New insights on diet variability revealed by DNA barcoding and high-throughput pyrosequencing: chamois diet in autumn as a case study

Characterizing the diet of large herbivores and the determinants of its variation remains a difficult task in wild species. DNA-based techniques have the potential to complement traditional time-consuming methods based on the microhistology of plant cuticle fragments in fecal or rumen samples. Recently, it has been shown that a short chloroplast DNA fragment, the P6 loop of the trnL (UAA) intron, can act as a minimalist barcode. Here, we used the trnL approach with high-throughput pyrosequencing to study diet from feces in a wild herbivore, the alpine chamois (Rupicapra rupicapra) and showed that the fine resolution in plant determination obtained with this method allows exploring subtle temporal shifts and inter-individual variability in diet composition. First, we built a DNA barcoding database of 475 plants species. Seventy-two percent of plant species can be unambiguously identified to species level, 79% to genus level and 100% to family level using the P6 loop. Second, we analysed 74 feces collected from October to November. Based on 47,896 P6 loop sequences, we identified a total of 110 taxa, 96 in October and 76 in November, with a clear diet shift between October and November. We recognized four and two clusters of feces composition in October and November, respectively, revealing different diet categories among individuals within each month. DNA-based diet analysis is faster and more taxonomically precise than studies based on microhistology, and opens new possibilities for analysing plant-herbivore interactions in the wild.     

The use of otoliths as indicators of Little Penguin Eudyptula minor diet

The validity of using otoliths from stomach contents quantitatively to determine the number and size of fish consumed was tested on Little Penguins. They were fed different meal sizes of known number and size of fish and the stomach contents were recovered after various time intervals. There were no differences in estimates of original fish size when calculated from otolith length or weight. Rate of digestion of otoliths tended to decrease with increased meal size but increased with time after ingestion. Digestion of otoliths proceeds rapidly and, if ignored, estimates of numbers of fish consumed and of original fish size can be significantly underestimated. This problem can be partially solved by inspection of otolith condition and restricting calculations of fish size to otoliths unaffected by digestion. Many factors introduce variations into rate of otolith degradation and further species-specific studies are required before appropriate correction factors can be applied.     

Cimetidine suspension as adjuvant to energy restricted diet in treating obesity.

OBJECTIVE--To study the effect of cimetidine suspension compared with placebo suspension on weight loss in moderately obese patients taking a 5 MJ/day diet supplemented with dietary fibre. To determine the relation between the effectiveness of the blinding and weight loss. DESIGN--Randomised double blind study with an eight week parallel group phase and a subsequent eight week crossover or continuation phase. SETTING--Outpatient clinic. SUBJECTS--60 patients (51 women) aged 18-60. MAIN OUTCOME MEASURE--Weight loss. RESULTS--After eight weeks of treatment the mean weight loss in the cimetidine group (5.7 kg) was similar to that of the placebo group (5.9 kg; p = 0.78, 95% confidence interval -2.0 to 1.5 kg). Body mass index, waist and hip measurements, waist-hip ratio, and systolic and diastolic blood pressures decreased similarly in the two groups. No association was found between weight loss and the patients'' ability to guess if they were being given drug or placebo. Correct guesses of current drug were more prevalent than expected by chance (25/37 correct, p = 0.05 for the parallel group phase; 26/30, p = 0.0001 for the crossover phase). CONCLUSIONS--Cimetidine had no effect on weight loss in moderately obese patients. The study underlines the potential problem that blinding of patients to treatment can be compromised.     

The use of otoliths as indicators of Little Penguin Eudyptula minor diet

The validity of using otoliths from stomach contents quantitatively to determine the number and size of fish consumed was tested on Little Penguins. They were fed different meal sizes of known number and size of fish and the stomach contents were recovered after various time intervals. There were no differences in estimates of original fish size when calculated from otolith length or weight. Rate of digestion of otoliths tended to decrease with increased meal size but increased with time after ingestion. Digestion of otoliths proceeds rapidly and, if ignored, estimates of numbers of fish consumed and of original fish size can be significantly underestimated. This problem can be partially solved by inspection of otolith condition and restricting calculations of fish size to otoliths unaffected by digestion. Many factors introduce variations into rate of otolith degradation and further species-specific studies are required before appropriate correction factors can be applied.     

