Detection of abnormally high amygdalin content in food by an enzyme immunoassay

Amygdalin is a cyanogenic glycoside compound which is commonly found in the pits of many fruits and raw nuts. Although amygdalin itself is not toxic, it can release cyanide (CN) after hydrolysis when the pits and nuts are crushed, moistened and incubated, possibly within the gastrointestinal tract. CN reversibly inhibits cellular oxidizing enzymes and cyanide poisoning generates a range of clinical symptoms. As some pits and nuts may contain unusually high levels of amygdalin such that there is a sufficient amount to induce critical CN poisoning in humans, the detection of abnormal content of amygdalin in those pits and nuts can be a life-saving measure. Although there are various methods to detect amygdalin in food extracts, an enzyme immunoassay has not been developed for this purpose. In this study we immunized New Zealand White rabbits with an amygdalin-KLH (keyhole limpet hemocyanin) conjugate and succeeded in raising anti-sera reactive to amygdalin, proving that amygdalin can behave as a hapten in rabbits. Using this polyclonal antibody, we developed a competition enzyme immunoassay for determination of amygdalin concentration in aqueous solutions. This technique was able to effectively detect abnormally high amygdalin content in various seeds and nuts. In conclusion, we proved that enzyme immunoassay can be used to determine the amount of amygdalin in food extracts, which will allow automated analysis with high throughput.     

昆虫寄生对栓皮栎坚果特征和萌发行为的影响

有多种昆虫常寄生于栎属植物的坚果中, 进而影响种子的质量、 萌发、 幼苗建成等植物的更新过程。为探讨昆虫寄生与上述过程之间的关系, 本研究于2007年和2008年在太行山济源地区调查了昆虫对栓皮栎Quercus variabilis坚果的寄生情况, 同时探讨了昆虫寄生对坚果单宁水平、 萌发和幼苗生长的影响; 并于2007年9月, 分别将完好的和昆虫寄生的栓皮栎坚果种植于土壤4 cm深处, 对坚果萌发情况、 幼苗出土时间、 叶片数量和生物量等进行了对比分析。结果表明: 1）2007年栓皮栎坚果的虫寄生率为30.04%, 显著低于2008年（47.68%）, 表现出年际变化; 2）虫寄生坚果中单宁酸含量（11.54%±1.36%）显著高于完好坚果（7.36%±1.31%）（P=0.004）; 3）虫寄生坚果的鲜重、 直径、 长度均小于完好坚果; 4）虫寄生坚果的霉烂率（28%）和不完全萌发率（28%）均高于完好坚果（霉烂率0%, 不完全萌发率2%）; 但虫寄生坚果幼苗建成率（56%）低于完好坚果（92%）; 虫寄生坚果幼苗出土持续时间（埋藏后35周）短于完好坚果（埋藏后37周）; 5）在坚果埋藏和幼苗萌出当年的冬季, 由虫寄生坚果和完好坚果建成的幼苗的高度、 叶片数间均无显著差异, 但在翌年的生长季节, 两项指标均出现显著性差异; 6）经过一个完整的生长周期（1年）之后, 由虫寄生坚果所建成幼苗的根长、 根重量和生物量3项指标显著低于完好坚果, 而叶片数、 茎长、 叶重和茎重指标在二者间无显著性差异。研究结果提示, 昆虫寄生会对栎类坚果的种子质量和萌发行为产生一定的影响, 这可能是栎类植物群落更新的适应性选择。     

Chimpanzees using stones to crack open oil palm nuts in Liberia

The use of stone-tools to open palm nuts (Elaeis guineensis) was studied in a group of 16 chimpanzees (Pan troglodytes) released from captivity to a natural island setting in Liberia. The behaviour was started by one female of the group; it then spread to 12 others over periods varying from a few seconds to a few weeks. Nut-cracking soon spread to three other sites, both spontaneously and with human encouragement. Both nuts and tools were carried distances of several hundred meters. Social interaction at cracking sites was rich and varied, ranging from fights over possession of tools to unsolicited sharing of nuts. Nut-crackers showed selectivity in schoosing “good” nuts and varied their methods according to the qualities of the nuts. Individual differences in technique emerged. These are the first detailed behavioural data on palm-nut-cracking, and they show many parallels with nut-cracking of other species by chimpanzees of the Tai Forest, Ivory Coast. This is another example of hammer-stone use from a limited region of West Africa: southeastern Guinea, eastern Liberia, and western Ivory Coast. This suggests limited cultural diffusion of the custom.     

