Spatial distribution of Heterodera trifolii in Chinese cabbage fields

Although Heterodera trifolii is commonly known as the clover cyst nematode, recently the nematode has been identified as a serious menace for Chinese cabbage growers in highland areas in Korea. Soil samples were collected from two Chinese cabbage fields highly infested with H. trifolii in highland areas of Korea, Jungsun and Samcheok, in 2014 and 2015, respectively. A total of 777 (2?×?2?m sampling area) and 414 (5?×?5?m area) soil samples were collected from Jungsun and Samcheok, respectively. The total cysts, cysts with eggs, number of eggs, and empty cysts were calculated for each sample. Distribution patterns for these variables were characterized using spatial analysis by distance indices (SADIE) and variogram model analysis. The aggregation index for cysts with eggs was higher in Jungsun (89.32) than Samcheok (3.63), which indicated that the cyst population density was higher. However, the spatial association of total cysts versus cysts with eggs was higher in Samcheok. The Gaussian model showed reasonable independent range of the nematode in Jungsun and Samcheok to be approximately 53.66?m and 48.54?m, respectively. The model suggested that each nematode sample should be taken at least 50?m apart in the given areas. Inclusion of this distribution pattern may significantly minimize the number of samples in future sampling methods, which could save time and labor, and initiate management practices by elucidating spatial variability factors that influence crop yield.     

食蚊鱼鳃上台湾棘带吸虫囊蚴内种群的月变化

1993年7月-1994年6月,作者对食蚊鱼鳃上台湾棘带吸虫囊蚴内种群的变化进行了为期12个月的研究,结果表明:食蚊鱼种群数量和种群重量的月变化相似,但两者与水温没有相关性;食蚊鱼种群的体长结构呈季节性动态变化规律;食蚊鱼的感染与其自身的生长紧密相关,感染率、感染强度表现出了与体长相似的变化趋势;中间的两组鳃片寄生的囊蚴较多,不同鳃片组囊蚴平均密度的月变化规律一致,食蚊鱼任何两个鳃片组分布的台湾棘带吸虫囊蚴的平均数差异都显著(P0.001);囊蚴在食蚊鱼鳃上的分布为聚集分布。       

THE CYST OF THE POTATO-ROOT EELWORM (HETERODERA ROSTOCHIENSIS WOLLENWEBER) AS A HATCHING UNIT

Two series of experiments are described: in one, the hatching of half and whole cysts is compared, in some cases after they have been subjected to tryptic digestion; in the second series, the hatching of free eggs is considered.
Total emergence from halved cysts was very much greater than that from intact cysts. Emergence from trypsin-treated halved cysts was greater than emergence from control halved cysts, suggesting that the enzyme affects the egg shell or the larva inside; it is possible, however, that the enzyme also affects the cyst wall, for treated whole cysts shrink more rapidly and to a greater extent when exposed to air.
Total emergence from trypsin-treated whole cysts did not differ significantly from that of untreated whole cysts, unlike previous findings (Ellenby, 1946 a); however, emergence began sooner from these cysts. Halving the cysts also resulted in earlier emergence, the biggest effect being produced by a combination of both trypsin and subsequent halving.
The variability shown by the intact cysts was considerably greater than that shown by the halved cysts; halving the cysts reduced the coefficient of variation from 0·55 for whole untreated cysts, to 0·19 for untreated halved cysts, and to 0·13 for trypsin-treated halved cysts. Apparently some of the variability in emergence, like the limitation of total emergence, is bound up with the nature of the cyst as a more or less closed system.
In two experiments the hatching of eggs freed from half-cysts was compared with that of eggs in their fellow intact halves. In one experiment there was no difference; in the other it was possible to show that a higher proportion of free eggs hatched. Evidently factors limiting hatching may operate among eggs in a mass, even though they are no longer completely enclosed in a cyst.     

