Amoebae and algae can prolong the survival of Campylobacter species in co-culture

Several species of free-living amoebae can cause disease in humans. However, in addition to the direct pathogenicity of e.g. Acanthamoebae and Naegleria species, they are recognized as environmental hosts, indirectly involved in the epidemiology of many pathogenic bacteria. Although several studies have demonstrated intracellular survival of many different bacteria in these species, the extent of such interactions as well as the implications for the epidemiology of the bacterial species involved, are largely unknown and probably underestimated. In this study, we evaluated eight different unicellular eukaryotic organisms, for their potential to serve as environmental hosts for Campylobacter species. These organisms include four amoebozoas (Acanthamoeba polyphaga, Acanthamoeba castellanii, Acanthamoeba rhysodes and Hartmanella vermiformis), one alveolate (Tetrahymena pyriformis), one stramenopile (Dinobryon sertularia), one eugoenozoa (Euglena gracilis) and one heterolobosea (Naegleria americana). Campylobacter spp. including Campylobacter jejuni, Campylobacter coli and Campylobacter lari are the most common cause of gastroenteritis in the western world. Survival and replication of these three species as well as Campylobacter hyointestinalis were assessed in co-cultures with the eukaryotic organisms. Campylobacter spp. generally survived longer in co-cultures, compared to when incubated in the corresponding growth media. The eukaryotic species that best promoted bacterial survival was the golden algae D. sertularia. Three species of amoebozoas, of the genus Acanthamoeba promoted both prolonged survival and replication of Campylobacter spp. The high abundance in lakes, ponds and water distribution networks of these organisms indicate that they might have a role in the epidemiology of campylobacteriosis, possibly contributing to survival and dissemination of these intestinal pathogens to humans and other animals. The results suggest that not only C. jejuni, but a variety of Campylobacter spp. can interact with different eukaryotic unicellular organisms.     

Effect of copper exposure on GST activity and on the expression of four GSTs under oxidative stress condition in the monogonont rotifer, Brachionus koreanus

Glutathione S-transferases (GSTs; EC 2.5.1.18) are major enzymes that function in Phase II detoxification reactions by catalyzing the conjugation of reduced glutathione through cysteine thiol. In this study, we cloned and sequenced four GST genes from the monogonont rotifer Brachionus koreanus. The domain regions of four Bk-GSTs showed a high similarity to those of other species. In addition, to evaluate the potential of GST genes as an early warning signal for oxidative stress, we exposed sublethal concentrations of copper (Cu) to B. koreanus and measured glutathione (GSH) contents and several antioxidant enzymes such as glutathione S-transferase (GST), glutathione peroxidase (GPx; EC 1.11.1.9), and glutathione reductase (GR; EC 1.8.1.7). The reactive oxygen species (ROS) at 12 h and 24 h after copper exposure increased significantly. GSH contents however did not increase significantly and even it decreased at 0.24 mg/L at 12 h. The activities of several antioxidant enzymes, particularly GPx and GR, showed a dramatic increase in 0.24 mg/L of CuCl₂. Messenger RNAs of each Bk-GST showed different patterns of modulations according to GST types, and particularly, Bk-GST-omega, Bk-GST-sigma, and Bk-GST zeta genes were highly sensitive to Cu. These results indicate that Bk-GSTs, functioning as one of the enzymatic defense mechanisms particularly in the early stage of oxidative stress response, were induced by Cu exposure. This also suggests that these genes and related enzymes have a potential as biomarkers for a more sensitive initial stress response.     

The identification of a new Giardia duodenalis assemblage in marine vertebrates and a preliminary analysis of G. duodenalis population biology in marine systems