Host diet as a determinant of parasite growth, reproduction and survival

Changes have been observed in the biology of protozoan and helminth parasites of small mammals during a variety of experimental nutritional manipulations ranging from deficiency in a specific nutrient to the general withholding of food from the host. Commonly observed effects include retardations in parasite growth rate, gametogenesis, fecundity, and asexual multiplication. The duration of patency and of the association of a parasite with its host have also been observed to be curtailed. The nutritional responses of parasites during host hibernation require investigation and much research will be needed to explain how perturbations in host dietary composition and intake may lead to the observed effects. Care should be exercised over applying the results from experimental studies to naturally occurring infections, but the experiments are justified because they indicate potential effects, if not their biological significance, and because causation can be determined.     

Dental sexual dimorphism and morphology in african colobus monkeys as related to diet

We considered the relationship between dental sexual dimorphism and diet in 542 specimens of olive, red, and black-and-white colobus. Using univariate statistical techniques, we examined 41 measurements of the maxillary and mandibular dentitions. The results reveal two trends of dental sexual dimorphism in black-and-white colobus wherein (i) maleColobus guereza andC. angolensis are generally larger than females throughout the dentition and (ii)C. satanas and, to a lesser degree,C. polykomos exhibit reduced sexual differences in the canine base and females are slightly larger than males in noncanine dimensions. Females of the red colobus,Procolobus (Piliocolobus) badius, are slightly larger than males in most noncanine measurements but canine sex differences are more pronounced than those of black-and-white colobus.Procolobus (Procolobus) verus, the olive colobus, is characterized by some of the largest canine sex differences, yet the sexes do not differ much in noncanine mean values. When patterns of sexual dimorphism are considered in terms of specific ecology and behavior, it is possible to relate sex differences, in part, to known dietary differences. For example, overall dental morphology and the trend of sexual differences inC. satanas andC. polykomos seem to be associated with the consumption of a diet rich in seeds. The pattern of dental sexual dimorphism inC. badius may also be influenced by dietary factors in that their patrilineal social organization could restrict female access to certain foods thereby affecting rates of attrition and creating selection pressure for larger teeth. Relatively less is known of the ecology and social organization ofP. verus but their dental sexual dimorphism is possibly less related to dietary factors than is the case for red or black-and-white colobus.     

SMXA-5 mouse as a diabetic model susceptible to feeding a high-fat diet

The SMXA-5 strain, a new mouse model for type 2 diabetes, is a recombinant inbred strain derived from non-diabetic SM/J and A/J strains. As dietary fat is a key component in the development of diabetes, we compared the glucose tolerance and diabetes-related traits among the SMXA-5, SM/J, and A/J strains while feeding a high-fat diet for 10 weeks. SMXA-5 fed on a high-fat diet showed an increased serum insulin concentration. Judging from the hyperinsulinemia in SMXA-5, this strain showed insulin resistance, an inability of peripheral tissues to respond to insulin, which was strengthened by feeding with a high-fat diet. When fed on a high-fat diet for 5 weeks, the SMXA-5 mice showed severely impaired glucose tolerance. On the other hand, SM/J showed mildly impaired glucose tolerance, even when fed on a high-fat diet for 10 weeks. These results indicate that SMXA-5 would be available for use as a diabetic model susceptible to a high-fat diet.     

Evaluation of long-chain alcohols as diet composition markers in goats grazing heathland areas

An experiment was conducted to assess the potential of long-chain alcohols (LCOH), in alternative or combined with alkanes and long-chain fatty acids (LCFA), as faecal markers to estimate the diet composition of goats grazing heathland vegetation with associated improved pastures. A total of seven diets were offered across the grazing season. The diets were composed of mixtures of herb species (Lolium perenne and Trifolium repens) and woody species (Erica spp., Calluna vulgaris and Ulex gallii) in an attempt to simulate diet selection of goats on these complex vegetation areas. The diet composition was estimated using LCOH markers alone or combined with alkanes, LCFA and alkanes+LCFA, by least square optimization procedures. The data showed large differences between plant species in their LCOH profile. Generally, plant species showed higher LCOH concentrations than those of alkanes and lower than LCFA markers. Faecal recovery of LCOH was incomplete and increased in a linear manner (P < 0.001) with the carbon-chain length, and was influenced by diet composition and its digestibility. The diet composition estimates based on LCOH alone were more accurate (P < 0.05) than those using alkanes or LCFA alone. Results showed that the combination of LCOH with alkanes, LCFA and alkanes+LCFA resulted in more accurate (P < 0.05) estimates of diet composition, indicating that LCOH provided different discriminatory information to that of alkanes and LCFA, helping in the discrimination of the plant species used in this experiment. Results indicate that correction of faecal LCOH concentrations to incomplete faecal recovery is necessary to obtain more accurate estimates of diet composition. Nevertheless, it seemed that the use of a less accurate recovery correction on LCOH markers had a lower impact on the accuracy of estimates than that of alkanes and LCFA.     

