Gastrointestinal and renal responses to variable water intake in whitebellied sunbirds and New Holland honeyeaters

Nectarivores face a constant challenge in terms of water balance, experiencing water loading or dehydration when switching between food plants or between feeding and fasting. To understand how whitebellied sunbirds and New Holland honeyeaters meet the challenges of varying preformed water load, we used the elimination of intramuscular-injected [14C]-L-glucose and 3H2O to quantify intestinal and renal water handling on diets varying in sugar concentration. Both sunbirds and honeyeaters showed significant modulation of intestinal water absorption, allowing excess water to be shunted through the intestine when on dilute diets. Despite reducing their fractional water absorption, both species showed linear increases in water flux and fractional body water turnover as water intake increased (both afternoon and morning), suggesting that the modulation of fractional water absorption was not sufficient to completely offset dietary water loads. In both species, glomerular filtration rate was independent of water gain (but was higher for the afternoon), as was renal fractional water reabsorption (measured in the afternoon). During the natural overnight fast, both sunbirds and honeyeaters arrested whole kidney function. Evaporative water loss in sunbirds was variable but correlated with water gain. Both sunbirds and honeyeaters appear to modulate intestinal water absorption as an important component of water regulation to help deal with massive preformed water loads. Shutting down glomerular filtration rate during the overnight fast is another way of saving energy for osmoregulatory function. Birds maintain osmotic balance on diets varying markedly in preformed water load by varying both intestinal water absorption and excretion through the intestine and kidneys

Mistletoebirds and xylose: Australian frugivores differ in their handling of dietary sugars

Carbohydrate-rich mistletoe fruits are consumed by a wide range of avian species. Small birds absorb a large portion of water-soluble nutrients, such as glucose, via the paracellular pathway. d-xylose, a pentose monosaccharide, is abundant in some nectars and mistletoe fruits consumed by birds, and it has been suggested that it is most likely absorbed via the paracellular pathway in birds. We measured apparent assimilation efficiency () and bioavailability (f) for d-xylose and d- and l-glucose in three frugivorous Australian bird species. Mistletoebirds, silvereyes, and singing honeyeaters showed significantly lower for d-xylose than for d-glucose. Across two diet sugar concentrations, silvereyes and singing honeyeaters significantly increased f of both l-glucose (a metabolically inert isomer of d-glucose commonly used to quantify paracellular uptake) and d-xylose on the more concentrated diet, probably because of increased gut processing time. By contrast, mistletoebirds (mistletoe fruit specialists) did not vary f of either sugar with diet concentration. Mistletoebirds also showed higher f for d-xylose than l-glucose and eliminated d-xylose more slowly than silvereyes and singing honeyeaters, demonstrating differences in the handling of dietary xylose between these species. Our results suggest that d-xylose may be absorbed by both mediated and nonmediated mechanisms in mistletoebirds

Sucrose digestion capacity in birds shows convergent coevolution with nectar composition across continents

The major lineages of nectar-feeding birds (hummingbirds, sunbirds, honey-eaters, flowerpiercers, and lorikeets) are considered examples of convergentevolution. We compared sucrose digestion capacity and sucrase enzymatic activ-ity per unit intestinal surface area among 50 avian species from the New World,Africa, and Australia, including 20 nectarivores. With some exceptions, nectari-vores had smaller intestinal surfaces, higher sucrose hydrolysis capacity, andgreater sucrase activity per unit intestinal area. Convergence analysis showedhigh values for sucrose hydrolysis and sucrase activity per unit intestinal surfacearea in specialist nectarivores, matching the high proportion of sucrose in thenectar of the plants they pollinate. Plants pollinated by generalist nectar-feedingbirds in the Old and New Worlds secrete nectar in which glucose and fructose arethe dominant sugars. Matching intestinal enzyme activity in birds and nectarcomposition in flowers appears to be an example of convergent coevolution be-tween plants and pollinators on an intercontinental scale.Todd J. McWhorter, Jonathan A. Rader, Jorge E. Schondube, Susan W. Nicolson, Berry Pinshow, Patricia A. Fleming, Yocelyn T. Gutie, rrez-Guerrero, and Carlos Martı, nez del Ri

A comparison of pharmacokinetic methods for in vivo studies of nonmediated glucose absorption

