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1.
An extracellular lethal toxin produced by Aeromonas salmonicida was purified by fast-protein liquid ion-exchange chromatography. The toxin is composed of glycerophospholipid:cholesterol acyltransferase (GCAT) (molecular mass, 25 kilodaltons) aggregated with lipopolysaccharide (LPS), the GCAT/LPS complex having a molecular mass of about 2,000 kilodaltons, estimated by gel filtration chromatography. The toxin is lethal for Atlantic salmon (Salmo salar L.) at a concentration of 0.045 micrograms of protein per g of body weight. The toxin is a hemolysin (T-lysin, active on fish erythrocytes), leukocytolysin, and cytotoxin. Antiserum to the purified toxin neutralized the lethal toxicity of the crude extracellular toxins, indicating this toxin to be the major lethal factor produced by A. salmonicida. In the crude extracellular products, small amounts of free GCAT were also present. This has been purified, and its activities and properties have been compared with those of the GCAT/LPS complex. The presence of LPS did not influence the GCAT activity of the enzyme with egg yolk or phosphatidylcholine (lecithin) as a substrate, but the specific hemolytic activity and lethal toxicity was about eightfold higher in the complexed form. Furthermore, the free GCAT was more susceptible to proteolytic and heat inactivation than was the GCAT/LPS complex. Recombination of LPS (phenol extracted from extracellular products of A. salmonicida) with free GCAT enhanced the hemolytic activity, lethal toxicity, and heat stability of the latter but did not influence its lecithinase activity. In native polyacrylamide gel electrophoresis, the GCAT/LPS complex and the recombined GCAT-LPS both showed a high-molecular-mass band which did not enter the gel, while the free GCAT produced a single band with low molecular mass. In isoelectric focusing gels, the GCAT/LPS and recombined GCAT-LPS produced a nonfocusing smear with pIs from pI 5.0 to 5.8, while the free GCAT produced a single band with pI 4.3. These data show that free GCAT can combine with LPS to produce a high-molecular-mass complex with enhanced toxicity and heat stability compared with those of free GCAT, similar to the preexisting GCAT/LPS complex, and indicate that the LPS moiety of the toxin plays an active role in toxicity.  相似文献   

2.
Growth of fathead minnow yolk sac larvae was characterized from changes in dry weight and total content and concentrations of RNA, DNA and protein in fish exposed to a sublethal level of HCN (58 micrograms/l) and in age matched controls. Cyanide toxicosis occurred within 24 hr of exposure as evidenced by significant reductions in protein and RNA content and RNA/DNA ratio of larvae. After 96 hr exposure to HCN, larvae exhibited the same growth rate and protein synthetic rate (RNA/DNA) as control fish. HCN toxicosis and recovery is rapid and at least partial tolerance to HCN develops within 96 hr of exposure in larval fathead minnows.  相似文献   

3.
The present study is based on two year experiments; it analyses swimming behaviour and swimming speed at different developmental stages of the herring. Yolk sac larvae tend to sink rather rapidly during resting phases. At the end of the yolk sac stage the sinking rate is at its minimum; it increases again with increasing larva size. During the phase of yolk sac resorption, vertical movements become gradually transformed into horizontal ones. Generally, three types of swimming can be distinguished: (1) “Abrupt swimming” consisting of very short periods of fast swimming; normally each dart is connected with a change in swimming direction. (2) “Normal swimming” characterized by steady movements for several seconds; it results in a winding path. (3) “Slow meandering” representing search swimming, a slowly winding locomotion with a large amplitude of each winding but with very little net progression of the larva. Swimming speed varies considerably in all size groups. The 8 to 11 mm (total length) larvae reach a mean swimming velocity (undulation) of 1.0 to 1.2 cm per second. Swimming speed, measured as the straight line distance between start and end points of a single swimming phase, attains mean values of 7 to 8 mm/sec in 8 to 11 mm larvae, 10 to 11 mm/sec in 11 to 15 mm larvae, 21 to 25 mm/sec in 19 to 24 mm larvae, and 40 to 50 mm/sec in 32 to 40 mm larvae. Swimming activity changes during larval development and seems to be influenced by food supply. The total distance travelled in 5 minutes by the head of a yolk sac larva is 1 to 3 m. About 8 days after hatching, sinking rate is low and “search swimming” (slow meandering movements) prevails. The path covered by the head within 5 minutes is 0.8 to 1.5 m.  相似文献   