Giardia duodenalis is an intestinal parasite of many vertebrates. The presence of G. duodenalis in the marine environment due to anthropogenic and wildlife activity is well documented, including the contributions from untreated sewage and storm water, agricultural run-off and droppings from terrestrial animals. Recently, studies have detected this protistan parasite in the faeces of marine vertebrates such as whales, dolphins, seals and shore birds. To explore the population biology of G. duodenalis in marine life, we determined the prevalence of G. duodenalis in two species of seal (Halichoerus grypus, Phoca vitulina vitulina and Phoca vitulina richardsi) from the east and west coasts of the USA, sequenced two loci from G. duodenalis-positive samples to assess molecular diversity and examined G. duodenalis distribution amongst these seals and other marine vertebrates along the east coast. We found a significant difference in the presence of G. duodenalis between east and west coast seal species. Only the zoonotic lineages of G. duodenalis, Assemblages A and B and a novel lineage, which we designated as Assemblage H, were identified in marine vertebrates. Assemblages A and B are broadly distributed geographically and show a lack of host specificity. Only grey seal (Halichoerus grypus) samples and one gull sample (Larus argentatus) from a northern location of Cape Cod, Massachusetts, USA, showed the presence of Assemblage H haplotypes; only one other study of harbour seals from the Puget Sound region of Washington, USA previously recorded the presence of an Assemblage H haplotype. Assemblage H sequences form a monophyletic clade that appears as divergent from the other seven Assemblages of G. duodenalis as these assemblages are from each other. The discovery of a previously uncharacterised lineage of G. duodenalis suggests that this parasite has more genetic diversity and perhaps a larger host range than previously believed.     

密度信号对两种臂尾轮虫种群增长的影响

为了更好的了解化学干扰对轮虫种间竞争的影响,分别用萼花臂尾轮虫(Brachionus calyciflorus)和角突臂尾轮虫(Brachionus angularis)研究了同种和异种种群密度信号对种群增长的影响。结果表明,密度信号对轮虫种群增长具有一定的影响。萼花臂尾轮虫的密度信号显著降低了本种和角突臂尾轮虫种群的环境容纳量K并提高了它们的种群增长率r(P0.05),角突臂尾轮虫的密度信号对本种和萼花臂尾轮虫的种群增长都没有显著影响(P0.05)。萼花臂尾轮虫种群增长受同种密度信号抑制十分显著,环境容纳量K降低了50%以上,可能主要受自毒作用的调节,但同种密度信号对高有性生殖的萼花臂尾轮虫种群的混交率影响不明显。角突臂尾轮虫几乎不受自身密度信号的影响。因此,在两种臂尾轮虫的竞争中,由于自身密度信号的抑制作用,萼花臂尾轮虫将更快被角突臂尾轮虫排除。据此可推测密度信号对轮虫的种间竞争具有重要影响。     

The effects of dietary supplementation of algae and synthetic astaxanthin on body astaxanthin, survival, growth, and low dissolved oxygen stress resistance of kuruma prawn, Marsupenaeus japonicus Bate

Natural carotenoids from astaxanthin containing alga Haematococcus pluvialis (H) and a non-astaxanthin carotenoid-containing alga Spirulina pacifica (S), and a synthetic astaxanthin Carophyll Pink (A) were supplemented in formulated diets at two concentrations, 50 (I) and 100 (II) mg kg⁻¹, resulting in seven pigmented diets HI, SI, AI, HII, SII, AII, and HS (H-50 mg kg⁻¹+S-50 mg kg⁻¹). Formulated diet without carotenoid supplementation served as a control (C). The different diets were fed to juvenile kuruma prawn Marsupenaeus japonicus for 9 weeks. Dietary carotenoid effects on survival, growth, and pigmentation were compared by the treatment individually or collectively. A low dissolved oxygen stress test was conducted 2 weeks later and prawns' survival time and oxygen consumption rate were also compared among treatments. After 9 weeks' rearing, C-fed prawn had significantly lower survival rate than the pigmented diets-fed prawns. No difference in weight gain was found among all prawns. C-fed prawn had 66.4% less flesh astaxanthin (FA) and 75.5% less shell astaxanthin (SA) than the pigmented diets-fed prawns. I-fed (AI, HI, and SI) prawns had 31.1% less FA and 29.6% less SA than II-fed (AII, HII, SII, and HS) prawns. No significant differences were found in the comparisons by other categories. The use of these three sources of carotenoids for pigmentation in crustacean was discussed along carotenoid conversion, deposition, digestibility, and absorption. When subjected to low dissolved oxygen stress, C-fed prawn had higher oxygen consumption rate (OCR) and shorter survival time (ST) than the prawns fed the pigmented diets. No differences in OCR or ST were found in the comparisons by other categories.     