Two pharmacokinetic methods are used primarily to assess systematic bioavailability of orally dosed water-soluble compounds in vivo, but there have been no direct comparisons of the estimates obtained. The "area under the curve" (AUC) method employs a single oral dose of probe compound(s) followed by multiple blood sampling to obtain plasma concentration time curves. Separate injection of probe(s) followed by multiple blood sampling is used to calculate fractional elimination rate (K-el) and distribution pool space (S). The "steady state feeding" method relies on ad lib. feeding of a marked diet, with a single blood sample taken to measure steady state feeding concentration of probe(s); K-el is estimated from the decline in probe concentration in excreta after injection, with a single blood sample taken to estimate S. We compared these methods directly in the Australian red wattlebird (Anthochaera carnunculata), measuring absorption of H-3-L-glucose. The K-el values estimated using the steady state feeding protocol were significantly higher, and estimates of S and bioavailability consequently lower, compared with the AUC protocol. The AUC method relies on fewer assumptions and allows simultaneous comparisons of absorption by mediated and nonmediated (i.e., paracellular) mechanisms but cannot be easily applied to freely feeding animals. The steady state feeding method allows work with smaller species and exploration of the effects of feeding on nutrient uptake but requires careful attention to the validity of assumptions that increase error in the calculations

A welfare approach for captive wild birds

Working with captive wild birds presents researchers with a multitude of challenges. Not least of these is appropriate cage size. Previous studies have highlighted some AEC concerns in this area. Our AEC has worked with a research group to ensure improved outcomes for captive wild birds in a specific study as well as for future studies. This involved the redesign of an outdoor aviary for the latest cohort of birds (n=8). The re-design includes 8 individual aviaries with sufficient space to allow flight for small birds (<150 g). The birds have been taught to feed in smaller cages within the aviaries so that they are easily re-caught and can be handled for the research. The capacity to reduce the aviary size for trial participation has also been incorporated, allowing researchers to conduct experiments with minimal handling of the birds. Current occupants (Silvereyes, ~10 g) appear to have adapted well. The AEC has also endeavoured to set some guidelines for the time space between the various components of the research so that the birds are provided with time frames free from research interaction in the aviaries. The student researcher has been proactive in including remote monitoring through cameras as well as through nearby windows, and has recently implemented a remote design to close the smaller cages. This session will discuss the process and evaluate its outcomes to date

Sugar preferences of avian nectarivores are correlated with intestinal sucrase activity

Nectar-feeding birds generally demonstrate preference for hexose solutions at low sugar concentrations, switching to sucrose/ no preference at higher concentrations. Species vary in the concentration at which the switch from hexose preference occurs; this could reflect physiological constraints that would also influence nectar selection when foraging. We recorded concentration- dependent sugar type preferences in three opportunistic/ generalist Australian nectarivorous species: Dicaeum hirundinaceum, Zosterops lateralis, and Lichenostomus virescens. All three preferred hexoses up to sugar concentrations of 0.25 mol L-1 and switched to sucrose/no preference for higher concentrations. Using these and literature records, we investigated physiological mechanisms that may explain the concentration dependence of sugar type preferences and compared diet preference data with foraging records.We measured sucrase activity in Z. lateralis and L. virescens as well as three specialized nectarivorous species (Anthochaera carunculata, Phylidonyris novaehollandiae, and Trichoglossus haematodus) for comparison with published concentration-dependent sugar preference data. Sucrase activity varied between these species (P = 0.006). The minimum diet concentration at which birds show no sugar preference was significantly correlated with sucrase activity for the 11 species analyzed (P = 0.005). Birds with the lowest sucrase activity showed hexose preference at higher diet concentrations, and birds with the greatest sucrase activity showed either no hexose preference or hexose preference on only the most dilute diets. Foraging data compiled from the literature also support the laboratory analyses; for example, T. haematodus (preference for hexose over a wide range of diet concentrations, low sucrase activity) also feed primarily on hexose nectars in the wild. Intestinal sucrase activity is likely to contribute to diet selectivity in nectarivorous bird species

Paracellular nutrient absorption in a gum-feeding new world primate, the common marmoset Callithrix jacchus