4.
A granulosis virus infecting the potato tuber moth, Phthorimaea operculella, has been identified, isolated and purified from diseased potato tuber moth larvae collected from potato fields in the highland basin Qa al-Boun of the Republic of Yemen. The granulosis virus was propagated by feeding potato tuber moth larvae with potatoes treated with a concentration of one granulosis virus-infected fourth instar larva to 5l of water (corresponding to an occlusion body (OB) concentration of 2 106 OB ml-1). Restriction enzyme analysis of viral DNA revealed that the isolated virus seemed to correspond to an isolate from the Lima region of Peru. A median LC 50 value of 7.3 104 OB ml-1 was calculated for a purified virus preparation. Different preparations of the granulosis virus were investigated for their persistence in the field on tubers and leaves. A purified virus preparation (PoGV) applied to potato tubers and exposed in the open had a half-life of 1.3 days. On leaves, the activity of granulosis virus spray deposits of an unpurified virus preparation (PoGV I) and of a glycerine-formulated preparation (PoGV II) also declined with exposure time. Mortalities of potato tuber moth larvae of 43% for PoGV I and 49% for PoGV II were recorded when first instar larvae were fed with potato leaves collected 2 days after treatment. After 8 days, 25.8% of the larvae had died from PoGV I treatment and 19.4% from PoGV II. Neither preparation displayed any effect 17 days after application.  相似文献   

5.
In presence of 7.5 microM of curcumin, no embryos or larva of zebrafish survived 3 days of incubation; however, coincubation with 144 microg/ml silymarin increased the survival rates of curcumin-treated embryos and larvae to about 70%. Moreover, in presence of 12.5 microM curcumin, all embryos died after 2 days of incubation; however, co-treatment with 144 microg/ml silymarin increased the survival rates of curcumin-treated embryos and larvae up to 60 and 50%, respectively. This protective effect was not found in the other phenolic compounds viz., ferulic acid, naringin, or crocin, tested. Finally, using a fluorescence microscope, accumulation of less curcumin has observed in the edema sac area of the larvae co-treated with curcumin and silymarin than in the larvae treated with curcumin only. The result suggests that the protective effects of silymarin may be due to a decreased accumulation of curcumin in the fish body.  相似文献   

6.
Early stages (neonate to 7- or 8-day-old larvae) of Spodoptera exigua and Pseudaletia unipuncta were exposed to the entomogenous nematode, Steinernema feltiae, at concentrations of 0, 10, 25, 60, 100, or 200 nematodes per larva. Larvae of both species were susceptible to nematode infections. However, neonate larvae of S. exigua were significantly less susceptible to nematode infection than 3- or 8-day-old larvae at or above 50 nematodes per larva. Mortalities of neonate larvae exposed to 50 or more nematodes ranged from 68 to 74% while mortalities of 3- and 8-day-old larvae ranged from 91 to 100%. The results with P. unipuncta showed similar trends as described for S. exigua, albeit at a lower mortality level and usually with no statistical differences. Mortalities of neonate larvae exposed to 50 or more nematodes ranged from 34 to 44% while mortalities of 7-day-old larvae ranged from 32 to 91%.  相似文献   

