Organochlorine and organophosphorus pesticide concentrations in water,sediment, and selected organisms in Lake Naivasha (Kenya)

Water, sediment, red swamp crayfish (Procambarus clarkii) and black bass (Micropterus salmoides) from Lake Naivasha were analyzed for selected organochlorine and organophosphorus pesticide residues. The mean p,p'-DDT, o,p'-DDT and p,p'-DDE residue levels recorded in black bass (28.3 (± 30.0), 34.2 (±54.0) and 16.1 (±16.1) g kg^–1, respectively) and crayfish (4.6 (±5.1), 3.2 (±2.8), and 1.4 (±1.1) g kg^–1, respectively), were higher than previously recorded. This indicated recent usage of technical DDT in the lake's catchment. Levels of p,p'-DDT, higher than those of p,p'-DDE further emphasized this. Mean lindane, dieldrin, -endosulfan and aldrin concentrations in black bass were 100.5, 34.6, 21.6 and 16.7 g kg^–1, respectively. The same residues were detected at lower concentrations in crayfish at 2.0, 2.0, 2.0 and 1.9 g kg^–1, respectively. The higher fat content (3.7 ± 2.7% SD) in black bass (compared to 0.6 ± 0.3% in crayfish) accounted for the significantly higher residue concentrations in black bass. Organophosphate pesticides were the most commonly used pesticides in the lake's catchment, but none was detected in any of the samples. The results indicate that there is need for further work to identify sources and fate of pesticide contaminants, as well as to improve monitoring of pesticide use throughout the catchment.     

Fever in the leech,Nephelopsis obscura (Annelida)

Summary Leeches were placed in a 0.8 m linear temperature gradient from 0°C to 40°C. The temperature selected by the leeches was recorded over 30 min intervals; series of animals then received an injection of either pure sterile water,Escherichia coli endotoxin (LPS) at doses of 0.05, 0.25 and 10 g·g^–1, alcohol, or prostaglandin E₁ (PGE₁) at doses of 2 and 4 g·g^–1. After a latency of about 90 min, the leeches developed a dose-dependent fever in response to LPS. Maximal preferred temperature was near 29.6±1.4 whereas the control was 20.5±0.6°C. PGE₁ also evoked dose-dependent fevers of the same magnitude as LPS (30.0±1.0°C), but without latency. Another series of leeches received an injection of LPS of 10 g·g^–1 while in water containing 1.6 mg acetaminophen per 20 l water. Acetaminophen prevented the development of LPS fever. It is concluded that annelids respond to infection with a behavioral fever analogous to that of vertebrates.     

Caecal water and electrolyte absorption and the effects of acetate and glucose, in dehydrated, low-NaCl diet hens

Summary In vivo electrolyte transport and water absorption from the caeca of dehydrated, low-NaCl diet hens are reported. In the absence of luminal glucose or acetate, net electrolyte transport rates and water absorption are small. When physiological concentrations of acetate (40 mM) are included in the perfusate, Na⁺ transport and water absorption increase significantly (P<0.01): 38±7 eqNa⁺/caecum kg·h and 256±33 l H₂O/caecum · kg · h.A similar increase in water absorption occurs with the inclusion of 15 mM glucose in the perfusate (219±30 l H₂O/caecum · kg · h), however both net Na⁺ and Cl^– absorption increase: 28±6 eq Na⁺/caecum · kg · h and 21±5 eq Cl^–/caecum kg · h.These pronounced increases in electrolyte and water absorption are not accompanied by any significant increase in transmural potential difference.The data presented establish caeca as important sites in the recuperation of water and electrolytes in dehydrated, low-NaCl diet hens.Abbreviations ECPD electrochemical potential difference - PD (transmural) potential difference - PEG polyethylene glycol     

Effect of acute and chronic epinephrine administration on clearance and metabolism of [3H]-epinephrine in trout (Oncorhynchus mykiss)

