THE LIFE CYCLE AND TOXICITY OF PFIESTERIA PISCICIDA REVISITED1

Despite use of excellent molecular techniques, Litaker et al. (2002) cannot provide insights about the life history of toxic Pfiesteria piscicida because they showed no data in support of having used toxic strains; rather they presented evidence that they used non‐inducible strains. Litaker et al. did not find amoeboid stages or a chrysophyte‐like cyst stage in several cultures and unequivocally concluded that the stages do not exist in all P. piscicida strains. Thus, they did not consider the tenet that absence of evidence does not constitute proof of absence. Apparent discrepancies between the research by Litaker et al. and previous research on Pfiesteria can be resolved as follows: First, Litaker et al. did not use toxic strains. We have reported findings (similar to Litaker et al.) showing few amoeboid transformations in non‐inducible strains, which manifest some but not all of the forms that have been documented in some toxic strains. We, and others, have documented active toxicity to fish, transformations to amoebae, and chrysophyte‐like cysts in some clonal toxic strains. Second, the data from several recent publications, which were available but not mentioned by Litaker et al. or by Coats (2002) in accompanying commentary, have verified P. piscicida amoebae, chrysophyte‐like cysts, and other stages in some toxic strains through a combination of approaches including PCR data from clonal cultures.     

黑斑蛙双睾吸虫的生活史研究

本文首次研究了黑斑蛙双睾吸虫的生活史。实验证明这种单殖吸虫的生活史具有两种不同的途径,即直接外生周期和直接内生周期。前一种发育途径为一般多盘吸虫所共有,后一种发育途径仅在少数多盘吸虫中存在。这两种发育途径的交替存在与其两栖动物宿主生态学有关,并且可能反映了这类多盘吸虫的一种种群调节机制。     

鸡沙氏住白虫的生活史研究

本文报告了鸡沙氏住白虫生活史及其全程发育。经人工实验和流行区调查证明,后宽绳蚋是该虫的自然传播媒介。孢子增殖在后宽绳蚋体内、经3—4天完成发育。无性裂殖体增殖在鸡的肝、肾、心、肺、脾、脑和肠等内脏器官组织细胞内进行,经4—9天完成发育,该裂殖体有二型,即肝裂殖体和巨噬细胞裂殖体。感染9—13天后,血细胞内的配子体发育成熟。感染后第15—25天末稍血液中出现虫体的高峰期。     

LIFE CYCLE OF THE HETEROTROPHIC DINOFLAGELLATE PFIESTERIA PISCICIDA (DINOPHYCEAE)1

The putatively toxic dinoflagellate Pfiesteria piscicida (Steidinger et Burkholder) has been reported to have an unusual life cycle for a free‐living marine dinoflagellate. As many as 24 life cycle stages were originally described for this species. During a recent phylogenetic study in which we used clonal cultures of P. piscicida, we were unable to confirm many reported life cycle stages. To resolve this discrepancy, we undertook a rigorous examination of the life cycle of P. piscicida using nuclear staining techniques combined with traditional light microscopy, high‐resolution video microscopy, EM, and in situ hybridization with a suite of fluorescently labeled peptide nucleic acid (PNA) probes. The results showed that P. piscicida had a typical haplontic dinoflagellate life cycle. Asexual division occurred within a division cyst and not by binary fission of motile cells. Sexual reproduction of this homothallic species occurred via the fusion of isogamous gametes. Examination of tanks where P. piscicida was actively feeding on fish showed that amoebae were present; however, they were contaminants introduced with the fish. Whole cell probing using in situ hybridization techniques confirmed that these amoebae were hybridization negative for a P. piscicida‐specific PNA probe. Direct observations of clonal P. piscicida cultures revealed no unusual life cycle stages. Furthermore, the results of this study provided no evidence for transformations to amoebae. We therefore conclude that P. piscicida has a life cycle typical of free‐living marine dinoflagellates and lacks any amoeboid or other specious stages.     

圈绒泡菌的生活史

利用基物培养、燕麦-琼脂培养技术及扫描电镜技术研究了圈绒泡菌的个体发育过程, 在燕麦琼脂培养基上完成了从孢子到孢子的生活史。结果表明,生活史包括单核的黏变形体或游动胞、多核的营养体原质团以及孢子形成阶段。琼脂培养基上获得的圈绒泡菌孢子与野生型相似,并具有可育性。     

针箍菌的生活史

利用基物和燕麦-琼脂技术研究了针箍菌的个体发育过程,在燕麦琼脂培养基上完成了从孢子到孢子的生活史。结果表明,生活史包括单核的黏变形体或游动胞、多核的营养体原质团以及孢子形成阶段。琼脂培养基获得的针箍菌孢子与野生型相似,并具有可育性。     

