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Although the determinants of sex differences in territoriality have been addressed for many species, the consequences of such differences in space use, particularly in southern hemisphere taxa, have received limited attention. The Namaqua rock mouse, Aethomys namaquensis, is a medium-sized, omnivorous, nocturnal murid with a wide distribution throughout southern Africa. Sex differences in space use behaviour were found: female captures were described by non-overlapping contiguous areas, whereas males were caught over 50% larger areas that overlapped spatially and temporally. Movements during breeding were characteristic of scramble-competition polygyny: a transient influx of non-resident males (coinciding with an increase in perforate females) skews the sex ratio towards males early in the breeding season. Whereas female condition improved with time, male body condition declined significantly over the breeding period (r²62 = 0.134, P = 0.002) and males had lower persistence in the trappable population. This study may therefore document a link between mobility (associated with a polygynous mating system) and reduced male survivorship and body condition for this murine rodent. [ABSTRACT FROM AUTHOR]

The paradox of secondary metabolites, toxic defence compounds produced by plants, in nectar and fruits is well known. Deterrence of feeding by nectarivorous and frugivorous birds is better understood than the effect of these chemicals on the digestive performance of birds. Digestive parameters such as transit time and sugar assimilation are important in assessing nutrient utilization and deterrence may be related to post-ingestive effects involving these parameters. Nectar and many fruits contain mainly sugars and water, and avian consumers compensate for low sugar content in their diet by increasing food intake: this may also increase their intake of secondary metabolites. We investigated how the alkaloid nicotine, naturally present in nectar of Nicotiana species, influences compensatory feeding and digestive performance of nectar-feeding birds. High nicotine concentration negatively affected compensatory feeding and apparent assimilation efficiency of white-bellied sunbirds Cinnyris talatala and Cape white-eyes Zosterops virens; but nicotine slowed gut transit time only in the latter species. In contrast, food intake and digestive performance of dark-capped bulbuls Pycnonotus tricolor was unaffected by nicotine up to a concentration of 50μM. Bulbuls are primarily frugivorous; hence, they are more exposed to secondary metabolites than sunbirds and possibly white-eyes. Because their diet is richer in toxins, frugivorous birds may have evolved more efficient detoxification strategies than those of specialist nectar-feeding birds., (Copyright © 2015 Elsevier Inc. All rights reserved.)

The effects of pesticides on honeybee larvae are less understood than for adult bees, even though larvae are chronically exposed to pesticide residues that accumulate in comb and food stores in the hive. We investigated how exposure to a plant alkaloid, nicotine, affects survival, growth and body composition of honeybee larvae. Larvae of Apis mellifera scutellata were reared in vitro and fed throughout development on standard diets with nicotine included at concentrations from 0 to 1000μg/100g diet. Overall mortality across all nicotine treatments was low, averaging 9.8% at the prepupal stage and 18.1% at the white-eyed pupal stage, but survival was significantly reduced by nicotine. The mass of prepupae and white-eyed pupae was not affected by nicotine. In terms of body composition, nicotine affected water content but did not influence either protein or lipid stores of white-eyed pupae. We attribute the absence of consistent negative effects of dietary nicotine to detoxification mechanisms in developing honeybees, which enable them to resist both natural and synthetic xenobiotics., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Protein-rich diets are known to promote ovarian and egg development in workers of the honeybee, Apis mellifera, even in the presence of a queen. Since the main source of protein for honeybees is pollen, its quality and digestibility might be important dietary factors determining reproductive capacity. We have compared the effect of two types of pollen-sunflower, Helianthus annuus, and aloe, Aloe greatheadii var davyana-on ovarian development in A. mellifera scutellata workers. Under queenright conditions in the field, worker bees exhibited greater ovarian development when feeding on aloe pollen than on sunflower pollen. In their midgut, we observed higher extraction efficiency for aloe (80%) than for sunflower (69%) pollen. This may be attributed to the morphology and size of the two kinds of pollen grains and explains, together with the high protein content of aloe pollen (32% dry mass in bee-collected pollen) compared to sunflower pollen (15%), why aloe pollen promoted higher ovarian development. However, in the laboratory workers sustained on aloe pollen had significantly less-developed ovaries and higher mortality than those fed sunflower pollen. These detrimental effects may be due to an unbalanced protein:carbohydrate ratio. We discuss the effects of unbalanced diets on the physiology and ecology of honeybee reproduction.