THE INFLUENCE OF SOIL MOISTURE ON THE EMERGENCE OF LARVAE FROM CYSTS OF THE BEET EELWORM, HETERODERA SCHACHTII SCHMIDT

The significance of soil moisture in relation to aeration and larval emergence from cysts of Heterodera schachtii Schmidt is discussed. The rate of larval emergence increased as oxygen concentration increased. A comparison of the moisture characteristics of a mass of cysts and of sand of about the same particle size showed that water was removed from between both the cysts and the sand particles at 12–16 cm. of water-pressure deficiency. There was an indication that water was removed from between the eggs within the cysts at 100–135 cm - of water-pressure deficiency. The rate of larval emergence showed a gradual decline is suction was increased beyond 20 cm. of water-pressure deficiency, approaching zero at 175 cm. of water-pressure deficiency. Results suggest that egg hatch is not directly dependent on pressure deficiency and that the presence of free larvae within the cyst inhibits further egg hatch. A technique is described for measuring larval motility in sand. There is a decline in motility when most of the water has been removed from the sand pore-spaces. It is suggested that the relatively low rates of larval emergence at high-pressure deficiencies are due to inhibition of larval migration from the cyst by the surface forces of the water film.     

草鱼生长与台湾棘带吸虫囊蚴内种群的关系

通过对草鱼鳃上台湾棘带吸虫囊蚴内种群的研究发现,随着草鱼的生长。其感染率,下蚴的平均密度都下降,其感染强度也呈下降的的趋势;草鱼鳃上囊蚴的频率分布,表明有大量的囊蚴寄生于少数草鱼鳃上,多数草鱼感染少量的囊蚴;囊蚴在草鱼不同组鳃片上的分布具有相同的变化规律;     

Effects of Inoculum Density and Egg Age on Establishment of Globodera rostochiensis Populations

The establishment of Globodera rostochiensis Rol populations was examined under greenhouse conditions. The probability of G. rostochiensis population establishment was calculated from the number of plants that produced new cysts with viable eggs following inoculation with various numbers of eggs of different ages. Probability of population establishment was positively correlated with inoculum density but was not affected by the age of eggs used in these experiments. The probability of G. rostochiensis establishment ranged from 5% at densities of 2 eggs/pot to 100% at densities of 25 eggs/pot or greater. At densities of 3 eggs/pot and beyond, there was no correlation between inoculum density and the number of viable eggs/new cyst. Also, the number of plants that produced new cysts was a function of inoculum density and not age of eggs. Juveniles from eggs 1 year old or older were equally as infective as were those from eggs in newly developed cysts (4 months old).     

THE EMERGENCE OF LARVAE FROM FREE EGGS OF THE POTATO-ROOT EELWORM HETERODERA ROSTOCHIENSIS (WOLL.)

A technique for conducting hatching experiments on eggs freed from cysts is described. The form of the hatching response was found to be similar to that of eggs contained within cysts, but the response of the free eggs to the hatching stimulus was slightly more rapid. Investigations into factors affecting free egg hatching showed that it was necessary to presoak cysts before extracting the eggs from them for hatching. Eggs taken from dry cysts or from cysts that had been opened or cracked before presoaking did not respond to diffusate. When free eggs and whole cysts were exposed to the same graded series of dilutions of diffusate, the L.A. values (i.e. log concentrations of hatching factor) derived from plotting the hatching curves were in very close agreement.     

Structure and Functioning of the “Egg Bank” of a Fairy Shrimp in a Temporary Pool: Chirocephalus ruffoi from Pollino National Park (Southern Italy) as a Case Study

To investigate their distribution and total numbers, resting eggs of the anostracan Chirocephalus ruffoi were collected from the bed of a temporary pool in southern Italy. Samples were taken at 0.5 m intervals along six transects oriented at 30° from each other, by means of a cylindrical core sampler. The horizontal distribution of intact resting eggs was extremely patchy, with cyst number per core ranging from 191 to 1,400 (CV = 32.7%), corresponding to a mean of between 0.8 and 4.3 cysts cm⁻³. Differences observed were related to core position and transect orientation, total cyst numbers being markedly higher in the leeward area of the pool compared to the windward area. Marked variation was also evident in vertical distribution, a significant, though weak correlation was recorded between egg density and sediment depth. Cyst‐bank size (± 95% confidence limits) of the pool bed, estimated from the mean cyst number cm⁻³ obtained for the 6 transects, ranged between 1.0 × 10⁸ and 1.3 × 10⁸ cysts. Hatching in the laboratory was very erratic. Despite significant differences in hatching, the observed variation was unrelated to most of the variables considered (position within sections, cores and transects, pre‐incubation treatment) and was explained only by initial sediment conditions (moist/dry). In none of the experimental conditions tested was synchronous hatching obtained. Possible causal factors (mixing of the bottom sediments by cattle, egg age, storage conditions, differential exposure to environmental cues as well as variability in hatching response even at clutch level) are discussed. (© 2004 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

