to water and probably urea, via beta-adrenergic receptors. To test the validity of the perfused head technique, water and urea efflux rates were compared with in vivo values.

Effect of environmental salinity change on osmotic permeability of the isolated gill of the eel, Anguilla anguilla L.

Net water fluxes in the isolated gills of Anguilla anguilla were studied during incubation in fresh water (FW) and in sea water (SW). When incubated in FW, water entry was greater in SW-adapted eels than in FW-adapted eels. In contrast, water loss in SW was less in SW-adapted eels than in FW-adapted eels. Rectification of osmotic water fluxes was observed for both FW and SW-adapted eels, net water fluxes in the mucosal-serosal (m-s) direction being greater than those in the opposite (s-m) direction. These results indicate that adaptation to a given external medium brings about a decrease in the osmotic permeability so that water gain in FW or water loss in SW is minimal.