Survival of Campylobacter jejuni in Water: Effect of Grazing by the Freshwater Crustacean Daphnia carinata (Cladocera)

Environmental studies of the human-pathogenic bacterium Campylobacter jejuni have focused on linking distributions with potential sources. However, in aquatic ecosystems, the abundance of C. jejuni may also be regulated by predation. We examine the potential for grazing by the freshwater planktonic crustacean Daphnia carinata to reduce the survival of C. jejuni. We use a system for measuring grazing and clearance rates of D. carinata on bacteria and demonstrate that D. carinata can graze C. jejuni cells at a rate of 7% individual⁻¹ h⁻¹ under simulated natural conditions in the presence of an algal food source. We show that passage of C. jejuni through the Daphnia gut and incorporation into fecal material effectively reduces survival of C. jejuni. This is the first evidence to suggest that grazing by planktonic organisms can reduce the abundance of C. jejuni in natural waters. Biomanipulation of planktonic food webs to enhance Daphnia densities offers potential for reducing microbial pathogen densities in drinking water reservoirs and recreational water bodies, thereby reducing the risk of contracting water-borne disease.     

Other environmental factors affect the size of crests developed by Daphnia carinata (Cladocera: Daphniidae) in response to notonectid predators

SUMMARY. 1. Crest size and body size was measured in Daphnia carinata King sampled from field enclosures in which environmental conditions and notonectid predator ( Anisops Hyperion Kirkaldy) levels had been manipulated, and from control treatments which had predator levels and other environmental factors equivalent to those in the natural environment.
2. Individuals of D. carinata developed larger crests in treatments with predators than in treatments without predators.
3. The environmentally manipulated treatment was considered to have lower levels of food but was also likely to have undergone other changes in physicochemistry and resource variability that may have affected crest size. The effect of the manipulation cannot be confirmed, therefore, as the result of variation in food levels. Animals in the treatments considered to have lower amounts of food had smaller crests than in the control treatment irrespective of whether predators were present. The effects of environmental manipulation and predators were additive.
4. Enclosures without predators, but in which water from the natural, predator-rich, environment was exchanged through the enclosure walls, showed no evidence of a chemical induction effect. Chemical induction effects may have been countered by a reduced food supply resulting from the higher D. carinata population densities that developed in these enclosures.
5. This work provides a second example of an alteration of the morphological response of daphnid prey to predators by environmental factors, probably food, and suggests this phenomenon may be general in cladoceran species displaying predator-induced changes in morphology.     

Mortality and growth responses of Daphnia carinata to increases in temperature and salinity

1.  Daphnia carinata King, a freshwater cladoceran, occurs in Lake Waihola, a tidal lake with seasonal fluctuations in temperature (4–21 °C), and salinity (30–2000 mg L^–1 Cl). We hypothesise that these fluctuations influence the seasonal changes in D. carinata abundance.
2. To test this hypothesis, adults and juveniles were exposed to combinations of temperature and salinity. We measured mortality of adults and juveniles, growth of juveniles over 20 days, and their age at first reproduction.
3. The salinity tolerance of adult D. carinata was temperature-dependent. Juveniles were more sensitive to salinity, but more tolerant of temperature increases.
4. Growth rates were higher at elevated temperatures, but reduced by elevated salinities. The onset of reproduction was earlier at elevated temperatures.
5. We conclude that seasonal changes in temperature and salinity contribute to seasonal population changes of D. carinata .     

上海滴水湖两种枝角类牧食对浮游植物群落影响的模拟研究

于2012年9月通过浮游动物添加实验,研究了大型溞(Daphnia magna)和隆腺溞(Daphnia carinata)对上海滴水湖浮游植物群落结构的影响.结果表明,实验组氨氮和可溶性活性磷浓度与空白组相比显著增加,大型溞组浮游植物密度较空白组降低了70.3％,隆腺溞组浮游植物密度较空白组降低了80.0％,叶绿素a浓度分别下降了80.4％和75.2％,叶绿素a与氨氮、可溶性活性磷呈显著相关性.浮游植物的群落结构较空白组也发生较大变化,硅藻、蓝藻和绿藻密度比例明显降低,隐藻、裸藻和甲藻在实验结束时没有检出,说明大型溞和隆线溞能有效控制水体中浮游植物密度,并能够对浮游植物群落结构产生影响.同时,附着藻类密度较空白组也有明显减少,说明大型溞和隆线溞对附着藻类也有一定的牧食作用.     