Several studies have measured the rate of catecholamine clearance, metabolism, and tissue accumulation in fish. However, no information is available on the effect of repeated stress or high circulating catecholamine levels on catecholamine clearance and metabolism. We measured the clearance and metabolism of [³H]-epinephrine (approximately 0.1 g·kg^-1) in SW-acclimated rainbow trout subjected to acute (five injections in 1 day) and chronic (4 days; five injections per day) administration of 4.0 g·kg^-1 epinephrine or saline. In addition, a saturation experiment, where 4.0 g·kg^-1 of unlabelled epinephrine was injected concurrently with [³H]-epinephrine, investigated whether catecholamine clearance and metabolism are affected by high circulatin levels. Neither the rate constants for catecholamine clearance, nor the post-injection proportions of unmetabolised [³H]-epinephrine, deaminated [³H]-epinephrine and O-methylated [³H]-epinephrine were affected by the acute or chronic injection protocols. The concurrent injection of [³H]-epinephrine and 4.0 g·kg^-1 of unlabelled epinephrine resulted in an elevated postinjection ³H:¹⁴C ratio, but increased proportions of O-methylated [³H]-epinephrine and reduced proportions of unmetabolised [³H]-epinephrine. We conclude that in fish (1) catecholamine clearance and metabolism are unlikely to be compromised by repeated exposure to acute stressors; (2) catecholamine extraction and/or metabolism is enhanced when circulating levels are high; and (3) there is a marked capacity to rapidly (minutes) clear and inactivate catecholamines that are released in response to stressful stimuli.Abbreviations CA catecholamines - dpm disintegrations per minute - MAO monoamine oxidase - COMT catecholamine-O-methyltransferase - MOPEG 3-methoxy-4-hydroxyphenylglycol - VMA 3-methoxy-4-hydroxymendelic acid - SW seawater - HPLC high-performance liquid chromatography     

Plasma glucose kinetics and tissue uptake in brown trout in vivo: effect of an intravascular glucose load

The object of the present study was to elucidate whether a glucose load modifies glucose uptake by tissues in brown trout in vivo. By the use of 2-[1,2-³H]-deoxyglucose, plasma glucose disappearance rate and tissue glucose uptake were measured after an intraaortic glucose load of 500 mg·kg^-1 (glucose load group) and under normoglycemic conditions (control). We also attempted to determine whether fasting modifies the glucose load disposal (fasted glucose load group). The procedure used to calculate 2-deoxyglucose uptake by tissues was evaluated, and the levels of 2-deoxyglucose uptake were compared with those of 2-deoxyglucose phosphorylation. Uptake and phosphorylation rates were similar in all tissues, except in brain and heart. In all the groups glucose uptake rates were highest in spleen, kidney, brain and gills, and lowest in red muscle, heart and white muscle. However, white muscle was the main site of glucose uptake on a whole tissue basis. The glucose load led to strong, long-lasting hyperglycemia, in spite of the increases observed in plasma insulin levels and in glucose uptake rate by the whole body (control: 4.9 mol·min^-1·kg^-1; glucose load group: 6.5 mol·min^-1·kg^-1). This higher rate was due to the higher glucose uptake only in white and red muscles (four- and threefold, respectively). Fasting halved the uptake of glucose by both red and white muscles in the load condition. In consequence the use of exogenous glucose decreased with fasting (fasted glucose load group: 5.1 mol·min^-1·kg^-1), causing still longer hyperglycemia.Abbreviations bw body weight - 2DG 2-[1,2-³H]-deoxyglucose - 2DG-P 2-[1,2-³H]-deoxyglucose phosphate - dpm disintegrations per min - FGL fasted glucose load group - GL glucose load group - G-6-Pase glucose-6-phosphatase - LG L-[1-¹⁴C]-glucose - MS-222 3-aminobenzoic acid ethyl ester methanesulphonate salt     

Secretory capacity of the lachrymal salt gland of hatchling sea turtles,Chelonia mydas

