小菜蛾幼虫对不同寄主的取食嗜好性及其适宜性

对不同寄主种类、不同寄主形态和不同寄主饲喂的小菜蛾(Plutella xylostella)幼虫之间的取食嗜好性比较试验表明,小菜蛾幼虫优先取食大白菜、萝卜或菜心幼苗,其次为油菜和甘蓝幼苗,在大白菜与油菜幼苗之间的取食选择比例是93.33%和6.67％;在甘蓝与菜心幼苗之间的取食选择比例是16.67%和83.33%.小菜蛾幼虫的取食嗜好性受饲喂寄主种类的影响,偏食大白菜或菜心幼苗.小菜蛾幼虫选择寄主取食的次序与寄主体内可溶性糖或淀粉含量没有明显关系,但与两者的相对量呈一定的负相关.取食大白菜或菜心幼苗的小菜蛾生长良好,单头取食达0.583～0.637 cm², 单头体重达2.07～2.18 mg, 与取食甘蓝或油菜幼苗的幼虫在取食量、个体发育方面有明显差异.小菜蛾幼虫也喜好取食已经被虫危害过的幼苗.     

虫害诱导的植株对小菜蛾取食和生长发育的影响

本文从菜田生态系统的角度出发,就黄曲条跳甲Phyllotreta striolata(Fabricius)取食诱导对小菜蛾Plutella xylostella(L.)造成的影响进行了研究。黄曲条跳甲取食诱导对小菜蛾取食的影响比较明显,一般来说,黄曲条跳甲少量或短时间的取食会刺激小菜蛾幼虫在相应叶片上的取食,反之则会抑制;而且不同的寄主,不同完整度的叶片(即是否被黄曲条跳甲取食过)上的承载能力不同,芥菜、白菜、菜心和萝卜的承载力弱于甘蓝和芥蓝,黄曲条跳甲取食过的叶片弱于未被黄曲条跳甲取食过的叶片;不同数量的黄曲条跳甲取食对小菜蛾的产卵量影响显著,对其余生物学参数影响不显著,少量的黄曲条跳甲取食会刺激小菜蛾的产卵,多数则会抑制。     

草履蚧不同寄主和地理种群遗传分化的RAPD分析

应用随机扩增多态性DNA(RAPD)技术对草履蚧保定、石家庄、邯郸16个不同寄主地理种群遗传多样性和种群分化进行研究,结果显示4个RAPD引物共扩增出41个多态性位点,多态位点比率为100%。遗传距离指数在0.701—0.4360,平均为0.2395。其中以邯郸枫杨和邯郸垂丝海棠为寄主的草履蚧种群遗传距离最小(0.0701);以石家庄紫叶李和邯郸木槿为寄主的种群遗传距离最大(0.4360)。遗传一致度系数在0.6466—0.9290。说明草履蚧不同种群遗传多样性丰富并存在遗传差异。聚类分析结果表明草履蚧种群遗传多样性同时受到寄主和地理因素的双重影响,且不同寄主草履蚧种群已产生明显的遗传分化。     

日本稻蝗、中华稻蝗和赤胫伪稻蝗地理种群的RAPD遗传分化研究

运用10个RAPD引物对日本稻蝗(Oxya japonica)、中华稻蝗(Oxya chinensis)和赤胫伪稻蝗(Pseudoxya diminuta)的种群遗传分化进行分析。10个随机引物共产生135条带,扩增谱带具有明显的属、种间多态性。Shannon信息指数表明中华稻蝗遗传多样性水平较高(2．693),日本稻蝗次之(2.319),赤胫伪稻蝗最低(1．042)。中华稻蝗和日本稻蝗的不同地理居群出现遗传分化,由Shannon信息指数估算的种群间遗传分化系数分别为20．7％,42．4％。用UPGMA和NJ法对Nei’s遗传距离作聚类分析,构建分子系统树。系统树显示：同一种群的不同个体优先相聚,而后,同一种的不同种群依次相聚;日本稻蝗广西南宁种群和广东广州种群首先聚为一支,陕西西安种群和浙江杭州种群聚为另一支,两支相聚后与中华稻蝗聚在一起,最后与赤胫伪稻蝗相聚。聚类结果表明：不同地域日本稻蝗亲缘关系的远近与地理距离呈现一定的相关趋势,日本稻蝗与中华稻蝗亲缘关系较近,Nei’S遗传距离平均为0．142,而赤胫伪稻蝗与它们关系较远,Nei’s遗传距离平均为0．451。聚类图所显示的物种间亲缘关系的远近程度与形态分类学和细胞分类学结果相一致。     