ULTRASTRUCTURE OF THE GAMETE OF PEDIASTRUM DUPLEX (CHLOROPHYCEAE)1

Gametes of Pediastrum duplex Meyen were investigated ultrastructurally, with emphasis on the flagellar apparatus. The cells are naked, biflagellate, and measure approximately 2.5 × 8 μm. Distinguishing them from zoospores is their possession of an eyespot and mating structure (the apical cap), and their lack of the peripheral band of cytoplasmic microtubules involved in colony formation. Four featurs of the flagellar apparatus are especially noteworthy: (1) the basal bodies are directly opposed and (2) are interconnected via their cores, (3) the central portion of the distal fiber is elaborated into an unusual ribbed structure which overlies the striated microtubule-associated component (SMAC) of the two-membered rootlets, and (4) the X-rootlets are dissimilar in microtubular number. The smaller X-rootlet consists of four microtubules in a three over one (3/1) configuration, whereas the larger has been found to be either 5 / 1, 6 / 1 or 7 / 1. The former rootlet extends past the nucleus whereas the latter extends down the opposite side of the cell, passing near the eyespot. The first two of these flagellar apparatus features have been previously noted in other motile cells of the Hydrodictyaceae. Although not specifically mentioned, published micrographs suggest the presence of the latter two as well, which may indicate that all four flagellar apparatus features are characteristics of all motile cells in the Hydrodictyaceae.     

STUDIES ON THE LIFE CYCLE AND TAXONOMY OF LIGNIERA VERRUCOSA

Miller , Charles E. (A. and M. College of Texas, College Station.) Studies on the life cycle and taxonomy of Ligniera verrucosa. Amer. Jour. Bot. 46(10): 725–729. Illus. 1959.—A study of the roots of Veronica persica Poir. and V. hederaefolia L. plants infected with Sorosphaera veronicae Schroeter revealed intracellular cystosori and zoosporangial sori of Ligniera verrucosa. The zoosporangial phase of this species has been heretofore unknown. The plasmodia of L. verrucosa occur in root hairs, and other epidermal and sub-epidermal cells of the roots. Zoosporangial and cystosoral plasmodia are indistinguishable until cleavage has started. It is thought that plasmodia produced during early infection develop into zoosporangia, while those produced later develop into resting spores. Zoospores discharged from zoosporangia may reinfect host cells developing there into zoosporangial or cystosoral plasmodia. No evidence for any sexual process was observed. The spherical zoosporangia making up a single zoosporangial sorus may be interconnected; a single discharge pore may serve to liberate zoospores from different zoosporangia. In the Plasmodiophorales the classical basis for generic distinction has been the arrangement of the resting spores in the sorus. Ligniera, because of the supposedly uncharacteristic nature of its cystosori, has been suggested as a host-variety of Sorosphaera. A comparative study of the cystosori and zoosporangia of Ligniera and Sorosphaera growing in a single host has led to the conclusion that these genera should be considered distinct.     

浮丝藻生活史及其亚显微结构特征的研究

通过观察浮丝藻（Ｐｌａｎｋｔｏｎｅｍａ）的生活史、显微和亚显微结构、细胞分裂方式,系统地研究了其藻丝体的生物学特性。浮丝藻为单列细胞不分枝的丝状体,细胞圆柱形或椭圆形,细胞之间主要靠筒状的纤维素细胞壁相互连接。藻细胞中含叶绿素ａ、ｂ和β－胡萝卜素。电镜下,细胞壁明显分层,绝大多数是２～３层;色素体片状、周生,通常２至多条类囊体重叠;１～２个蛋白核在色素体体上,蛋白核外常有淀粉鞘,有时１至多条类囊体重叠;１～２个蛋白核在色素体上,蛋白核外常有淀粉鞘,有时１至多条类囊体侵入蛋白核内;１～２个线粒体位于色素体旁,嵴不分枝;高尔基复合体靠近细胞周边,分泌许多潴泡;细胞核１个,球形或不规则方形,常靠近细胞的一端;光镜下细胞两端的折光亮点在电镜下是１至多个泡囊,泡囊内通常含些被染色很深的物质。浮丝藻的形态学、细胞分裂和生活史观察结果都证明,浮丝藻的细胞在整个生活史中没有营养分裂,细胞分裂时母细胞产生两个有独立、完整细胞壁的似亲孢子,一系列似亲孢子通过存留的、不连续的筒状或杯状母细胞壁连接成形状上象丝状体的结构。因此,藻丝体形成和成长的过程是一个无性繁殖过程,这样的藻丝体本质上是一种假丝体。     

REPLY TO COMMENT ON THE LIFE CYCLE AND TOXICITY OF PFIESTERIA PISCICIDA REVISITED1

Free‐living, marine dinoflagellates are typified by a well‐defined, haplontic life cycle with relatively few stages. The most unusual departure from this life cycle is one reported for the heterotrophic dinoflagellate Pfiesteria piscicida Steidinger et Burkholder. This species is alleged to have at least 24 life cycle stages including amoebae and a chrysophyte‐like cyst form ( Burkholder et al. 1992 , Burkholder and Glasgow 1997a ) not previously known in free‐living marine dinoflagellates. Litaker et al. (2002) redescribed the life cycle of P. piscicida from single‐cell isolates and found only life cycle stages typical of free‐living marine dinoflagellates. The discrepancy between these observations and the life cycle reported in the literature prompted a rigorous study to resolve the life cycle of P. piscicida. Burkholder and Glasgow (2002) took exception to this study, arguing that Litaker et al. (2002) misunderstood the life cycle of P. piscicida and ignored recent publications. We present a rebuttal of their criticisms and suggest a simple way to resolve the discrepancies in the P. piscicida life cycle.     