三种甲壳纲浮游动物与萼花臂尾轮虫的种间关系

为研究隆线溞(Daphnia carinata)、锯缘真剑水蚤(Eucyclops serrulatus)及中华薄壳介(Dolerocypris sinensis)3种甲壳纲动物种群与萼花臂尾轮虫(Brachionus calyciflorus)种群的相互关系,作者分别将其以不同接种密度与萼花臂尾轮虫(接种密度为350 ind/L)进行混合培养。轮虫种群密度增长率自混合培养后的第3或4天开始下降,且随着甲壳纲动物接种密度的增加轮虫种群受到的抑制作用增大。当轮虫接种密度为350 ind/L时,3种甲壳纲动物的种群变化有所不同:(1)当隆线溞起始密度低于150 ind/L时,其种群生长受到轮虫的明显抑制;而当隆线溞起始密度达到350 ind/L时,其在混合培养与单独培养下的种群变化基本一致,这表明轮虫对隆线溞种群生长无抑制作用。(2)与隆线溞不同,锯缘真剑水蚤和中华薄壳介在混合培养下的种群数量持续增长。但与锯缘真剑水蚤相比,中华薄壳介在混合培养下的种群增长速度较慢。     

Grazing of two toxic Planktothrix species by Daphnia pulicaria: potential for bloom control and transfer of microcystins

The role of zooplankton in the control of cyanobacterial bloomsand the transfer of cyanotoxins to higher trophic levels areof great importance to the management of water resources. Manystudies have focused on the cyanobacterium Microcystis, butfew have examined the interactions between zooplankton and filamentouscyanobacteria. In this study, we provide experimental evidencefor the potential grazing of two toxic strains of filamentouscyanobacteria, Planktothrix rubescens and P. agardhii, by Daphniapulicaria, and for transfer of toxins in the planktonic foodchain. We determined clearance rates (CRs) by adult and juvenileD. pulicaria of the two Planktothrix strains, Scenedesmus acutusand a mixture of S. acutus cells with P. rubescens culture filtrate.Filament lengths were analyzed, and microcystin (MCY) presencein Daphnia was assessed using the Protein Phosphatase-2A (PP-2A)Inhibition Assay. The two Planktothrix strains were equallygrazed by D. pulicaria, but at lower CRs than S. acutus. Potentialanti-grazer toxins in P. rubescens filtrate did not inhibitDaphnia grazing. Small P. rubescens (<100 µm) filamentswere preferentially grazed by adult D. pulicaria, suggestingtheir limited ability to control a Planktothrix population duringa bloom. Large quantities of MCYs were found in unstarved Daphniapreviously exposed to Planktothrix, whereas quantities weresignificantly smaller in individuals starved for 24 h beforepreservation. This indicated a potential for transfer of toxinsin the food chain by Daphnia, especially immediately after ingestionof toxic cyanobacteria.     

武汉东湖透明溞和隆线溞一亚种的种群变动和生产量

本文报道了1980—1982年东湖二种优势枝角类:透明溞(Daphnia hyalina)和隆线溞一亚种(Daphnia carinata ssp.)的种群变动和生产量的研究结果。分析了影响种群变动和生产量的生态因子。尽管这二种枝角类在一年中有一段共存时间,但高峰期从来是错开的。一般来说,透明溞的高峰期出现在春季或冬季;而隆线溞一亚种则出现在夏季或秋季源沓ㄋ蘑蛘纠此?1980—1982年透明溞的种群密度的年平均值为28.43(17.59—38.50)个/升;隆线溞一亚种为4.64(3.79—5.31)个/升。种群的瞬时出生率(b),;通常大于瞬时增长率(r);瞬时死亡率(d)与b的季节变动在大多数情况下是一致的。透明溞的年平均生物量为0.218(0.163—0.315)毫克干重/升,生产量为9.181(4.877—13.428)毫克干重/升·年,P/B系数为42.38(27.76—56.70)。隆线溞一亚种的年平均生物量为0.046(0.039—0.060)毫克干重/升,生产量为3.417(2.752—3.829)毫克干重/升·年,P/B系数为77.04(61.24—99.74)。       