水稻生育期对褐飞虱和白背飞虱卵巢发育及起飞行为的影响

研究了取食分蘖初期、拔节期和孕穗抽穗期稻株上的褐飞虱和白背飞虱成虫的卵巢发育和起飞情况。其中,水稻生育期对褐飞虱的影响相对较小,不同生育期稻株上试虫的卵巢发育级别羽化后的4d内均无显著差异,羽化后第5天的褐飞虱在孕穗抽穗期卵巢发育最快,分蘖初期最慢,拔节期居中。褐飞虱的起飞率在各个生育期均无显著差异。白背飞虱卵巢发育、起飞率均受水稻生育期的显著影响,在水稻分蘖初期,卵巢发育最快,孕穗抽穗期则卵巢发育最慢,拔节期居中,水稻分蘖初期,起飞率最低,孕穗抽穗期起飞率最高,拔节期居中。结果表明,水稻生育期对两种飞虱卵巢发育和起飞行为的影响明显不同。认为水稻生育期对两种飞虱卵巢发育与起飞行为的不同影响,应是水稻田间白背飞虱发生较早、褐飞虱发生较晚的一个重要原因。     

Minor effects of long-term ozone exposure on boreal peatland species Eriophorum vaginatum and Sphagnum papillosum

The effects of long-term ozone fumigation on two common peatland plant species, a sedge Eriophorum vaginatum L. and a moss Sphagnum papillosum Lindb., were studied applying peatland microcosms. The peat cores with intact vegetation were cored from an oligotrophic pine fen and partially embedded into the soil of an open-air experimental field for four growing seasons. The open-air ozone exposure field consists of eight circular plots of which four were fumigated with elevated ozone concentration (doubled ambient) and four were ambient controls. The results showed that E. vaginatum and S. papillosum can tolerate ozone better than expected. Elevated ozone concentration did not affect overall relative length growth of E. vaginatum or S. papillosum. The leaf cross-section area of E. vaginatum leaves was 8% bigger in the ozone treatment compared to that in the ambient control. Ultrastuctural variables did not show any significant treatment effect in E. vaginatum or in S. papillosum. Total chlorophyll (a + b) concentration tended to increase in early growing season under ozone exposure. During the first growing season, elevated ozone concentration decreased methanol-extractable, UV-absorbing compounds in E. vaginatum. The results suggest that E. vaginatum and S. papillosum are ozone tolerant plant species and are likely able to cope with expected increase in tropospheric ozone concentration.     

A new species of ferret-badger, Genus Melogale, from Vietnam

Ferret-badgers, genus Melogale, are distributed in the Indochinese region, Java, Bali and NE-Borneo. There are currently four species described each having very similar phenotypes. In March 2005, a living ferret-badger of a different phenotype was confiscated by rangers from Cuc Phuong National Park, Vietnam. This individual died and the carcass was not preserved. In January 2006, a newly deceased individual of the same phenotype was found at the Endangered Primate Rescue Center, Cuc Phuong National Park. Due to several different characteristics these individuals vary greatly from the current species. Thus, we describe an additional species, M. cucphuongensis sp. nov. from northern Vietnam, which occurs sympatrically with M. moschata and M. personata, but differs from both species clearly in skull morphology and other features.Based on a 423 bp-long fragment of the mitochondrial cytochrome b gene, M. cucphuongensis sp. nov. is a member of the genus Melogale and represents a sister lineage to a clade consisting of M. personata and M. moschata.     

Zooid size and growth rate of the bryozoan Cryptosula pallasiana Moll in relation to temperature, in culture and in its natural environment

The size of cheilostome bryozoan zooids has been widely discussed for its potential in inferring palaeotemperatures, based on correlations between zooid size and temperature. Studies in both the natural environment and under experimental laboratory conditions have shown that an increase in temperature significantly decreases zooid size in a range of bryozoan taxa. In order to test the effect of temperature on zooid size, the cheilostome bryozoan Cryptosula pallasiana was for the first time successfully cultured under laboratory conditions. C. pallasiana was grown at 14 °C and 18 °C using Rhodomonas sp. as a food organism. Zooid size, tentacle number and growth rate were measured over a period of 26 days. For comparison, zooids from colonies of C. pallasiana collected from the natural environment were measured in winter and summer months. Results showed that colonies grown in laboratory culture had significantly longer and wider zooids at 14 °C than at 18 °C. The specific growth rate of C. pallasiana doubled from 14 °C to 18 °C. Comparison of tentacle number in culture showed a significantly higher value at lower temperatures. This may be related to differing food availability, longer polypide life spans, or a shift of energy use at colder temperatures. In nature the zooids were significantly longer in colonies sampled in July than in January, a clear difference from laboratory results. The utility of cheilostome Bryozoa as indicators of environmental change and their potential for studies of paleotemperature are highlighted.     

