Evaluation of lead exposition length on the liver and intestinal histoarchitecture of eared dove (Zenaida auriculata).

Lead (Pb) is a metal that can produces irreversible damage in living organisms. Some studies had reported that Pb produces histophysiological alterations in the digestive system (mainly liver) of birds; however, the effect of this metal on small intestine has not been fully examined. Additionally, little information is available on Pb disturbances in native birds of South America. The present study aimed to evaluate the effect of different Pb exposure times on blood δ-aminolevulinic acid dehydratase (δ-ALAD) activity and on the histological and morphometric characteristics of the digestive system (liver and proximal intestine) of eared doves (Zenaida auriculata). A decrease of the blood δ-ALAD activity, dilatation of blood vessels and leukocyte infiltrates in intestinal submucosa and muscular layers, and reduction of the enterocyte nuclear diameter and Lieberkühn crypts area were observed. In liver were noted steatosis, proliferation of bile ducts, dilated sinusoids, leukocyte infiltrates, and melanomacrophage centers. The portal tract area and the thickness of the portal vein wall were increased. In conclusion, the results showed that Pb produces histological and morphometric alterations on the liver and small intestine according to the exposure time, which should be considered when the dangerousness of environmental pollutants is evaluated in wild animals.

Effects of corticosterone administration on the body condition and blood parameters of the house sparrow, Passer domesticus.

Glucocorticoids are steroid hormones produced by the adrenal glands and released into the bloodstream in response to stressful situations. In birds, corticosterone (CORT) is the main glucocorticoid released under stress. Throughout their lives, animals in nature are continually exposed to noxious stimuli known as stressful events. Any alteration of homeostatic stability is a stressful situation and this alteration triggers physiological changes to restore homeostasis. Glucocorticoids are one of the components of the complex set of physiological and behavioral responses to stress. In this work, we use CORT supplied in drinking water to evaluate its effect in blood parameters and physiology in house sparrows (Passer domesticus). To accomplish this, P. domesticus were administered three different doses of CORT in drinking water for 72 h. Body mass, organ mass, pectoral muscle mass, leg muscle mass, and blood parameters (CORT, triglycerides, glucose and uric acid, heterophils/lymphocytes ratio, hematocrit, and serum protein profile) were determined before and after CORT treatment. A 15% decrease in body mass with a significant decrease in pectoral mass were observed after the higher CORT treatment, as well as a decrease in the plasma concentration of uric acid. Lastly, we found a reversal of the heterophils/lymphocytes ratio and a decrease in hematocrit. It was possible to establish first data for baseline and CORT-alteration values in serum protein profile for P. domesticus. Baseline and altered values of blood parameters and serum protein profile could be an important tool in field ecology because they provide important data to assess the physiological condition in wild birds.

Daily expression of sodium-dependent glucose cotransporter-1 protein in jejunum during rat ontogeny.

It is widely known that intestinal capacities such as the enzymatic hydrolysis of carbohydrates, lipids and proteins, and the subsequent absorption of the hydrolyzed products, are evolutionary matched to dietary loads and feeding behaviors. In this study, we demonstrate that the protein expression of apically located sodium-dependent glucose cotransporter-1 (SGLT-1) throughout rat ontogeny is daily adjusted to afford glucose uptake when the load of this metabolically essential monosaccharide in the intestinal lumen is maximum. The jejunal expression of SGLT-1 protein in 14 one-day-old suckling pups was found to increase at dark and early light phase (P < 0.05), when they have a better access to mother milk. In weaning 21-d-old and juvenile 28-d-old rats, the cotransporter expression was high throughout the entire day (P < 0.05). Finally, adult 90-d-old rats showed a well-developed circadian rhythm for SGLT-1 protein (P < 0.05), whose expression increased at late light and dark phase when the highest intestinal glucose load was achieved. To our knowledge, these results are the first reporting the daily profile of SGLT-1 expression during rat early developmental stage and may contribute to understand the biological significance of a well-established molecular capacity to deal with the crucial increase of glucose load in the diet during the weaning process.

House sparrow biomarkers as lead pollution bioindicators. Evaluation of dose and exposition length on hematological and oxidative stress parameters.