Measuring the multi-scale integration of mechanical forces during morphogenesis

The elaborate changes in morphology of an organism during development are the result of mechanical contributions that are a mixture of those generated locally and those that influence from a distance. We would like to know how chemical and mechanical information is transmitted and transduced, how work is done to achieve robust morphogenesis and why it sometimes fails. We introduce a scheme for separating the influence of two classes of forces. Active intrinsic forces integrate up levels of scale to shape tissues. Counter-currently, extrinsic forces exert influence from higher levels downwards and feed back directly and indirectly upon the intrinsic behaviours. We identify the measurable signatures of different kinds of forces and identify the frontiers where work is most needed.     

Theoretical study of the acidic strength of amino acid side chains

The acidic strengths in gas phase of three groups (NH₄⁺, H₂S, and HCOOH) that mimic the most common amino acid side chains of enzymes are studied by means of quantum mechanical methods. The results demonstrate that in gas phase the acidities of such groups change drastically with respect to those reported in aqueous phase. Moreover, the dependence between the energetics of the proton-transfer process and the distance separating the acid and base groups is stated. The biological implications of these results are discussed.     

Long-term variation in fruiting and the food habits of wild Japanese macaques on Kinkazan Island, northern Japan

We studied the relationship between the food habits of Japanese macaques (Macaca fuscata) and the availability of nuts (Fagus crenata, Zelkova serrata, Carpinus spp., and Torreya nucifera) on Kinkazan Island, northern Japan, from 1997 to 2005 to examine the long-term variations in both food habits and availability. The food habits of the monkeys showed clear seasonal changes: the staple foods were woody leaves and flowers in spring (May and June), woody leaves and seeds or fruits other than nuts in summer (July and August), nuts and seeds or fruits other than nuts in fall (September-November), and herbaceous plants in winter (December-April). The availability of nuts, combinations of masting species, and energy production varied among years. Food habits varied among years, but the magnitude of variability of food habits differed among seasons, with large variability during summer and winter, and small variability in spring. Food availability was poor in summer and winter, but in several years the monkeys were able to consume nuts during those seasons. We emphasize the importance of conducting long-term studies on both food availability and the food habits of animals in the temperate zone.     

Aspergillus on tree nuts: incidence and associations

California exports tree nuts to countries where they face stringent standards for aflatoxin contamination. Trade concerns have stimulated efforts to eliminate aflatoxins and Aspergillus flavus from almonds, pistachios and walnuts. Incidence of fungi on tree nuts and associations among fungi on tree nuts were studied. Eleven hundred pistachios, almonds, walnuts and brazil nuts without visible insect damage were plated on salt agar and observed for growth of fungi. Samples came both from California nut orchards and from supermarkets. To distinguish internal fungal colonization of nuts from superficial colonization, half the nuts were surface-sterilized before plating. The most common genera found were Aspergillus , Rhizopus and Penicillium . Each species of nut had a distinct mycoflora. Populations of most fungi were reduced by surface sterilization in all except brazil nuts, suggesting that they were present as superficial inoculum on (rather than in) the nuts. In general, strongly positive associations were observed among species of Aspergillus ; nuts infected by one species were likely to be colonized by other species as well. Presence of Penicillium was negatively associated with A. niger and Rhizopus in some cases. Results suggest that harvest or postharvest handling has a major influence on nut mycoflora, and that nuts with fungi are usually colonized by several fungi rather than by single species. This revised version was published online in June 2006 with corrections to the Cover Date.     