The effects of light and nutrients on Caulerpa taxifolia and growth

Caulerpa taxifolia, an invasive species elsewhere in the world, is native to Moreton Bay where its distribution has been increasing in recent years. In Australia, dense beds of C. taxifolia are predominantly found in areas of low light and high nutrients (low water quality). Monitoring data from Moreton Bay suggests that native C. taxifolia is not directly replacing seagrass, but that there is a successional trend of seagrass loss and subsequent C. taxifolia colonization. The current study examined responses of C. taxifolia in relation to changes in environmental conditions using ambient water quality and a light/nutrient manipulative experiment. In the ambient water quality experiment we found that C. taxifolia grew significantly faster in areas with higher light (lower turbidity). The manipulative experiment demonstrated that nutrients stimulate C. taxifolia growth, however, light availability and seasonality appear to influence the response of C. taxifolia growth to nutrients in Moreton Bay. These findings suggest that C. taxifolia is unlikely to colonize seagrass beds in areas with high light and low nutrients; however, in areas with moderate light and moderate to high nutrients C. taxifolia and seagrass are likely to coexist.     

脂肪酸对中华哲水蚤摄食两种海洋微藻的指示作用

在室内以饥饿培养为对照,以海洋原甲藻(Prorocentrum micans)和中肋骨条藻(Skeletonema costatum)培养中华哲水蚤(Calanus sinicus),研究了各脂肪酸标记对中华哲水蚤摄食不同饵料的指示作用。结果显示,海洋原甲藻中18 ∶ 4ω3、22 ∶ 6ω3含量较高,中肋骨条藻中16 ∶ 1ω7、20 ∶ 5ω3的含量较高。二者分别表现出典型的甲藻门和硅藻门的脂肪酸组成特征。中华哲水蚤的脂肪酸组成有两个特点:(1)20 ∶ 5ω3和22 ∶ 6ω3的含量均较高;(2)其体内表征桡足类浮游植物食性、由桡足类自身合成的20 ∶ 1和22 ∶ 1脂肪酸占有相当的比例。虽然中华哲水蚤对不同脂肪酸的吸收和转化效率不同,但以脂肪酸作为标记还是成功的指示了中华哲水蚤对微藻的摄食。在饥饿培养中,首先消耗的是那些浮游动物自身不能合成的多不饱和脂肪酸,而结构脂肪酸都表现出了较高的保守性。结合各脂肪酸标记变化趋势和Pearson相关性分析的结果认为,18 ∶ 4ω3、18 ∶ 4ω3/16 ∶ 1ω7、∑18/∑16能较好的指示中华哲水蚤对海洋原甲藻的摄食,仅16 ∶ 1ω7/18 ∶ 4ω3能指示中华哲水蚤对中肋骨条藻的摄食。     

Inhibitory effects of leachate from Eupatorium adenophorum on germination and growth of Amaranthus retroflexus and Chenopodium glaucum

