Getting to the root of neuronal signalling in plant-parasitic nematodes using RNA interference

A variety of genes expressed in preparasitic second-stage juveniles (J2) of plant-parasitic nematodes appear to be vulnerable to RNA interference (RNAi) in vitro by coupling double-stranded (ds)RNA soaking with the artificial stimulation of pharyngeal pumping. Also, there is mounting evidence that the in planta generation of nematode-specific double-stranded RNAs (dsRNAs) has real utility in the control of these pests. Although neuronally-expressed genes in Caenorhabditis elegans are commonly refractory to RNAi, we have discovered that neuronally-expressed genes in plant-parasitic nematodes are highly susceptible to RNAi and that silencing can be induced by simple soaking procedures without the need for pharyngeal stimulation. Since most front-line anthelmintics that are used for the control of nematode parasites of animals and humans act to disrupt neuromuscular coordination, we argue that intercellular signalling processes associated with neurons have much appeal as targets for transgenic plant-based control strategies for plant-parasitic nematodes. FMRFamide-like peptides (FLPs) are a large family of neuropeptides which are intimately associated with neuromuscular regulation, and our studies on flp gene function in plant-parasitic nematodes have revealed that their expression is central to coordinated locomotory activities. We propose that the high level of conservation in nervous systems across nematodes coupled with the RNAi-susceptibility of neuronally-expressed genes in plant-parasitic nematodes provides a valuable research tool which could be used to interrogate neuronal signalling processes in nematodes.

Flatworm nerve-muscle: structural and functional analysis

Platyhelminthes occupy a unique position in nerve-muscle evolution, being the most primitive of metazoan phyla. Essentially, their nervous system consists of an archaic brain and associated pairs of longitudinal nerve cords cross-linked as an orthogon by transverse commissures. Confocal imaging reveals that these central nervous system elements are in continuity with an array of peripheral nerve plexuses which innervate a well-differentiated grid work of somatic muscle as well as a complexity of myofibres associated with organs of attachment, feeding, and reproduction. Electrophysiological studies of flatworm muscles have exposed a diversity of voltage-activated ion channels that influence muscle contractile events. Neuronal cell types are mainly multi- and bi-polar and highly secretory in nature, producing a heterogeneity of vesicular inclusions whose contents have been identified cytochemically to include all three major types of cholinergic, aminergic, and peptidergic messenger molecules. A landmark discovery in flatworm neurobiology was the biochemical isolation and amino acid sequencing of two groups of native neuropeptides: neuropeptide F and FMRFamide-related peptides (FaRPs). Both families of neuropeptide are abundant and broadly distributed in platyhelminths, occurring in neuronal vesicles in representatives of all major flatworm taxa. Dual localization studies have revealed that peptidergic and cholinergic substances occupy neuronal sets separate from those of serotoninergic components. The physiological actions of neuronal messengers in flatworms are beginning to be established, and where examined, FaRPs and 5-HT are myoexcitatory, while cholinomimetic substances are generally inhibitory. There is immunocytochemical evidence that FaRPs and 5-HT have a regulatory role in the mechanism of egg assembly. Use of muscle strips and (or) muscle fibres from free-living and parasitic flatworms has provided baseline information to indicate that muscle responses to FaRPs are mediated by a G-protein-coupled receptor, and that the signal transduction pathway for contraction involves the second messengers cAMP and protein kinase C.

