Puddling: from natural history to understanding how it affects fitness

The term ‘puddling’ includes feeding on (dried) mud and various excrements and secretions of vertebrates, and carrion. It is thought to be a form of supplementary feeding, not targeted at obtaining energy. Although the natural history of the puddling phenomenon in herbivorous arthropods becomes better known, it is still largely unclear how puddling (in particular for sodium) affects fitness despite the growing knowledge of insect physiology at the cellular level. If we follow the definition used for puddling in Lepidoptera, representatives of a wide range of herbivorous and detrivorous terrestrial arthropods (Lepidoptera, Orthoptera, Blattodea, Hymenoptera, Hemiptera, Diptera, and Diplopoda) have been observed to puddle. It appears that those species with diets low in sodium (e.g., folivorous larvae) puddle for sodium whereas those with diets low in nitrogen (e.g., detritivores) puddle for nitrogen. Sex differentials in puddling behavior can usually be explained by transfers of nutrients from males to females during mating. Puddling is rare or absent in immature stages and there is some evidence that nutrients from puddles increase female reproductive success. Strong evidence for the widely cited hypothesis that sodium from puddles is used to enhance neuromuscular activity is still lacking. High mobility and long life spans could be associated with puddling behavior, whereas insects that are concealed or well defended are less likely to puddle (e.g., beetles). The role that risks of pathogen and parasite infection as well as predation at puddling substrates may play in the evolution of puddling remains virtually unexplored.     

Relative contribution of neutral and deterministic processes in shaping fruit‐feeding butterfly assemblages in Afrotropical forests

The unified neutral theory of biodiversity and biogeography has gained the status of a quantitative null model for explaining patterns in ecological (meta)communities. The theory assumes that individuals of trophically similar species are functionally equivalent. We empirically evaluate the relative contribution of neutral and deterministic processes in shaping fruit‐feeding butterfly assemblages in three tropical forests in Africa, using both direct (confronting the neutral model with species abundance data) and indirect approaches (testing the predictions of neutral theory using data other than species abundance distributions). Abundance data were obtained by sampling butterflies using banana baited traps set at the forest canopy and understorey strata. Our results indicate a clear consistency in the kind of species or species groups observed at either the canopy or understorey in the three studied communities. Furthermore, we found significant correlation between some flight‐related morphological traits and species abundance at the forest canopy, but not at the understorey. Neutral theory's contribution to explaining our data lies largely in identifying dispersal limitation as a key process regulating fruit‐feeding butterfly community structure. Our study illustrates that using species abundance data alone in evaluating neutral theory can be informative, but is insufficient. Species‐level information such as habitat preference, host plants, geographical distribution, and phylogeny is essential in elucidating the processes that regulate biodiversity community structures and patterns.     

Nutrients in fruit increase fertility in wild-caught females of large and long-lived Euphaedra species (Lepidoptera, Nymphalidae)

Fruit-feeding butterflies can experience a more nutrient rich adult diet than nectar-feeding species, and can be expected to use these nutrients for egg production. Here we compare life span, and reproduction parameters of wild-caught females of large and long-lived species on either a sucrose or a mashed banana diet. With small sample sizes per species, but rich longitudinal data for each individual, we examined the longitudinal reproduction pattern, egg size and hatchability of these butterflies in captivity. Diet significantly affected mortality in captivity in a time-dependent manner. On average, we found that butterflies fed mashed banana laid 1.855 times more eggs than those fed sugar. They laid significantly more eggs when they laid and conserved egg size with age while butterflies fed sucrose showed significantly declining egg sizes. Egg hatchability was not significantly affected by diet. Long pre-oviposition periods, significantly smaller first eggs, and absence of age at capture effects on intensity of reproduction indicate low reproduction rates in the field that are due to low food availability. With our small sample sizes, we did not detect significant differences between the species in their response to the diet treatments.     

