Carbaryl-induced behavioural and reproductive abnormalities in the earthworm Metaphire posthuma: a sensitive model

Carbaryl, an N-methyl carbamate insecticide, is used in India to control foliar insects, but, due to soil contamination, it also adversely affects non-target organisms such as earthworms. This paper deals with the toxic effects of carbaryl on the behavioural and reproductive profiles of the earthworm, Metaphire posthuma. Locomotion and geotaxis were significantly affected, even after a 20-minute exposure to 0.125ppm carbaryl. The hatching of cocoons was altered at 0.5ppm, whereas cocoon production was retarded even at 0.125ppm carbaryl. No cocoon production was observed at 2.0ppm carbaryl. Sperm head abnormalities were reported even at the lowest test concentration of 0.125ppm. Wavy head abnormalities were observed at 0.125ppm carbaryl, whereas at 0.25ppm and 0.5ppm, the sperm heads became amorphous and the head nucleus was turned into granules deposited within the wavy head. It is concluded that the earthworm could be used as an ecosystem model for the initial toxicity testing of environmental pollutants.     

Cloning, expression, and characterization of cadmium-induced metallothionein-2 from the earthworms Metaphire posthuma and Polypheretima elongata

In this study we report the sequences of MT-2 cDNA from two species of Megascoleidae earthworms, Metaphire posthuma and Polypheretima elongata, by mRNA differential display after exposure of the organisms to cadmium. Complementary (c)DNA was verified as the MT-2 gene by the characteristics of its predicted translation product, namely a high cysteine content, conserved CXC motifs, and a molecular weight of around 8 kDa. Amino acid sequence alignment revealed a conserved TKCCG in the cloned MT-2 of both megascolecid earthworms instead of the corresponding conserved TQCCG found in lumbricid earthworms. The cDNAs corresponding to the two megascolecid MT-2 genes were expressed, and the MT-2 proteins were purified for biochemical characterization. The binding of Cu2+ exhibited monophasic kinetics and those of Zn2+ and Cd2+ biphasic kinetics. The proteins bound more tightly to Cd2+ than to Zn2+ and more tightly still to Cu2+. Zn-MT and apo-MT were the most effective at scavenging free radicals, followed by Cd-MT. In conclusion, MT-2s from M. posthuma and P. elongata showed unique sequence features compared to those of lumbricid earthworms. These earthworms could be used to evaluate heavy-metal pollution in soil due to the inducible MT-2 by cadmium exposure.     

Molecular systematics and phylogeography of the gigantic earthworms of the Metaphire formosae species group (Clitellata, Megascolecidae)

The earthworms of the Metaphire formosae species group distributed in Taiwan are members of the Pheretima complex within the Megascolecidae. In this study, the systematics and phylogeography of this species group were investigated using DNA sequences of mitochondrial cytochrome c oxidase subunit I (COI), 16S ribosomal (r)RNA, and NADH dehydrogenase subunit 1 (ND1). The results indicated that the 13 taxa of the M. formosae species group form a clade, including a cryptic species discovered in this study. In addition, Metaphire hengchunensis (James, S.W., Shih, H.-T., Chang, H.-W., 2005. Seven new species of Amynthas (Clitellata: Megascolecidae) and new earthworm records from Taiwan. J. Nat. Hist. 39, 1007-1028) should be regarded as a subspecies of Metaphire paiwanna Tsai, C.-F., Shen, H.-P., Tsai, S.-C., 2000a. Native and exotic species of terrestrial earthworm (Oligochaeta) in Taiwan with reference to Northeast Asia. Zool. Stud. 39, 285-294, Tsai, C.-F., Tsai, S.-C., Liaw, G.-J., 2000b. Two new species of pontandric pheretimoid earthworms belonging to the genus Metaphire (Megascolecidae: Oligochaeta) from Taiwan. J. Nat. Hist. 34, 1731-1741, and Metaphire bununa glareosa Tsai et al. 2000 should be elevated to specific status. Phylogeographical inferences showed that allopatric speciation occurred in this species group during the rapid uplift of the main island of Taiwan between 5.0 and 2.5 million years ago. Our analysis exposes non-monophyly within each of the genera Amynthas and Metaphire, and more generally within the Pheretima complex. Further revisions of this speciose complex are urgently needed.     