Basiepidermal nervous system in Nemertoderma westbladi (Nemertodermatida): GYIRFamide immunoreactivity

The Nemertodermatida are a small group of microscopic marine worms. Recent molecular studies have demonstrated that they are likely to be the earliest extant bilaterian animals. What was the nervous system (NS) of a bilaterian ancestor like? In order to answer that question, the NS of Nemertoderma westbladi was investigated by means of indirect immunofluorescence technique and confocal scanning laser microscopy. The antibodies to a flatworm neuropeptide GYIRFamide were used in combination with anti-serotonin antibodies and phalloidin-TRITC staining. The immunostaining revealed an entirely basiepidermal NS. A ring lying outside the body wall musculature at the level of the statocyst forms the only centralisation, the

Evolution of the nervous system in Paraphanostoma (Acoela)

According to recent molecular studies, the Acoela are the earliest extant bilaterian group. Their nervous system displays a striking variety of patterns. The aim of the present investigation was to study the variability of the nervous system in a monophyletic group of the Acoela. Six species of Paraphanostoma were chosen for the study. Using immunocytochemical methods and confocal scanning laser microscopy, the immunoreactive patterns of serotonin (5-HT) and the neuropeptide GYIRFamide were described in detail. The study has demonstrated that the brains in Paraphanostoma species, although diverse in detail, still follow the same general pattern. 18S rDNA sequences were used to generate a hypothesis of the phylogeny within the group. Characters of the nervous system revealed in this study were coded and analysed together with 18S rDNA data. Several synapomorphies in the nervous system characters were identified. However, numerous parallelisms in the nervous system evolution have occurred. Data obtained demonstrate that the genus Paraphanostoma is closely related to Childia and should belong to the same family, Childiidae.

Seasonal patterns in activity and habitat use by bats (Pipistrellus spp. and Nyctalus leisleri) in Northern Ireland, determined using a driven transect

The seasonal activity of Leisler's bat Nyctalus leisleri and pipistrelle bats Pipistrellus spp. with respect to minimum bat numbers and habitat use were investigated in County Down, Northern Ireland using a driven transect from April 1998 to October 1998. Data were collected in lowland farmland near Belfast, Northern Ireland using two BatBox III bat detectors tuned to detect both species and species groups simultaneously. The number of bats/km increased during April, May and June, peaking in July and tailed off after this period. The main peak in July is assumed to reflect the occurrence of newly volant young. An increase in the number of pipistrelle social calls during August and September probably represented mating activity. Bat activity correlated with temperature in both N. leisleri and Pipistrellus spp., although bat numbers were independent of temperature after the middle of June. There was significant variation in habitat use by pipistrelle bats along roads over the study period. Pipistrelle bats were observed in greater numbers in areas of tree-line, cut hedge (

A review of the status and decline in abundance of the Irish Hare (Lepus timidus hibernicus) in Northern Ireland

The Northern Ireland Hare Survey documented the distribution of the Irish Hare (Lepus timidus hibernicus). Historical game bag records and other, more contemporary, records of hare distribution were examined. These data indicate how numbers of L t. hibernicus may have changed over the last 140 years. The results of the Northern Ireland Hare Survey suggested that L. t. hibernicus was widespread throughout Northern Ireland. Current average densities are no more than 0.65 hares/km(2). Game bag records indicate that hare densities may have been much higher in the past, with a maximum of 138 hares/km(2) recorded on Crom Estate, Co. Fermanagh, in 1864. Evidence from hare distribution recorded during the Northern Ireland Rabbit Survey indicates that hare numbers declined between 1984 and 1994. Evidence from all sources suggests that L. t. hibernicus has declined in abundance substantially, with present total population estimates for Northern Ireland ranging from 8250 to 21000 individuals. Flushing data indicate that rushes and hedgerows are important diurnal resting areas for hares. While the principal reason for the decline in numbers of L. t. hibernicus in Northern Ireland is not clear, more species-rich pasture and provision of areas of cover, such as rushes, may arrest further declines, or indeed promote numbers of hares, particularly in lowland areas.

The diet and landclass affinities of the Irish hare Lepus timidus hibernicus

The diet of the Irish hare Lepus timidus hibernicus was investigated in Northern Ireland. Faecal pellets were collected at regular intervals from three contrasting study sites and from a number of randomly selected sites from known land classes. Microhistological techniques were employed to analyse the pellets. Grass species occurred in high percentage frequencies in all samples of faecal pellets and a wide variety of plant species were represented (up to 26 species in one study site). In a stratified, random survey of the presence of hares, areas of lowland intensive agriculture had fewer positive records than expected. It is contended that L. t. hibernicus may not be able to sustain viable populations in areas of intensive agriculture, which are almost exclusively composed of ryegrass Lolium spp.