Amaranthus retroflexus L. and Chenopodium glaucum L. are two widely distributed destructive weeds. Their strong adaptability and massive seed production make them the hardest weeds to deal with. This present study intended to investigate the effect of leachate from Eupatorium adenophorum on the growth of these weeds and explore the potential to develop an environmental friendly strategy to use the leachate to control the weeds. Seeds of A. retroflexus L. and C. glaucum L. were soaked in solutions containing 0%, 0.6%, 1.25%, 2.5%, and 5% leachate from E. adenophorum leaves. A. retroflexus and C. glaucum seedlings grown in pots were sprayed with leachate solutions in the same concentration range. The effects of these leachate solutions on membrane permeability and germination of seeds, and growth and physiological characteristics of the seedlings were investigated. The highest concentration of leachate (5%) caused significant damage to the cell membrane of seeds of both weed species, whereas lower concentrations (0.6%) promoted repair of the membrane system, as reflected by higher and lower than control in relative conductivity (RC), respectively. Different concentrations of leachate showed distinct allelopathic inhibitory effects on the two weed species; lower concentrations showed weak inhibitory or even positive effects, whereas higher concentrations showed stronger inhibitory effects. Higher concentrations of leachate (2.5% and 5%) delayed germination and significantly decreased the emergence rate of the seeds, survival rate, and dry matter accumulation of the seedlings. When treated by 5% leachate, the emergence date of A. retroflexus was delayed by 3.6 d, emergence rate of the seeds and survival rate was 69.1% and 70.6% of the control, respectively, seedling dry matter was 48.6% less than the control; In the case of C. glaucum, the emergence date was delayed by 2.7 d, emergence rate of the seeds and survival rate was 45.1% and 58.6% of the control, respectively, seedling dry matter was 44.7% less than the control. There were significant interactions among the different concentrations of leachate and the length of treatment period with respect to activities of antioxidant enzymes, malondialdehyde (MDA) contents, and chlorophyll contents. Seedlings treated with 0.6%, 1.25%, or 2.5% leachate solution for 24–72 h showed increased superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities. When seedlings were treated with leachate solutions for 96 h, antioxidant enzyme activities and chlorophyll content decreased in A. retroflexus, but only CAT activity decreased in C. glaucum. When seedlings of the two weed species were treated with 5% leachate solution, CAT activity and chlorophyll content decreased and MDA content gradually increased with longer treatment times (from 24 to 96 h). The two weed species showed different allelopathic responses to E. adenophorum; A. retroflexus was more sensitive than C. glaucum. Based on the investigation, it could be speculated that the delayed germination and low germination rate of the weeds after treatment by leachate could be due to the fact that leachate damaged the membrane system of the seeds. By delaying germination, lowering the germination rate of the weeds and inhibiting seedling growth, leachate from E. adenophorum could provide an effective way of controlling the weeds.     

Comparative studies on the allelopathic effects of two different strains of Ulva pertusa on Heterosigma akashiwo and Alexandrium tamarense

Allelopathic effects of fresh tissue and dry powder of a nonsexual and a sexual strain of the macroalga Ulva pertusa on the growth of the microalgae Heterosigma akashiwo and Alexandium tamarense were evaluated using long-term coexistence culture systems in which several concentrations of macroalga fresh tissue and dry powder were used. The effects of macroalga culture medium filtrate on the two HAB algae were also investigated. Moreover, isolation co-culture systems were built to confirm the existence of allelochemicals and preclude the growth inhibition by direct contact. Short-term algicidal effect assays of macroalgae on H. akashiwo were carried out to measure the rate of algal cell lysis. The results of the coexistence assays showed that the growth of H. akashiwo and A. tamarense was strongly inhibited by fresh tissue and by dry powder of both strains of U. pertusa. The allelochemicals were lethal to H. akashiwo at relatively higher concentrations. The macroalga culture medium filtrate exhibited no apparent growth inhibitory effect on the two HAB algae under initial or semicontinuous filtrate addition, which suggested that continuous release of small quantities of rapidly degradable allelochemicals from the fresh tissue of both strains of U. pertusa was essential to effectively inhibit the growth of H. akashiwo and A. tamarense.     

Koralionastetales, a new order of marine Ascomycota in the Sordariomycetes

Based on molecular studies using 1760 bp of the nuSSU and 604 bp of the nuLSU rRNA genes and using morphological characters, the genera Koralionastes and Pontogeneia are assigned to the new order Koralionastetales, family Koralionastetaceae, class Sordariomycetes. Koralionastetales is a sister group to Lulworthiales; differences in morphological characters are expressed in the ascospores and the presence/absence of periphyses and paraphyses. A new species of Pontogeneia, P. microdictyi from Microdictyon sp. in the Bahamas, is described.     