Summary The lachrymal salt glands ofChelonia mydas were functional when hatchlings emerged from the nest. Osmotic concentrations up to 720 mosmol kg^–1 were recorded in spontaneously produced tears (salt gland secretions). When injected with a Na⁺ load (1500–2700 mol (100 g)^–1) newly emerged hatchlings produced tears ranging in osmotic concentration from 1000–1900 mosmol kg^–1 with Na⁺ secretion rates from single glands of 200–475 mol (100 g·h)^–1. In these circumstances the rate of sodium excretion, via the salt glands, was equivalent to the sodium content of 0.2 to 0.5 ml of sea water per hour. Since the apparent drinking rate of hatchlings within the first two days of entering sea water was approximately 0.5 to 1.7 ml per day, the excretion of Na⁺ imbibed by drinking is well within the secretory capacity of the lachrymal salt glands.In feeding hatchlings extraordinarily high Na⁺ secretion rates were induced by Na⁺ loading. Hatchlings which were loaded with Na⁺ by injection (1500–5400 mol (100 g)^–1) produced tears having osmotic concentrations between 1500 and >2000 mosmol kg^–1. The Na⁺ secretion rates from single glands were 750–4185 mol (100 g·h)^–1 with extremely high short term rates of 10700 mol (100 g·h)^–1 (50 mol min^–1 for 28 g hatchlings).In terms of gland mass the highest long term secretion rate translates into 21 mmol of Na⁺ per gram of salt gland per hour and is the highest secretion rate yet recorded for a reptilian salt gland. This rate is almost three times the highest rate recorded for sea snakes (8 mmol g·h^–1) and is similar to rates commonly observed in avian salt glands (25 mmol g·h^–1).Secretion by the lachrymal salt glands was initiated by increased blood concentrations of Na⁺ or K⁺, K⁺ being as effective as Na⁺ but with the composition of the teras being virtually unchanged compared to tears from Na⁺ stimulated hatchlings. Preliminary experiments indicated that secretion was not initiated by increased Cl^– concentration in the blood or by increased volume or osmotic concentration of the blood.Abbreviation O.P. osmotic pressure     

Sodium influx in isolated gills of the freshwater mussel,Ligumia subrostrata

Summary Isolated gills of the freshwater mussel,Ligumia subrostrata, accumulate Na from a pondwater bathing medium. The rate of Na transport by the isolated gill is 13.2±1.1 mol (g dry gill·10 min)^–1 which equals or exceeds the estimated Na transport rate of intact animals. Sodium influx is saturable with aV _max of 13.6±1.2 mol (g dry gill·10 min)^–1 and an affinity (K _s) of 0.17 mM Na/l. The isolated gills survive prolonged exposure to pondwater with a constant of 890 l O₂ (g dry gill·h)^–1 over a 4 h period. Sodium transport in the isolated gills is stimulated 80% above control values by 10^–4 M serotonin, 60% by 0.5 mM cAMP and 60% by 12.5 g/ml nystatin. Sodium influx is inhibited by 0.5 mM amiloride and 1 mM lithium.     

Mediation of serotonin-induced hyperventilation via 5-HT3-receptor in European eel Anguilla anguilla

The effects of serotonin (5-hydroxytryptamine) on ventilation were investigated by continuous measurements of intrabuccal pressure in unrestrained eel. Intravenous administration of 5-hydroxytryptamine (30 g·kg^-1) caused a large increase in ventilatory frequency (+100%) and amplitude (+140%). The 5-hydroxytryptamine-induced hyperventilation was blocked by the 5-HT₃-receptor antagonists metoclopramide (1.0 mg·kg^-1) or MDL72222 (1.0 mg·kg^-1), and was insensitive to the 5-HT_1/2-receptor antagonist methysergide (3.0 mg·kg^-1) and to the 5-HT₄-receptor antagonist DAU 6285 CL (3.0 mg·kg^-1). The hyperventilatory response to 5-hydroxytryptamine could be mimicked by the 5-HT₃ receptor agonist 1-phenylbiguanide (300 g·kg^-1). These results strongly implicate the 5-HT₃-receptor as the mediator of the 5-hydroxytryptamine-induced hyperventilation in eel.Abbreviations a.u. arbitrary units - 5-HT 5-hydroxytryptamine - SEM standard error of mean - VA ventilatory amplitude - VF ventilatory frequency - RBI 1-phenylbiguanide     

Decrease of polypeptides in the PS I antenna complex with increasing growth irradiance in the red alga Porphyridium cruentum

