Artemia cysts as an alternative food for the predatory bug Macrolophus pygmaeus

The suitability of cysts of the brine shrimp Artemia sp. as a factitious food for the predator Macrolophus pygmaeus Rambur was investigated. The influence of decapsulation time and hydration of the cysts on the performance of the predator were studied in the absence of plant material. A longer time of decapsulation had a positive influence on the development of the predator. Hydration of cysts had a significant impact on nymphal survival when cysts where non‐decapsulated or poorly decapsulated. An experiment in which nymphs were switched from a diet of hydrated cysts to non‐hydrated cysts showed that in the absence of plant material the relative importance of hydrating the cysts decreased with nymphal age. Especially, the first instar and to a lesser extent the second instar appear to be susceptible to water shortage. Effects of prolonged rearing on development and reproduction on brine shrimp cysts from different origins were tested in the presence of plant material. Rearing M. pygmaeus on Artemia sp. (Jingyu Lake) cysts yielded similar survival, development, adult weight and fecundity in the fourth as in the second generation. In contrast, for Artemia franciscana cysts, an increase in nymphal development was notable. Biochemical analyses showed that total amino acid content and the concentration of the different amino acids did not differ among diets and generations. There were, however, differences in total fatty acid content between the different diets and generations and in the concentration of certain fatty acids, indicating that insects fed brine shrimp cysts may show nutritional deficiencies compared to those reared on a diet of Ephestia kuehniella eggs. Our results indicate that decapsulated brine shrimp cysts are an economically viable alternative food source in at least part of the rearing process for M. pygmaeus.     

Effects of UV-Radiation on Kinetics of Encystment and on Morphology of Resting Cysts of the Ciliate Laurentiella acuminata1

The effects of ultraviolet radiation (λ= 254 nm) on the kinetics of encystment of the hypotrichous ciliate Laurentiella acuminata and the structure of resting cysts obtained from irradiated precystic cells are reported. High doses of UV-radiation caused a delay of encystment with a linear increase in the average time for obtaining 50% of encystment (EN₅₀). Resting cysts with abnormal cyst walls were obtained when precystic cells were irradiated in the exposure range 720 to 960 J/m². The cystic layer (mesocyst) was approximately twice as thick (6.5 μ m) as normal (3.7 μ m). Microscopical observations of abnormal cysts revealed the presence of two complete mesocysts, and the absence of the spines characteristic of the ectocyst. The UV-dependent effects on the cyst wall were gradually corrected in successive generations of the irradiated cells.     

Oviposition preference and development of Maruca vitrata (Fabricius) (Lepidoptera: Crambidae) on different radiofrequency fields

Maruca vitrata is considered as one of the major impediments in boosting up the production of tropical legumes. Since chemical pesticides have an adverse effect on human health and environment, there is an urgent need for developing a non‐chemical based method to replace chemical insecticides for controlling of pests. We investigated the oviposition, development and adult performances of M. vitrata at five radiofrequency levels of 0 (control without frequency), 5, 10, 20, and 30 kHz on artificial diet. In addition, the effects were also studied in three successive generations (Parent, F1‐ and F2‐ generations) of M. vitrata. The results showed that radiofrequency application had significant effects on the life‐variables of M. vitrata. Radiofrequency exposures on M. vitrata not only impacted on oviposition, developmental time, adult longevity, and adult weight but negative effects were also transmitted to the successive generations. Oviposition was found to reduce as generation passed after exposed to radiofrequencies. Total developmental times from egg to adult emergence were longer with shorter longevity in case of radiofrequency treatment than untreated. Decreasing the radiofrequency level further increased time for development of adults as generation passed. The shortest adult longevity occurred at 5 kHz treatment. The outcomes of this study are discussed in terms of targeting susceptibilities to radiofrequency as an alternative to chemical treatments.     

Cytology of lepidoptera. III. Giant cysts: A morphological trait of apyrene spermatogenesis in an Ephestia kuehniella strain