The common marmoset is one of the few callitrichid species that is not threatened or endangered in the wild, and is widely used in biomedical research, yet relatively little is understood about its digestive physiology. Dietary specialization on plant exudates has lead to relatively reduced small intestines, yet the common marmoset has exceptional dietary breadth, allowing it to successfully utilize a variety of habitats. We predicted that passive, paracellular nutrient absorption would be used by the common marmoset to a greater extent than in other non-flying mammals. We measured the bioavailability and rates of absorption of two metabolically inert carbohydrates not transported by mediated pathways (L-rhamnose and cellobiose, molecular masses of 164 and 342, respectively) to measure paracellular uptake, and of a non-metabolized D-glucose analog (3-O-methyl-D-glucose) to measure total uptake by both mediated and paracellular pathways. We found high bioavailability of 3-O-methyl-D-glucose (83 ± 5%), and much higher bioavailability of the paracellular probes than in similarly sized non-flying mammals (30 ± 3% and 19 ± 2% for L-rhamnose and cellobiose, respectively). Passive, paracellular nutrient absorption accounts for around 30% of total glucose absorption in common marmosets and intestinal permeability is significantly higher than in humans, the only other species of primate measured to date. This may allow the common marmoset to maintain high digestive efficiency when feeding on higher quality foods (fruit, arthropods, gums with higher proportions of simple sugars), in spite of relatively reduced small intestines correlated with adaptations for fermentative digestion of plant gums. We find no evidence to support, in primates, the hypothesis that reliance on paracellular nutrient absorption should increase with body size in mammals, but suggest instead that it may be associated with small body size and/or taxon-specific adaptations to diet

Mechanism and rate of glucose absorption differ between an Australian honeyeater (Meliphagidae) and a lorikeet (Loriidae)

Efficient mechanisms of glucose absorption are necessary for volant animals as a means of reducing mass during flight: they speed up gut transit time and require smaller volume and mass of gut tissue. One mechanism that may be important is absorption via paracellular (non-mediated) pathways. This may be particularly true for nectarivorous species which encounter large quantities of sugar in their natural diet. We investigated the extent of mediated and non-mediated glucose absorption in red wattlebirds Anthochaera carunculata (Meliphagidae) and rainbow lorikeets Trichoglossus haematodus (Loriidae) to test the hypothesis that paracellular uptake accounts for a significant proportion of total glucose uptake in these species. We found that routes of glucose absorption are highly dynamic in both species. In lorikeets, absorption of L-glucose (non-mediated uptake) is slower than that of D-glucose (mediated and non-mediated uptake), with as little as 10% of total glucose absorbed by the paracellular pathway initially (contrasting previous indirect estimates of ∼80%). Over time, however, more glucose may be absorbed via the paracellular route. Glucose absorption by both mediated and non-mediated mechanisms in wattlebirds occurred at a faster rate than in lorikeets, and wattlebirds also rely substantially on paracellular uptake. In wattlebirds, we recorded higher bioavailability of L-glucose (96±3%) compared with D-glucose (57±2%), suggesting problems with the in vivo use of radiolabeled D-glucose. Further trials with 3-O-methyl-D-glucose revealed high bioavailability in wattlebirds (90±5%). This non-metabolisable glucose analogue remains the probe of choice for measuring uptake rates in vivo, especially in birds in which absorption and metabolism occur extremely rapidly

Mechanistic bases for differences in passive absorption

Increasing evidence indicates that small birds have more extensive non-mediated, paracellular intestinal absorption of hydrosoluble compounds than do mammals, although studies have not employed uniform methodologies or demonstrated differences at the tissue level. The mechanistic bases behind apparent species differences are poorly understood. We show using uniform methodology at the whole-animal level that intact, unanesthetized pigeons had significantly higher absorption of L-arabinose and L-rhamnose, two water-soluble compounds used to measure paracellular absorption, than similarly sized laboratory rats. The species differences were also evident using perfused isolated loops of duodenum, showing that the difference in paracellular absorption occurred at the tissue level, even when D-glucose absorption rates (transcellular+paracellular) were similar between the two species. The greater absorption of these probes in pigeons could not be explained by mediated uptake of the putative paracellular probes, or by increased nominal surface area, increased villus area or increased number of tight junctions. Rats and pigeons had comparable absorption of larger probes, which is consistent with similar effective pore size of the tight junction between enterocytes. The elimination of these mechanistic explanations might suggest that pigeon intestine has relatively higher paracellular solvent drag, but pigeon duodenal segments did not have higher net water absorption than rat duodenal segments. Whatever the exact mechanism(s), the paracellular pathway of both species limits substantial (>5%) fractional absorption to molecules smaller than about 4.8 Å (Mr ca. 350), and permeability to smaller molecules at the tissue level is higher in pigeons than in rats