中国二化螟不同地理种群的AFLP遗传分析

利用AFLP标记对中国11省13个二化螟Chilo suppressalis(Walker)地理种群进行遗传多态性分析,以揭示不同区域种群间遗传分化,为建立较准确的二化螟发生区划及区域治理对策提供新的依据。选取3对引物,共扩增出445个位点,其中多态性位点数为386,多态性比率为86.8%。种群间分化系数(Gst)为0.74,基因流仅为0.176;13个地理种群间遗传一致度较高,均大于0.67。遗传距离聚类分析表明,二化螟13个地理种群可区分为4类,其中淮北平原类(赣榆、阜宁、阜南)、东南山区类(鄞县、闽侯)、荆湘川类(荆州、邵阳、德阳)三类种群分别具有较一致的地理气候特征或相对较近的距离。研究还表明,与其他测试种群的遗传距离普遍较大、相对孤立的地理种群,其聚类结果有一定的随机性,因此,取样点较多、且样点分布较均匀的聚类分析结果才有较高可信度。     

15个芥蓝的表型多样性和SCoT多态性评价

芥蓝(Brassica oleracea L.var.alboglabra Bailey)是起源于我国南方的甘蓝性蔬菜。为探讨和评价芥蓝种质资源的遗传多样性,本研究选用15个典型的芥蓝品种进行了7个性状的表型多样性和SCo T多态性分析。结果发现齐口期、株高、茎粗、开展度、叶重、薹茎重和分枝数的变异系数分别为21.60%、30.32%、23.21%、69.77%、65.16%、48.71%和43.04%。只有薹茎重与叶重的线性相关达到显著水平。表型分析的遗传距离变化范围是22.865 5~269.316 5,平均值是113.817 3,在122.7处将15个品种分为3类。SCo T分析中29条引物扩增出119条带,平均每条引物4.1条带,多态性条带93条,多态性百分率为78%。在各个品种中平均扩增79条。分子遗传距离变化范围为0.154 2~0.516 4,平均值是0.339 9,在0.20处将15个品种分为3类。10个品种的表型和分子聚类结果相同,但mantel test检验表明表型多样性和SCo T多态性相关不显著。本研究结果表明SCo T可以作为一种鉴定芥蓝种质资源遗传多样性的简单的常规分子标记。     

柴达木地区野生黑果枸杞种群遗传多样性的AFLP分析

采用扩增片段长度多态性(AFLP)分子标记技术对青海省柴达木地区5个野生黑果枸杞(Lycium ruthenicum)种群的120份样品的遗传多样性进行分析。结果表明: 柴达木地区野生黑果枸杞具有很高的遗传多样性, 9对选扩引物共得到1691条清晰条带, 其中多态性条带1678条, 多态性变异率为99.23%, 种群间的有效等位基因数为1.4712, Nei’s基因多样性为0.3245, Shannon信息指数为0.4367。分子方差分析(AMOVA)结果表明: 柴达木地区5个黑果枸杞种群的遗传变异主要存在于种群内部(92%), 种群间的遗传分化较小(8%, 遗传分化系数0.08)。黑果枸杞种群间的遗传相似系数介于0.9709-0.9922之间, 平均值为0.9835。种群间的聚类及Mantel检验(γ = 0.3368, p = 0.8064)均表明柴达木地区黑果枸杞种群地理距离与遗传距离之间的相关性不明显; 黑果枸杞个体间的聚类表明同一种群的个体不能完全聚在一起。对同一种源的遗传多样性分析发现, 诺木洪奥斯勒草场的种源内部的遗传变异更为丰富, 这或许可以推断诺木洪可能为柴达木地区野生黑果枸杞种质资源的中心产区。     

十字花科蔬菜对小菜蛾实验种群的影响

通过组建4种十字花科蔬菜上小菜蛾实验种群生命表发现,在菜心、芥蓝、芥菜和小白菜4种蔬菜上繁殖的小菜蛾实验种群增长趋势指数分别为41．0、69．5、38．2、52．4,影响种群增长的重要因子为幼虫的自然死亡,不同蔬菜种类对小菜蛾雌蛾的总产卵量和寿命无影响,但芥蓝繁殖的小菜蛾日平均产卵量显著高于其它几种蔬菜。在芥蓝和小白菜上繁殖的小菜蛾幼虫具有较高的存活概率,是其种群具有较大增长潜力的重要原因。试验结果表明,在4种蔬菜中,芥蓝提供的营养最适宜于小菜蛾种群生长、发育和繁殖,是最佳的寄主,其次为小白菜、菜心,而芥菜尽管被报道为小菜蛾成虫最喜欢产卵的寄主,由于所繁殖的小菜蛾种群在幼虫期有较高的死亡率,是最不适宜于小菜蛾生长发育的寄主。     