This paper reports the effects of different diuretic factors on the Malpighian tubules of beetles. Calcitonin (CT)-like peptides from silkmoth and mosquito increase fluid secretion in a dose-dependent manner in the tubules of Tenebrio molitor, but the cockroach CT-like peptide, Dippu-DH(31), has no effect. Thapsigargin induces a small but significant increase in tubule secretion rates. The interactions between different factors in mealworm tubules were explored by testing CT-like peptides, thapsigargin and the mealworm CRF-related diuretic factor Tenmo-DH(37) in various combinations, but no synergistic effects were observed. C-terminal fragments of the CRF-related diuretic peptides Locmi-DH(46) and Dippu-DH(46) fail to increase fluid secretion in mealworm tubules, unlike their corresponding whole peptides. Cross-reactivity of factors between beetle species was investigated using the scarabaeid Onthophagus gazella. Tenmo-DH(37) increases fluid secretion in isolated tubules of O. gazella in a dose-dependent manner, revealing a high degree of cross-reactivity in this distantly related beetle species. However, homogenates of O. gazella brains inhibited fluid secretion in mealworm tubules.

The main objective of this study was to determine whether or not the renal outputs of the osmoregulatory hormones arginine vasotocin (AVT) and aldosterone (ALDO) reflect the osmotic status of whitebellied sunbirds (Nectarinia talatala). The birds were fed a range of sucrose concentrations (from 0.07 to 2.5 mol/l, with osmolalities of 70 to approximately 5,800 mosM/kg), and adjusted their intakes so that they drank large volumes of dilute diets and small volumes of concentrated diets. Renal fluid outputs were appropriately regulated so that large volumes of cloacal fluid (CF) were voided on the dilute diets and small volumes on the concentrated diets. Accordingly, plasma AVT concentrations increased with increasing sugar concentration; however, AVT outputs in CF did not change in a similar manner, rather they decreased as dietary concentration increased. It was not possible to measure plasma ALDO concentrations in the small sunbirds because of insufficient blood samples available; however, ALDO outputs in CF did vary with the sucrose diets and renal function, being highest on the most concentrated diet. In addition ALDO output in CF fell markedly when sodium was added to the 0.5 mol/l sucrose diet. We conclude that in sunbirds fed increasingly concentrated sucrose solutions, changes in CF outputs of ALDO, but not AVT, appear to reflect the water flux and hydration state of these birds.

Gut sterilization via the oral administration of antibiotics facilitates physiological studies of the nutritionally important relationship between intestinal microflora and the host. However, the composition of gut flora is extremely variable, and as a result, the efficacy of antibiotics in achieving gut sterilization varies considerably between species. We tested the effectiveness of three antibiotic cocktails in sterilizing the gut of a rodent pollinator, the Namaqua rock mouse (Aethomys namaquensis). The cocktails were (1) streptomycin sulfate and bacitracin (previously used with domestic mice and rats), (2) chloramphenicol and bacitracin (based on antibiotic screening tests performed on faecal flora) and (3) Baytril 10% oral solution (a veterinary antimicrobial agent containing enrofloxacin). We tested for antibiotic inactivation by determining bacterial viability through fluorescence staining of faecal samples. We also tested techniques to maintain sterility during antibiotic treatment without the benefit of a laminar flow cabinet. Antibiotics were administered orally in food and water consumed ad libitum over 4 consecutive days. Antibiotic effectiveness was assessed by culturing anaerobic bacteria from faecal samples collected before and after each antibiotic treatment. Treatment with Baytril 10% oral solution eliminated or significantly reduced faecal flora, whereas other antibiotics did not. This study clearly demonstrates the importance of testing the effectiveness of antibiotics before their use in studies that involve antibiotic-treated subjects, particularly if these are species previously untested.