Plant-mediated effects of different Salix species on the performance of the braconid parasitoid Perilitus brevicollis

The blue willow beetle, Phratora vulgatissima, is considered to be the most damaging herbivorous pest in Salix short-rotation coppices throughout Europe. The braconid parasitoid Perilitus brevicollis is an important natural enemy of Phratora. As several different Salix species are used in coppices, I investigated the bottom-up (tritrophic) effects of Salix on the parasitoid. Three host plants were studied: the introduced fast-growing S. viminalis, which is highly susceptible to the beetle; S. dasyclados, which is introduced and moderately-resistant to the beetle; and the native slow-growing Salix cinerea, which is not currently used in coppices. The identity of the host-plant species had significant effects on parasitoid larval development time; parasitoids developed rapidly on the susceptible S. viminalis and slowly on the moderately resistant S. dasyclados. Increased development time resulted in reduced adult longevity. Host-plant species identity also affected larval survival; 57%, 64%, and 49% of the parasitoids successfully completed larval development in beetles fed S. viminalis, S. cinerea, and S. dasyclados, respectively. Parasitoid development was also correlated with the body size of their beetle host, but this effect was independent of the identity of the host-plant species. The results of this study suggest that the parasitoid has higher survival and growth rates when it parasitizes beetles feeding on the common coppice species S. viminalis, but the performance of the parasite is reduced when the beetle feeds on the moderately-resistant S. dasyclados. Conversely, the omnivorous biocontrol agents sometimes used in these systems appear to perform better on S. dasyclados compared to S. viminalis. The results of this study suggest that Perilitus parasitoids and omnivorous beetle predators may provide complementary protection to Salix and therefore be useful in coppice management.     

Isolation and distribution of iridescent Cellulophaga and other iridescent marine bacteria from the Charente-Maritime coast,French Atlantic

An intense colored marine bacterium, identified as Cellulophaga lytica, was isolated previously from a sea anemone surface on the Charente-Maritime rocky shore (Atlantic Coast, France), and iridescence of its colonies under direct light was recently described. In addition, iridescence intensities were found to differ strongly between C. lytica strains from different culture collections. However, importantly, the occurrence and distribution of iridescent bacteria in the marine environment were still unknown. Therefore, in this study, a search was undertaken for marine iridescent bacterial strains in different biotopes of the Charente-Maritime coast. Various marine samples (water, sediment, macroalgae, other macroorganisms and detritus) were collected from seven biotopes using a direct plate inoculation method. As a result, 34 iridescent strains related to the genus Cellulophaga, as well as 63 iridescent strains affiliated to the genera Tenacibaculum and Aquimarina, were isolated. Iridescent colors were different according to the genera but iridescent marine bacteria were widely distributed. However, a majority of strains were isolated from rocky shores and, in particular, red seaweed surfaces and mollusks. The data from the study suggested that isolates with iridescent properties were well conserved in stressful environments such as the coastal shoreline. This origin may provide an insight into the ecological and biological functions of iridescence.     

Effects of juvenile non-indigenous Carcinus maenas on the growth and condition of juvenile Cancer irroratus

The Atlantic rock crab, Cancer irroratus, is a commercially fished species and a critical prey item for the American lobster, Homarus americanus, in Atlantic Canada. The recent invasion of European green crab, Carcinus maenas, may have significant effects on the growth and condition of native C. irroratus, because both species overlap spatially and temporally and have similar habitat and dietary requirements. To examine such potential effects, we measured the growth of juvenile C. irroratus in the presence of juvenile C. maenas over a period of 4 months (growing season), under the following species combinations: (1) one C. irroratus (10-25 mm CW); (2) two C. irroratus (10-25 mm CW); (3) one C. irroratus (10-25 mm CW) and one C. maenas (10-15 mm CW). Morphological measurements included pre- and post-molt carapace width, chela height, abdomen width (mm), weight (g), and estimates of molt increment (%) and intermolt duration (days). Analysis of the hepatopancreas for % lipid content at the end of the experiment provided an estimate of physiological condition. The effect of the presence of C. maenas on the growth of C. irroratus shifted from negative to positive, when C. irroratus reached CW of 19-22 mm and gained a presumably significant size advantage over C. maenas. The positive effect resulted from increased energy intake through crab consumption. In the absence of crab consumption, the presence of a second crab (conspecific or C. maenas) had no effect on growth. C. irroratus consumed crabs more frequently when the second individual was a green crab than a conspecific. Consumption of C. maenas had a pronounced effect on the growth rate of C. irroratus, resulting in shorter intermolt periods and larger percent molt increments than in the presence of a conspecific. Therefore, the presence of juvenile C. maenas does not appear to have a prolonged negative effect on the growth of C. irroratus; rather, it may provide an additional food item as rock crabs grow, as long as encounters between the two species occur at high enough rates.