7.
The physiological and physicochemical bases for the effect of 5, 10, 50, or 100 micrograms of Cd and Zn ml-1 on the attachment and detachment interactions of Pseudomonas fluorescens H2 with glass substrata were determined. Attachment and detachment varied with the type and concentration of metal and the time at which cells were exposed to the metal. The largely inhibitory effect of the metals on bacterial motility and physiological activity did not directly influence attachment. The amount of Cd or Zn accumulated by the cells increased with metal concentration and was greater for free than for attached cells. The hydrophobicity and negative and positive charges of the bacterial surfaces (measured by hydrophobic and electrostatic interaction chromatography) were increased by cell exposure to the metals, particularly after Cd treatment. Cells exposed to Cd prior to attachment showed increased adhesion. Zinc-treated cells did not. There was a positive correlation between adhesion and Cd concentration in the attachment solution. No such relationship existed for Zn. P. fluorescens H2 exposed to Cd prior to attachment desorbed similarly to untreated controls. Zinc pretreatment resulted in decreased desorption. Cells attached in 5 or 10 micrograms of Cd or Zn ml-1 detached less than those attached in 50 or 100 micrograms of Cd or Zn ml-1. The presence of Cd or Zn during detachment had little effect on desorption. The dominant influence of Cd and Zn on attachment and detachment appears to be through modification of the bacterial surface. In natural ecosystems, heavy metals may influence the distribution of bacteria between the solid and liquid phases.  相似文献   

8.
The larval ontogeny of Hypseleotris galii is described and illustrated. 'Premature' yolk sac larvae hatch with unpigmented eyes and no mouth. 'Late' yolk sac larvae hatch with pigmented eyes and a functional mouth. Hatching glands are distributed on the head and ventral surface of the body. The yolk is absorbed and the larvae begin feeding 6 days after hatching. Larval development is completed 74 days after hatching. Hypseleotris galii larvae have six pairs of naked neuromasts: two pairs on the head and four pairs on the body. The significance of these results to developmental strategies in Hypseleotris species is discussed.  相似文献   

9.
Eggs of Oryzius latipes in the blastula stage were exposed to M/100 artificial sea water which contained cadmium at the concentrations of 0.1, 1.0, 10.0, 20.0 or 50.0 mg 1−1. The 96 h TL50, value for cadmium was estimated to be 20 5 mg 1−1. When the eggs were incubated for 24 h in the M/100 sea water with 10.0 mg Cd 1−1 and then rinsed in glycine buffer solution (pH; 2.0), the cadmium content of the egg decreased markedly. Cadmium levels were determined in parts of the embryonic body, the chorion and the yolk sac. The most cadmium was detected in the chorion (94.6%). Prolonged cadmium exposure revealed that most of the cadmium was absorbed by the chorion and little was detected in the embryonic body and the yolk sac.  相似文献   

10.
Eggs and larvae of the carangid fish, Caranx mate (Cuv. & Valenc.), were incubated at various temperature (17.2 to 33.1 °C) and salinity (10 to 42 ‰) combinations in five experiments. The following rates were directly proportional to temperature: embryonic development, yolk absorption, eye and jaw development, and increase in length. Unfed C. mate larvae attained a maximum size at 25 °C and 20 ‰ Eyes and jaws of larvae were functional by the end of the yolk sac stage at all temperature and salinity levels tested.Hatching success and larval survival at the end of the yolk sac stage were generally greater than 50 % between 22° and 32°C. Hatching success and larval survival at the end of the yolk sac stage were reduced at salinity extremes, especially in low temperature-low salinity and high temperature-high salinity combinations. The frequency of morphological abnormalities was also high at extreme temperatures and salinities.The incipient upper thermal TLm for unfed C. mate larvae acclimated to 23.8°C increased from 31.5°C for newly hatched larvae, to 34.2°C for 72 h larvae, but decreased to 32.0°C for starving larvae after the exhaustion of the yolk supply.  相似文献   

11.
Splenic antibody-producing cells were produced by rainbow trout that had been exposed to O-antigens extracted from Yersinia ruckeri and Aeromonas salmonicida by adding the concentrated antigen preparation directly into the water of the tank holding the fish for a flush exposure. This method was compared with the proven techniques of exposure: intraperitoneal injection or a 2 minute immersion of the fish in the antigen preparation. Dosage experiments showed that the production of antibody-producing cells was induced by the immersion of trout for 2 minutes in water with 5.0 μg/ml-1 (or more) with the Y. ruckeri O-antigen, or 500 μg ml-1 (or more) of the A. salmonicida O-antigen. Similar differences were evident when the respective antigens were added directly to the water.  相似文献   