Nectarivores may be required to switch between water conservation and water excretion as rapidly as they change food plants in nature. We examined the rehydration response in Whitebellied Sunbirds (Nectarinia talatala) that had been fed a concentrated sucrose diet (2.5 mol/l) for 2 days and then were switched to a diet 10 x less concentrated (0.25 mol/l) on the morning of the third day. We measured water gain as well as cloacal fluid (CF) volume hourly over 12 h, and analysed CF osmolality and calculated osmotic excretion. CF was also assayed for the osmoregulatory hormone aldosterone (ALDO). As in most water-deprived birds, whitebellied sunbirds cope with water shortage when fed a concentrated sugar diet by almost completely ceasing to void CF. Although osmolality of CF is high, volumes are not sufficient to maintain a steady rate of excretion and the birds retain osmolytes. Immediately upon switching to dilute diet, sunbirds produce copious volumes of CF and osmotic excretion is elevated and maintained at high levels over the first 6 h of rehydration. This stabilises by the afternoon at levels expected for hydrated birds. Some 2-3 h after peak osmotic excretion, there is a peak in the discrepancy between water intake and output, mirrored in an increase in ALDO output. These data suggest that excretion of retained osmolytes is undertaken as soon as water is available, with changes in the body fluid composition occurring subsequently. This study vindicates the use of CF to obtain repeated measurements of changes in the osmoregulatory steroid ALDO in small birds.

The Malpighian tubules of Tenebrio molitor provide a model system for interpreting the actions of endogenous diuretic and antidiuretic peptides. The effects of diuretic (Tenmo-DH(37)) and antidiuretic (Tenmo-ADFa) peptides and their respective second messengers (cyclic AMP and cyclic GMP) on basolateral (V(bl)) and transepithelial (V(te)) potentials of Tenebrio Malpighian tubules were determined using conventional microelectrodes. In the presence of 6 mmol l(-1) Ba(2+), Tenmo-DH(37) (100 nmol l(-1)) reversibly hyperpolarized V(bl) and depolarized V(te). A similar response was seen with the addition of 1 mmol l(-1) cyclic AMP; however, the apical membrane potential (V(ap)) then showed a hyperpolarization, whereas a depolarization of V(ap) was observed with Tenmo-DH(37). Bafilomycin A(1) (5 micromol l(-1)) inhibited fluid secretion of stimulated tubules and reversed the hyperpolarization of V(bl) in response to Tenmo-DH(37). In response to 100 nmol l(-1) Tenmo-ADFa or 1 mmol l(-1) cyclic GMP, V(bl) and V(te) depolarized, although cyclic GMP affected membrane potentials somewhat differently by causing an initial hyperpolarization of V(bl) and V(te). In high [K(+)]-low [Na(+)] Ringer, 1 mmol l(-1) amiloride decreased fluid secretion rates, and depolarized both V(bl) and V(te). Amiloride significantly decreased luminal pH in paired experiments, indicating the presence of a K(+)/nH(+) exchanger in tubule cells of Tenebrio. The results suggest that the endogenous factors and their second messengers stimulate/inhibit fluid secretion by acting on the apical V-ATPase, basolateral K(+) transport, and possibly Cl(-) transport.

We investigated the mechanism and efficiency of digestion of two types of pollen, maize, Zea mays, and sunflower, Helianthus annuus, by the spotted maize beetle, Astylus atromaculatus (Melyridae). We found similar and high extraction efficiencies, but different mechanisms of digestion. Osmotic shock was apparently involved in digestion of the large and thin-walled maize pollen grains. In the anterior midgut most maize pollen grains were already ruptured, in contrast with the intact exines of sunflower pollen, which suggests another mechanism of digestion for the latter, such as enzymatic action. We investigated the effect of osmotic shock on maize pollen in vitro by looking at the behavior of pollen grains in varying osmotic concentrations. Maize pollen grains burst in both distilled water and sugar solutions of various concentrations, and the amount of rupturing decreased with an increase in sugar concentration. Digestion of maize pollen was much slower in honeybees than in spotted maize beetles. Maize pollen bursts early in the midgut of maize beetles, but remains intact in honeybees: this suggests that osmotic shock may not be as important for honeybees as previously suggested.