日本稻蝗部分种群遗传多样性研究

采用随机引物扩增多态性分析方法,研究我国长江以南地理距离跨度较大的6个不同种群日本稻蝗Oxya japonica的种群遗传学关系.10条随机引物对57个日本稻蝗个体共扩增出131个片段,其中多态性位点为128个,总的多态位点百分率是97.7%.种群问的遗传分化系数表明:日本稻蝗的遗传变异大部分存在于种群内.Nei's遗传距离显示:日本稻蝗种群问的遗传距离存在较大差异,河北平山种群和云南种群遗传距离最大,为0.1715,广东广州种群和广西南宁种群遗传距离最小,为0.0770.聚类结果分析表明,地理上相距较近的种群优先相聚,遗传距离、地理距离和亲缘关系间呈现出一定的相关趋势.     

不同干扰生境下河口莲座蕨遗传多样性分析

该研究利用 ISSR 分子标记,对不同生境下的6个河口莲座蕨种群进行遗传多样性分析。结果表明：从44条ISSR引物中筛选出8条能够扩增出清晰、稳定条带的引物,对6个河口莲座蕨种群进行基因组 DNA扩增,共扩增出144条带,其中多态性条带119个,多态性条带比率为93.7％,Nei’s遗传多样性指数(H)为0.296,Shannon多样性指数(I)为0.457,遗传分化指数(Gst)为0.1520,种群遗传一致度和遗传距离分别为0.9138~0.9548和0.0584~0.0901;UPGMA聚类分析表明,种群间遗传距离与空间距离和生境类型有关。河口莲座蕨在不同干扰程度的生境中,种群具有高水平的遗传多样性,一定强度或频率的干扰生境中,种群遗传多样性较高,与海拔、坡度和坡向无相关性。     

The effect of chronic ethanol ingestion on protein metabolism in type-I- and type-II-fibre-rich skeletal muscles of the rat.

1. The effects of chronic ethanol feeding on muscles containing a predominance of either Type I (aerobic, slow-twitch) or Type II (anaerobic, fast-twitch) fibres were studied. Male Wistar rats, weighing approx. 90 g or 280 g, were pair-fed on a nutritionally complete liquid diet containing 36% of total energy as ethanol, or isovolumetric amounts of the same diet in which ethanol was replaced by isoenergetic glucose. After 6 weeks feeding, fractional rates of protein synthesis were measured with a flooding dose of L-[4-(3)H]-phenylalanine and muscles were analysed for protein, RNA and DNA. 2. Ethanol feeding decreased muscle weight, protein, RNA and DNA contents in both small and large rats. Type-II-fibre-rich muscles showed greater changes than did Type-I-fibre-rich muscles. Changes in protein paralleled decreases in DNA. 3. The capacity for protein synthesis (RNA/protein), fractional rates of protein synthesis and absolute rates of protein synthesis were decreased by ethanol feeding in both small and large rats. The amounts of protein synthesized relative to RNA and DNA were also decreased. Changes were less marked in Type-I than in Type-II-fibre-rich muscles. Loss of protein, RNA and DNA was greater in small rats, but protein synthesis was more markedly affected in large rats. 4. It was concluded that chronic ethanol feeding adversely affects protein metabolism in skeletal muscle. Fibre composition and animal size are also important factors in determining the pattern of response.     