Long-term changes in crustacean zooplankton and water quality in a shallow, eutrophic Chinese lake densely stocked with fish

From surveys made in 1962–1963, 1973–1974, 1979–1996 at two Stations in Lake Donghu, a shallow eutrophic water body near Wuhan, P. R. China, the authors, derive long-term changes in species composition, standing crop and body-size of planktonic crustaceans. The species number decreased from the 1960s to the 1990s. The cladocerans dropped from 46 (1960s) to 26 (1980s) to 13 (1990s); the copepods decreased from 14 (1960s) to 10 (1980s) to 7 (1990s). From the mid-1980s on, the dominant crustaceans also changed: Daphnia hyalina and D. carinata ssp. were replaced by Moina micrura and Diaphanosoma brachyurum at Stations 1 and 2, respectively; Cyclops vicinus replaced Mesocyclops leuckarti.Densities and biomass of Cladocera decreased markedly after 1987. Annual average densities and biomass of cladocerans were statistically differences between 1962–1986 and 1987–1996 (P > 0.01). Annual average densities of Daphnia (Station 1 + Station 2) were negatively correlated with fish yield .Since the 1980s, annual average body length of Cladocera and Calanoida decreased, while annual average body length of Cyclopoida increased. In the same years, average body length of copepods was lower during May–October than during January–April and November–December.A 12-yr data analysis showed annual average concentration of chlorophyll^-a (Chl^-a) to be negatively correlated with annual average density of Daphnia, whilst lake transparency was positively correlated with annual average densities of Daphnia. The results imply that, since Daphnia feeds efficiently on phytoplankton, it could decrease concentration of Chl^-a, and enhance water transparency.     

Carbon load in aquatic ecosystems affects the diversity and biomass of water biofilm consortia and the persistence of the pathogen Campylobacter jejuni within them

The influence of carbon load on autochthonous water microflora population distribution and diversity, and on the persistence of Campylobacter jejuni, was examined with a two-stage aquatic biofilm model. Serine was chosen since it is a carbon source utilised by C. jejuni and concentrations were chosen to reflect upper limits of amino acid load reported in surface water. The total viable count of the autochthonous biofilm microflora increased with increasing serine concentration (10-fold and 20-fold with 5 nM and 5 μM serine, respectively), as did the counts of the microflora in the planktonic phase. Differences in biofilm species distribution as determined by culture were small with changes in temperature or the addition of serine; but was markedly affected by serine as determined by light microscopy, becoming more luxuriant and dominated by long filamentous cells. The addition of serine to the water significantly and progressively reduced the persistence of C. jejuni, which decreased by 25% and 50% with serine concentrations of 5 nM and 5 μM respectively. We have demonstrated that carbon load affects the species diversity and density of both the planktonic and biofilm phase of aquatic autochthonous microflora. Although the survival of C. jejuni in water in a culturable form was sufficient for this to be an important vehicle for its transmission, carbon load significantly influenced survival; an increase in serine concentration significantly reduced survival.     

若干种枝角类卵鞍表面亚显微结构的比较研究

以扫描电镜对发头裸腹溞、蚤状溞、隆线溞指名亚种以及隆线溞东湖一亚种的卵鞍分别进行了比较研究,发现卵鞍表面的亚显微结构有明显的差异,但亚种之间差异较小。隆线溞指名亚种与隆线溞东湖一亚种二者的卵鞍在光学显微镜下看不出有什么差异,然而亚显微结构却互不相同。这就为东湖一新亚种的确定提供了一项可靠的依据,同时也进一步证实卵鞍的超微结构确可作为枝角类分类的表征。     

