Acarologic risk of exposure to Borrelia burgdorferi spirochaetes: long-term evaluations in north-western California, with implications for Lyme borreliosis risk-assessment models

Over a 5-year period (1997-2001) the population densities of Ixodes pacificus Cooley & Kohls (Acari: Ixodidae) nymphs infected with spirochaetes of Borrelia burgdorferi sensu lato (s.l.) were evaluated in areas of 2000 ha at two localities (CHR, nine sites; HREC, seven sites) 25 km apart in Mendocino County, north-western California. The 5-year median density of infected nymphs was significantly higher at CHR than at HREC (0.51 vs. 0.09 per 100 m(2) and site-specific yearly densities exceeding one infected nymph per 100 m2 were 10-fold more likely to occur at CHR than at HREC. The importance of long-term data in acarologic risk assessment was demonstrated by significantly higher median yearly densities of infected nymphs at CHR from 1997 to 1999, whereas both areas had similar densities during 2000-2001. Overall, the causative agent of Lyme borreliosis in North America, B. burgdorferi Johnson et al. sensu stricto (s.s.) accounted for 76% of 46 genetically characterized B. burgdorferi s.l. infections from I. pacificus nymphs. Tremendous variability in acarologic risk was recorded within both areas: yearly densities of infected nymphs varied 11-97-fold between sites at CHR and 8-30-fold at HREC. Part of this variation could be explained by environmental traits, most notably deer usage. However, correlations between environmental factors and density of infected nymphs (for CHR and HREC combined) did not necessarily apply when these areas were considered separately. Thus, a Lyme borreliosis ecology model developed in one of these areas needs testing in the other area.     

日本海太平洋褶柔鱼生物学特征的年际变化

根据2010~2013年连续4个年度在日本海进行鱿钓生产采集的样本,利用SSPS软件统计拟合生长和Logistic曲线的方法,分析了太平样褶柔鱼(Todarodes pacificus)的胴长、体重、性别、性成熟度和摄食强度等生物学特征,以及日本海太平洋褶柔鱼群体结构和组成的年际变化。太平洋褶柔鱼总样本数量为388尾,其中雌、雄个体的平均胴长分别为231 mm和230 mm,优势胴长组为220~250 mm。各年度样本的雄雌性比均接近1︰1,总性比为0.89。胴长和体质量关系呈幂函数变化。Logistic曲线拟合个体初次性成熟胴长,其中雌性为216.04 mm,雄性为216.71 mm。样本以性成熟个体为主,成熟率(Ⅲ、Ⅳ和Ⅴ期)高达78.35%。摄食强度主要分布在Ⅰ、Ⅱ、Ⅲ等级,从胃含物组成的出现频率来看,以鱼类最高且主要是沙丁鱼类和灯笼鱼类,频率达70%以上;头足类次之,甲壳类较少。日本海太平洋褶柔鱼各年度的群落结构稍有差异,推测其生长发育与海洋环境因子有关,如温度、海流、盐度、饵料丰度等不同变化造成。     

RNAi mediated gene knockdown and transgenesis by microinjection in the necromenic Nematode Pristionchus pacificus

Although it is increasingly affordable for emerging model organisms to obtain completely sequenced genomes, further in-depth gene function and expression analyses by RNA interference and stable transgenesis remain limited in many species due to the particular anatomy and molecular cellular biology of the organism. For example, outside of the crown group Caenorhabditis that includes Caenorhabditis elegans, stably transmitted transgenic lines in non-Caenorhabditis species have not been reported in this specious phylum (Nematoda), with the exception of Strongyloides stercoralis and Pristionchus pacificus. To facilitate the expanding role of P. pacificus in the study of development, evolution, and behavior, we describe here the current methods to use microinjection for making transgenic animals and gene knock down by RNAi. Like the gonads of C. elegans and most other nematodes, the gonads of P. pacificus is syncitial and capable of incorporating DNA and RNA into the oocytes when delivered by direct microinjection. Unlike C. elegans however, stable transgene inheritance and somatic expression in P. pacificus requires the addition of self genomic DNA digested with endonucleases complementary to the ends of target transgenes and coinjection markers. The addition of carrier genomic DNA is similar to the requirement for transgene expression in Strongyloides stercoralis and in the germ cells of C. elegans. However, it is not clear if the specific requirement for the animals' own genomic DNA is because P. pacificus soma is very efficient at silencing non-complex multi-copy genes or that extrachromosomal arrays in P. pacificus require genomic sequences for proper kinetochore assembly during mitosis. The ventral migration of the two-armed (didelphic) gonads in hermaphrodites further complicates the ability to inject both gonads in individual worms. We also demonstrate the use of microinjection to knockdown a dominant mutant (roller,tu92) by injecting double-stranded RNA (dsRNA) into the gonads to obtain non-rolling F(1) progeny. Unlike C. elegans, but like most other nematodes, P. pacificus PS312 is not receptive to systemic RNAi via feeding and soaking and therefore dsRNA must be administered by microinjection into the syncitial gonads. In this current study, we hope to describe the microinjection process needed to transform a Ppa-egl-4 promoter::GFP fusion reporter and knockdown a dominant roller prl-1 (tu92) mutant in a visually informative protocol.     