Thylakoids isolated from cells of the red alga Porphyridium cruentum exhibit an increased PS I activity on a chlorophyll basis with increasing growth irradiance, even though the stoichiometry of Photosystems I and II in such cells shows little change (Cunningham et al. (1989) Plant Physiol 91: 1179–1187). PS I activity was 26% greater in thylakoids of cells acclimated at 280 mol photons · m^–2 · s^–1 (VHL) than in cells acclimated at 10 mol photons · m^–2 · s^–1 (LL), indicating a change in the light absorbance capacity of PS I. Upon isolating PS I holocomplexes from VHL cells it was found that they contained 132±9 Chl/P700 while those obtained from LL cells had 165±4 Chl/P700. Examination of the polypeptide composition of PS I holocomplexes on SDS-PAGE showed a notable decrease of three polypeptides (19.5, 21.0 and 22 kDa) in VHL-complexes relative to LL-complexes. These polypeptides belong to a novel LHC I complex, recently discovered in red algae (Wolfe et al. (1994a) Nature 367: 566–568), that lacks Chl b and includes at least six different polypeptides. We suggest that the decrease in PS I Chl antenna size observed with increasing irradiance is attributable to changes occurring in the LHC I-antenna complex. Evidence for a Chl-binding antenna complex associated with PS II core complexes is lacking at this point. LHC II-type polypeptides were not observed in functionally active PS II preparations (Wolfe et al. (1994b) Biochimica Biophysica Acta 1188: 357–366), nor did we detect polypeptides that showed immunocross-reactivity with LHC II specific antisera (made to Chlamydomonas and Euglena LHC II).Abbreviations Bis-Tris bis(2-hydroxyethyl)imino-tris(hydroxymethyl)methane - DCPIP 2,6-dichlorophenol indophenol - -dm dodecyl--d-maltoside - HL high light of 150 mol photons · m^–2 · s^–1 - LGB lower green band - LHC I light-harvesting complex of PS I - LHC II light-harvesting complex of PS II - LL low light of 10 mol photons · m^–2 · s^–1 - ML medium light of 50 mol photons · m^–2 · s^–1 - MES 2-(N-morpholino) ethanesulfonic acid - P700 reaction center of PS I - PFD photon flux density - Trizma tris(hydroxymethyl)aminomethane - UGB upper green band - VHL very high light of 280 mol photons · m^–2 · s^–1     

Comparison of the effects of volatile anesthetics in varying concentrations on brain energy metabolism with brain ischemia in rats

The effects of varying concentrations and types of volatile anesthetics on neurochemical sequelae of brain ischemia were evaluated in the rat. Rats were assigned to treatment defined by a 3×3 design (anesthetic type and dose) with 5 rats/cell. Each group received halothane, enflurane, or isoflurane 0.5, 1.0, or 2.0 MAC (minimal alevolar concentration). This was followed by preischemic plasma glucose sampling, 5 min hypotension (30 mmHg) and 5 min decapitation cerebral ischemia. Preischemia plasma glucose increased with increasing anesthetic concentration and was highest in the isoflurane groups, varying from a low (±SD) of 7.19±1.79 mol/ml in the 0.5 MAC halothane group to a high of 12.68±3.65 mol/ml in the 2.0 MAC isoflurane group. End-ischemic brain lactate correlated with preischemic plasma glucose (r=0.5, =0.5). We conclude that increasing concentration of volatile anesthesia with iv phenylephrine blood pressure support produces higher levels of plasma glucose and brain lactate with cerebral ischemia.     

Effect of age on acetylcholinesterase and other hemolymph proteins in Aplysia

Summary Hemolymph was examined in young (ca. 86 days old), mature (ca. 163 days old), and old (ca. 294 days old) Aplysia for age-related changes in constituent proteins. In young, mature, and old animals protein concentrations were 1.6±0.27, 1.41±0.53, and 1.45±0.43 mg·ml^-1, respectively. The copper-containing respiratory protein, hemocyanin, measured by determining the copper concentration, was found to increase significantly from young (0.98±0.51 g·ml^-1) to mature (2.02±0.95 g·ml^-1) Aplysia, with little change between mature and old (1.92±0.43 g·ml^-1) animals. These findings were consistent with the results obtained when hemocyanin was directly measured by spectrophotometric absorption at 340 nm. Acetylcholinesterase (AChE) was present in the hemolymph of Aplysia. Its activity was highest in mature animals (3121±1627 units·mg^-1) and least in old animals (1463±599 units·mg^-1). Young animals had intermediate levels (2080±762 units·mg^-1). SDS-PAGE revealed a distinct pattern of protein bands for hemolymph from each age group; hemolymph from the young group contained six prominent protein bands with molecular weights (MW) from 13 to 300 kDa. Hemolymph of mature and old animals exhibited four and three prominent protein bands, respectively, with MW between 45 and 300 kDa. A prominent band at 97 kDa was present in samples from the mature group, but was faint in samples from the old group and absent in samples from the young group. Under non-denaturing conditions the hemolymph protein band patterns for each group differed from the others, thereby demonstrating that the age-dependent differences in the protein profiles are intrinsic to hemolymph in vivo. Isoelectric focusing of the hemolymph samples revealed that the proteins were all acidic (pI ca. 3.0–6.5). The hemolymph from the young differed from the other two groups in having an additional acidic protein (pI ca. 4.0). A possible link between age-related changes in hemolymph proteins and age-related changes in the nervous system is proposed.Abbreviations 2-ME 2-mercaptoethanol - AChE acetylcholinesterase - FMRFamide amidated tetrapeptide containing phenylalanine, methionine, arginine and phenylalanine - MW molecular weight - PAS periodic acid-Schiff - SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis - TRIS tris-(hydroxymethyl)-aminomethane     