Spatial and temporal variation in butterfly biodiversity in a West African forest: lessons for establishing efficient rapid monitoring programmes

Because tropical forests face serious threats and are usually situated in developing countries, cheap and easy Rapid Biodiversity Assessment (RBA) seems an ideal strategy for documenting changes in habitat quality and setting conservation priorities. Fruit‐feeding butterflies are often used as focal taxa to document biodiversity trends in tropical forests. However, the choice of inappropriate sampling strategies may nullify its value. More extensive data sets that sample multiple months and seasons and incorporate the vertical dimension are needed as a baseline for designing reliable sampling regimes. Therefore, we performed a butterfly trapping study in Ghana, using both understorey and canopy traps, and sampling seven periods covering dry and wet seasons. Both individual numbers and species richness were on average three to four times higher in the understorey than in the canopy traps with strikingly different species composition (only 11% overlap in species). The number of species and individual abundance recorded in a month varied extensively. This study underlined the importance of taking into account temporal variation and vertical stratification when designing RBAs of fruit‐feeding butterflies in West African forests. We recommend the use of both understorey and canopy traps and inclusion of both ‘wet’ and ‘dry’ seasons into RBA sampling regimes.     

<Emphasis Type="Italic">NIPTeR</Emphasis>: an R package for fast and accurate trisomy prediction in non-invasive prenatal testing

Background

Various algorithms have been developed to predict fetal trisomies using cell-free DNA in non-invasive prenatal testing (NIPT). As basis for prediction, a control group of non-trisomy samples is needed. Prediction accuracy is dependent on the characteristics of this group and can be improved by reducing variability between samples and by ensuring the control group is representative for the sample analyzed.

Results

NIPTeR is an open-source R Package that enables fast NIPT analysis and simple but flexible workflow creation, including variation reduction, trisomy prediction algorithms and quality control. This broad range of functions allows users to account for variability in NIPT data, calculate control group statistics and predict the presence of trisomies.

Conclusion

NIPTeR supports laboratories processing next-generation sequencing data for NIPT in assessing data quality and determining whether a fetal trisomy is present. NIPTeR is available under the GNU LGPL v3 license and can be freely downloaded from https://github.com/molgenis/NIPTeR or CRAN.

     

Quantifying income breeding: using geometrid moths as an example

Although the general concept of capital vs. income breeding has become widely used in insect ecology, finding easy‐to‐measure indices for quantifying the role of larval‐ vs. adult‐derived nutrients in egg production has remained a challenge. When searching for possible candidates for this task, we evaluated the applicability of three morphometric ratios to be measured on freshly eclosed adults: (1) relative size of female abdomen, (2) sexual size dimorphism, and (3) proportion of volume of mature eggs relative to total volume of female abdomen. We report the values of these indices in five species of geometrid moths [Hypomecis punctinalis Scopoli, Ematurga atomaria L., Selenia tetralunaria Hufnagel, Semiothisa clathrata L., and Epirrhoe alternata Müller (all Lepidoptera: Geometridae)] and compare them to the degrees of income breeding measured directly by oviposition experiments. As a comparison, we also recorded the ovigeny index and the effect of income on lifespan. All morphometric indices varied considerably between the species studied, and, as predicted, the values of all three proposed indices were higher in the more capital‐breeding species. The only exception to this pattern was a low proportion of mature eggs in abdomens in one primarily capital‐breeding species, S. tetralunaria. Unlike other capital breeders, this species also proved to be largely synovigenic, indicating that capital breeding and pro‐ovigeny are not strictly linked. The reproductive traits measured concord with ecological niches occupied by the species studied: the oligophagous meadow dwellers were largely income breeding and synovigenic, whereas the polyphagous forest dwellers were capital breeders. The high and predictable among‐species variation in egg‐production strategies suggests that geometrid moths form a promising target group for comparative studies on respective traits. Simple anatomical indices, in turn, appear to be applicable as easy‐to‐record proxies of egg production strategies.     

