What the hen can tell about her eggs: egg development on the basis of energy budgets

By a simple model involving the state variables size and storage, it is possible to describe a wide variety of observations on the feeding, growth, energy storage and reproduction of animals. The model is based on the assumption that reproduction, growth as well as maintenance depend on the stored energy only and not directly on feeding. If an egg is thought of as a non-feeding animal, the model predicts the respiration ontogeny and growth of the embryo inside the egg. These predictions seem to hold well for published data on the development of eggs of fish and ratite, precocial and altricial birds. The latter two are known to follow different respiration ontogenies, but both are described well, differing only in one (compound) parameter value. The model explains why the incubation times of eggs of different species tend to increase linearly with egg size to the power 1/4, and why kiwis and petrels, which lay relatively large eggs, have to brood them much longer than larger birds with eggs of the same size. Conversely, it explains why the small eggs of the (parasitic) European cuckoo, hatch earlier than the still smaller eggs of their tiny hosts.Furthermore, it has been shown how the maintenance rate constant, which frequently appears in the microbial literature, can be obtained from measurements on the respiration and weight ontogeny in embryos, so linking independent lines of research. Application of the model shows an increase of the maintenance rate constant from bacteria, crustaceans, up to fish and birds, and a decrease from bacteria to green algae, suggesting lines of evolutionary development.     

Components and patterns in the behaviour of the nematode Caenorhabditis elegans

The behavioural activities during movement, feeding and defaecation have been recorded and measured in adult females of Caenorhabditis elegans . The postures and components of recognizable wave forms are described. Stress has been laid on the mechanism of antagonistic interaction of backward and forward movement, and the rates and characteristics of "spontaneous" and "induced" reversal periods. During feeding, rapid rates of pharyngeal activity are invariably related to low rates of somatic muscle wave propagation. Head oscillations are considered to be separate events not directly linked with feeding or foraging. The combination of certain wave forms, together with other measurements have been used to develop a hypothesis to describe a co-ordinating mechanism applicable to the nematode level of organization.     

kin-18, a C. elegans protein kinase involved in feeding.

TAO1 and TAO2 are recently described protein kinases whose initial characterization has placed them at the mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) kinase kinase (MEKK) level of stress-responsive MAPK pathways. Because their physiological roles have not been identified, we sought to study their C. elegans homolog to learn more about their functions. kin-18 encodes a previously uncharacterized protein in C. elegans whose catalytic domain shares over 60% identity with TAO1 and TAO2. We demonstrate that KIN-18 is a protein of 120 kDa whose promoter is active in the pharynx and intestine of C. elegans. To learn more about TAO/KIN-18 function, we studied how expression of constitutively active forms of TAO1 or KIN-18 would affect the physiology of intact worms. Strains of C. elegans expressing active forms of TAO1 or KIN-18 exhibit altered pharyngeal electrophysiology as measured by electropharyngeogram. These worms grow more slowly and lay fewer eggs, phenotypes that could result from reduced feeding. We have also identified a C. elegans gene that encodes a protein kinase similar to mammalian MAPK/ERK Kinase (MEK) 4 whose promoter is active in the pharynx. It is phosphorylated by TAO1 in vitro and physically interacts with TAO1.     

A decline in p38 MAPK signaling underlies immunosenescence in Caenorhabditis elegans

The decline in immune function with aging, known as immunosenescence, has been implicated in evolutionarily diverse species, but the underlying molecular mechanisms are not understood. During aging in Caenorhabditis elegans, intestinal tissue deterioration and the increased intestinal proliferation of bacteria are observed, but how innate immunity changes during C. elegans aging has not been defined. Here we show that C. elegans exhibits increased susceptibility to bacterial infection with age, and we establish that aging is associated with a decline in the activity of the conserved PMK-1 p38 mitogen-activated protein kinase pathway, which regulates innate immunity in C. elegans. Our data define the phenomenon of innate immunosenescence in C. elegans in terms of the age-dependent dynamics of the PMK-1 innate immune signaling pathway, and they suggest that a cycle of intestinal tissue aging, immunosenescence, and bacterial proliferation leads to death in aging C. elegans.     