House sparrows (Passer domesticus) have been proposed as a key ecological indicator of urban pollution. Remarkably, we lack knowledge about the physiological effects of lead on this bird species. Therefore, this study was aimed to evaluate the effect of Pb on several physiological parameters in house sparrows exposed to environmental Pb concentrations. In a first experiment, birds were exposed to Pb sub-lethal doses (from 1.3 to 14.0 µg of Pb/g animal/day) during 5 days, which resulted in a dose response increase of blood Pb levels and decrease of blood ALAD activity. However, at the higher doses tested (> 7 µg of Pb/g animal/day) the blood ALAD activity inhibition (~82%) remained constant. Hematocrit and hemoglobin were significantly reduced only at the highest-doses, and the stress indicator, heterophils to lymphocyte (H/L) ratio, did not show apparent changes. In a second experiment, house sparrows were exposed to Pb in drinking water (12.3 ppm) during either 15 or 30 days. Pb concentration used in this study was enough to produce blood lead levels equivalents to those found recently in house sparrows inhabiting urban areas, reduced blood ALAD activity and inversion of the H/L ratio. Decreasing blood ALAD activities were correlated with increasing blood Pb levels. In addition, Pb exposure produced modification in the levels of hepatic antioxidant enzymes, increased GST activity and decreased CAT activity, without lipid peroxidation. In conclusion, our results suggest that blood ALAD activity is a reliable and sensitive biomarker for environmental Pb exposure in house sparrows, additionally chronic exposure produce physiological stress (H/L inversion) and small changes in antioxidant enzyme activity. Finally, this specie could be considered a bioindicator for monitoring the urban Pb contamination.

Modulation of digestive enzyme activities in the avian digestive tract in relation to diet composition and quality.

In nature, birds are faced with variable food sources that may differ in composition (protein vs. carbohydrates) and quality (highly digestible material vs. indigestible fiber). Studies in passerine birds and some commercial poultry demonstrate that the gastrointestinal tract can respond to varying diet composition and quality by changing morphology and/or activities of digestive enzymes. However, studies in additional avian species are warranted to understand generalities of these trends. We first fed juvenile mallards (Anas platyrhynchos), chickens (Gallus gallus), and quails (Coturnix coturnix) on either high-carbohydrate or high-protein diets. For the most part, birds fed the high-carbohydrate diet had higher small intestinal and cecal disaccharidase activities (maltase and sucrase). However, only mallards exhibited higher small intestinal aminopeptidase-N (APN) activities when fed the high-protein diet. These results differ from passerine birds, which largely modulate small intestinal proteases, but not disaccharidases. In another trial, we fed Canada geese (Branta canadensis) diets that varied in both their protein and fiber concentrations for approximately 3.5 months. Birds fed the high-fiber diets had significantly longer small intestines and caeca compared to those fed low-fiber diets. Additionally, geese fed the high-fiber diets exhibited lower mass-specific activities of small intestinal sucrase, and higher activities of APN when summed across the small intestine and ceca. Similar to the avian species above, geese fed the high-protein diets did not exhibit flexibility in their small intestinal APN activities. Overall, these experiments demonstrate that responsiveness of the avian digestive tract to diet composition may have phylogenetic or ecological constraints. Studies on other avian taxa are needed to understand these patterns.

Paracellular absorption in laboratory mice: Molecule size-dependent but low capacity.

Water-soluble nutrients are absorbed by the small intestine via transcellular and paracellular processes. The capacity for paracellular absorption seems lower in nonfliers than in fliers, although that conclusion rests largely on a comparison of relatively larger nonflying mammals (>155g) and relatively smaller flying birds (<155g). We report on paracellular absorption in laboratory mice, the smallest nonflying mammal species studied to date. Using a standard pharmacokinetic technique, we measured the extent of absorption (fractional absorption=f) of inert carbohydrate probes: L-arabinose (M(r)=150.13Da) and cellobiose (342.3) that are absorbed exclusively by the paracellular route, and 3-O-methyl D-glucose (3OMD-glucose) (M(r)=194) absorbed both paracellularly and transcellularly. f was measured accurately in urine collection trials of 5-10h duration. Absorption of 3OMD-glucose by mice was essentially complete (f=0.95±0.07) and much higher than that for L-arabinose (f=0.21±0.02), indicating that in mice, like other nonflying mammals, >80% of glucose is absorbed by mediated process(es) rather than the passive, paracellular route. As in all other vertebrates, absorption of cellobiose (f=0.13±0.02) was even lower than that for L-arabinose, suggesting an equivalent molecular size cut-off for flying and nonflying animals and thus a comparable effective TJ aperture. An important ecological implication is that smaller water-soluble plant secondary metabolites that have been shown to be absorbed by the paracellular path in cell culture, such as phenolics and alkaloids, might be absorbed in substantial amounts by bats and small birds relative to nonflying mammals such as mice.

The capacity for paracellular absorption in the insectivorous bat Tadarida brasiliensis.