The gastrointestinal tract as the portal of entry for foreign macromolecules: fate of DNA and proteins

The gastrointestinal tract (GIT) of mammals is the main portal of entry for foreign DNA and proteins. We have documented the fate of orally administered DNA or protein in the GIT of the mouse. The gene for the Green Fluorescent Protein (GFP) (4.7 kb) and the genomes of bacteriophage M13 (7.25 kb) and adenovirus type 2 (Ad2; 35.9 kb) were used as test DNAs. Persistence of these DNAs in the GIT was monitored by Southern hybridization and fluorescent in situ hybridization (FISH) or by PCR. For studies on proteins, recombinant glutathione-S-transferase was fed to mice. Survival of the protein in the GIT was then assessed by Western blotting. Depending on feeding schedules and food regimens, but irrespective of mouse strain or DNA length, fragments of the GFP gene or other DNAs were detectable for up to 18 h after feeding by Southern blot analysis. The GFP DNA could be visualized by FISH in cecal epithelia. A high fiber diet reduced the time required for food to pass through the GIT, and foreign DNA was cleared more rapidly. A high fat diet or complexing of the foreign DNA with protamine or lipofectin did not extend DNA persistence times. Undegraded GST protein was detected only in foregut contents up to 30 min after feeding. At 15 and 30 min post feeding, trace amounts of GST were found in extracts of the kidney. The GIT is constantly exposed to highly recombinogenic fragments of foreign DNA and to intact foreign proteins. Our data have implications for studies on carcinogenesis and mutagenesis, and on the pathogenicity of infectious proteins such as prions.The first two authors contributed equally to this work     

Effects of starvation and hormones on DNA synthesis in silk gland cells of the silkworm,Bombyx mori

Silk gland cells of silkworm larvae undergo multiple cycles of endomitosis for the synthesis of silk proteins during the spinning phase. In this paper, we analyzed the endomitotic DNA synthesis of silk gland cells during larval development, and found that it was a periodic fluctuation, increasing during the vigorous feeding phase and being gradually inhibited in the next molting phase. That means it might be activated by a self‐regulating process after molting. The expression levels of cyclin E, cdt1 and pcna were consistent with these developmental changes. Moreover, we further examined whether these changes in endomitotic DNA synthesis resulted from feeding or hormonal stimulation. The results showed that DNA synthesis could be inhibited by starvation and re‐activated by re‐feeding, and therefore appears to be dependent on nutrition. DNA synthesis was suppressed by in vivo treatment with 20‐hydroxyecdysone (20E). However, there was no effect on DNA synthesis by in vitro 20E treatment or by either in vivo or in vitro juvenile hormone treatment. The levels of Akt and 4E‐BP phosphorylation in the silk glands were also reduced by starvation and in vivo treatment with 20E. These results indicate that the activation of endomitotic DNA synthesis during the intermolt stages is related to feeding and DNA synthesis is inhibited indirectly by 20E.     

Effect of Chronic Caloric Restriction on the Circadian Regulation of Physiological and Behavioral Variables in Old Male B6C3F1 Mice

The circadian rhythms of food and water consumption, the number of feeding and drinking episodes, oxygen consumption, carbon dioxide production, respiratory quotient, gross motor activity, and body temperature were measured in male B6C3F, mice that were fed ad libitum (AL) or fed a caloric-restricted diet (CR). The CR regimen (60% of the normal AL consumption) was fed to mice during the daytime (5 hr after lights on). CR animals exhibited fewer feeding episodes but consumed more food per feeding bout and spent more total time feeding than AL mice. It appears that CR caused mice to change from their normal “nibbling behavior” to meal feeding. Compared to AL animals, the mean body temperature was reduced in CR animals, while the amplitude of the body temperature rhythm was increased. Spans of reduced activity, metabolism, and body temperature (torpor) occurred in CR mice for several hours immediately before feeding, during times of high fatty acid metabolism (low RQ). The acute availability of exogenous substrates (energy supplies) seemed to modulate metabolism shifting metabolic pathways to promote energy efficiency. CR was also associated with lower DNA damage, higher DNA repair, and decreased proto-oncogene expression. Most of the circadian rhythms studied seemed to be synchronized primarily to the feeding rather than the photoperiod cycle. Night-time CR feeding was found to be better than daytime feeding because the circadian rhythms for AL and CR animals were highly synchronized when this regimen was used.     

Effect of Chronic Caloric Restriction on the Circadian Regulation of Physiological and Behavioral Variables in Old Male B6C3F1 Mice