An Assessment of the Impact of Chromium-Amended Sediment on a Marine Nematode Assemblage Using Microcosm Bioassays

Microcosms were used to assess the impact of chromium on free-living marine nematodes. Nematodes were exposed to three chromium concentrations (500 ppm (dm, dry mass), 800 ppm (dm) and 1,300 ppm (dm)), and effects were examined after 4 weeks. Results showed significant differences between univariate measures of control nematodes and those from medium- and high-chromium microcosms. Most, decreased significantly with increasing level of chromium contamination. The medium-chromium treatment seems to be the minimal concentration that could has a negative effect on nematodes. Results from multivariate analyses demonstrated that responses of nematode species to chromium treatments were varied: Leptonemella aphanothecae was eliminated at all doses tested and seemed to be intolerant species to chromium; Daptonema normandicum and Sabatieria longisetosa which significantly increased at 500 ppm chromium (dm) appeared to be “opportunistic” species at this dose whereas the two Bathylaimus species (Bathylaimus capacosus and Bathylaimus tenuicaudatus) which increased at all the doses tested seemed to be “chromium resistant”. As we think such “opportunistic” and “resistant” species could be used as sensitive indicators of unsafe marine food. The use of microcosms has allowed the effects of the chromium on nematodes to be assessed individually, which was not possible in the field.     

Bacterial removal of chromium (VI) and (III) in a continuous system

The capacity of Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans to reduce different concentrations of hexavalent chromium in shake flask cultures has been investigated. A. ferrooxidans reduces 100% of chromium (VI) at concentrations of 1, 2.5 and 5 ppm, but in the presence of 10 ppm only 42.9% of chromium (VI) was reduced after 11 days of incubation. A. thiooxidans showed a lower capacity to reduce this ion and total reduction of chromium (VI) was only obtained for concentrations of 1 and 2.5 ppm, whereas 64.7% and 30.5% was reached for 5 and 10 ppm, respectively, after 11 days. A continuous flow mode system was subsequently investigated, in which A. thiooxidans was immobilized on elemental sulphur and the acidic medium obtained was employed to solubilize chromium (III) and to reduce chromium (VI) present in a real electroplating waste [30% of chromium (III) and 0.1% of chromium (VI)]. The system enabled the reduction of 92.7% of hexavalent chromium and represents a promising way to treat this type of waste in the industry.     

Gas Chromatography- Mass Spectrometry Based Metabolomic Approach for Optimization and Toxicity Evaluation of Earthworm Sub-Lethal Responses to Carbofuran

Despite recent advances in understanding mechanism of toxicity, the development of biomarkers (biochemicals that vary significantly with exposure to chemicals) for pesticides and environmental contaminants exposure is still a challenging task. Carbofuran is one of the most commonly used pesticides in agriculture and said to be most toxic carbamate pesticide. It is necessary to identify the biochemicals that can vary significantly after carbofuran exposure on earthworms which will help to assess the soil ecotoxicity. Initially, we have optimized the extraction conditions which are suitable for high-throughput gas chromatography mass spectrometry (GC-MS) based metabolomics for the tissue of earthworm, Metaphire posthuma. Upon evaluation of five different extraction solvent systems, 80% methanol was found to have good extraction efficiency based on the yields of metabolites, multivariate analysis, total number of peaks and reproducibility of metabolites. Later the toxicity evaluation was performed to characterize the tissue specific metabolomic perturbation of earthworm, Metaphire posthuma after exposure to carbofuran at three different concentration levels (0.15, 0.3 and 0.6 mg/kg of soil). Seventeen metabolites, contributing to the best classification performance of highest dose dependent carbofuran exposed earthworms from healthy controls were identified. This study suggests that GC-MS based metabolomic approach was precise and sensitive to measure the earthworm responses to carbofuran exposure in soil, and can be used as a promising tool for environmental eco-toxicological studies.     