A comparative investigation of testicular eupyrene cysts (in larvae) and apyrene cysts (in pupae) of Ephestia kuehniella laboratory strains was conducted using light and electron microscopy. Eupyrene cysts in the first meiotic division contained 64 spermatocytes, which showed only moderate asynchrony. In one of the strains, a wild-type strain, L, normal-sized cysts occurred together with abnormally large cysts. These are called giant cysts in this article. One of the premeiotic cysts, early giant cysts, studied in detail, contained approximately a fourfold number of cells compared with the number in a eupyrene cyst of the same stage. In cysts harboring spermatocytes and spermatids, late giant cysts, cell differentiation was highly asynchronous. Failure in one of two control mechanisms in early cyst development may have caused the appearance of the cysts. Control of cell division might have been sloppy in apyrene spermatogonia. Hence, the spermatogonia within the cyst could have passed through additional division cycles. Alternatively, the giant cysts may have originated from more than one predefinitive gonial cell enclosed in a common envelope of sheath cells. As a third possibility, giant cysts could have arisen by fusion of normal cysts at a later stage. In either case, this is evidence that separation of eupyrene and apyrene pathways is earlier than was previously expected. In two other Ephestia strains, apyrene sperm development proceeded without formation of giant cysts. One was a mutant strain, a, and the other one was a recently established wild-type strain, Sbr. Apyrene sperm development is considered an example of degenerate evolution in which enhanced variability between species and even between populations of one species is a common phenomenon.     

Development,morphological characteristics and viability of temporary cysts of Pyrodinium bahamense var. compressum (Dinophyceae) in vitro

Pellicle or temporary cysts of Pyrodinium bahamense var. compressum (Böhm) Steidinger, Tester & F.J.R. Taylor and their role in bloom dynamics have not yet been adequately characterized and understood. We investigated the role of temperature- and nutrient-mediated stress as factors that could induce pellicle formation in batch cultures. Cellular features and their implications for temporary cyst viability were examined using confocal laser scanning microscopy (CLSM). Our data suggest that temperature change is one of the key factors influencing pellicle formation, preserving viability at low temperature (i.e. 13°C). Hypnocysts (resting cysts) were not observed. During pellicle formation, motile cells generally undergo ecdysis, extrusion of cytoplasmic materials and bacteria, compaction of the nucleus and non-motility. The outermost covering of the temporary cysts shows red autofluorescence and it contains lower concentrations of chlorophyll (chl) a and no detectable chl c. The nuclear region is surrounded by transitional red bodies and other unidentified cellular structures. Temporary cysts can immediately revert back to the motile state upon exposure to optimum conditions. This is accompanied by the expansion of the nuclear region, regeneration of the chloroplasts and enlargement of the cell. Developmental changes during reversal of temporary cysts to motile forms were also observed to cause breaks in the cell covering that could serve as sites for bacterial entry. Though observed in vitro, such behaviour may also be occurring in nature especially as a response to drastic short-lived environmental changes. This is the first detailed report on the characteristics of temporary cysts of P. bahamense var. compressum.     

Encystment of Peridinium gatunense: occurrence, favourable environmental conditions and its role in the dinoflagellate life cycle in a subtropical lake

1. The abundance of cysts of the bloom‐forming dinoflagellate Peridinium gatunense in the sediments of Lake Kinneret and the effects of environmental conditions on encystment were studied in relation to bloom dynamics. Peak cyst formation coincided with the highest growth rate of the population, prior to bloom peak. 2. Peridinium cysts were counted in water and sediment corer samples from 2000 to 2003 and in archived sediment trap samples collected during 1993–94. The cyst data were examined in relation to ambient temperature and nutrient records, and revealed no direct correlation. 3. In laboratory encystment experiments with Peridinium cells collected from the lake, 0.2–3% of the vegetative cells encysted. Temperature, light and cell density had no significant effect on the percentage of encystment. 4. Cysts were always present in the lake sediments but their abundance in ‘non Peridinium’ years was much lower than after a massive bloom. Vegetative cells were always present in the water column after the collapse of the annual dinoflagellate bloom, potentially serving as the inoculum for the next bloom. We propose that the hardy cysts serve as an emergency ‘gene bank’ to initiate population build up following catastrophic die outs.     

Selection of nematodes for zero cyst phenotypes with soybean

Definitive genetic studies of the soybean cyst nematode (Heterodera glycines)-soybean (Glycine max) relationship are complicated by the use of soybean lines with many genes for resistance to heterogeneous, amphimictic nematode populations. Inbreeding and artificial selection of cyst nematodes decreased their ability to form cysts on specific soybean lines assumed to have few genes for resistance. In the initial sib selection, single cysts were used to inoculate two-seedling units consisting of a seedling of one soybean line to evaluate the nematodes' ability to form cysts on it and a seedling of another line for comparison and maintenance of the nematode inbreds being developed. This method permitted some selection but was slow and labour-intensive. A better method of artificial selection for fewer cysts on specific soybean lines was to inbreed nematodes for about four generations, then to evaluate the inbreds for ability to form cysts on the soybean lines before selecting particular inbreds with poor ability to form cysts for the development of more inbreds. This increased the frequencies of specific nematode genes for avirulence or alleles for incompatibility with soybean lines containing the interacting [gene-for-gene] alleles for incompatibility (genes for resistance). The artificial selection apparently was for at least two nematode alleles for incompatibility since at least two soybean alleles were indicated in the segregation from crosses of soybean lines.     