Diet-dependent effects of gut bacteria on their insect host: the symbiosis of Erwinia sp. and western flower thrips

Studies on bacteria in the gut of insect species are numerous, but their focus is hardly ever on the impact on host performance. We showed earlier that Erwinia bacteria occur in the gut of western flower thrips, most probably acquired during feeding. Here, we investigate whether thrips gain a net benefit or pay a net cost because of these gut bacteria. On a diet of cucumber leaves, the time to maturity is shorter and the oviposition rate is higher in thrips with bacteria than in thrips without (aposymbionts). When fed on cucumber leaves and pollen, aposymbionts develop faster and lay more eggs. So Erwinia bacteria benefit or parasitize their thrips hosts depending on the diet, which is in accordance with theoretical predictions for fitness of organisms engaged in symbiotic interactions. Possibly, the transmission of gut bacteria has not become strictly vertical because of this diet-dependent fitness variability.     

A liquid-based method for the assessment of bacterial pathogenicity using the nematode Caenorhabditis elegans

Caenorhabditis elegans has previously been used as an alternative to mammalian models of infection with bacterial pathogens. We have developed a liquid-based assay to measure the effect of bacteria on the feeding ability of C. elegans. Using this assay we have shown that Pseudomonas aeruginosa strain PA14, Burkholderia pseudomallei and Yersinia pestis were able to inhibit feeding of C. elegans strain N2. An increase in sensitivity of the assay was achieved by using C. elegans mutant phm-2, in place of the wild-type strain. Using this assay,P. aeruginosa PA01 inhibited the feeding of C. elegans mutant phm-2. Such liquid-based feeding assays are ideally suited to the high-throughput screening of mutants of bacterial pathogens.     

基于16S rDNA基因序列的泽兰实蝇幼虫肠道细菌多样性分析

【目的】探究泽兰实蝇 Procecidochares utilis 幼虫肠道细菌的多样性。【方法】利用Illumina HiSeq技术对泽兰实蝇幼虫肠道细菌的16S rDNA-V6变异区序列进行测序,应用USEARCH和QIIME等软件整理和统计样品序列数目和操作分类单元（operational taxonomic unit, OTU）数量,分析物种的丰度和Alpha多样性。【结果】共获得1 593 506 对reads,拼接为1 579 372条tags,经过滤后得到的1 572 860条tags聚类为1 341个OTU。总共注释到13个门,4个纲,6个目,7个科,10个属和4个种。其中变形菌门(Proteobacteria)的细菌为优势菌,占99%;在属分类阶元上,沃尔巴克氏体属 Wolbachia 占45%,是优势属。【结论】泽兰实蝇幼虫肠道细菌多样性丰富。相关的种类和丰度信息为后期研究揭示肠道细菌介导泽兰实蝇寄主植物专化性奠定了基础。     

“Male factors” in ticks: their role in feeding and egg development

Summary

Female ticks of the family Ixodidae utilize their salivary glands as the major organs for fluid balance, secreting back into the host a dilute saliva. Feeding is composed of three phases: a preparatory phase (1–2 days) during which the tick establishes the feeding lesion, a slow phase (~7 days) during which body weight increases 10-fold, and a rapid phase (~1 day) in which body weight increases a further 10-fold. Following engorgement, the salivary glands are resorbed by an autolytic process triggered by an ecdysteroid hormone. If a female is removed from the host prior to repletion, her subsequent behaviour depends mostly on two factors: the degree of engorgement achieved and whether or not she has mated. If removed during the preparatory or slow phase of engorgement, the salivary glands are not resorbed, the tick will lay virtually no eggs and she will reattach to a host if given the opportunity, all of this irrespective of whether she is virgin or mated. If removed during the rapid phase of engorgement, however, mated females will not reattach to a host even if given the opportunity. Instead, they will resorb the salivary glands within 4 days post-removal and lay a batch of eggs. Virgin females removed after exceeding 10-fold the unfed weight likewise refuse to resume feeding if given the opportunity, but salivary gland reabsorption is delayed (to 8 days post-removal); if any eggs are laid, they are infertile. A number of chemical “factors” entering the female during copulation influence her feeding behaviour and egg development. Here we discuss the complexities of these interactions and suggest how they might be adaptive to ticks in nature.     