Winter hatching in New England populations of invasive pheretimoid earthworms <Emphasis Type="Italic">Amynthas agrestis</Emphasis> and <Emphasis Type="Italic">Amynthas tokioensis</Emphasis>: a limit on population growth,or aid in peripheral expansion?

Some Asian megascolecid earthworms, e.g. Amynthas agrestis and Amynthas tokioensis, are highly invasive and have only recently been reported from natural ecosystems in the northeastern USA. There, they are annual earthworms that survive the winter as cocoons (an egg in a tough covering). Hatching occurs in mid-April when temperatures rise consistently above 10 °C. In some years, winter temperatures also reach 10 °C during short warming episodes, but hatchlings then are likely to die when temperatures drop again. To test this hypothesis, soil was collected on 7 sampling dates during January–May 2016 at one site in the Champlain Valley, Vermont and extracted for the cocoons. Both hatched and unhatched cocoons were enumerated and identified to species by both size and sequencing of the mitochondrial cytochrome c oxidase I gene (COI). A regression model on the number of unhatched cocoons over time predicted that ~ a quarter (A. agrestis) and ~ a third (A. tokioensis) would hatch before mid-May; no hatchlings were observed. Thus, hatching during warming periods in winter seemingly resulted in high mortality. Such winter hatching, and loss, may increase with climate warming in the region because winter warming periods should become more common. Climate warming could therefore reduce ability of the invasive earthworms to persist in northern habitats. Conversely, rapid hatching when the soil warms in spring could have an ecological benefit by allowing opportunities to expand at the thermal edge of the range as the climate continues to warm.     

A comparative study on heavy metal biosorption characteristics of some algae

The biosorption of copper(II), nickel(II) and chromium(VI) from aqueous solutions on dried (Chlorella vulgaris, Scenedesmus obliquus and Synechocystis sp.) algae were tested under laboratory conditions as a function of pH, initial metal ion and biomass concentrations. Optimum adsorption pH values of copper(II), nickel(II) and chromium(VI) were determined as 5.0, 4.5 and 2.0. respectively, for all three algae. At the optimal conditions, metal ion uptake increased with initial metal ion concentration up to 250 mg l⁻¹. Experimental results also showed the influence of the alga concentration on the metal uptake for all the species. Both the Freundlich and Langmuir adsorption models were suitable for describing the short-term biosorption of copper(II), nickel(II) and chromium(VI) by all the algal species.     

Acute and chronic effects of an anionic surfactant on some freshwater tubificid species

We report the results of research on acute and chronic effects of linear alkylbenzensulfonate (LAS) on two tubificid species. 96 h LC₅₀ assay values were estimated at 10° for Limnodrilus hoffmeisteri and Branchiura sowerbyi exposed to different concentrations of LAS dissolved in water, both with and without sediment. The presence of sediments modified LAS toxicity and increased values: NOEC and LOEC resulted in values 2.5 times higher for Branchiura sowerbyi and 4–4.5 times for Limnodrilus hoffmeisteri, when the sediments were present. The chronic effects induced by a long exposure to LAS were evaluated for different stages of the biological cycle of Branchiura sowerbyi. Using concentrations between the NOEC and LOEC (0.5, 2.5, and 5 ppm), with control, we could observe that: 1) at 5 ppm the cocoons were laid precociously compared to controls, 2) in all treated series the number of cocoons was lower than in controls, 3) the mean number of oocytes per cocoon was lower for the worms submitted to LAS, compared to the control, 4) the period of embryonic development was similar for all used concentrations and for control, and 5) the number of degenerated cocoons was unchanged by the LAS treatment.     