An investigation of allelopathic effects of Daphnia

• 1 The hypothesis is tested that large daphnids are able to suppress their own and other species' feeding and reproduction by means of excreting an inhibitory chemical (or chemicals).
• 2 In laboratory experiments with an Australian species, Daphnia carinata, water preconditioned with 3–67 daphnids 1-^?1 for 30 h had the effect of reducing feeding rates of D. carinata and D. lumholtzi provided with Selenastrum capricornutum.
• 3 For the two Daphnia species, there were highly significant negative correlations between feeding rate and the preconditioning density of D. carinata.
• 4 Water preconditioned with 20–30 daphnids 1-^?1 for 1–2 weeks reduced the grazing rates of Daphnia, Moina, and Diaphanosoma 2–3-fold.
• 5 Moina kept in such water for 2 days stopped feeding. Conditioned water kept for 3 days without animals still inhibited grazing by Moina. Hearing to 100°C removed the inhibitory effect.
• 6 Given excess food, and in non-renewed water, a gradient of D. carinata densities developed a strong negative correlation between clutch size and daphnid density after a 6-day time lag. This result may help explain the direct density-dependent regulation of cladoceran reproduction observed earlier in a subtropical lake.

     

Survival of Campylobacter jejuni in waterborne protozoa

The failure to reduce the Campylobacter contamination of intensively reared poultry may be partially due to Campylobacter resisting disinfection in water after their internalization by waterborne protozoa. Campylobacter jejuni and a variety of waterborne protozoa, including ciliates, flagellates, and alveolates, were detected in the drinking water of intensively reared poultry by a combination of culture and molecular techniques. An in vitro assay showed that C. jejuni remained viable when internalized by Tetrahymena pyriformis and Acanthamoeba castellanii for significantly longer (up to 36 h) than when they were in purely a planktonic state. The internalized Campylobacter were also significantly more resistant to disinfection than planktonic organisms. Collectively, our results strongly suggest that protozoa in broiler drinking water systems can delay the decline of Campylobacter viability and increase Campylobacter disinfection resistance, thus increasing the potential of Campylobacter to colonize broilers.     

基于16S rDNA和COⅠ基因序列探讨四种溞类的系统关系和分类地位

为了检验已记录的4个溞属种类（(隆线溞Daphnia carinata、透明溞D. hyalina、蚤状溞D. pulex和大型溞D. magna）的系统分类, 用试剂盒法分别提取4种溞类的基因组DNA。利用特异性引物, 通过PCR扩增了4种溞属种类的16S rDNA和COⅠ基因部分序列, 并与来自GenBank中每个种类相似度较高的同种序列进行分析。结果表明, 基于16S rDNA和COⅠ基因, 4个溞属种类的平均种间相似度分别为85.56%和80.67%, 碱基中A+T含量均明显高于G+C含量。基于COⅠ基因, 巢湖隆线溞与来自GenBank上的拟同形溞（(Daphnia similoides AB549199）)相似度为99%, 分歧度为0.4%0.9%。就16S rDNA和COⅠ基因而言, 巢湖透明溞更接近于盔形溞（Daphnia galeata）。基于16S rDNA和COⅠ基因构建的NJ树和贝叶斯树, 两者的主要分枝基本一致。结果暗示, 巢湖隆线溞与拟同形溞为一个种, 透明溞应属于盔形溞。由于缺乏核基因的研究, 有关巢湖溞属的分类地位还需进行更深入的探讨。       

Top-down control of natural phyto- and bacterioplankton prey communities by Daphnia magna and by the natural zooplankton community of the hypertrophic Lake Blankaart