大豆胞囊线虫的休眠

以前研究发现,辽宁地区大豆生长期间及收获期土壤中胞囊孵出的二龄幼虫量很少,推测线虫卵的休眠与大豆生长时期或季节相关。为明确该地区大豆胞囊线虫的休眠特点,2002-2003年采用田间随机多点取样、室内分离及模拟自然条件孵化等方法对大豆胞囊线虫的休眠进行深入研究。结果表明:在生长季节,感病品种辽豆10根围土壤中的白色雌虫、卵囊及褐色的胞囊均可孵出二龄幼虫,且孵化持续时间较长,第21d仍有幼虫孵出,白色雌虫及卵囊内的卵孵化率高于褐色胞囊;不同作物对其根围土壤中胞囊内卵的孵化影响不大,寄主作物大豆、非寄主作物玉米根围及休闲地土壤中的胞囊在条件适宜均可孵出二龄幼虫;季节对胞囊内卵的孵化有较大的影响,出苗期孵化率最高,收获期最低,2周时平均1个胞囊孵出幼虫分别为83.8和9.7条;胞囊皮对线虫卵的孵化有显著的影响。表明沈阳地区大豆胞囊线虫在正常和逆境条件下均有部分卵表现休眠。     

长枝木霉对禾谷胞囊线虫寄生和致死作用的显微观察及测定

通过室内试验测定长枝木霉分生孢子悬浮液对小麦禾谷胞囊线虫胞囊的寄生和致死作用.结果表明： 不同浓度（1.5×10⁵～1.5×10⁷ cfu·mL^-1）长枝木霉分生孢子悬浮液对小麦禾谷胞囊线虫胞囊具有明显的寄生和致死作用,并且不同浓度的长枝木霉分生孢子悬浮液之间存在显著差异.第18天浓度为1.5×10⁷ cfu·mL^-1的长枝木霉分生孢子悬浮液对胞囊的寄生率为96.7%,第22天对胞囊孵化的相对抑制率为91.2%.显微观察表明,侵染初期长枝木霉分生孢子附着在胞囊体表,并且萌发产生大量的菌丝寄生于胞囊体表,使〖JP2〗胞囊胚胎发育停止和内容物凝集,甚至有的胞囊出现畸形和表面形成深褐色的小液泡.侵染后期大量菌丝穿透胞囊体表,胞囊破裂,内容物外渗,有的胞囊体表菌丝形成分生孢子梗,其上着生卵圆形的分生孢子.表明长枝木霉可作为一种高效的生防制剂防治小麦禾谷胞囊线虫的发生与危害.     

Hymenolepis nana: Maturation in an immunosuppressed unnatural rat host

When eggs of the dwarf tapeworm Hymenolepis nana, cycled exclusively and directly through mice for more than 10 years, were inoculated into previously uninfected inbred Fischer (F344) strain rats, they failed to mature in the rat intestinal lumen. Eggs of H. nana inoculated into the rat developed normally into cysticercoids (cysts) in the intestinal tissue, but thereafter failed to mature in the lumen except when the host was treated with cortisone acetate from the day of cyst maturation. The Fischer rat initially given eggs of H. nana became completely immune to egg challenge within 2 days of egg inoculation; no cysts derived from challenge eggs were found in the immunized rat. Immunosuppression, assessed by the success of cyst recovery in the tissue 4 days after egg challenge, had no promotive effect on the recovery of adult worms derived from eggs initially inoculated. Rats initially given eggs and immunosuppressed by cyclophosphamide or antithymocyte serum did not harbor any adult worms. Cortisone acetate treatment which was sufficient for eggs inoculated to mature (a total of 75 or even 200 mg, from Day 5 of egg inoculation) had no effects of immunosuppression, whereas cortisone acetate treatment which was sufficient for immunosuppression (a total of 150 mg from Day -2, two days prior to the initial egg inoculation) induced some adult formation as well. In addition, when mouse-derived cysts were inoculated into the rat instead of eggs, they also failed completely to mature even when the rat was treated with cyclophosphamide or antithimocyte serum. However, when the rat was treated with cortisone acetate from the day of cyst inoculation, the cysts developed into adult worms. Therefore, these results indicate that the Fischer rat clearly differs in its susceptibility to the tissue phase of egg inoculation and to the lumen phase of cyst inoculation of H. nana, and strongly suggest that the failure of maturation of H. nana in the unnatural host Fischer rat is not attributed to innate and/ or acquired immunity of the rat but to other nonimmunological mechanisms.     