Biomonitoring of chromium(VI) deposited in pulmonary tissues: Pilot studies of a magnetic resonance imaging technique in a post-mortem rodent model

The biomonitoring of individuals exposed to chromium(VI) by inhalation is often based on determinations of chromium in body fluids such as blood, plasma or urine, or on assessments of DNA damage in non-lung surrogate tissues such as peripheral blood lymphocytes. These techniques are of some use as biomarkers of internal exposure or biological effect, mainly in the case of soluble chromium(VI) compounds, but they provide at best only indirect information about chromium(VI) concentrations in the main target organ of interest – the lung. An urgent need exists for a non-invasive technique to permit the visualization and quantification of chromium(VI) in the lung of exposed humans. This study details the development of a lung imaging technique based on the detection of paramagnetic chromium using magnetic resonance imaging (MRI). The intracellular reductive conversion of chromium(VI) is a crucial bioactivation step in its carcinogenicity, and the MRI method described here relies on the conversion of non-paramagnetic (MRI ‘silent’) chromium(VI) to detectable paramagnetic species such as chromium(III). Initial studies with chromium(III) revealed that a range of 2.5–5 μg chromium(III) instilled in rat lung is considered to be the lower limit of detection of this method. It was possible to demonstrate the presence of 30 μg chromium(VI) in our post-mortem rat model. The ultimate objective of this work is to determine whether this technique has applicability to the biomonitoring of chromium(VI) inhalation exposures that result in internalized lung doses in human subjects.     

Biomonitoring of chromium(VI) deposited in pulmonary tissues: Pilot studies of a magnetic resonance imaging technique in a post-mortem rodent model

The biomonitoring of individuals exposed to chromium(VI) by inhalation is often based on determinations of chromium in body fluids such as blood, plasma or urine, or on assessments of DNA damage in non-lung surrogate tissues such as peripheral blood lymphocytes. These techniques are of some use as biomarkers of internal exposure or biological effect, mainly in the case of soluble chromium(VI) compounds, but they provide at best only indirect information about chromium(VI) concentrations in the main target organ of interest - the lung. An urgent need exists for a non-invasive technique to permit the visualization and quantification of chromium(VI) in the lung of exposed humans. This study details the development of a lung imaging technique based on the detection of paramagnetic chromium using magnetic resonance imaging (MRI). The intracellular reductive conversion of chromium(VI) is a crucial bioactivation step in its carcinogenicity, and the MRI method described here relies on the conversion of non-paramagnetic (MRI 'silent') chromium(VI) to detectable paramagnetic species such as chromium(III). Initial studies with chromium(III) revealed that a range of 2.5-5 μg chromium(III) instilled in rat lung is considered to be the lower limit of detection of this method. It was possible to demonstrate the presence of 30 μg chromium(VI) in our post-mortem rat model. The ultimate objective of this work is to determine whether this technique has applicability to the biomonitoring of chromium(VI) inhalation exposures that result in internalized lung doses in human subjects.     

Bioaccumulation of copper(II), lead(II) and chromium(VI) by growing Aspergillus niger

The effect of copper(II), lead(II) and chromium(VI) ions on the growth and bioaccumulation properties of Aspergillus niger was investigated as a function of initial pH and initial metal ion concentration. The optimum pH values for growth and metal ion accumulation were determined as 5.0, 4.5 and 3.5 for copper(II), lead(II) and chromium(VI) ions, respectively. Although all metal ion concentrations caused an inhibition effect on the growth of A. niger, it was capable of removing of copper(II) and lead(II) with a maximum specific uptake capacity of 15.6 and 34.4 mg g⁻¹ at 100 mg dm⁻³ initial copper(II) and lead(II) concentration, respectively. Growth of A. niger was highly effected by chromium(VI) ions and inhibited by 75 mg dm⁻³ initial chromium(VI) concentration since some inhibition occurred at lower concentrations.     