Elemental diet as primary treatment of acute Crohn's disease: a controlled trial.

Acute exacerbations of Crohn''s disease are usually treated with prednisolone or potentially more toxic immunosuppressive drugs or by surgery. In pilot studies replacing the normal diet by a protein free elemental diet also induced remission. A controlled trial was therefore conducted in which 21 patients acutely ill with exacerbations of Crohn''s disease were randomised to receive either prednisolone 0.75 mg/kg/day or an elemental diet (Vivonex) for four weeks. Assessment at four and 12 weeks showed that the patients treated with the elemental diet had improved as much as and by some criteria more than the steroid treated group. Elemental diet is a safe and effective treatment for acute Crohn''s disease.     

Mustard oil glucosides as feeding stimulants for Pieris brassicae larvae in a semi-synthetic diet

Nine glucosides tested as feeding stimulants for Pieris brassicae larvae were effective. The tetraacetates of three of them were not. Two of the glucosides (glucocapparin and glucoiberin) were active at 0.333×10^–5 molar in the diet and at 0.333×10^–4 molar all were effective. Usually all the larvae in a test do not feed and the increase in effectiveness with concentration, which is observed in groups, is largely brought about by a progressive increase in the number of larvae which actually feed, though with some glucosides an increase in the average frass production of the feeding larvae was found. The relative effectiveness of the glucosides as feeding stimulants changes with concentration. At a concentration of 0.333×10^–5 molar, glucocapparin gave the best result while at 0.333×10^–3 molar concentrations glucotropaeolin was most effective. Neither of these glucosides has been reported to occur in cabbage leaves.

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Senfölglukoside als frasstimulantien für pieris brassicae-raupen bei einer halbsynthetischen diät

Zusammenfassung Neun als Fraßstimulatoren für Kohlweißlingsraupen geprüfte Glukoside erwiesen sich als wirksam. Die Tetraacetate von drei von ihnen waren es nicht. Zwei der Glukoside (Glukokapparin und Glukoiberin) waren 0,333×10^–6 und 0,333×10^–4 molar in der Diät wirksam. Gewöhnlich fressen in einem Versuch nicht alle Raupen und der Anstieg der Wirksamkeit mit der Konzentration, die gruppenweise beobachtet wird, läßt sich hauptsächlich durch ein zunehmendes Anwachsen der Anzahl von Larven feststellen, die wirklich fressen, obwohl bei einigen Glukosiden ein Anwachsen der Kotproduktion der fressenden Larven gefunden wurde. Die relative Wirksamkeit der glukoside als Fraßanregungsmittel ändert sich mit der Konzentration. Bei einer molaren Konzentration von 0,333×10^–5 ergab Glukokapparin den besten Erfolg, während Glukotropaeolin bei 0,333×10^–3 molarer Konzentration am wirksamsten war. Keines dieser Glukoside ist bisher in Kohlblättern nachgewiesen.

     

Rice bran as a diet for culturing Streptocephalus proboscideus (Crustacea: Anostraca)

We cultured Streptocephalus proboscideus using rice bran as a food source, and compared its growth, mortality, fertility, and cyst hatching with published data on animals fed other food sources. We also analysed the fatty acid profile of animals and cysts. The performance of rice-bran fed shrimp was indistinguishable from that of shrimp fed most other formulated diets, but there are indications that some previously applied diets were either defective in essential fatty acids, or toxic.     

Morphological traits as predictors of diet and microhabitat use in a diverse beetle assemblage

We explored how morphological traits can complement phylogenetic information to extend our predictions of the ecology of a diverse beetle assemblage. We analysed ten morphological traits from an assemblage of 239 species from 35 families, and identified three axes of morphological variation that were independent of body length: (1) relative robustness; (2) relative appendage length; and (3) relative abdomen length. The trait associations defining these axes of morphological variation did not change after adjusting for family‐level phylogeny. We detected significant differences in morphological variation across the beetle assemblage according to diet and microhabitat use, and these patterns were only partially influenced by family membership. Further analysis within dominant families showed that species of Carabidae, Curculionidae, Scarabaeidae and Staphylinidae had greater body length in open versus tree litter microhabitat, and species of Carabidae and Curculionidae had greater relative robustness, but shorter relative appendage length, in open versus tree litter microhabitat. Although it is clear that family‐level phylogeny and morphology share some explanatory power for predicting the diet and microhabitat use by beetles, we demonstrate that body length, robustness and appendage length are correlated significantly with microhabitat use when comparing members of the same family. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 102 , 301–310.