RESTING CYST FORMATION OF EUTREPTIELLA GYMNASTICA (EUGLENOPHYCEAE) IN THE NORTHERN COASTAL BALTIC SEA1

Resting cyst formation of Eutreptiella gymnastica Throndsen was observed during a mesocosm experiment, where nutrient enrichment had induced almost a unialgal bloom. Cells and resting cysts of E. gymnastica were examined in scanning (SEM) and transmission electron microscopy (TEM) and light microscopy. Mature cysts were spherical, with a smooth thick mucilaginous cover that appeared layered when observed with the TEM. Intermediate forms were spherical and lacking flagella and a mucilaginous cover; the euglenoid pellicular striation and canal opening were easily visible. The volume of these intermediate spherical cells and mature cysts was estimated to have increased threefold compared to flagellated cells and contained many paramylon grains. When the cells were grazed by zooplankton, the paramylon grains passed the gut intact and were packed into fecal pellets. Intact undigested paramylon grains were observed in SEM after the breaking up of the pellets.     

Artificial and factitious foods support the development and reproduction of the predatory mite Amblyseius swirskii

The generalist predatory mite Amblyseius swirskii Athias-Henriot (Acari: Phytoseiidae) was reared on Ephestia kuehniella Zeller eggs (Lepidoptera: Pyralidae), decapsulated dry cysts of the brine shrimp Artemia franciscana Kellogg (Anostraca: Artemiidae), and on meridic artificial diets (composed of honey, sucrose, tryptone, yeast extract, and egg yolk) supplemented with pupal hemolymph of the Chinese oak silkworm Antheraea pernyi (Guérin-Méneville) (Lepidoptera: Saturniidae) (AD1), with E. kuehniella eggs (AD2) or with A. franciscana cysts (AD3). Development, reproduction and predation capacity of the predatory mites were assessed in the first (G1) and sixth generation (G6) of rearing on the different diets. Immature survival rates in G1 were similar on all diets (96.8–100 %). After six generations, however, survival of A. swirskii was significantly reduced on all diets except on A. franciscana cysts. Oviposition rates did not differ between generations when females were fed on E. kuehniella, AD2 or AD3. The total number of deposited eggs was similar among diets except in G6 where the females fed on A. franciscana cysts produced more eggs than those maintained on E. kuehniella eggs. On most diets the intrinsic rates of increase in G1 were superior to those in G6, except for predators supplied with A. franciscana cysts where no differences were observed among generations. Female mites did not lose their capacity to kill first instar Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) after six generations on the different diets, but predation rates in G6 on E. kuehniella were lower than in G1. In conclusion, the different factitious and artificial diets tested in the present study supported the development and reproduction of A. swirskii for a single generation but fitness losses occurred to a varying degree after several generations on E. kuehniella eggs or the artificial diets. Artificial diet enriched with A. franciscana cysts yielded better results than the other artificial diets. Amblyseius swirskii performed best on decapsulated Artemia cysts indicating their potential for use in the mass production of the predator or to sustain its populations in the crop after release.     

MORPHOLOGY AND LIFE CYCLE OF A NEW CHYTRID WITH AERIAL SPORANGIA

An investigation of the life cycle of Caulochytrium gloeosporii Voos and Olive, a parasite of Gloeosporium, has confirmed the existence of an alternation of generations. Zoospores from sessile sporangia may develop vegetatively into similar sporangia or they may function as isogametes, fusing in pairs to produce cysts (zygotes) that give rise to slender-stalked aerial sporangia. Meiosis is believed to occur in the aerial sporangium, which at germination liberates eight asexual zoospores. The species is homothallic.     

Raunsgaardispermum lusitanicum gen. et sp. nov., a new seed with in situ pollen from the Early Cretaceous (probably Berriasian) of Portugal: Further support for the Bennettitales‐Erdtmanithecales‐Gnetales link

Assemblages of Recent and sub-Recent dinoflagellate cysts are described from the southern Beaufort Sea, offshore Mackenzie Delta region, Canadian Arctic, and include the new taxa Spiniferites frigidus Harland &; Reid, Multispinula minuta Harland &; Reid, and Rottnestia amphicavata Dobell &; Norris. These assemblages, which appear to be unique to Arctic waters, are dominated by Operculodinium centrocarpum, and compared to assemblages found in north-temperate seas, also dominated by Operculodinium centrocarpum, are ten times less diverse. The three new species described herein may prove to be indicators of the Arctic environment. Several genera of Recent cysts that were not originally supplied with Latin diagnoses are validated.     