Effect of Added Heavy Metal Ions on Biotransformation and Biodegradation of 2-Chlorophenol and 3-Chlorobenzoate in Anaerobic Bacterial Consortia

The effect of added Cd(II), Cu(II), Cr(VI), or Hg(II) at 0.01 to 100 ppm on metabolism in anaerobic bacterial consortia which degrade 2-chlorophenol (2CP), 3-chlorobenzoate (3CB), phenol, and benzoate was examined. Three effects were observed, including extended acclimation periods (0.1 to 2.0 ppm), reduced dechlorination or biodegradation rates (0.1 to 2.0 ppm), and failure to dechlorinate or biodegrade the target compound (0.5 to 5.0 ppm). 3CB biodegradation was most sensitive to Cd(II) and Cr(VI). Biodegradation of benzoate and phenol was most sensitive to Cu(II) and Hg(II), respectively. Adding Cr(VI) at 0.01 ppm increased biodegradation rates of phenol (177%) and benzoate (169%), while Cd(II) and Cu(II) at 0.01 ppm enhanced biodegradation rates of benzoate (185%) and 2CP (168%), respectively. Interestingly, with Hg(II) at 1.0 to 2.0 ppm, 2CP and 3CB were biodegraded 133 to 154% faster than controls after an extended acclimation period, suggesting adaptation to Hg(II). Metal ions were added at inhibitory, but sublethal, concentrations to investigate effects on metabolic intermediates and end products. Phenol accumulated to concentrations higher than those in controls only in the 2CP consortium with added Cu(II) at 1.2 ppm but was subsequently degraded. There was no effect on benzoate, and little effect on acetate intermediates was observed. In most cases, methane yields were reduced by 23 to 97%. Thus, dehalogenation, aromatic degradation, and methanogenesis in these anaerobic consortia showed differential sensitivities to the heavy metal ions added. These data indicate that the presence of heavy metals can affect the outcome of anaerobic bioremediation of aromatic pollutants. In addition, a potential exists to use combinations of anaerobic bacterial species to bioremediate sites contaminated with both heavy metals and aromatic pollutants.     

Effect of hexavalent chromium on the activated sludge process and on the sludge protozoan community

The objectives of this study were the determination of chromium effects to the performance of an activated sludge unit and the investigation of the response of the activated sludge protozoan community to Cr(VI). Two bench scale activated sludge reactors were supplied with synthetic sewage containing Cr(VI), at concentrations from 1 up to 50 mg L(-1). Protozoan species were identified and were related to the system efficiency. Variations in the abundance and diversity of the protozoan species were observed under various chromium concentrations. High removal rates of organics and nutrients were observed after the acclimatization of the activated sludge, which were related to the initial chromium(VI) concentration. Chromium(VI) removal efficiency was high in all cases. The protistan community was affected by the influent chromium content. Dominance of sessile species was observed in the reactor receiving 5 mg L(-1) influent chromium, whereas co-dominance of sessile and carnivorous species was observed in the reactors receiving higher chromium concentrations.     

A Confocal microscopic study on colony morphology and sporulation of Bacillus sp.

Hexavalent chromium Cr(VI) is one of the dominant oxidation states of chromium that exist in the environment and is highly toxic to all forms of life. In the present study, we employ a confocal laser scanning microscope (CLSM) and investigate the effect of Cr(VI) on colony morphology of a Bacillus sp. isolated from soil exposed to tannery effluent. The colonies grown at chromium concentrations, control and 100 ppm are found to be opaque and beyond 200 ppm the colonies were translucent thus exhibiting phase variation. CLSM studies on colonies grown on control plates showed significant increase in height and in biovolume as a function of time whereas, the translucent colonies showed very little change in height and biovolume corresponding to the colony growth. Exopolymeric substance (EPS) content of translucent colonies was lesser than that of opaque colonies, indicating that EPS also plays a role in the observed phenomenon of phase variation. Studies on the effect of Cr(VI) on spore formation showed that Cr(VI) concentrations up to 100 ppm favored spore formation, while concentrations beyond 100 ppm showed a steady decline in spore formation.     