啮齿动物对印度栲种子扩散的效率

从2007 年11 月到2009 年11 月,在西双版纳热带雨林中选取三棵母树,在每棵树下每年释放标记印度栲种子200 粒(三年共计1 800 粒),并追踪其命运。通过调查啮齿动物搬运和分散贮藏印度栲种子的比例,以及
查贮藏种子的微生境、贮藏点大小和扩散距离,分析贮藏种子的存活情况,进而评估啮齿动物对印度栲种子扩散的效率。结果表明,啮齿动物搬运了69.3% 的印度栲种子,被搬运的种子中18% 被分散贮藏。所有被分散贮藏的种子均被埋于落叶下或埋于土壤表层,并且大部分贮藏点仅含一粒种子。种子的扩散距离从0.5 m 到43.8 m,平均距离为7.1 m,扩散距离在年间没有显著差异。2007 年(种子密度低,啮齿动物密度高) 没有种能最终存活到实验结束,而2008 和2009 年(种子密度高,啮齿动物密度低)分别有0.3% 和1.5% 的种子存活。研究表明,啮齿动物是印度栲有效的种子扩散者,但其扩散效率很大程度上取决于森林中种子的密度和啮齿动物的丰富度。     

Predation and dispersal of large and small seeds of a tropical palm

Seed size may vary greatly among individuals within plant species. What effects the extremes of this variation have for seeds taken by small mammals are poorly understood. Not all seeds removed by small mammals are necessarily eaten. Small rodents are common seed predators, but they may disperse a significant proportion of seeds by scatter hoarding them via burial. Size-dependent predation and dispersal of seeds has not been directly tested within a plant species for tropical rodents. This study tested whether or not large and small nuts of Astrocaryum mexicanum (Palmae) differed in their fates due to handling by the spiny pocket mouse Heteromys desmarestianus (Heteromyidae). Exclosures were used to give small rodents exclusive access to A. mexicanum nuts. H. desmarestianus preferentially consumed large over small A. mexicanum nuts, but cached (in burrows and by scatter hoarding) similar proportions of these nuts by size. Small nuts tended to be buried farther away from exclosures than large nuts. Although sample sizes of buried nuts were small, the rodents retrieved all buried large nuts, but 30% of the small nuts remained buried long enough to germinate. I also examined predispersal predation by insects and found that insects appear to have no size preference for A. mexicanum nuts, but insect predation appears to hinder nut development. Thus, nuts attacked by insects develop to be significantly smaller, with a low proportion of undamaged endosperm, than uninfested nuts. It is hypothesized that the preferential predation of large A. mexicanum nuts by H. desmarestianus is a response by these rodents to insect predation.     

Micromanipulator for Yeast Genetic Studies

An inexpensive mechanical micromanipulator, designed primarily for separating yeast ascospores, can be assembled from commercially available components and without extensive custom machining.     

Nut-like oil seeds: food for monkeys, chimpanzees, humans, and probably ape-men

The hypothetical hyperrobust australopithecine gnathic nutcracker adaptation is reexamined in light of ecobotanical information on edible wild nuts provided by the flora of tropical and subtropical Africa. The nut producing species are tree-forms. Those of the forest region do not as a rule produce fruits with edible mesocarps. In contrast, the woodland savanna species (particularly in the Zambezian region) characteristically provide an important whole fruit, i.e., a nutritious mesocarp in addition to edible oil-rich nut seeds. These fruits drop from the tree before they are fully mature and go through the final ripening phase on the ground. They are important seasonal foods for a variety of vertebrates, including primates, elephants, and antelope. Altogether the nuts exhibit a broad range of toughness values, measured here as strength under compression. The woodland nuts are not as tough (177-934 kg force, breaking load) as those of the tropical forest (192-1,673 kg force). The seed-predators of the woodland species include squirrels, baboons, warthogs, and parrots. Paleoecological analyses indicate that it was the woodland nuts that were probably available to Australopithecus boisei and A. robustus. Preliminary estimates of adult male gnathic nut-cracking capabilities suggest that A. boisei could have orally cracked a significant portion of the woodland nuts. In spite of this, ecobotanical data indicate that we can probably reject the hypothesis that these hominids were year-round gnathic nut-cracking specialists. Both the indirect and direct evidence support this conclusion.     