VerÄnderte Anordnung der Cysten beiAcetabularia mediterranean

Zusammenfassung Die Cysten vonAcetabularia mediterranea sind in den Hüten normalerweise radiÄrsymmetrisch angeordnet. In einigen Zellen unseres Standard-Kulturmaterials fanden sich jedoch Hüte mit bilateralsymmetrischer Cystenanordnung. Die Nachkommenschaft dieser Zellen (bisher zwei Folgegenerationen) zeigt das gleiche morphologische Merkmal. Das beschriebene Merkmal könnte daher auf einer genetischen VerÄnderung beruhen. Dies würde die erste erfolgreiche Isolierung und Kultur einer Mutante beiAcetabularia bedeuten.

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Changed arrangement of cysts inAcetabularia mediterranea

Summary Cysts ofAcetabularia mediterranea are normally arranged within the cap in a radial symmetric manner. In one population of our standard laboratory material, however, some cells were found with cysts arranged in a bilateral symmetric manner. The progeny of these cells (two following generations, so far) shows the same morphological characteristic, indicating that the different arrangement of cysts might be the result of genetical differences. This would mean the first successful isolation and cultivation of a mutant ofAcetabularia.

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Herrn Prof. J.HÄmmerling zur Vollendung seines 75. Lebensjahres gewidmet.     

Virulence of Septoria nodorum as Affected by Passage Through Detached Leaves of Wheat and Barley Cultivars1

Abstract Three genetically marked, single–spore isolates of Septoria nodorum from wheat were passed through detached leaves of wheat cvs Blueboy and Coker 747 and the barley cv. Boone to produce three sub–isolates per original isolate. Each sub–isolate was cultured for three pycnidiospore generations on its respective host. Virulence of each sub–isolate on detached leaves of Blueboy, Caldwell, Coker 747, and NK81W701 wheat, and Boone and Surry barley was compared with that of the original single–spore isolate from which it was derived. In most cases, sub–isolates passed through wheat were significantly more virulent than the originals on wheat cultivars. They also were more virulent to barley than the original isolates but they were less virulent to barley than to wheat cultivars. Isolate × cultivar interactions were statistically significant (P < .0001) for isolates passed through wheat or barley and were greater than isolate × cultivar interactions among the original isolates. In seven of eight isolates passed through wheat or barley, only the original genetic marker was recovered after three generations, indicating that cross–contamination could not account for the observed change of virulence. In the single case of apparent contamination, of a sub–isolate, virulence declined.     

Hatching from cysts and egg sacs of Heterodera cruciferae and effects of temperature on hatching and development on oilseed rape

Juveniles hatched readily from field cysts and very readily from eggs in egg sacs of Heterodera cruciferae, when exposed to oilseed rape root diffusate. They hatched very poorly, however, from white or brown females from which the above egg sacs had been removed. Some hatching occurred at 8 ^oC but much more occurred at 12 , 16, 20 and 24 ^oC, with most at 16 ^oC. Development of juveniles in roots of oilseed rape occurred throughout the range 8–24 ^oC, and proceeded faster the higher the temperature. The basal development temperature was taken as 5 ^oC and the number of day degrees above this temperature required to reach each stage of development was calculated. From invasion of roots to the hatching of F₁ juveniles required an average of 680 day degrees, but only 210 day degrees were required for the first appearance of egg sacs on adult females. On this basis, two consecutive generations of H, cruciferae would be possible on autumn-sown oilseed rape in southern England, but the second would mature fully only after the crop was harvested. In Scotland, two consecutive generations could also occur but the second would be much less mature by harvest: only about 850 day degrees are available compared to almost 1100 in southern England. In practice, however, overlapping generations probably occur due to flushes of hatching of juveniles (i) at sowing, (ii) when soils warm up after winter and (iii) when the first generation completes its development. The proportion of eggs found in egg sacs was never more than 37% and some field cysts contained about 220 eggs; their egg sacs may, therefore, have contained as many as 150 eggs. Any study of population dynamics or damage assessment will require a quantification of the contribution of eggs in egg sacs to population density. Oilseed rape is direct drilled and may, therefore, be more sensitive to a given population density of the nematode than host crops which are transplanted.     