CETACEANS OF THE WESTERN TROPICAL INDIAN OCEAN: DISTRIBUTION, RELATIVE ABUNDANCE, AND COMPARISONS WITH CETACEAN COMMUNITIES OF TWO OTHER TROPICAL ECOSYSTEMS

We conducted a cetacean survey in the pelagic western tropical Indian Ocean (WTIO) aboard an 85-m research vessel from March to July 1995, covering 9,784 linear km. Using 25 × binoculars and line-transect methods, we recorded 589 sightings of 21 species. Stenella longirostris was the most abundant cetacean, in terms of number of individuals sighted, by an order of magnitude above any other species, while Physeter macrocephalus was the most frequently sighted, in terms of number of schools. Twelve species were widespread, seven were rare, and two were localized; our sightings include new distributional records for 12 species. Significant observations included the following: (1) Delphinus cf. tropicalis was abundant off the coast of Oman (16 sightings) and readily distinguishable in the field from D. delphis and D. capensis, (2) Balaenoptera musculus was fairly common and localized in the area of the Maldives (17 sightings), and (3) three sightings were made of an unidentified bottlenose whale tentatively referred to as Indopacetus (i. e., Mesoplodon) pacificus. We recorded 26 mixed-species cetacean schools, 43 schools with which seabirds associated, and 17 schools associated with tuna. Notable among these were mixed aggregations of Stenella attenuata, S. longirostris, yellowfin tuna, and seabirds. The cetacean community of the WTIO was similar to that of the eastern tropical Pacific (ETP) and the Gulf of Mexico (GM) in several respects. First, differences in abundance rank of individual species were small, with the result that common species were common and rare species were rare, regardless of ocean. Second, these differences in abundance were due primarily to differences in encounter rate, which varied with ocean by as much as 3,000%, and less so to school size, which generally varied less than 100%. Third, regardless of ocean, three species comprised the majority of cetaceans in the community, Stenella attenuata, S. longirostris, and S. coeruleoalba, representing 62%-82% of all individuals for all species. However, the rank order of abundance for these three species differed with ocean. Most notably, S. attenuata was abundant in the ETP and GM (abundance rank = 2 and 1, respectively) but much less common in the WTIO (abundance rank = 6). Although habitat preferences for S. attenuata appear to overlap considerably with those of S. longirostris in the ETP, our results suggest there may actually be significant differences between these two species. Detailed analysis of oceanographic correlates of distribution will be necessary in order to understand fully the habitat requirements of these pelagic dolphins, often the most conspicuous elements of tropical cetacean communities around the world.     

Transmission cycles of Borrelia burgdorferi and B. bissettii in relation to habitat type in northwestern California

This study was undertaken to determine which rodent species serve as primary reservoirs for the Lyme disease spirochete Borrelia burgdorferi in commonly occurring woodland types in inland areas of northwestern California, and to examine whether chaparral or grassland serve as source habitats for dispersal of B. burgdorferi‐ or B. bissettii‐infected rodents into adjacent woodlands. The western gray squirrel (Sciurus griseus) was commonly infected with B. burgdorferi in oak woodlands, whereas examination of 30 dusky‐footed woodrats (Neotoma fuscipes) and 280 Peromyscus spp. mice from 13 widely‐spaced Mendocino County woodlands during 2002 and 2003 yielded only one infected woodrat and one infected deer mouse (P. maniculatus). These data suggest that western gray squirrels account for the majority of production by rodents of fed Ixodes pacificus larvae infected with B. burgdorferi in the woodlands sampled. Infections with B. burgdorferi also were rare in woodrats (0/47, 0/3) and mice (3/66, 1/6) captured in chaparral and grassland, respectively, and therefore these habitats are unlikely sources for dispersal of this spirochete into adjacent woodlands. On the other hand, B. bissettii was commonly detected in both woodrats (22/47) and mice (15/66) in chaparral. We conclude that the data from this and previous studies in northwestern California are suggestive of a pattern where inland oak‐woodland habitats harbor a B. burgdorferi transmission cycle driven primarily by I. pacificus and western gray squirrels, whereas chaparral habitats contain a B. bissettii transmission cycle perpetuated largely by I. spinipalpis, woodrats, and Peromyscus mice. The dominant role of western gray squirrels as reservoirs of B. burgdorferi in certain woodlands offers intriguing opportunities for preventing Lyme disease by targeting these animals by means of either host‐targeted acaricides or oral vaccination against B. burgdorferi.     