Water-soluble nutrients are absorbed by the small intestine via transcellular and paracellular processes. The capacity for paracellular absorption seems greater in fliers than in nonfliers, although that conclusion rests mainly on a comparison of flying birds and nonflying mammals because only two frugivorous bat species have been studied. Furthermore, the bats studied so far were relatively large (>85 g, compared with most bat species which are <20 g) and were not insectivores (like about 70 % of bat species). We studied the small (11 g) insectivorous bat Tadarida brasiliensis and tested the prediction that the capacity for paracellular absorption would be as high as in the other bat and avian species studied so far, well above that in terrestrial, nonflying mammals. Using standard pharmacokinetic technique, we measured the extent of absorption (fractional absorption = f) of inert carbohydrate probes: L-arabinose (MM = 150.13) absorbed exclusively by paracellular route and 3OMD-glucose (MM = 194) absorbed both paracellularly and transcellularly. As predicted, the capacity of paracellular absorption in this insectivorous bat was high (L-arabinose f = 1.03 ± 0.14) as in other frugivorous bats and small birds. Absorption of 3OMD-glucose was also complete (f = 1.09 ± 0.17), but >80 % was accounted for by paracellular absorption. We conclude that passive paracellular absorption of molecules of the size of amino acids and glucose is extensive in this bat and, generally in bats, significantly higher than that in nonflying mammals, although the exact extent can be somewhat lower or higher depending on molecule size, polarity and charge.

Isolation of epithelial cells, villi and crypts from small intestine of pigeons (Columba livia).

The isolation of viable enterocytes, villi and crypts from the small intestine of a feral bird (Columba livia) is important for performing physiological experiments in ecologically relevant processes of membrane transport. The effectiveness of mechanical disruption, enzymatic digestion and chelating agents were compared. The objectives were to isolate enterocytes, villi and crypts from the small intestine of young pigeons; to evaluate the viability of the isolated intestinal epithelial cells isolated; and to verify the integrity of enterocytes by biochemical features. Enzymatic and mechanical methods yielded both elongated columnar and spherical cells. With the chelating method villi and crypts were obtained. All methods produced a high yield of intestinal epithelial cells with about 50% viability. Brush border enzymes (sucrase-isomaltase and alkaline phosphatase) activities were high and, as reported in chickens, they did not differ along the intestinal villus-crypt axis. Although the three methods have good viabilities, the enzymatic technique gives the best yield in cell number, while the chelating method provides the highest populations of morphologically distinctive villi and crypts.

The digestive adaptation of flying vertebrates: high intestinal paracellular absorption compensates for smaller guts.

Anecdotal evidence suggests that birds have smaller intestines than mammals. In the present analysis, we show that small birds and bats have significantly shorter small intestines and less small intestine nominal (smooth bore tube) surface area than similarly sized nonflying mammals. The corresponding >50% reduction in intestinal volume and hence mass of digesta carried is advantageous because the energetic costs of flight increase with load carried. But, a central dilemma is how birds and bats satisfy relatively high energy needs with less absorptive surface area. Here, we further show that an enhanced paracellular pathway for intestinal absorption of water-soluble nutrients such as glucose and amino acids may compensate for reduced small intestines in volant vertebrates. The evidence is that l-rhamnose and other similarly sized, metabolically inert, nonactively transported monosaccharides are absorbed significantly more in small birds and bats than in nonflying mammals. To broaden our comparison and test the veracity of our finding we surveyed the literature for other similar studies of paracellular absorption. The patterns found in our focal species held up when we included other species surveyed in our analysis. Significantly greater amplification of digestive surface area by villi in small birds, also uncovered by our analysis, may provide one mechanistic explanation for the observation of higher paracellular absorption relative to nonflying mammals. It appears that reduced intestinal size and relatively enhanced intestinal paracellular absorption can be added to the suite of adaptations that have evolved in actively flying vertebrates.

Electroaffinity in paracellular absorption of hydrophilic D-dipeptides by sparrow intestine.

We previously demonstrated size selectivity in the absorption of nonelectrolyte hydrosoluble probes in birds, presumably by the paracellular pathway. Our goal in this study was to determine the charge selectivity in the absorption of hydrosoluble D-dipeptides, because there have been no studies of the electroaffinity of this absorption pathway in birds. For this purpose isosmotic solutions with two hydrophilic D-dipeptides: serine-lysine (positive at pH 7.4) and serine-aspartic (negative at pH 7.4) were gavaged into the stomach in nonanesthetized house sparrows (Passer domesticus), and injected into the pectoralis with a syringe in different trials. Fractional absorption was calculated as F = [AUC by gavage)]/[AUC by injection] (AUC = area under the curve of plasma probe concentration vs. time). Fractional absorption was significantly higher for the positively charged than negatively charged dipeptide (respectively, F=0.30+/-0.05 vs. F=0.17+/-0.03). These findings give the first evidence of cation selectivity by the paracellular route in the absorption of hydrosoluble solutes in the small intestine in birds.