Cold hardiness and geographic distribution of earthworms (Oligochaeta,Lumbricidae, Moniligastridae)

Cold hardiness of 12 species and 2 subspecies of earthworms from Northern Eurasia was studied. Supercooling temperatures, the water content and the thresholds of tolerated temperatures of worms and their cocoons were determined. The threshold values varied within ?1…?35°C for worms and within ?1…?196°C for cocoons. Earthworms of 4 species and 2 subspecies survived freezing. Cocoons of all species except Eisenia fetida possessed a protective dehydration mechanism which prevented their freezing. During wintering at subzero temperatures, earthworms lost up to 20% of water, cocoons up to 37%. Species of the same life form can overwinter at different phases and have different cold hardiness values. On the whole, epigeic and epi-endogeic species (except for Eisenia fetida) were more resistant to cold than endogeic ones. The following preliminary classification of earthworms according to their tolerance to negative temperatures is proposed: (1) both onthogenetic phases are tolerant; (2) only cocoons are tolerant; (3) both onthogenetic phases are intolerant. The geographic distribution of all the studied species (except for Eisenia nordenskioldi nordenskioldi) is partially or completely limited by insufficient resistance of the worms to negative temperatures. A significant cold hardiness of cocoons of most species is nonadaptive, since the worms hatched from the eggs in spring die without having enough time to reach maturity and to lay cocoons before the onset of subzero temperatures. Only 3 species (Eisenia nordenskioldi nordenskioldi, Eisenia atlavinyteae, and Dendrobaena octaedra) can live in permafrost regions; this is the main reason for a drastically reduced diversity of earthworm assemblages in eastern Siberia except for its southern, mountain parts. In general, the reasons for the impoverishment lie in the modern climatic conditions correlated with the ecophysiological capacities of earthworms.     

Life cycle of the earthworm Octodrilus complanatus (Oligochaeta, Lumbricidae)

The earthworm Octodrilus complanatus (Dugès, 1828) is a large-sized species often found in grasslands and pastures, with a wide distribution area in Europe and North Africa. In order to determine the characteristics of its life cycle, earthworms of this species were cultured from hatching until they reached maturity. Development rates, cocoon production, and incubation period were recorded under controlled conditions. On average, individuals of O. complanatus reared in isolation from hatching reached maturity at day 150 and body weight at maturity ranged between 6 and 8 g. The cocoon incubation period was 66 days, and only one individual hatched from each cocoon, with a mean hatchability of 55%. Reproductive rate showed by O. complanatus after mating was of 52 cocoons per earthworm per year. The results showed that the life cycle of O. complanatus was characterized by both low growth rates and low fecundity. Specific features in life history traits of O. complanatus were compared with those of species of the genus Octodrilus and other related earthworms.     

Effects of Selected Chemical Pesticides on Agamermis unka (Nematoda: Mermithidae), a Parasite of the Brown Plant Hopper,Nilaparvata lugens