THE INFLUENCE OF CLIMATIC SEASONALITY ON THE LIFE CYCLE OF THE PRIBILOF NORTHERN FUR SEAL

Weather conditions recorded from 1956 to 1986 on St. Paul Island, Alaska, were probed to establish their influence upon the northern fur seal's life cycle ( Callorhinus ursinus ). Air temperatures, wind speeds, and relative humidity levels were seasonally decomposed and compared with the timing of pupping and migration. Most pups are born in early July when air temperatures and relative humidity approach their highest annual levels and wind speeds are at their lowest. Weather conditions favor growth and survival of pups from July to September but are unfavorable in June. A rapid deterioration in weather through October and November corresponds with the fall migration of pups and lactating females. The data suggest the pivotal event in the fur seal's life cycle is the timing of birth and survival of nursing pups. As such, the ultimate determinant of the precisely timed fur seal life cycle appears to be climatic seasonality during the breeding season.     

MACROCYSTS IN THE LIFE CYCLE OF THE DICTYOSTELIACEAE. II. GERMINATION OF THE MACROCYSTS

Macrocyst germination was demonstrated in the five species of the Dictyosteliaceae known to produce these structures. The morphological changes that occurred during germination appeared to be identical in all of the strains examined, showing the following stages: (1) swelling of the dark, contracted content of the dormant cysts, (2) gradual loss of color and reappearance of cells within what previously appeared as a homogeneous protoplasmic mass, and (3) rupture of the heavy cellulosic cyst wall to liberate the myxamoebae. The age of the macrocyst appeared to be the most critical factor in determining whether or not germination would occur, since the cysts in many of the strains needed to age for several weeks or months before germination could be demonstrated. In Dictyostelium mucoroides strain DM-7, upon which the current study was centered, light was necessary to stimulate germination of young macrocysts—a requirement that gradually diminished as the cysts aged. The rate of germination and the temperature permitting germination were also age dependent: older macrocysts germinated more rapidly and at considerably higher temperatures than did young cysts. Although light was not essential for germination in every strain, the results obtained with strain DM-7 seem to be generally applicable to the germination process.     

MACROCYSTS IN THE LIFE CYCLE OF THE DICTYOSTELIACEAE. I. FORMATION OF THE MACROCYSTS

Macrocysts, a morphogenetic phase that is alternative to sorocarp formation in the life cycle of some cellular slime molds, are known for two genera and five species of the Dictyosteliaceae. In all of these macrocyst formation was found to be strongly affected by four factors: light, temperature, moisture, and the composition of the medium. In general, macrocyst formation was suppressed and sorocarp formation was enhanced by exposure to light, by incubation at temperatures lower than 20 C, by buffering nutrient media with phosphates, and by reducing atmospheric moisture through the use of clay covers on Petri dishes. The extent to which these environmental factors, singly or in combination, inhibited the production of macrocysts varied among the different strains and species.     

PHAGOTROPHIC FEEDING AND ITS IMPORTANCE TO THE LIFE CYCLE OF THE HOLOZOIC DINOFLAGELLATE,GYMNODINIUM FUNGIFORME1

The holozoic dinoflagellate, Gymnodinium fungiforme Anissimova, has been observed in both asexually and sexually reproducing cultures. Asexual reproduction is characterized by zoosporangium formation and subsequent new cell release. Sexuality is gametic, and planozygotes and hypnozygotes are present. The life cycle is highly dependent on feeding, and in food-depleted cultures the swimming cells rapidly disappear. These are replaced with resistant long-term resting cysts. Despite its small size (8.5–19 μm), G. fungiforme can feed on prey as large as the ciliated protozoan, Condylostoma magnum Spiegel (600–1000 μm in length), or small injured metazoans, and has been cultured phagotrophically with the chlorophyte, Dunaliella salina Teodoresco as a food source. Eleven additional species of algae including 1 chlorophyte, 7 chrysophytes and 3 rhodophytes, however, were not suitable as food sources. Feeding is characterized by the formation of ‘dynamic aggregations’ of hundreds of dinoflagellates that attach to the surface of a prey organism by a peduncle. G. fungiforme ingests the cytoplasm or body fluids of its prey and a feeding aggregation can ingest a C. magnum in 20–30 minutes.