Influence of nematicide, resistant and susceptible potato cultivars and bare fallow on the population dynamics of Globodera rostochiensis Woll. Rol under field conditions in Spain

The susceptible potato cultivar Désirée and the resistant cv. Diamant were cultivated on untreated and nematicide-treated plots on two fields in northern Spain infested with Globodera rostochiensis Rol. Initial and final infestations of cysts and viable eggs were assessed. Multiplication rates differentiated more accurately the effects of the different treatments than final number of cysts and eggs, due to the elimination of some of the errors associated with variability of the initial infestation. Many new cysts were observed on all plots where susceptible potatoes had been grown, whether they had been treated with nematicide or not, although significantly fewer cysts were actually found on the nematicide-treated plots. With the resistant cultivar no significant effects of the nematicide were detected even though there was a noticeable reduction of viable cyst contents over and above the effect of the resistant cultivar.     

Hymenolepis nana: Protective immunity against mouse-derived cysticercoids induced by initial inoculation with eggs

Mice were almost completely resistant to a mouse-derived cysticercoid (cyst) challenge after 6 to 10 months following an initial immunizing inoculation with eggs of Hymenolepis nana. Previously uninfected control mice of the same age became infected with the cyst-derived tapeworms. There was no age resistance to H. nana in mice. Immunity against the cyst challenge was acquired by initial egg inoculation and blocked by injecting cortisone acetate just prior to the challenge. However, the number of worms recovered from mice given cortisone was significantly less than that from nonimmunized controls. Unexpected evidence was obtained that a few of the egg-derived tapeworms can survive for 6 or more months in some of the immunized mice, which are resistant to both egg and cyst challenges. The relative immunogenicity of oncospheres and cysts is discussed. It is strongly suggested that the cysts are different from the oncospheres in their immunogenicity, and, because of this, H. nana can complete its life cycle in the same immunized host. It is also suggested that the host possesses at least two separate immune responses: One is an early response directed exclusively against the oncosphere and/or the early postoncospheral stage (s) acquired within 2 days of egg inoculation, and the other is a late response against the cyst acquired after a time lag of unknown duration.     

Flow Cytometric Analysis and Sorting of Heterodera glycines Eggs

A nondestructive technique was developed to characterize and separate eggs of soybean cyst nematode, Heterodera glycines, by developmental stage using flow cytometry. Eggs from cysts cultured on susceptible soybean roots were suspended in 0.1% xanthan gum or 59% sucrose and loaded into either a Coulter EPICS 752 or EPICS 753 flow cytometer. Eggs were analyzed and sorted according to forward angle and 90° light scatter, flow cytometric parameters that are relative measures of object size and granularity, respectively. Mature eggs containing vermiform juveniles were less granular and slightly larger than eggs in earlier stages of embryogeny, allowing for separation of mature eggs from immature eggs. The effectiveness of flow cytometric sorting was evaluated by comparing the developmental stages of subpopulations of unsorted and sorted eggs. Of a subpopulation of unsorted eggs, 62% contained vermiform juveniles, whereas 85 to 95% of sorted subpopulations of larger, less granular eggs contained vermiform juveniles. Suspending H. glycines eggs in 0.1% xanthan gum or 59% sucrose for flow cytometric analysis had no effect on subsequent egg hatch in vitro. This technique is an efficient and effective means to collect large, relatively homogeneous quantities of H. glycines eggs in early or late embryogeny, and would likely be useful for analyzing and sorting eggs of other nematode species for use in developmental, genetic, or physiological research, or for identification and collection of parasitized eggs.     