野生和人工饲喂条件下甘肃鼢鼠肠道细菌多样性比较

【目的】肠道微生物在宿主代谢和健康方面发挥着重要的作用,已有报道表明甘肃鼢鼠肠道菌群和地域、性别以及季节等因素有关,但对于野生和人工饲喂条件下,取食不同食物的甘肃鼢鼠肠道菌群是否存在差异尚不明确。【方法】本研究选取苜蓿、樟子松根、油松根和云杉根对甘肃鼢鼠进行分组饲喂,同时以野生个体作为对照,采用16S rRNA V3-V4高通量测序技术对野生和人工饲喂甘肃鼢鼠肠道细菌进行比较。【结果】野生和人工饲喂条件下,优势菌门均为Firmicutes和Bacteroidetes,但含量却存在显著性差异。在人工饲喂条件下,樟子松根、油松根和云杉根饲喂的三组甘肃鼢鼠肠道细菌群落结构相似度较高,而与苜蓿组和野生组之间均存在较大差异,樟子松组、油松组和云杉组甘肃鼢鼠肠道细菌多样性高于苜蓿组和野生组。【结论】野生和人工饲喂条件下甘肃鼢鼠肠道菌群组成和结构存在显著性差异。     

Salmonella typhimurium proliferates and establishes a persistent infection in the intestine of Caenorhabditis elegans

Genetic analysis of host-pathogen interactions has been hampered by the lack of genetically tractable models of such interactions. We showed previously that the human opportunistic pathogen Pseudomonas aeruginosa kills Caenorhabditis elegans, that P. aeruginosa and C. elegans genes can be identified that affect this killing, and that most of these P. aeruginosa genes are also important for mammalian pathogenesis. Here, we show that Salmonella typhimurium as well as other Salmonella enterica serovars including S. enteritidis and S. dublin can also kill C. elegans. When C. elegans is placed on a lawn of S. typhimurium, the bacteria accumulate in the lumen of the worm intestine and the nematodes die over the course of several days. This killing requires contact with live bacterial cells. The worms die with similar kinetics when placed on a lawn of S. typhimurium for a relatively short time (3-5 hours) before transfer to a lawn of E. coli. After the transfer to E. coli, a high titer of S. typhimurium persists in the C. elegans intestinal lumen for the rest of the worms' life. Furthermore, feeding for 5 hours on a 1:1000 mixture of S. typhimurium and E. coli followed by transfer to 100% E. coli, also led to death after several days. This killing correlated with an increase in the titer of S. typhimurium in the C. elegans lumen, which reached 10,000 bacteria per worm. These data indicate that, in contrast to P. aeruginosa, a small inoculum of S. typhimurium can proliferate in the C. elegans intestine and establish a persistent infection. S. typhimurium mutated in the PhoP/PhoQ signal transduction system caused significantly less killing of C. elegans.     

Ovarian control of blood meal retention the the mosquito Anopheles freeborni

Female Anopheles freeborni begin to defaecate the haem containing residue of a blood meal about the time their ova mature, 40–45 hr after feeding. Ovariectomized females begin to defaecate 15 hr prematurely, while ovariectomized females treated with 20-hydroxyecdysone retain blood meals longer, proportional to dose, than do those untreated. It is concluded that the production of ecdysteroids by the ovary initiates a mechanism for retention of the blood meal in the mosquito.     

An altered method of feeding RNAi that knocks down multiple genes simultaneously in the nematode Caenorhabditis elegans

In reverse genetics, RNA interference (RNAi) which is substitutable for gene-disruption, is an outstanding method for knockdown of a gene's function. In Caenorhabditis elegans, feeding RNAi is most convenient, but this RNAi is not suitable for knockdown of multiple genes. Hence, we attempted to establish an efficient method of feeding RNAi for multiple knockdown. We produced bacteria yielding three distinct double-stranded RNAs bound to one another, and fed those bacteria to C. elegans. Quantitative RT-PCR and observation of phenotypes indicated that our method is much more efficient than the traditional one. Our method is useful for investigating genes' functions in C. elegans.     

Egg and larval load assessment and its influence on oviposition behaviour of the leaf beetle Galerucella nymphaeae