Effects of Pb2+, Ni2+, Hg2+ and Se4+ on cultured cells. Analysis of uptake, toxicity and influence on radiosensitivity

Effects of Pb²⁺, Ni²⁺, Hg²⁺ and Se⁴⁺ on cultured human glioma U-343MG cells were investigated considering uptake, toxicity and, in combination with radiation, clonogenic cell survival. The cells were exposed to 0-100 m of the metals for a week before the evaluation. The tests showed a tendency to toxicity with 10 m nickel although not significant (P > 0.05). Selenium, lead and mercury exerted a significant toxicity (P < 0.05) at 2.5 m, 10 m and 1 m, respectively. To challenge the clonogenic cell survival capacity, the cells were irradiated with⁶⁰Co photons after being exposed to the highest nontoxic concentration of the different metals. The clonogenic cell survival tests, after irradiation, showed no significant change if the cells were exposed to 5 m nickel, 0.5 m selenium or 5 m lead compared with those not exposed. Mercury, 0.1 m, gave a relative reduction in survival compared with only irradiated cells of 58 ± 17%. Thus, only mercury affected the radiation-induced damage and/or repair. When exposed to the highest nontoxic concentrations of the different metals, the cultures did not display a significant uptake ratio (metal concentration ratio of exposed cells to control cells) of nickel (3.1 ± 3.3), only a small uptake ratio of selenium (4.0 ± 0.4), while there was a large uptake ratio of both lead (2.6 ± 1.7) x 10² and mercury (1.5 ± 0.2) x 10¹. The results indicated that nickel was neither especially toxic nor influenced the clonogenic cell survival after irradiation. Mercury was more toxic and also influenced the radiation sensitivity. Lead was taken up strongly but did not influence the radiation sensitivity. Selenium accumulated but gave no detectable effect on the radiation sensitivity.     

Biological tertiary treatment of urban wastewaters with chitosan-immobilizedPhormidium

Summary Chitosan:Phormidium aggregates (chitosan: algae=1:2, dry weight basis) were used as a biological tertiary treatment to remove the nitrogen (NH ₄ ⁺ , NO ₂ ^- , NO ₃ ^- ) and phosphorus (PO ₄ ^3- ) from a secondary effluent. In a batch system, 71 and 92% of P–PO ₄ ^3- were removed after 6 and 24 h, respectively. The orthophosphate removal rate was identical for all three concentrations of algae-chitosan tested (3.3, 4.6, 5.9 g d. wt.·l^-1), and was 90 g±2 g P–PO ₄ ^3- ·l^-1·h^-1, for a 90% removal. Under control conditions (chitosan flakes only added to the effluent) 73 and 78% of PO ₄ ^3- were removed after 6 and 24 h respectively. A 95% removal of inorganic nitrogen (NH ₄ ⁺ , NO ₂ ^- , NO ₃ ^- ) was attained after 4–6 h withPhormidium immobilized on chitosan flakes, as compared to 30% with chitosan flakes alone (5 g d. wt.·l^-1). The system gave a similar performance when operated semi-continuously over 5 days at a daily retention time of 1.0. In the presence of chitosan-immobilized algae, medium P–PO ₄ ^3- levels were reduced by 87.3%±6.4% after 24 h (61.1 g±7.0 g P·l^-1·h^-1). The reduction of inorganic nitrogen in the medium was 98% after 24 h (370 g±50 g N·l^-1·h^-1). In the presence of chitosan alone, some 60% orthophosphate removal was recorded, whereas no reduction of nitrogen was observed. Disappearance of orthophosphate was attributed to its co-precipitation with calcium released from the chitosan by abrasion. The presence of the algae protected the chitosan from abrasion andPhormidium directly assimilated the orthophosphate and inorganic nitrogen, thus reducing their levels in the effluent.     