The circadian rhythms of food and water consumption, the number of feeding and drinking episodes, oxygen consumption, carbon dioxide production, respiratory quotient, gross motor activity, and body temperature were measured in male B6C3F, mice that were fed ad libitum (AL) or fed a caloric-restricted diet (CR). The CR regimen (60% of the normal AL consumption) was fed to mice during the daytime (5 hr after lights on). CR animals exhibited fewer feeding episodes but consumed more food per feeding bout and spent more total time feeding than AL mice. It appears that CR caused mice to change from their normal “nibbling behavior” to meal feeding. Compared to AL animals, the mean body temperature was reduced in CR animals, while the amplitude of the body temperature rhythm was increased. Spans of reduced activity, metabolism, and body temperature (torpor) occurred in CR mice for several hours immediately before feeding, during times of high fatty acid metabolism (low RQ). The acute availability of exogenous substrates (energy supplies) seemed to modulate metabolism shifting metabolic pathways to promote energy efficiency. CR was also associated with lower DNA damage, higher DNA repair, and decreased proto-oncogene expression. Most of the circadian rhythms studied seemed to be synchronized primarily to the feeding rather than the photoperiod cycle. Night-time CR feeding was found to be better than daytime feeding because the circadian rhythms for AL and CR animals were highly synchronized when this regimen was used.     

Effects of feeding regime on food consumption, growth rates and tissue nucleic acids in juvenile Arctic charr, Salvelinm alpinus, with particular respect to compensatory growth

Juvenile Arctic charr responded to a change from restricted to satiation feeding by showing a growth spurt (compensatory growth). During this period of rapid growth the fish became hyper-phagic and in the days immediately following transfer from restricted to satiation feeding showed improved food conversion efficiency compared to their counterparts raised on a liberal feeding regime. Tissue (liver and muscle) nucleic acid concentrations were influenced by feeding regime, and RNA : DNA ratios were low in both starved fish and those fed restricted rations. Following transfer from restricted to satiation feeding, tissue RNA : DNA ratios were rapidly restored to initial levels. The uses of tissue RNA: DNA ratios both in evaluating nutritional status and as growth indices are discussed.     

Comparative analysis of three methods from dried blood spots for expeditious DNA extraction from mosquitoes; suitable for PCR based techniques

The objective of this work was to compare the quality, purity and quantity of DNA isolated from dried blood spots (DBS) by three methods (Chelex-100, QIAamp DNA mini kit, and TE (Tris EDTA)-Buffer). Sample collection was performed in six districts in Odisha, India and screened for cases of clinical malaria and dengue and vector density. Mosquito abdomens were spotted on Whatman 3MM (MERCK) Filter paper and dried for 10 min at room temperature. DNA was isolated from DBS using three methods (Chelex-100, QIAamp DNA mini kit, and TE-Buffer), and PCR was used to determine the feeding behaviours of vector mosquitoes. DNA was quantified using a UV-spectrophotometer, and q-PCR was used to determine the target gene copy number to compare the methods. The QIAamp DNA mini kit method was used as the reference method. The yield and purity of DNA extracted with Chelex-100 and TE were 14–72 ng/µl and 1.51–1.85 and 9–50 ng/µl and 1.68–2.1, respectively. DNA extracted using the Chelex-100 method was stored for over 1 month at ? 20 °C and was suitable for later use. The Chelex-100 method had a sensitivity of 99.5% and specificity of 78%. A Bland–Altman plot suggested that the Chelex-100 method was similar to the QIAamp DNA mini kit method for determining the feeding behaviours of vector mosquitoes. The Chelex-100 method is simple, cost-effective, and safe and requires minimal time for DNA extraction from dried blood spots. In malaria and dengue research, detecting the feeding behaviours from mosquito DNA from dried blood spots on filter paper by PCR is an easy, minimally invasive and inexpensive molecular technique that can be performed in remote areas.

     

Effect of season and artificial diet on amino acids and nucleic acids in gonads of green sea urchin Strongylocentrotus droebachiensis

The content of total and free amino acids (FAA) in green sea urchin (Strongylocentrotus droebachiensis) gonads varied with the season and feeding on an artificial diet. Glycine was the dominant amino acid in each season contributing 12.9-16.6% to the total amino acid (TAA) content, peaking in the spring. In the FAA profile, glycine accounted for 30.3-61.4% in different seasons. A grain-based artificial diet had noticeable effects on the total and FAA compositions of S. droebachiensis. Although, glycine was the dominant amino acid in the TAA profile during early harvesting, tyrosine in gonads became more dominant on week 9 of feeding. Furthermore, glycine was the dominant amino acid in the FAA pool after feeding the artificial diet. The total FAA content in the gonads increased significantly (P<0.05) from 20.6 on week 0 to 180.6 mg/g dry mass tissue on week 3. There were no significant (P<0.05) changes between week 6 and week 9. Deoxyribonucleic acid (DNA) content exceeded that of ribonucleic acid (RNA) in each season, while in cultured urchins, RNA content exceeded that of DNA only on week 6. The RNA/DNA ratio was significantly increased in the summer, whereas this ratio was increased up to week 6 followed by a decrease on week 9 in cultured counterparts.     