Ovariole number - a predictor of differential reproductive success among worker subfamilies in queenless honeybee (Apis mellifera L.) colonies

A honeybee queen normally mates with 10-20 drones, and reproductive conflicts may arise among a colony's different worker patrilines, especially after a colony has lost its single queen and the workers commence egg laying. In this study, we employed microsatellite markers to study aspects of worker reproductive competition in two queenless Africanized honeybee colonies. First, we determined whether there was a bias among worker patrilines in their maternity of drones and, second, we asked whether this bias could be attributed to differences in the degree of ovary activation of workers. Third, we relate these behavioral and physiological factors to ontogenetic differences between workers with respect to ovariole number. Workers from each of three (colony A) and one (colony B) patrilineal genotypes represented less than 6% of the worker population, yet each produced at least 13% of the drones in a colony, and collectively they produced 73% of the drones. Workers representing these genotypes also had more developed follicles and a greater number of ovarioles per ovary. Across all workers, ovariole development and number were closely correlated. This suggests a strong effect of worker genotype on the development of the ovary already in the postembryonic stages and sets a precedent to adult fertility, so that

Variation in shell morphology of Calliostoma zizyphinum (Gastropoda : Trochidae)

Differences in shell morphology in the intertidal prosobranch mollusc Calliostoma zizyphinum were studied from a number of sites within four geographical reo ions of the British Isles with varying exposures to wave action and crab predation. Mean values of damage scarring were highest in shells sampled from Strangford Lough, Northern Ireland, and lowe;t in individuals from the Atlantic Coast. Shells collected front the Isle of Man were smaller than those from either the Atlantic coast of Ireland or Strangford Lough. Shells front Strangford Lough had taller shells (higher aspect ratio) than shells from the County Down and Atlantic Coast and shells from the County Down coast were more squat (lower aspect ratio) than those from all other areas. Shells from Strangford Lough were significantly thinner than shells from all other geographical areas. The relationships between shell damage scarring and shell size and tallness were not consistent among areas. Shells from Strangford Lough are notable in showing a steep, positive relationship between shell size and scarring and a steep, negative relationship between scarring and tallness. The unusual shell phenotypes observed in shells from Strangford 1,Lough may be explained by rapid shell growth, which would not only allow Calliostoma to attain a size refuge from crab predators but also to recover successfully from repeated crab attacks on the shell lip. Such a strategy would result in larger, thinner shells with a high number of damage scars.

Water requirements as a bottleneck in the reintroduction of European roe deer to the southern edge of its range

Success rates of reintroduction programs are low, often owing to a lack of knowledge of site-specific ecological requirements. A reintroduction program of European roe deer (Capreolus capreolus (L., 1758)) in a dry Mediterranean region in Israel provides an opportunity to study the bottleneck effect of water requirements on a mesic-adapted species. Four does were hand-reared and released in a 10 ha site consisting of an early succession scrubland and a mature oak forest. We measured daily energy expenditure (DEE) and water turnover (WTO) using the doubly labeled water technique during summer and winter. DEE was similar in the summer and winter, but there was a significant difference in WTO and in the source of gained water. In winter, WTO was 3.3 L/day, of which 67% was obtained from vegetation. In summer, WTO dropped to 2.1 L/day, of which only 20% was obtained from the diet and 76% was gained from drinking. When the water source was moved to a nonpreferred habitat, drinking frequency dropped significantly, but water consumption remained constant. In a dry Mediterranean environment, availability of free water is both a physiological contraint and a behavioral constraint for roe deer. This study demonstrates the importance of physiological and behavioral feasibility studies for reintroduction programs.