Chloride cells and chloride exchange in the skin of a sea-water teleost,the shanny (Blennius pholis L.)

Summary The fine structure of the skin and its importance in chloride outfluxes were investigated in a sea-water teleost, the shanny (Blennius pholis L.).The epidermis is composed of three cells types: epithelial cells, mucous cells and chloride cells. These chloride cells typically contain a great number of mitochondria and an extensive agranular reticulum extending through the whole cell body. They open at the surface of the epidermis into an apical pit. An undifferentiated small cell is often observed near these chloride cells and probably corresponds to the adjacent chloride cell.The values of chloride outfluxes through the skin and the gills are respectively 5333±884 Eq·h^–1·kg^–1 and 4479±2521 Eq·h^–1·kg ^–1; n=6; t=13±0.5°C. Thus the ratio between skin chloride outflux and total chloride outflux is 64.7±9.3%.     

Cardiovascular and metabolic responses to noradrenaline in men acclimatized to cold baths

The purpose of this study was to see whether artificial acclimatization to cold would reduce the pressor response to noradrenaline (NA) as natural acclimatization has been shown to do, and whether it would induce nonshivering thermogenesis. Three white men were infused with NA at four dosage levels between 0.038 and 0.300 g·kg^–1·min^–1 (2–23 g·min^–1), before and after artificial acclimatization to cold and again 4 months later when acclimatization had decayed. Acclimatization was induced by ten daily cold (15°Q baths of 30–60 min followed by rapid rewarming in hot (38–42°C) water, and was confirmed by tests of the subjects responses to whole-body cooling in air. Three control subjects also underwent the first and third tests. Acclimatization substantially reduced the pressor response to NA at 0.150 and 0.300 g·kg^–1·min^–1, confirming earlier findings by the same technique in naturally acclimatized men, and its decay increased this response to beyond its initial levels (P<0.05 for both changes). Acclimatization did not change the response to NA of heart rate, subjective impressions, skin temperature of finger and toe, pulmonary ventilation, or plasma free fatty acids and ketone bodies. At no time did NA increase oxygen consumption, or increase skin temperature or heat flow over reported sites of brown fat. These findings would seem to show that acclimatization to cold reduces sensitivity to the pressor effect of NA but does not induce nonshivering thermogenesis, and that the reduced sensitivity is replaced by a hypersensitivity to NA when acclimatization decays.     

Effect of osmotic gradient on ADH-induced intramembranous particle aggregates in toad bladder

Summary Paired toad urinary bladders were prepared without or with an osmotic gradient (175 mosm) across them, stimulated for 2.5 (n=6), 5 (n=6), 30 (n=6) or 60 (n=6) min with ADH (20 mU/ml), and studied by freeze-fracture electron microscopy. Water permeability at these times was assessed in additional bladders (n=6 for each case) after tissue fixation according to the technique of Eggena. After both 60 and 30 min of ADH stimulation, the presence of a gradient compared with the absence of one was associated with fewer aggregates (242±35vs. 382±14 ×235 m^–2 at 60 min,P<0.01; 279±36vs. 470±51 ×235 m^–2 at 30 min,P<0.01) and lower water permeability (8.4±1.1vs. 18.8±1.8g×min^–1×cm^–1 ×mosm ^–1 at60min,P<0.005; 9.2±1.0vs. 22.0±2.1 g ×min^–1×cm^–2×mosm ^–1 at 30 min,P<0.001). In addition, with a gradient both maximum water permeability and maximum aggregate frequency were reached nearly together; a similar correspondence occurred without a gradient. We conclude that in the presence of an osmotic gradient both the ADH-associated aggregates and the water permeability response to ADH are prevented from reaching the higher levels observed in bladders not exposed to a gradient.     