Biomanipulation, through the reduction of fish abundance resulting in an increase of large filter feeders and a stronger top-down control on algae, is commonly used as a lake restoration tool in eutrophic lakes. However, cyanobacteria, often found in eutrophic ponds, can influence the grazing capacity of filter feeding zooplankton. We performed grazing experiments in hypertrophic Lake Blankaart during two consecutive summers (1998, with and 1999, without cyanobacteria) to elucidate the influence of cyanobacteria on the grazing pressure of zooplankton communities. We compared the grazing pressure of the natural macrozooplankton community (mainly small to medium-sized cladocerans and copepods) with that of large Daphnia magna on the natural bacterioplankton and phytoplankton prey communities. Our results showed that in the absence of cyanobacteria, Daphnia magna grazing pressure on bacteria was higher compared to the grazing pressure of the natural zooplankton community. However, Daphnia grazing rates on phytoplankton were not significantly different compared to the grazing rates of the natural zooplankton community. When cyanobacteria were abundant, grazing pressure of Daphnia magnaseemed to be inhibited, and the grazing pressure on bacteria and phytoplankton was similar to that of the natural macrozooplankton community. Our results suggest that biomanipulation may not always result in a more effective top-down control of the algal biomass.     

Impact of zooplankton grazing on the excystation, viability, and infectivity of the protozoan pathogens Cryptosporidium parvum and Giardia lamblia

Very little is known about the ability of the zooplankton grazer Daphnia pulicaria to reduce populations of Giardia lamblia cysts and Cryptosporidium parvum oocysts in surface waters. The potential for D. pulicaria to act as a biological filter of C. parvum and G. lamblia was tested under three grazing pressures (one, two, or four D. pulicaria grazers per 66 ml). (Oo)cysts (1 x 10(4) per 66 ml) were added to each grazing bottle along with the algal food Selenastrum capricornutum (6.6 x 10(4) cells per 66 ml) to stimulate normal grazing. Bottles were rotated (2 rpm) to prevent settling of (oo)cysts and algae for 24 h (a light:dark cycle of 16 h:8 h) at 20 degrees C. The impact of D. pulicaria grazing on (oo)cysts was assessed by (i) (oo)cyst clearance rates, (ii) (oo)cyst viability, (iii) (oo)cyst excystation, and (iv) oocyst infectivity in cell culture. Two D. pulicaria grazers significantly decreased the total number of C. parvum oocysts by 52% and G. lamblia cysts by 44%. Furthermore, two D. pulicaria grazers significantly decreased C. parvum excystation and infectivity by 5% and 87%, respectively. Two D. pulicaria grazers significantly decreased the viability of G. lamblia cysts by 52%, but analysis of G. lamblia excystation was confounded by observed mechanical disruption of the cysts after grazing. No mechanical disruption of the C. parvum oocysts was observed, presumably due to their smaller size. The data provide strong evidence that zooplankton grazers have the potential to substantially decrease the population of infectious C. parvum and G. lamblia in freshwater ecosystems.     

Daphnia predation on the amphibian chytrid fungus and its impacts on disease risk in tadpoles

Direct predation upon parasites has the potential to reduce infection in host populations. For example, the fungal parasite of amphibians, Batrachochytrium dendrobatidis (Bd), is commonly transmitted through a free‐swimming zoospore stage that may be vulnerable to predation. Potential predators of Bd include freshwater zooplankton that graze on organisms in the water column. We tested the ability of two species of freshwater crustacean (Daphnia magna and D. dentifera) to consume Bd and to reduce Bd density in water and infection in tadpoles. In a series of laboratory experiments, we allowed Daphnia to graze in water containing Bd while manipulating Daphnia densities, Daphnia species identity, grazing periods and concentrations of suspended algae (Ankistrodesmus falcatus). We then exposed tadpoles to the grazed water. We found that high densities of D. magna reduced the amount of Bd detected in water, leading to a reduction in the proportion of tadpoles that became infected. Daphnia dentifera, a smaller species of Daphnia, also reduced Bd in water samples, but did not have an effect on tadpole infection. We also found that algae affected Bd in complex ways. When Daphnia were absent, less Bd was detected in water and tadpole samples when concentrations of algae were higher, indicating a direct negative effect of algae on Bd. When Daphnia were present, however, the amount of Bd detected in water samples showed the opposite trend, with less Bd when densities of algae were lower. Our results indicate that Daphnia can reduce Bd levels in water and infection in tadpoles, but these effects vary with species, algal concentration, and Daphnia density. Therefore, the ability of predators to consume parasites and reduce infection is likely to vary depending on ecological context.     