Long-term patterns and feeding sites of southern green stink bug (Hemiptera: Pentatomidae) in Hawaii macadamia orchards, and sampling for management decisions

Southern green stink bug (Nezara viridula, Hemiptera: Pentatomidae) is a pest of macadamia nuts, causing pitting to kernels by feeding. In spite of its pest status, many aspects of the ecology of this insect in macadamia orchards are poorly understood. This study analyzes long-term N. viridula damage to macadamia nuts and investigates the extent to which damage to nuts occurs in the tree canopy, prior to nut-drop. We show that there are distinct seasonal peaks in damage detected after harvest and that, over six years of data collection, mean damage levels were fairly low, albeit with spikes in damage levels recorded. Sampling nuts at peak harvest periods from different strata in the trees and from the ground showed that incidence of damaged nuts within the canopy was typically half as high as on the fallen nuts. Damage to fallen nuts may have occurred prior to nut-drop, and continued to accumulate after nut-drop. These results show that management of N. viridula within macadamia canopies, as opposed to only on fallen nuts, is important. A sampling procedure and predictive model for estimating late-season damage based on early-season damage samples is provided. The model uses January and March damage measurements (based on samples with set level of accuracy), mean temperature and month of the year for which damage is predicted. Early-season damage of 6-10% predicts late-season damage levels that should justify N. viridula suppression based on the nominal threshold (13% damage) used by kernel processors to reject nuts based on damage.     

Contamination of pine nuts by fumonisin produced by strains of Fusarium proliferatum isolated from Pinus pinea

AIMS: To test the ability of Fusarium proliferatum strains isolated from Pinus pinea to synthesize fumonisin B(1) (FB(1)) in pine nuts. METHOD AND RESULTS: Eleven strains were inoculated in moist sterile shelled pine nuts and whole pine nuts, and incubated for 28 days. Moist sterile maize was inoculated in parallel as an optimum substrate for FB(1) production by Fusarium species. CONCLUSIONS: Six of the strains produced FB(1) in shelled pine nuts, and two of them did so in whole pine nuts. IMPACT AND SIGNIFICANCE OF THE STUDY: F. proliferatum is able to grow and produce FB(1) in the husk of pine nuts and the mycotoxin can be found in the nut once shelled. Consequently, FB(1) may be an important hazard in pine nuts.     

Linkage map construction involving a reciprocal translocation

This paper is concerned with a novel statistical–genetic approach for the construction of linkage maps in populations obtained from reciprocal translocation heterozygotes of barley (Hordeum vulgare L.). Using standard linkage analysis, translocations usually lead to ‘pseudo-linkage’: the mixing up of markers from the chromosomes involved in the translocation into a single linkage group. Close to the translocation breakpoints recombination is severely suppressed and, as a consequence, ordering markers in those regions is not feasible. The novel strategy presented in this paper is based on (1) disentangling the “pseudo-linkage” using principal coordinate analysis, (2) separating individuals into translocated types and normal types and (3) separating markers into those close to and those more distant from the translocation breakpoints. The methods make use of a consensus map of the species involved. The final product consists of integrated linkage maps of the distal parts of the chromosomes involved in the translocation.     

Chinese chestnut—A promising new orchard crop

This chestnut, resistant to the annihilating blight of the American species, gives promise, in selected varieties, of providing an American source of edible nuts comparable to those now imported from. Europe and threatened with extermination by the same blight.     