Malpighian tubules of the mealworm Tenebrio molitor were isolated for intracellular measurement of basolateral (V(bl)) and, indirectly, apical (V(ap)) membrane potentials. In control Ringer (50 mmol l(-1) K(+), 140 mmol l(-1) Na(+)), V(bl) was 24 mV, cell negative, and V(ap) was 48 mV, cell negative with reference to the lumen. Ion substitution experiments involving K(+) and Na(+) indicated that both V(bl) and V(ap) were sensitive to the bathing K(+) concentration, with the change in V(ap) being 60-77% that of V(bl). A 10-fold drop in bath [K(+)] irreversibly decreased fluid secretion rates from 6.38+/-0.95 nl x min(-1) (mean +/- S.E.M.) to 1.48+/-0.52 nl x min(-1) (N=8). In the presence of 6 mmol l(-1) Ba(2+), a blocker of basal K(+) channels, fluid secretion rates reversibly decreased and the hyperpolarization of both V(bl) and V(ap) seen in 50 mmol l(-1) and 140 mmol l(-1) K(+) indicated a favourable electrochemical gradient for basal K(+) entry. In 5 mmol l(-1) K(+), Ba(2+) induced two different responses: V(bl) either hyperpolarized by approximately 10 mV or depolarised by approximately 14 mV, according to the electrochemical gradient for K(+), which was either inward or outward in low bath [K(+)]. Rubidium, a 'permeant' potassium substitute, caused a hyperpolarization of V(bl), indicating the specificity of K(+) channels found in Tenebrio tubule cells. Other possible K(+) uptake mechanisms located in the basolateral membrane were investigated. Blocking of the putative electroneutral Na(+)/K(+)/2Cl(-) cotransporter by 10 micromol l(-1) bumetanide reversibly decreased fluid secretion rates, with no detectable change in membrane potentials. Ouabain (1 mmol l(-1)), an Na(+)/K(+)-ATPase inhibitor, irreversibly decreased fluid secretion rates but had no effect on electrical potential differences either in the absence or presence of Ba(2+). The results implicate K(+) channels, the Na(+)/K(+)/2Cl(-) contransporter and the Na(+)/K(+)-ATPase in basal K(+) and fluid transport of Tenebrio tubule cells.

Although the mealworm Tenebrio molitor inhabits very dry environments, it has at least two diuretic peptides, which increase fluid secretion by the free portions of the Malpighian tubules. Unlike other insect corticotropin-releasing factor (CRF)-related peptides isolated to date, these are non-amidated peptides. The immunocytochemical localization of Tenmo-DH(37) was investigated using antisera raised against this hormone. Immunoreactive neurosecretory cells were found in the brain and abdominal ganglia with immunoreactive processes projecting to the peripheral nervous system. Intense staining of the neurohaemal release site, the corpora cardiaca, was observed. In addition, neurosecretory cells immunoreactive to Tenmo-DH(37) were found in the posterior midgut and a network of immunoreactive nerve processes extended over the surface of the midgut. Tenmo-DH(37) is widely distributed and its staining pattern resembles that found for other, amidated CRF-related diuretic peptides.

Fluid secretion by insect Malpighian tubules is controlled by haemolymph-borne factors. The mealworm Tenebrio molitor provides the first known example of antagonistic interactions between endogenous neuropeptides acting on Malpighian tubules. The two corticotropin-releasing-factor (CRF)-related diuretic peptides previously isolated from Tenebrio molitor, Tenmo-DH(37) and Tenmo-DH(47), were found to stimulate Tenebrio molitor tubules in vitro in a dose-dependent manner with EC(50) values of 0.12 nmol l(-1) and 26 nmol l(-1) respectively. However, no synergistic or additive effect was observed when these two peptides were tested simultaneously. We then investigated antagonism between second messengers: dose-response curves were constructed for stimulation of Tenebrio molitor tubules by cyclic AMP and their inhibition by cyclic GMP. When both cyclic nucleotides were included in the bathing Ringer, the stimulatory effect of cyclic AMP was neutralised by cyclic GMP. Similarly, the stimulatory effect of Tenmo-DH(37) was reversed on addition of an antidiuretic peptide (Tenmo-ADF), which was recently isolated from Tenebrio molitor and acts via cyclic GMP. The cardioacceleratory peptide CAP(2b), originally isolated from Manduca sexta, also increases intracellular cyclic GMP levels and inhibited fluid secretion by Tenebrio molitor tubules, with an EC(50) value of 85 nmol l(-1). This inhibitory effect was reversed by Tenmo-DH(37). Endogenous diuretic and antidiuretic peptides, effective at low concentrations and acting via antagonistic second messengers, have the potential for fine control of secretion rates in the Malpighian tubules of Tenebrio molitor.