Differences in resting metabolic rates of two southern African tortoises: Psammobates oculiferus and Geochelone pardalis

Energy metabolism varies considerably between different groups of endotherms, yet there is little or no reported variation among extant groups of reptiles. We measured lower resting metabolic rates (RMRs) in Kalahari tent tortoises (Psammobate oculiferus) than in sympatric Leopard tortoises (Geochelone pardalis). G pardalis also had RMR values that were higher than the allometric prediction for reptiles whereas P oculiferus had RMR values that were not significantly different from the prediction. Differences in RMR between the two species may have occurred because of the large differences in body mass, differing body temperatures, differences in growth rates, or because there may have been differences in the heat increment of feeding.

Individual trade-offs between nutrition and risk of interspecific transmission of disease by grazing: cows, badger latrines and bovine tuberculosis

1. We examine whether various measures of herbivore current physiological state (age, breeding and immune status) and genetic potential can be used as indicators of exposure to and risk from disease. We use dairy cattle and the risks of tuberculosis (TB) transmission posed to them by pasture contaminated with badger excreta (via the fecal-oral route) as a model system to address our aim.

Comparative seasonal acclimatization of food and energy consumption in adjacent populations of common spiny mice (Acomys cahirinus)

Animals inhabiting environments with low productivity and food availability commonly have reduced energy demands and increased digestive efficiencies. The dry matter intake (DMI), apparent digestible dry matter (ADDM), digestible efficiency (DE) and digestible energy intake (DEI) of two populations of common spiny mouse Acomys cahirinus were compared during both winter and summer under conditions of simulated water stress. Mice were captured from the north- and south-facing slopes (NFS and SFS) of the same canyon that represent mesic and xeric habitats, respectively. Measured variables were also compared between F-1 mice that had been born to either NFS or SFS mice, and raised in the laboratory. SFS mice were able to assimilate energy more efficiently than NFS mice during the summer. By comparison, NFS mice were able to assimilate more energy during the winter. During winter, NFS mice assimilated more energy at low levels of water stress, whereas SFS mice assimilated more energy at higher levels. Differences were also apparent in F-1 mice. It is therefore suggested that local climatic conditions can impose physiological adaptations that are retained in succeeding generations, creating unique meta-populations.

Seasonal acclimatization of non-shivering thermogenesis in common spiny mice (Acomys cahirinus) from different habitats

We compared non-shivering thermogenesis between two adjacent populations of freshly captured common spiny mice (Acomys cahirinus) during both winter and summer. Mice were captured from north- and south-facing slopes (NFS and SFS) of the same valley that represent 'Mediterranean' and 'Desert' habitats, respectively. Oxygen consumption and body temperature responses to an injection of exogenous noradrenaline (NA) were higher during the winter than during the summer. in addition, SFS mice had a lower body temperature response to NA during the summer than the other groups of mice. This suggests that heat dissipation is likely to have been greatest in SFS mice during the summer. Overall this study shows that seasonal acclimatization of NST mechanisms is an important trait for small mammals that inhabit the Mediterranean ecosystem. Differences in physiological capabilities can occur temporally within populations across seasons, and spatially between populations that are only a short distance (200-500 m) apart.

Risk of disease from wildlife reservoirs: Badgers, cattle, and bovine tuberculosis

Livestock face complex foraging options associated with optimizing nutrient intake while being able to avoid areas posing risk of parasites or disease. Areas of tall nutrient-rich swards around fecal deposits may be attractive for grazing, but might incur fitness costs from parasites. We use the example of dairy cattle and the risks of tuberculosis transmission posed to them by pastures contaminated with badger excreta to examine this trade-off. A risk may be posed either by aerosolized inhalation through investigation or by ingestion via grazing contaminated swards. We quantified the levels of investigation and grazing of 150 dairy cows at badger latrines (accumulations of feces and urine) and crossing points (urination-only sites). Grazing behavior was compared between strip-grazed and rotation-grazed fields. Strip grazing had fields subdivided for grazing periods of