Ingested foreign (phage M13) DNA survives transiently in the gastrointestinal tract and enters the bloodstream of mice

Is the epithelial lining of the mammalian gastrointestinal (GI) tract a tight barrier against the uptake of ingested foreign DNA or can such foreign DNA penetrate into the organism? We approached this question by pipette-feeding circular or linearized double-stranded phage M13 DNA to mice or by adding M13 DNA to the food of mice whose fecal excretions had previously been shown to be devoid of this DNA. At various post-prandial times, the feces of the animals was tested for M 13 DNA sequences by Southern or dot blot hybridization or by the polymerase chain reaction (PCR). On Southern blot hybridization, the majority of M13 DNA fragments were found in the size range between < 200 and 400 by (base pairs). For the PCR analysis, synthetic oligodeoxyribonucleotide primers were spaced on the M13 DNA molecule such that the sizes of the persisting M13 DNA fragments could be determined. We also extracted DNA from whole blood or from sedimented blood cells of the animals at different times after feeding M t3 DNA and examined these DNA preparations for the presence of M13 DNA by dot blot hybridization or by PCR. M13 DNA fragments were found between 1 and 7 h postprandially in the feces of mice. By PCR analysis, fragments of 712, 976, and 1692 by in length were detected. In DNA from blood, M13 DNA fragments of up to 472 by were found by PCR between 2 and 6 h after feeding. Dot blot or Southern blot hybridization revealed M13 DNA at 2 and 4 h, but not at 1, 8 or 24 h after feeding. This DNA was shown to be DNase sensitive. M13 DNA was found both in blood cells and in the serum. A segment of about 400 by of the DNA amplified by PCR from feces or blood was analyzed for its nucleotide sequence which was found to be identical to that of authentic M13 DNA, except for a few deviations. M13 DNA could not be detected in the feces or in the blood of the animals prior to feeding or prior to 1 h and later than 7 h after feeding. These controls attest to the validity of the results and also argue against the possibility that the murine GI tract had been colonized by phage M13. Moreover, M13 DNA-positive bacterial colonies were never isolated from the feces of animals that had ingested M13 DNA. The results of reconstitution experiments suggested that 2 to 4% of the orally administered M13 DNA could be detected in the GI tract of mice. A proportion of about 0.01% to 0.1% of the M13 DNA fed could be retrieved from the blood.     

Neotrombicula autumnalis (Acari, Trombiculidae) as a vector for Borrelia burgdorferi sensu lato?

Larvae of the trombiculid mite Neotrombicula autumnalis were collected at 18 sites in and around Bonn, Germany, to be screened for infection with Borrelia burgdorferi s.l. by means of PCR. Questing larvae numbering 1380 were derived from the vegetation and 634 feeding ones were removed from 100 trapped micromammals including voles, mice, shrews and hedgehogs. In a laboratory infection experiment, a further 305 host-seeking larvae from the field were transferred onto Borrelia-positive mice and gerbils, and examined for spirochete infection at various intervals after repletion. In three cases borrelial DNA could be amplified from the mites: (1) from a larva feeding on a wild-caught greater white-toothed shrew (Crocidura russula), (2) from a pool of four larvae feeding on a B. garinii-positive laboratory mouse, and (3) from a nymph that had fed on a B. afzelii-positive laboratory gerbil as a larva. In the first case, borrelial species determination by DNA hybridization of the PCR product was only possible with a B. burgdorferi complex-specific probe but not with a species-specific one. In the second case, probing showed the same borrelial genospecies (B. garinii) as the laboratory host had been infected with. In the latter case, however, DNA hybridization demonstrated B. valaisiana while the laboratory host had been infected with B. afzelii. Subsequent DNA sequencing confirmed much higher similarity of the PCR product to B. valaisiana than to B. afzelii indicating an infection of the mite prior to feeding on the laboratory host. The negligible percentage of positive mites found in this study suggests that either the uptake of borrelial cells by feeding trombiculids is an extremely rare event or that ingested spirochetes are rapidly digested. On the other hand, the results imply a possible transstadial and transovarial transmission of borreliae once they are established in their trombiculid host. However, unless the transmission of borreliae to a given host is demonstrated, a final statement on the vector competence of trombiculid mites is not possible.