12.
Seven chemotherapeutic agents (dimetridazole, metronidazole, pyrimethamine, albendazole, fenbendazole, mebendazole and magnesium sulfate) were examined for growth inhibition on the cultivation of Spironucleus vortens. Dimetridazole and metronidazole were effective in inhibiting the parasite's growth. At concentrations of 1 microgram ml-1 or higher, both dramatically decreased numbers of parasites. At 24 h exposure, 33% of parasites were inhibited when exposed to dimetridazole or metronidazole at concentrations of 2 and 4 micrograms ml-1, respectively. Dimetridazole at 4 micrograms ml-1 or higher concentrations decreased the number of organisms to 50% or less after 48 h exposure. During the same period of time, the numbers of parasites decreased to 50% or less when exposed to metronidazole at 6 micrograms ml-1 or higher. Pyrimethamine at concentrations of 1 to 10 micrograms ml-1 was not effective in inhibiting the parasite's growth. Albendazole and fenbendazole at concentrations of 0.1 and 0.5 microgram ml-1 were similar in inhibiting the growth of the organism. Both compounds suppressed parasite growth at concentrations of 1.0 microgram ml-1 or higher after 24 h exposure. Mebendazole inhibited the parasite's growth at concentrations of 0.5 microgram ml-1 or higher. At 72 h exposure, 45 to 50% of the parasites were inhibited when exposed to mebendazole at concentrations higher than 0.5 microgram ml-1. Magnesium sulfate at concentrations of 70 mg ml-1 or higher also suppressed the growth of parasites after 24 h exposure. These results indicate that dimetridazole, metronidazole and mebendazole are the most effective chemotherapeutic agents in vitro at inhibiting the growth of S. vortens.  相似文献   

13.
Preliminary evaluation of bare or polymer-coated colloidal monoclinic zirconia of nominal particle size 100 nm indicated that it is an effective adsorbent for pyrogenic lipopolysaccharides (LPS) as measured by chemical and Limulus amebocyte lysate (LAL) assays. Zirconia at 50 micrograms ml-1 adsorbed 99.95% of added E. coli O128 LPS. Residual LPS levels below 0.1 ng ml-1 were easily attained. Colloidal zirconia was able to remove LPS from solution in the presence of bovine albumin (BSA). Some LPS contaminating BSA lacked affinity for zirconia. Preadsorption of phosphate onto bare zirconia blocked LPS adsorption. However, phosphated-oligomeric glycidyl (epoxy) pentaerythritol-coated colloidal zirconia could be derivatized with imidazole-containing ligands to produce an LPS-binding surface. Preliminary results of adsorption of LPS by the coated particles indicated a reduced level of LPS binding compared to bare zirconia, probably because the particles aggregated during the derivatization process, reducing the effective surface available for LPS adsorption.  相似文献   

14.
This study describes the morphological development and early growth of laboratory-raised chub, Leuciscus cephalus (L. 1758), larvae. Larvae hatched with relatively large yolk sacs, stayed motionless at the tank bottom and exhibited short and sudden bursts from time to time. They were olive in colour and with complete absence of melanophores on the body. Larvae were transparent but showed brownish eye pigment. Intense body pigmentation first appeared on day 4 after hatching. By day 8 the yolk sac was fully absorbed; only 30% of the initial population of larvae successfully established exogenous feeding. Fin rays first appeared on the dorsal and anal fins of larvae around day 12. Growth during the yolk sac stage was initially fast but slowed down with the increasing size of larvae at the time of yolk absorption. The specific growth rate (SGR) of larvae declined with time, although the total observation period was relatively short (12 days). SGR values ranged from 5.18 (day 2) to 2.97 (day 12). Only a negligible egg mortality was observed during the period of early endogenous feeding (between days 1 and 6), and about 45% towards the end of endogenous feeding and immediately after the yolk sac phase (between days 7 and 9). During the exogenous feeding period (between days 10 and 12) deaths were negligible.  相似文献   