胡瓜钝绥螨对太平洋细须螨的捕食效能初步研究

胡瓜钝绥螨Amblyseius cucumeris Oudemans能捕食为害蝴蝶兰的害螨---太平洋细须螨Tenuipalpus pacificus Baker的卵、幼螨、若螨和成螨。室内研究了胡瓜钝绥螨雌成螨对太平洋细须螨卵、幼螨、若螨的捕食功能反应。结果表明,随着太平洋细须螨密度的增加,胡瓜钝绥螨雌成螨对太平洋细须螨的捕食数量也随之上升,捕食功能反应曲线符合Holling II型方程。胡瓜钝绥螨雌成螨对太平洋细须螨卵的控制能力显著强于对幼螨和若螨的控制能力,a/Th值分别为165.8、79.2和63.8。在猎物密度一定时,由于胡瓜钝绥螨个体间存在相互竞争和相互干扰作用,随着捕食螨密度的提高,捕食螨平均捕食量逐渐减少。     

Vulva formation in Pristionchus pacificus relies on continuous gonadal induction

One of the best known features of vulva development in Caenorhabditis elegans is the induction of vulval precursor cells by the gonadal anchor cell. Induction is crucial for the initiation of pattern formation within the C. elegans vulva equivalence group, and it is therefore surprising to find that this aspect of vulva formation, in particular, varies greatly among nematodes. In some species which form vulvae in the posterior body region, no gonadal signal is necessary for vulva induction. In other nematodes, such as Panagrolaimus, Oscheius, and Rhabditella, vulva formation depends on two temporally distinct gonadal inductions which specify the different cell fates. Here we report our analysis of vulva induction in Pristionchus pacificus, a specieswhich has recently been used as a genetic system to analyze the evolution of vulva development. Cell ablation studies in P. pacificus show that another mode of vulva induction exists. P. pacificus vulva formation depends on a continuous gonadal induction that starts several hours after hatching and continues until the birth of the anchor cell, some 20 h later. Mutations defective in gonadal induction result in the absence of vulva differentiation, suggesting that only one signaling system is involved in the gonadal-epidermal interaction. This new mode adds further to the great variety of gonadal inductions among nematode species. Received: 25 February 1999 / Accepted: 20 April 1999     

Development of a treatment threshold for sucking insects in determinate Bollgard II transgenic cotton grown in winter production areas

Abstract  Little is known about the impact of sucking insects on cotton grown in tropical production areas. To examine this, an experiment was conducted at Katherine in northern Australia to determine the impact of mirids ( Creontiades dilutus (Stål) and Creontiades pacificus (Stål), green vegetable bug ( Nezara veridula (L.)) and red-banded shield bug ( Piezodorus hybneri (Gmelin)) on the yield and fibre quality of the transgenic cotton variety DP50bx containing genes for the expression of Cry1A(c) and Cry2A(b) δ-endotoxins of Bacillus thuringiensis Berliner variety kurstaki . The trial examined the impact of sucking insect populations in the range 0.5–2.0/m². Yields of unsprayed plots were approximately 30% of those obtained in plots sprayed on a threshold of 0.5/m² which did not differ significantly from plots sprayed regularly to exclude sucking insects. The increase in yield in the low pest density treatments was the result of improved fruit retention in the middle part of the plant. Fibre quality was similar in all treatments. However, damage ratings to individual bolls did differ between treatments. Bolls in the high insect density treatments received more damage than those in the low density treatments. As a result of this study, a tentative treatment threshold of 0.5 sucking insect/m² is recommended in determinate Bollgard II cotton varieties grown in winter production areas.     

Natural variation in chemosensation: lessons from an island nematode

All organisms must interact with their environment, responding in behavioral, chemical, and other ways to various stimuli throughout their life cycles. Characterizing traits that directly represent an organism's ability to sense and react to their environment provides useful insight into the evolution of life‐history strategies. One such trait for the nematode Pristionchus pacificus, chemosensation, is involved in navigation to beetle hosts. Essential for the survival of the nematode, chemosensory behavior may be subject to variation as nematodes discriminate among chemical cues to complete their life cycle. We examine this hypothesis using natural isolates of P. pacificus from La Réunion Island. We select strains from a variety of La Réunion beetle hosts and geographic locations and examine their chemoattraction response toward organic compounds, beetle washes, and live beetles. We find that nematodes show significant differences in their response to various chemicals and are able to chemotax to live beetles in a novel assay. Further, strains can discriminate among different cues, showing more similar responses toward beetle washes than to organic compounds in cluster analyses. However, we find that variance in chemoattraction response is not significantly associated with temperature, location, or beetle host. Rather, strains show a more concerted response toward compounds they most likely directly encounter in the wild. We suggest that divergence in odor‐guided behavior in P. pacificus may therefore have an important ecological component.