The oviposition behaviour of the water-lily beetle Galerucella nymphaeae was examined. This species is a specialist herbivore on the floating leaves of nymphaeids Nymphaeaceae and especially on the yellow water-lily, Nuphar lutea. Females lay their eggs in clutches on the leaves, and after hatching, the larvae feed on the leaves. The quality of the leaves decreases quickly after the larvae hatch, and eventually the leaves will sink below the water surface, whereupon the eggs, 1st-instar larvae and pupae are killed by drowning. The influence of conspecific eggs, larvae and feeding tracks on the oviposition preferences of the beetles was tested. Females were allowed to choose between fresh leaves and leaves with conspecific eggs and larvae as well as between leaves with larvae and leaves with feeding tracks but no larvae. An attempt was also made to determine whether eggs and larvae affect the oviposition rate of females when they are not given the opportunity to oviposit on untouched leaves. The results indicate that females tended to avoid leaves with conspecific larvae or to exhibit a decreased oviposition rate on such leaves. Females also avoided conspecific eggs, although the oviposition rate was not influenced by the presence of conspecific eggs. When females were allowed to choose between leaves with larvae and leaves with feeding tracks, possible discrimination against leaves with larvae just fails to reach the 5% level.     

Oogenesis in the date stone beetle, Coccotrypes dactyliperda, depends on symbiotic bacteria

Abstract.  It has been suggested that sex ratio distorting symbionts are involved in the sex determination and female-biased sex ratios observed in strongly inbred scolytid beetles. Coccotrypes dactyliperda (Coleoptera: Scolytinae) is a species in which mother-son- and sib-mating occur inside the date seeds it inhabits, and the sex ratios produced are highly skewed toward females. In the present study, polymerase chain reaction (PCR) techniques and antibiotic treatments are applied to determine the possible role of Bacteria in this system. PCR with primers specifically designed to target the 16S rDNA gene in all Bacteria reveals the presence of Wolbachia and Rickettsia in control beetles, but not in antibiotic-treated individuals. Virgin females fed with antibiotics lay no eggs, and no sign of oogenesis is detected compared with all-male progeny of virgin control females. Mated females fed with antibiotics lay significantly fewer eggs than control females, with a strong effect of female age at the time of antibiotic feeding on the number of eggs laid. The study suggests that symbiotic bacteria are not involved in female-biased sex ratios but are required for oogenesis in C. dactyliperda . The specific role each of the bacteria ( Wolbachia and Rickettsia ) plays in the oogenesis remains to be determined.     

A potential tradeoff between feeding rate and aversive learning determines intoxication in a Caenorhabditis elegans host-pathogen system

Despite being the first line of defense against infection, little is known about how host-pathogen interactions determine avoidance. Caenorhabditis elegans can become infected by chemoattractant-producing bacteria through ingestion. The worms can learn to associate these chemoattractants with harm through aversive learning. As a result, the worms will avoid the pathogen. Evolutionary constraints have likely shaped the attraction, intoxication and learning dynamics between bacteria and C. elegans, but these have not been explored. Using bacteria engineered to express an acylhomoserine lactone chemoattractant and a nematicidal protein, we explored how manipulating the amount of attractant produced by the bacteria affects learning and intoxication in mixed stage populations of C. elegans. We found that increasing the production rate of the chemoattractant increased the feeding rate in C. elegans, but decreased the time required for C. elegans to learn to avoid the chemoattractant. Learning generally coincided with a decreased feeding rate. We also observed that the percentage of intoxicated worms was maximized at intermediate production rates of the attractant. We propose that interactions between attractant driven feeding rate and aversive learning are likely responsible for this trend. Our results increase our understanding of behavioral avoidance in C. elegans and have implications in understanding host-pathogen dynamics that shape avoidance.     

Evolution of parental care in dart poison frogs (Amphibia: Anura: Dendrobatidae)

Dendrobatid frogs perform a unique mode of parental care, ranging from egg attendance and tadpole transport to tadpole feeding. It is hypothesized that a behaviour in which the egg attending parent remains with the cluch is the most primitive condition. In more advanced forms or parental care, the male is able to attend several clutches of eggs. Tadpole attendance and feeding, finally, started as deceit; the male induced the female to lay eggs into a bromeliad leaf axil already occupied by a larva. Costs and benefits of the different modes of parental care and possible alternative reproductive tactics are discussed.     

Towards a virtual C. elegans: a framework for simulation and visualization of the neuromuscular system in a 3D physical environment

The nematode C. elegans is the only animal with a known neuronal wiring diagram, or "connectome". During the last three decades, extensive studies of the C. elegans have provided wide-ranging data about it, but few systematic ways of integrating these data into a dynamic model have been put forward. Here we present a detailed demonstration of a virtual C. elegans aimed at integrating these data in the form of a 3D dynamic model operating in a simulated physical environment. Our current demonstration includes a realistic flexible worm body model, muscular system and a partially implemented ventral neural cord. Our virtual C. elegans demonstrates successful forward and backward locomotion when sending sinusoidal patterns of neuronal activity to groups of motor neurons. To account for the relatively slow propagation velocity and the attenuation of neuronal signals, we introduced "pseudo neurons" into our model to simulate simplified neuronal dynamics. The pseudo neurons also provide a good way of visualizing the nervous system's structure and activity dynamics.     