Material affects attack rates on dummy caterpillars in tropical forest where arthropod predators dominate: an experiment using clay and dough dummies with green colourants on various plant species

Predation can be one of the key factors that determine abundance in insect herbivore communities, and drive evolution of body size, and anti‐predator traits, including crypsis. Population dynamics and selection pressures will depend on the identity of dominant predators in the system, and these may vary substantially among habitats. Arthropods emerge as chief predators on caterpillars in the understorey of non‐montane tropical forest, whereas birds dominate elsewhere. In a tropical forest in Uganda, Africa, we evaluated marks on dummy caterpillars that differed in size, material (clay vs. dough), colourant, and plant species on which dummy caterpillars were exposed. We included live caterpillars to estimate the extent to which studies using artificial caterpillars reflect actual levels of predation. Ants and wasps were the most important damagers of dummy caterpillars, whereas bug and beetle damage was very rare, and no bird or small mammal damage was observed. Daily attack rates did not differ significantly from apparent mortality of live caterpillars (daily mortality = 12.1%), but dummy caterpillars made from dough were attacked more frequently (daily attack rate = 18.4%) than those from clay (daily attack rate = 6.9%). Caterpillars of different colour and size, and caterpillars exposed on different plant species had the same chances to be predated. This is in contrast to results from temperate area studies where birds dominate and are not affected by dummy caterpillar material, but prefer larger caterpillars. Our results are consistent with dominant predators on tropical forest caterpillars being invertebrates that are more chemically than visually oriented, so that: (1) material used for dummy caterpillars is important, (2) background matching is relatively unimportant, and (3) being large may have less of a cost. These patterns in predation might facilitate polyphagy and evolution of large body size in tropical Lepidoptera.     

Maternal Investment and Infant Survival in Gray-Cheeked Mangabeys (Lophocebus albigena)

Differences among females in infant survival can contribute substantially to variance in fitness. Infant survival is a product of external risk factors and investment by kin, especially the mother, and is thus closely tied with the evolution of behavior and life history. Here we present a 9-yr study (2004–2012) of infant survival and sex ratio relative to age and dominance ranks of mothers and the presence of immigrant males in a free-ranging population of gray-cheeked mangabeys (Lophocebus albigena) in Kibale National Park, Uganda. We consider immigrant males because they are known to increase infant mortality in several other species. We found that infants of older mothers had higher survival than those of younger mothers but that high rank did not confer a significant benefit on infant survival. Female infants had higher survival than male infants. Young, low-ranking females had more male infants than young, high-ranking females, which had slightly more daughters, but this difference declined as females aged because low-ranking females had more daughters as they aged. With limited data, we found a significant relationship between the presence of male immigrants and infant mortality (falls and unexplained disappearances) to 18 mo. Our results suggest that infant survival in gray-cheeked mangabeys is most precarious when mothers must allocate energy to their own growth as well as to their infants, that sons of young mothers are at greatest risk, and that immigrant males can negatively affect infant survival.     

Adult diet affects lifespan and reproduction of the fruit‐feeding butterfly Charaxes fulvescens

Fruit‐feeding butterflies are among the longest lived Lepidoptera. While the use of pollen‐derived amino acids by Heliconius butterflies has been interpreted as important for the evolution of extended lifespans, very little is known about the life‐history consequences of frugivory. This issue is addressed by investigating effects of four adult diets (sugar, sugar with amino acids, banana, and moistened banana) on lifespan and reproduction in the fruit‐feeding butterfly Charaxes fulvescens Aurivillius (Lepidoptera: Nymphalidae). Female butterflies were collected from Kibale National Park, Uganda, and kept individually in cages near their natural habitat and data were collected on lifespan, oviposition, and hatching of eggs. Lifespan in captivity was longer for the sugar and the amino acid cohort, than for the banana cohorts. The longitudinal pattern of oviposition was erratic, with many days without oviposition and few periods with high numbers of eggs laid. Butterflies typically did not lay eggs during their 1st week in captivity and the length of the period between capture and first reproduction was significantly shorter for butterflies fed moistened banana. The length of the reproduction period (first reproduction–last reproduction in captivity) and the reproduction rate (total number of eggs/length of the reproduction period) did not differ significantly between the diet treatments. Those fed with amino acid and moistened banana had significantly higher egg hatchability than those fed with sugar and banana. We found no evidence for a lifespan cost of reproduction. Our results show that (1) female C. fulvescens can use amino acids in their diet for laying fertile eggs, (2) more wing‐wear does correlate with lower survival in captivity (indicating aging in the wild), but not with intensity of reproduction (providing no evidence for reproductive aging), and (3) fruit‐feeding butterflies may be dietary restricted in the field.