Herbivorous animals can mitigate unfavourable ratios of energy and material supplies by enhancing nutrient recycling

Recent evidence shows that high supply ratios of light and nutrients limit planktonic herbivore growth by lowering the nutritional quality of algae. Over longer time scales, however, grazers may ameliorate this effect by their impact on nutrient cycling. We examine this possibility using two species of the herbivorous zooplankter Daphnia and its algal prey under different light intensities and low phosphorus supply in laboratory microcosms. At high light, Daphnia biomass was limited for a substantial period because of low P content of algal cells. However, a gradual increase in Daphnia density eventually improved food quality through grazing and nutrient cycling and via a novel process involving positive density dependence. Competitive exclusion of one of the two Daphnia species occurred under low light but not under high light when algae were nutritionally unsuitable. Such stoichiometrically mediated interactions among herbivorous animals may represent important mechanisms that affect community structure and material flows in ecosystems.     

Evidence of phosphorus-limited individual and population growth of Daphnia in a Canadian Shield lake

We performed bag experiments in a Canadian Shield lake with generally high seston (suspended food particles mainly composed of algae) carbon (C):phosphorus (P) ratios, and investigated the responses of individual and population growth of herbivorous Daphnia dentifera on their abundance with (+P) and without (−P) a phosphorus enrichment to lake water. In both treatments, increased abundance of D. dentifera reduced seston C concentration and was accompanied by decreases in population and individual growth rates. However, P-enrichment increased seston P concentration and then reduced seston C:P ratio from 400–700 to ca 100 (by atoms). As a result, both individual and population growth rates were significantly higher in the +P treatment at all animal abundances even though seston C concentrations were similar between the treatments. The magnitude of the growth enhancement by the P-enrichment was independent of animal abundance. Stepwise regression analyses revealed that 71 and 90% of the variance in the population and individual growth rates, respectively, were explained by seston C and P concentrations, and that the contribution of the seston P concentration was roughly the same as that of seston C. Such joint effects of seston C and P indicate that food quality (P content) as well as food quantity (C concentration) can influence Daphnia not only at the level of individual growth but also at the level of population dynamics in P-limited lakes. Our results thus strongly corroborate the hypothesis that the population development of a key herbivore Daphnia in P-limited Canadian Shield lakes is inhibited by the direct effects of P-limited food on individual growth, which weaken the strength of trophic cascading interactions starting from piscivorous fish through planktivorous fish and zooplankton to algae.     

温度及pH对隆线溞生长和生殖的影响

研究了隆线溞(Daphnia carinata)在不同温度(15、20、25、30℃)和pH(6、7、8、9、10)影响下的内禀增长率(rm)、周限增长率(λ)、平均世代周期(T)、净生殖率(R0)、平均寿命、第一怀卵日、平均生殖间隔期和总生殖量.结果表明,温度对隆线溞的生命参数有显著影响,30℃组的内禀增长率(rm)...     

Impact of livestock grazing on abundance of Miridae and Reduviidae (Hemiptera) in crested wheatgrass pastures

Miridae of the genera Labops and Irbisia, collectively referred to as "black grass bugs," can cause significant damage to wheatgrasses (Poaceae) of several genera on western North American rangeland. Another mirid in the same area, Capsus cinctus (Kolenati), causes damage to bluegrass (Poa spp.). Previous studies suggest that grazing management may reduce mirid populations on rangeland by eliminating preferred oviposition sites and reducing accumulations of litter that provide diurnal refuges for nymphs. We tested the hypothesis that grazing reduces mirid populations, along with those of Reduviidae, during a controlled grazing experiment. Densities of mirids and reduviids declined with increasing intensity of grazing, even though grazing occurred after the peak of mirid abundance each year. This suggests that declines in hemipteran densities resulted from grazing that occurred during previous years, perhaps because the most heavily grazed plots had the least plant litter. The results further confirm that grazing has the potential to control black grass bug populations, although the benefits could be potentially offset by negative impacts on beneficial insects such as reduviids.