Measuring the Effects of Bacteria on C. Elegans Behavior Using an Egg Retention Assay

C. elegans egg-laying behavior is affected by environmental cues such as osmolarity¹ and vibration². In the total absence of food C. elegans also cease egg-laying and retain fertilized eggs in their uterus³. However, the effect of different sources of food, especially pathogenic bacteria and particularly Enterococcus faecalis, on egg-laying behavior is not well characterized. The egg-in-worm (EIW) assay is a useful tool to quantify the effects of different types of bacteria, in this case E. faecalis, on egg- laying behavior.EIW assays involve counting the number of eggs retained in the uterus of C. elegans⁴. The EIW assay involves bleaching staged, gravid adult C. elegans to remove the cuticle and separate the retained eggs from the animal. Prior to bleaching, worms are exposed to bacteria (or any type of environmental cue) for a fixed period of time. After bleaching, one is very easily able to count the number of eggs retained inside the uterus of the worms. In this assay, a quantifiable increase in egg retention after E. faecalis exposure can be easily measured. The EIW assay is a behavioral assay that may be used to screen for potentially pathogenic bacteria or the presence of environmental toxins. In addition, the EIW assay may be a tool to screen for drugs that affect neurotransmitter signaling since egg-laying behavior is modulated by neurotransmitters such as serotonin and acetylcholine^5-9.     

Effects of the novel anthelmintic emodepside on the locomotion, egg-laying behaviour and development of Caenorhabditis elegans

Emodepside, a cyclooctadepsipeptide, is a broad-spectrum anthelmintic previously shown to paralyse body wall muscle and pharyngeal muscle in the model nematode Caenorhabditis elegans. We demonstrate that wild-type C. elegans L4 are less sensitive than adults to emodepside in two independent assays of locomotor behaviour: body bend generation on agar (adult IC(50) 3.7 nM, L4 IC(50) 13.4 nM) and thrashing behaviour in liquid (thrashing behaviour as a % of controls after 1h in 10 microM emodepside: adults 16%, L4 worms 48%). We also show that continuous exposure of wild-type C. elegans to emodepside throughout the life-cycle from egg onwards, slows worm development, an effect that is emodepside concentration-dependent. The rate of worm-hatching from eggs on agar plates containing emodepside was not significantly different from controls, suggesting that it is development post-hatching rather than hatching itself that is affected by the drug. Emodepside also inhibits wild-type C. elegans egg-laying, with acute exposure to the drug at 500 nM resulting in an almost total inhibition within the first hour. However, the rate of egg production was not inhibited and therefore emodepside-treated worms became bloated with eggs, eventually rupturing. This suggests that the effect of emodepside on reproduction is not due to an inhibition of egg production but rather a paralytic effect on the egg-laying muscles. These results, when coupled with previous research, suggest that emodepside interferes with signalling at the neuromuscular junction on the body-wall muscles (Willson et al., 2003), pharynx (Willson et al., 2004) and egg-laying muscles and thus inhibits three important physiological functions: locomotion, feeding and reproduction.     

柳蓝叶甲的生物学特性室内观察

在实验室条件下(27±0.5℃,12 D:12 L)研究了柳蓝叶甲Plagiodera versicolora的生物学特性。结果表明,幼虫每隔2~3 d蜕皮1次,共蜕皮2次。成虫产卵前期4 d,世代周期17 d。产卵具有一定的周期性,约10 d为1个周期。叶龄对幼虫的生长发育和成虫的产卵行为有极显著的影响。当幼虫以成熟叶为食料时,其生长发育速率明显减慢,各龄期延长,蛹重较轻;成虫以成熟叶或老叶为食料时产卵几乎完全被抑制,而以幼叶为食料时几乎每天均能产卵。此外,柳蓝叶甲产出卵表面化合物对产卵行为有极为显著的影响。卵表面的有机提取物对成虫产卵具有显著的引诱作用;相反,卵表面的水提取物对产卵有一定的驱避作用。