Dispersal of sweet pignut hickory in a year of low fruit production,and the influence of predation by a curculionid beetle

Summary The rate at which fallen hickory nuts are removed from beneath the parent tree, and the effect on this rate of the seed predatorConotrachelus affinis, was studied in an oak-hickory forest in southeastern Michigan, USA, during a year in which few nuts were produced. The trees responded toConotrachelus, which destroyed half the nut crop, by aborting inviable nuts during the summer. The seed dispersers, mostly gray squirrels, removed fallen nuts rapidly, showing the ability to distinguish viable nuts and remove them preferentially. The number of nuts removed in a week varies directly with the number available, and removal rate increases when many viable nuts are falling. The death of most seeds before dispersal, and the squirrels' efficiency at foraging on nuts and recovering them after burial, imply that successful hickory reproduction takes place only in years of heavy nut production.     

秦岭南坡短柄枹栎和锐齿槲栎的种子产量和种子大小及其与昆虫寄生的关系

【目的】为了解昆虫寄生特征与短柄枹栎 Quercus glandulifera 和锐齿槲栎 Quercus aliena var. acuteserrata 的种子产量及种子大小的关系。【方法】本研究于2011和2012年在秦岭南坡佛坪国家自然保护区,对短柄枹栎和锐齿槲栎种群的种子产量和昆虫寄生特征进行了野外调查研究。【结果】2年中短柄枹栎、锐齿槲栎种子雨密度和种子产量均差异明显,2011年短柄枹栎种子雨密度和种子产量分别为145.76±24.56粒/m²和74.97±11.56粒/m²,分别显著高于2012年的64.09±9.61粒/m²和34.30±3.51粒/m²;2011年锐齿槲栎种子雨密度和种子产量分别为238.88±43.97粒/m²和117.34±18.76粒/m²,分别显著高于2012年的112.00±19.20粒/m²和55.96±19.20粒/m²。2011年短柄枹栎和锐齿槲栎的种子产量中完好种子所占比例分别为49.73%和50.73%,分别显著高于2012年的38.69%和44.28%;而2011年虫蛀短柄枹栎和锐齿槲栎种子所占比例分别为28.05%和19.31%,分别低于2012年的39.77%和26.63%,但并无显著差异。2011和2012年,短柄枹栎和锐齿槲栎虫蛀种子个体均显著大于完好种子,且2011年二者虫蛀种子所含幼虫数与种子大小的相关性显著,2012年二者相关性不显著。【结论】短柄枹栎和锐齿槲栎种子的虫蛀率与种子产量有一定的关系,即在种子产量较低年份,虫蛀率有升高趋势,反之有降低趋势。同时,昆虫对这两种栎树种子中个体大的种子有寄生选择偏好,同时虫蛀种子所含幼虫数与种子大小也有一定的关系,即种子越大,所含昆虫幼虫数就越多。     

The evolutionary ecology of nut dispersal

A variety of nut-producing plants have mutualistic seed-dispersal interactions with animals (rodents and corvids) that scatter hoard their nuts in the soil. The goals of this review are to summarize the widespread horticultural, botanical, and ecological literature pertaining to nut dispersal inJuglans, Carya, Quercus, Fagus, Castanae, Castanopsis, Lithocarpus, Corylus, Aesculus, andPrunus; to examine the evolutionary histories of these mutualistic interactions; and to identify the traits of nut-bearing plants and nut-dispersing rodents and jays that influence the success of the mutualism. These interactions appear to have originated as early as the Paleocene, about 60 million years ago. Most nuts appear to have evolved from ancestors with wind-dispersed seeds, but the ancestral form of dispersal in almonds (Prunus spp.) was by frugivorous animals that ingested fruit. Nut-producing species have evolved a number of traits that facilitate nut dispersal by certain rodents and corvids while serving to exclude other animals that act as parasites of the mutualism. Nuts are nutritious food sources, often with high levels of lipids or proteins and a caloric value ranging from 5.7 to 153.5 kJ per propagule, 10–1000 times greater than most wind-dispersed seeds. These traits make nuts highly attractive food items for dispersers and nut predators. The course of nut development tends to reduce losses of nuts to insects, microbes, and nondispersing animals, but despite these measures predispersal and postdispersal nut mortality is generally high. Chemical defenses (e.g., tannins) in the cotyledons or the husk surrounding the nut discourage some nut predators. Masting of nuts (periodic, synchronous production of large nut crops) appears to reduce losses to insects and to increase the number of nuts dispersed by animals, and it may increase cross-pollination. Scatter hoarding by rodents and corvids removes nuts from other sources of nut predation, moves nuts away from source trees where density-dependent mortality is high (sometimes to habitats or microhabitats that favor seedling establishment), and buries nuts in the soil (which reduces rates of predation and helps to maintain nut viability). The large nutrient reserves of nuts not only attract animal dispersers but also permit seedlings to establish a large photosynthetic surface or extensive root system, making them especially competitive in low-light environments (e.g., deciduous forest) and semi-arid environments (e.g., dry mountains, Mediterranean climates). The most important postestablishment causes of seedling failure are drought, insufficient light, browsing by vertebrate herbivores, and competition with forbs and grasses. Because of the nutritional qualities of nuts and the synchronous production of large nut crops by a species throughout a region, nut trees can have pervasive impacts on other members of ecological communities. Nut-bearing trees have undergone dramatic changes in distribution during the last 16,000 years, following the glacial retreat from northern North America and Europe, and the current dispersers of nuts (i.e., squirrels, jays, and their relatives) appear to have been responsible for these movements.     