Cryphodera kalesari,a new heteroderid nematode species from Haryana,India

Cryphodera kalesari n. sp., recorded on Colebrookea oppositifolia Sm. from the Kalesar forests (Haryana), India, is described. Females: swollen, saccate, with two to three lip annules, with vulval lips protruding. Second stage juveniles: length 384±19 m, spear=26.5±1 m, tail 46.5±6 m, hyaline portion 21.5±2.5 m, en face dorsal and ventral arcs of head skeleton bifurcate. Males: not found.     

Ion transport across leech integument

The dorsal skin of the leech Hirudo medicinalis was used for electrophysiological measurements performed in Ussing chambers. The leech skin is a tight epithelium (transepithelial resistance = 10.5±0.5 k· cm^-2) with an initial short-circuit current of 29.0±2.9 A·cm^-2. Removal of Na⁺ from the apical bath medium reduced short-circuit current about 55%. Ouabain (50mol·l^-1) added to the basolateral solution, depressed the short-circuit current completely. The Na⁺ current saturated at a concentration of 90 mmol Na⁺·l^-1 in the apical solution (K _M=11.2±1.8 mmol·l^-1). Amiloride (100 mol·l^-1) on the apical side inhibited ca. 40% of the Na⁺ current and indicated the presence of Na⁺ channels. The dependence of Na⁺ current on the amiloride concentration followed Michaclis-Menten kinetics (K _i=2.9±0.4 mol·l^-1). The amiloride analogue benzamil had a higher affinity to the Na⁺ channel (K _i=0.7±0.2 mol·l^-1). Thus, Na⁺ channels in leech integument are less sensitive to amiloride than channels known from vertebrate epithelia. With 20 mmol Na⁺·l^-1 in the mucosal solution the tissue showed an optimum amiloride-inhibitable current, and the amiloride-sensitive current under this condition was 86.8±2.3% of total short-circuit current. Higher Na⁺ concentrations lead to a decrease in amiloride-blockade short-circuit current. Sitmulation of the tissue with cyclic adenosine monophosphate (100 mol·l^-1) and isobutylmethylxanthine (1 mmol·l^-1) nearly doubled short-circuit current and increased amiloride-sensitive Na⁺ currents by 50%. By current fluctuation analysis we estimated single Na⁺ channel current (2.7±0.9 pA) and Na⁺ channel density (3.6±0.6 channels·m^-2) under control conditions. After cyclic adenosine monophosphate stimulation Na⁺ channel density increased to 5.4±1.1 channels·m^-2, whereas single Na⁺ channel current showed no significant change (1.9±0.2 pA). These data present a detailed investigation of an invertebrate epithelial Na⁺ channel, and show the similarities and differences to vertebrate Na⁺ channels. Whereas the channel properties are different from the classical vertebrate Na⁺ channel, the regulation by cyclic adenosine monophosphate seems similar. Stimulation of Na⁺ uptake by cyclic adenosine monophosphate is mediated by an increasing number of Na⁺ channels.Abbreviations slope of the background noise component - ADH antidiuretic hormone - cAMP cyclic adenosine monophosphate - f frequency - f _c coner frequency of the Lorentzian noise component - Hepes N-hydroxyethylpiperazine-N-ethanesulphonic acid - BMX isobutyl-methylxanthine - i _Na single Na⁺ channel current - I _Na max, maximal inhibitable Na⁺ current - I _SC short circuit current - K _i half maximal blocker concentration - K _M Michaelis constandard error of the mean - S _(f) power density of the Lorentzian noise component - S ₀ plateau value of the Lorentzian noise component - TMA tetramethylammonium - Trizma TRIS-hydroxymethyl-amino-methane - V _max maximal reaction velocity - V _T transepithelial potential - K half maximal blocker concentration     

Secretion of electrolytes,protein and urea by the mandibular gland of the common wombat (Vombatus ursinus)