CONTROL OF GERMINATION OF ALEXANDRIUM TAMARENSE (DINOPHYCEAE) CYSTS FROM THE LOWER ST. LAWRENCE ESTUARY (CANADA)

Cysts of the toxic dinoflagellate Alexandrium tamarense (Lebour) Balech 1992 from the lower St. Lawrence estuary were used in a test of the following hypotheses: (1) cyst germination is triggered by a change in temperature, and (2) germination rate varies throughout the year and is controlled by a circannual internal biological clock. Results show that cyst germination was not affected significantly by temperature of incubation over the range 1°–16° C, and light showed no significant stimulation of germination. This is supported by the lack of effect of cyst incubation conditions during evaluation of the seasonal changes in germination rate (two temperatures: 4° and 15° C, and two light conditions: darkness and 150 μmol photons·m^?2·s^?1). Thus, direct environmental control through short-term increases in temperature and exposure to light has no effect on the germination of the cysts tested. The rate of germination, observed monthly over a 16-month period, showed low germination (<20%) over most of the period tested, except for a maximum reaching more than 50% germination in August to October of the second year of the experiment. This pattern was observed for cysts both from monthly field collections and from laboratory-stored cysts kept under constant environmental conditions (4° C, in the dark). The peak in germination observed under constant environmental conditions (in the laboratory), the almost coincidental increase in cyst germination observed for the field-collected cysts, and the absence of effects of temperature and light during incubation could be explained either by a temperature-controlled cyst maturation period (the time-temperature hypothesis of Huber and Nipkow 1923) or by the presence of an internal biological clock. However, the large decline in the rate of germination 2 months after the maximum provides strong support for the biological clock hypothesis. The ca. 12-month maturation (dormancy) period observed for the laboratory-stored cysts is the longest reported for this species to our knowledge; this might be related to the low storage temperature (4° C), which is close to bottom temperatures generally encountered in this environment (0° to 6° C). Similar field and laboratory storage temperatures could explain the coincidental increase in germination rate in the fall of the second year if cyst maturation is controlled by temperature. A fraction of the laboratory-stored cysts did not follow a rhythmic pattern: A rather constant germination rate of about 20% was observed throughout the year. This continuous germination of likely mature cysts may supplement the local blooms of this toxic dinoflagellate, as these often occur earlier than peak germination observed in late summer. It seems that two cyst germination strategies are present in the St. Lawrence: continuous germination after cyst maturation, with temperature controlling the length of the maturation period, and germination controlled by a circannual internal rhythm.     

Population dynamics of a host-specific root-feeding cyst nematode and resource quantity in the root zone of a clonal grass

Recent studies have suggested that root-feeding nematodes influence plant community dynamics, but few studies have investigated the population dynamics of the nematodes. In coastal foredunes, feeding-specialist cyst nematodes ( Heterodera spp.) are dominant in the soil nematode community and greenhouse studies pointed at bottom-up control by their host plant. Here we examine the population dynamics of H. arenaria in the field in relation to resource quantity in the root zone of Ammophila arenaria (marram grass).
Ammophila arenaria is a pioneer grass in mobile coastal foredunes of northwestern and Mediterranean Europe. Every year, the plant expands clonally into freshly deposited layers of wind-blown sand, followed by dispersal and build-up of the nematode population into the new root zone. In a newly developing root zone the first H. arenaria cysts were observed one month after the first new roots were detected, indicating that nematode dispersal is not limiting the initial establishment of new populations. Throughout the growth season, the numbers of cysts, as well as the numbers of eggs and juveniles within cysts, were related to the quantity of roots. However, cyst density varied between years. Therefore, we conclude that in new root layers of A. arenaria cyst nematodes are bottom-up controlled by resource quantity, but that other factors, for example resource quality, influence the relation between nematode abundance and resource quantity.
In deeper root zones the nematode abundance declines over time. Here, numbers of cysts were not related to root biomass, while numbers of eggs and juveniles inside the cysts were weakly related. This points at other factors than resource quantity, for example the quality of the roots or unsuitable abiotic environmental conditions that have a stronger influence on cyst numbers than resource quantity. We discuss how bottom-up control of cyst nematodes may indirectly protect the plant against harmful root knot nematodes.     