Selected commercial and technical grade pesticides were tested against the egg, preparasite and adult stages of Agamermis unka , a nematode parasite of the brown planthopper, Nilaparvata lugens . The commercial insecticide, diazinon (LC = 0.37 ppm), was most toxic to the 50 preparasites, followed by phenthoate (LC = 0.43 ppm), BPMC (LC = 0.44 ppm), IBP 50 50 (LC = 0.46 ppm), cartap hydrochloride (LC = 0.82 ppm) and buprofezin + isoprocarb 50 50 (LC = 1.11 ppm). The least toxic commercial pesticide tested was the fungicide, pencycuron 50 (LC = 2.19 ppm). Out of 12 technical grade insecticides tested, phenthoate, monocrotophos, 50 diazinon and carbofuran (LC = 0.37-0.46 ppm) were highly toxic to the preparasites, followed by 50 buprofezin, BPMC and fenitrothion (LC = 0.74-0.86 ppm). Fenthion, etofenprox, chlorpyrifos, 50 imidacloprid and MIPC (LC = 1.11-2.19 ppm) were the technical grade insecticides least toxic 50 to the preparasites. Most preparasites survived for up to 24 h at the low insecticide concentrations (0.63 and 0.31 ppm). Preparasites that were exposed to BPMC for 24 h at concentrations as high as 5.0 ppm and survived the treatments infected brown planthopper nymphs. Four selected insecticides-chlorpyrifos, BPMC, imidacloprid and carbofuran-had significant adverse effects on A. unka egg hatching. Eggs that were in the insecticide solution for 168 h fared poorly with imidacloprid having the best survival ( > 2% of the eggs hatching at 0.04 ppm). No eggs hatched from the other insecticide treatments. Three selected insecticides, BPMC, imidacloprid and chlorpyrifos, tested against adult A. unka showed that most adults survived the exposure to the insecticides between 0.31 and 2.5 ppm. At 5.0 ppm of BPMC or chlorpyrifos none of the adults survived, whereas with imidacloprid 70% of the adults survived. Egg deposition by the surviving adults was greatly reduced in those treated with the insecticides compared with those in the controls. Imidacloprid had some negative impact on the preparasites' ability to infect BPH nymphs, but it had the least detrimental effect of the insecticides tested on preparasite survival and on the eggs and adults of A. unka .     

体重、年龄和应激刺激对菲牛蛭繁殖性能的影响

采用单因素方差分析等方法探讨人工养殖条件下体重、年龄和环境应激对菲牛蛭繁殖性能的影响。结果表明,体重对卵茧大小和湿质量及幼蛭体长、体宽和湿体质量无显著影响(P>0.05),而亲蛭性腺系数、平均产茧数、孵化率和平均孵化数在各组间差异显著(P<0.05),其中体重10～15g组明显高于其他组(P<0.05);年龄对亲蛭平均产茧数、孵化率、平均孵化数和卵茧大小影响显著(P<0.05),其中2～3年龄亲本具有最高的平均产茧数、孵化率和平均孵化数,8月龄亲本所产卵茧个体明显大于其他年龄组(P<0.05),而卵茧湿质量及幼蛭体长、体宽和湿体质量在各年龄组间均无显著差异(P>0.05)。环境应激对亲蛭的平均产茧数、孵化率和平均孵化数影响显著(P<0.01),其中1～2次操作应激下的平均产茧数和孵化数最高,无操作刺激下的孵化率略高于1～2次操作刺激(P>0.05),而卵茧大小和湿质量及幼蛭体长、体宽和湿体质量在各应激处理组间无显著差异(P>0.05)。本研究获得了菲牛蛭繁殖的最佳体重和年龄,建议繁育生产中挑选10～15g和2～3年龄的个体作为亲本,期间进行1～2次操作刺激可提高其繁殖性能。     

The influence of body size on immediate reproductive success in Dugesia gonocephala (Tricladida,Paludicola)

The individual reproductive output of the stream-dwelling flatworm Dugesia gonocephala was investigated. Various measures of reproductive success were related to body size. (I) For the first 30 days in the laboratory small individuals produced no cocoons, individuals of intermediate size produced unfertilized cocoons and large individuals usually produced fertilized cocoons. (II) In individuals that produced a cocoon, no correlation was found between the number of cocoons produced in one month and body size. (III) Large individuals, however, produced larger cocoons. This was not due to the fact that unfertilized cocoons were smaller. (IV) Large cocoons tended to contain more young. (V) The average size of young hatching from large cocoons was larger. (VI) Large individuals produced their first cocoon soon after their arrival in the laboratory and seemed to have a higher chance of producing a fertilized first cocoon. (VII) A trade-off existed between producing many small versus few large young.