Distribution patterns of coconut mite, Aceria guerreronis, and its predator Neoseiulus aff. paspalivorus in coconut palms

Distribution patterns and numerical variability of the coconut mite Aceria guerreronis Keifer (Acari: Eriophyidae) and its predator Neoseiulus aff. paspalivorus DeLeon (Phytoseiidae) on the nuts of 3- to 7-month-old bunches of coconut palms were studied at two sites in Sri Lanka. At the two sites, coconut mites were present on 88 and 75% of the nuts but no more than three-quarters of those nuts showed damage symptoms. N. aff. paspalivorus was found more on mature nuts than on immature nuts. Spatial and temporal distribution of coconut mites and predatory mites differed significantly. The mean number of coconut mites per nut increased until 5-month-old bunches and declined thereafter. The densities of predatory mites followed a similar trend but peaked 1 month later. Variability in the numbers of mites among palms and bunches of the same age was great, but was relatively low on 6-month-old bunches. The results indicate that assessment of infestation levels by damage symptoms alone is not reliable. Sampling of coconut and/or predatory mite numbers could be improved by using several nuts of 6-month-old bunches. The effect of predatory mites on coconut mites over time suggests that N. aff. paspalivorus could be a prospective biological control agent of A. guerreronis.     

Wild Bearded Capuchin Monkeys (Sapajus libidinosus) Strategically Place Nuts in a Stable Position during Nut-Cracking

Humans can use hand tools smoothly and effectively in varying circumstances; in other words, skillfully. A few other species of primates crack encased foods using hammer tools and anvils. Are they skilled? Positioning the food on the anvil so that it does not fall off when struck is a component of skilled cracking. We discovered that bearded capuchin monkeys deliberately place palm nuts in a relatively stable position on the anvil before striking them. In the first experiment, we marked the meridians of palm nuts where they stopped when rolled on a flat surface (“Stop meridian”). We videotaped monkeys as they cracked these nuts on an anvil. In playback we coded the position of the Stop meridian prior to each strike. Monkeys typically knocked the nuts on the anvil a few times before releasing them in a pit. They positioned the nuts so that the Stop meridian was within 30 degrees of vertical with respect to gravity more often than expected, and the nuts rarely moved after the monkeys released them. In the second experiment, 14 blindfolded people (7 men) asked to position marked nuts on an anvil as if to crack them reliably placed them with the Stop meridian in the same position as the monkeys did. In the third experiment, two people judged that palm nuts are most bilaterally symmetric along a meridian on, or close to, the Stop meridian. Thus the monkeys reliably placed the more symmetrical side of the nuts against the side of the pit, and the nuts reliably remained stationary when released. Monkeys apparently used information gained from knocking the nut to achieve this position. Thus, monkeys place the nuts skillfully, strategically managing the fit between the variable nuts and pits in the anvil, and skilled placement depends upon information generated by manual action.