棉田棉铃虫卵估值抽样的研究

由于棉铃虫卵在棉株上的空间分布结构属负二项分布,并且它在棉株上的垂直分布特征系上部卵量明显较下部为多,这样,本文应用了二阶抽样技术代替当前国内采用的简单随机抽样法,在允许误差为10％时,每株抽取上部15个叶蕾,每块棉田共抽60株棉株。 抽样单位的大小对于抽样的时间消费与抽样精确度亦有直接的影响,因此,确定最适的抽样单位也是很重要的。本文根据棉铃虫卵在田间个体群平均占据的面积大小,提出一次调查6抹(2行×3株/行)棉株较单株为优。即在棉田中每次调查6株,共调查10次 新的抽样方法较当前国内应用抽样法,提高效率2.2—2.6倍。     

Fungal parasitism of the cyst nematode Heterodera schachtii: infection and enzymatic activity

Abstract Fungal egg parasites isolated from eggs of the cyst nematode Heterodera avenae in Sweden were investigated with respect to their ability to infect cyst nematode eggs of H. schachtii in vitro. The infection was studied by interference phase contrast microscopy of whole cysts and of cryosections of cysts exposed to the fungi on agar plates.
Verticillium suchlasporium was the most effective parasite, infecting 53% of the nematode eggs, while V. chlamydosporium infected 12% of the eggs. The fungi Paecilomyces lilacinus, Cylindrocarpon destructans or Fusarium oxysporum did not parasitize nematode eggs; nor did Arthrobotrys oligospora , a nematode trapping fungus nor Penicillium viridicatum which served as a control fungus.
The ability of the fungi to infect eggs was correlated with their lytic enzyme activity. Fungi that readily infected eggs also showed chitinase activity and presence of proteolytic activity. The Verticillium species had an activity between 3.7 and 14.6 μmol N -acetyl-glucosamine per mg protein per hour (CU) while it was 4.5 CU or lower for P. lilacinus . Other isolates did not shown any chitinase activity.     

Multivariate Analysis of Selected Edaphic Factors and Their Relationship to Heterodera glycines Population Density

The influence of selected soil physical and chemical factors on population density of Heterodera glycines was investigated in 1988 and 1989 in two different locations of a soybean (Glycine max) field. Soil variables of a Norfolk loamy sand were measured after planting soybeans susceptible to H. glycines. Cyst and egg populations were determined after harvest. Nematode population density was found to be clustered. Up to 91% of the eggs were parasitized by a sterile fungus. Principal component analysis with orthogonal VARIMAX rotation grouped 12 variables into five uncorrelated factors in 1988 and three in 1989. In 1988, the factor "pH and Mg" was positively correlated (P < 0.001) with cyst and egg population density. Also, the factor "fine texture and Cu" was negatively correlated (P < 0.05) with egg population density. In 1989, the factor "pH, Mg and Cu" was positively correlated (P < 0.05) with levels of cysts and percentage of parasitized eggs, but not with total egg number. Across 2 years, factors containing soil pH and Mg were positively associated with cyst nematode population density. Copper appeared to be negatively associated with populations of H. glycines.     

Specific cross-immunity between Hymenolepis nana and H. diminuta: immunization with heterologous and homologous light infections

The consequences of previous and concurrent infection with two related species of cestodes, Hymenolepis nana and H. diminuta, were studied in CD1 mice. A H. diminuta infection strongly affected the establishment and the survival of a secondary H. nana egg or cyst infection administered 30 days later. An infection of 20 H. nana eggs strongly protected against a 5-cyst H. diminuta challenge, whereas an infection of 10 H. nana cysts was ineffective; 20 H. nana eggs also protected against a challenge with 5 cysts of H. diminuta administered 5 days later. No effects were observed in either parasite during a concurrent infection established by administration of cysts. An H. nana egg-infection was unable to affect the establishment of a secondary H. nana cyst-infection given 1 month later; however a significant decrease in growth was found. Similar results were found when a primary H. nana egg-infection was followed 5 days later by the homologous cyst-infection. But an infection with 5 H. nana cysts was unable to protect against a homologous challenge of 5 cysts or 200 eggs. The reciprocal cross immunity between the heterologous parasites and the failure of protection of homologous challenges are discussed in relation to light infections.