Saliva was collected from the mandibular glands of anaesthetized common wombats (Vombatus ursinus) to ascertain maximal flow rates, salivary compostion and possible adaptations, particularly PO₄ ^3- secretion, to assist digestion. After temporary catheterization of the main duct through its oral opening, salivary secretion was evoked at flow rates ranging from 0.02±0.002 (±SEM) ml·min^-1 (0.7±0.07 l·min^-1·kg body weight^-1) to 0.4±0.05 ml·min^-1(14±1.9 l·min^-1·kg body weight^-1) by ipsilateral intracarotid infusion of acetylcholine. The [Na⁺] (15±5.1 to 58±8.6 mmol·l^-1) and [HCO₃ ^-] (35±1.9 to 60±1.9 mmol·l^-1) were positively correlated with salivary flow rate. The [K⁺] (58±5.2 to 30±2.4 mmol·l^-1), [Ca²⁺] (10.4±1.67 to 4.1±0.44 mmol·l^-1), [Mg²⁺] (0.94±0.137 to 0.17±0.032 mmol·l^-1), [Cl^-] (71±9.2 to 45±6.0 mmol·l^-1), [urea] (9.3±0.79 to 5.1±0.54 mmol·l^-1), H⁺ activity (29±1.6 to 17±1.6 nEq·l^-1) and amylase activity (251±57.4 to 92±23.3 kat·l^-1) were negatively correlated with flow. Both concentration and osmolality fell with increasing flow at the lower end of the flow range but osmolality always increased again by maximal flow whereas the relation between protein and flow was not consistent at the higher levels of flow and stimulation. Salivary [PO₄ ³⁺] was not correlated with flow and at 3–14% of the plasma concentration was extremely low. Thus, in contrast to its nearest relative, the koala (Phascolarctos cinereus), the wombat secretes little PO₄ ³⁺ presumably because it does not need high levels of PO₄ ³⁺ in its saliva to facilitate microbial digestion of plant fibre.Abbreviations bw body weight - ww wet weight     

The effect of carbonic anhydrase inhibitors on secretion by the parotid and mandibular glands of red kangaroos Macropus rufus

Summary The effects of carbonic anhydrase inhibitors on secretion by macropodine parotid and mandibular glands were investigated using anaesthetized red kangaroos. In the parotid gland, acetazolamide (500 mol·l^-1) reduced a stable acetylcholine-evoked, half-maximal flow rate of 2.02±0.034 to 0.27±0.023 ml·min^-1 (87% reduction). Concurrently, salivary bicarbonate concentration and secretion fell (129.4±1.46 to 80.9±1.63 mmol·l^-1 and 264.8±7.96 to 22.3±2.30 mol·min^-1, respectively), phosphate and chloride concentrations rose (14.0±0.79 to 27.6±0.85 mmol·l^-1 and 5.6±0.25 to 27.5±1.32 mmol·l^-1, respectively), sodium concentration and osmolality were unaltered, and potassium concentration fell (8.8±0.33 to 6.4±0.29 mmol·l^-1). High-rate cholinergic stimulation during acetazolamide blockade was unable to increase salivary flow beyond 11±0.9% of that for equivalent unblocked control stimulation. However, superimposition of isoprenaline infusion on the acetylcholine stimulation caused a three-fold increase in the blocked flow rate. These treatments were accompanied by small increases in salivary phosphate and chloride concentrations but not bicarbonate concentration. Methazolamide infusion caused similar changes in parotid secretion. In the mandibular gland, acetazolamide infusion had no effect on salivary flow rate during either low- or high-level acetylcholine stimulation. Acetazolamide caused no alterrations in salivary electrolyte secretion at low flow rates, but curtailed the rise in bicarbonate concentration associated with high-level acetylcholine stimulation. Acetazolamide administration did not affect the increase in salivary flow rate associated with isoprenaline infusion, but did block the concomitant increase in bicarbonate concentration and secretion substantially. It was concluded that neither cholinergic nor adrenergic stimulation of mandibular fluid secretion depends on secretion of bicarbonate derived from catalysed hydration of CO₂, but a substantial proportion of the increase in bicarbonate secretion during isoprenaline administration, which is probably ductal in origin, is so dependent. In contrast to other salivary glands, including the ovine parotid, fluid secretion by the kangaroo parotid gland during cholinergic stimulation is largely dependent (about 90%) on secretion of bicarbonate derived from hydration of CO₂ catalysed by glandular carbonic anhydrase. Fluid secretion during adrenergic stimulation is not bicarbonate dependent.Abbreviations b.w. body weight - PAH p-aminohippurate - PCO₂ partial pressure carbon dioxide - PCO₂ partial pressure of oxygen