DETECTION OF RICKETTSIA TSUTSUGAMUSHI FROM VECTOR TROMBICULID MITE BY DNA AMPLIFICATION USING POLYMERASE CHAIN REACTION TECHNIQUE*

Abstract With reference to the Sta 58 major antigen gene of Rickettsia tsutsugamushi (Karp strain), we had designed and synthesized a pair of DNA primers; and a kilobase fragment of Sta 58 is amplified for using in polymerase chain reaction (PCR) with the primers. This is the first report on detection of R. tsutsugamushi in trombiculids by using PCR technique. The experimental results indicated that the time for which R. tsutsugamushi could exist in the adult of Leptotromhidium deliense when inoculated into its abdomenal cavity was 360 days at least and R. tsutsugamushi could be transovarially transmitted to the offsprings for 4 generations; and the time for which R. tsutsugamushi could exist in L. deliense after biting the infective mouse was 270 days at least and R. tsutsugamushi could be transmitted transovarially to the offsprings for 2 generations. The results also showed that the PCK technique was highly specific and sensitive for detection of R. tsutsugamushi; and the amplification of gene outside body to detect the rickettsiae in the body of trombiculid and mouse can be used as a new method for investigation of scrub typhus epidemiology.     

Change in shoot proliferation with repeated in vitro subculture of shoots of woody species of Rosaceae

Shoots of 6 ornamental species and cultivars of Rosaceae were repeatedly subcultured in vitro for 9 generations on Linsmaier and Skoog (1965) medium with the addition of BA. Shoot proliferation increased over the first few generations and then gradually declined in all species and at all BA concentrations tested with the exception of Chaenomeles japonica in which a decline in shoot formation occurred only at 5.0 mg 1^-1 BA. A decrease in shoot length and leaf size and an increase in the incidence of callus formation was observed after several subcultures. This apparently irreversible decline could be due to either genetic or epigenetic change resulting from repeated fluxes in cytokinin, nutrient status or sucrose, or to elimination of seasonal environmental fluctuations.     

Functional Ecology of the Ciliate Glaucomides bromelicola,and Comparison with the Sympatric Species Bromeliothrix metopoides

We investigated the ecology and life strategy of Glaucomides bromelicola (family Bromeliophryidae), a very common ciliate in the reservoirs (tanks) of bromeliads, assessing its response to food quality and quantity and pH. Further, we conducted competition experiments with the frequently coexisting species Bromeliothrix metopoides (family Colpodidae). In contrast to B. metopoides and many other colpodean ciliates, G. bromelicola does not form resting cysts, which jeopardizes this ciliate when its small aquatic habitats dry out. Both species form bactivorous microstomes and flagellate‐feeding macrostomes. However, only G. bromelicola has a low feeding threshold and is able to adapt to different protist food. The higher affinity to the local bacterial and flagellate food renders it the superior competitor relative to B. metopoides. Continuous encystment and excystment of the latter may enable stable coexistence of both species in their natural habitat. Both are tolerant to a wide range of pH (4–9). These ciliates appear to be limited to tank bromeliads because they either lack resting cysts and vectors for long distance dispersal (G. bromelicola) and/or have highly specific food requirements (primarily B. metopoides).     

Notes on the morphology,ecology and life cycles of Fukaurahydra anthoformis and Hataia parva (Hydrozoa,Athecata)

Fukaurahydra anthoformis and Hataia parva are solitary athecate hydroids occurring in northern Japan. New information on the external morphology, nematocysts, ecology, and life cycles of these species is presented. It is noteworthy that H. parva bears stenoteles, which are generally not found among the families of Filifera. Neither species produces free medusae. The eggs are fertilized in the female gonophores, from which unciliated larvae are released. These larvae do not swim and soon attach to a substrate. After attachment the larvae become covered by a sheath to form cysts. The cysts rest on a substrate without any outer change for several months. As the water temperature drops in autumn to early winter the cysts begin to hatch, forming tiny polyps after the larva creeps out from the chitinous sheath. Cyst formation proves to be common also in other solitary hydroids, most of which are inhabitants of cool or cold waters.     

Morphology of the Cyst of Giardia microti by Light and Electron Microscopy1

ABSTRACT. Cysts of Giardia microti, isolated from feces and intestinal contents of Microtus ochrogaster, were examined by light and electron microscopy. These cysts differed morphologically from cysts of other G. duodenalis morphological types in that these cysts often contained two apparently differentiated trophozoites with mature ventral discs. Cysts more closely resembling those reported for G. lamblia and G. muris were in greater abundance in preparations made from intestinal contents and were interpreted as immature cysts. “Multiple fission” cysts, reported in G. muris and G. microti by earlier workers, were not observed; however, endosymbiotic bacteria were found in the cysts of G. microti and may have been responsible for reports of multiple fission in the cysts of Giardia.