15.
Efficacy of thiamethoxam (Cruiser) and imidacloprid (Gaucho) were evaluated as seed treatments for controlling European corn borer, Ostrinia nubilalis (Hübner) and Indianmeal moth, Plodia interpunctella (Hübner) larvae in stored grain. At approximately 22-26 degrees C, all fifth instar European corn borers died after two or 4 d of exposure to corn treated with 250 and 500 ppm thiamethoxam, respectively, while mortality of larvae exposed for two and 4 d on corn treated with 6.3-937.5 ppm imidacloprid did not exceed 48% at any concentration. At 29 degrees C, all nondiapausing fifth instars were killed after 3, 4, and 6-d exposure to 400, 300 and 200-ppm thiamethoxam, respectively, while survival increased at successively lower concentrations of 100, 50, 25, and 12.5 ppm. At 29 degrees C, the LC50 decreased from 85.9 to 7.2 ppm as the duration of exposure on treated corn increased from 2 to 6 d. All second and third instar Indianmeal moth larvae died after a 5 d exposure period to corn grain treated with thiamethoxam at 50 ppm or higher, but as the larvae aged, higher concentrations and longer exposure periods were required to give 100% mortality of each larval instar. Similar results were obtained when larval Indianmeal moths were exposed on corn treated with imidacloprid, or on sorghum treated with thiamethoxam. Mature wandering phase fifth instars were the most tolerant larval stage of the Indianmeal moth.  相似文献   

16.
The dynamics of the steroidogenic response of nonprimate gonadal cells to gonadotropins suggests that the biologic action of pituitary LH differs from that of placental CG. To compare the response to LH and CG in primate species, luteinized granulosa cells (LGCs) obtained from rhesus monkeys following follicle stimulation were cultured in vitro. The pattern and levels of progesterone (P) produced during culture was influenced by the concentration (0-10%) and type (fetal bovine or macaque) of serum in the medium and whether LGCs were plated on plastic or extracellular matrix from bovine corneal endothelial cells. After 2-3 days of culture, LGCs were exposed acutely (15-30 min) or chronically (6 h) to 1 or 100 ng/ml human LH (hLH, NIH 1-2) or hCG (CR123), 50 micrograms/ml ovine LH (oLH, NIH-oLH-25), or incubated in the absence of gonadotropins (controls). After the first 15-30 min, the media were changed at 30-min intervals. Both acute and chronic exposure to hLH, hCG, and oLH increased (p less than 0.05) P concentrations above control levels within 15-30 min. There were no differences in the patterns or levels of P elicited by hLH or hCG over time for each treatment condition. Chronic exposure to 1 and 100 ng/ml hLH or hCG and 50 micrograms/ml oLH sustained P levels above that of controls for the 6-h interval. Acute exposure to 1 ng/ml hLH or hCG failed to maintain elevated P levels throughout the experiment.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

17.
This study investigated effects of microalgae (Rhodomonas baltica) and heterotrophic protists (Oxyrrhis marina) on the daily growth, activity, condition and feeding success of Atlantic herring (Clupea harengus) larvae from hatch, through the end of the endogenous (yolk sac) period. Yolk sac larvae were reared in the presence and absence of microplankton and, each day, groups of larvae were provided access to copepods. Larvae reared with microalgae and protists exhibited precocious (2 days earlier) and ≥ 60% increased feeding incidence on copepods compared to larvae reared in only seawater (SW). In the absence and presence of microalgae and protists, life span and growth trajectories of yolk sac larvae were similar and digestive enzyme activity (trypsin) and nutritional condition (RNA-DNA ratio) markedly declined in all larvae directly after yolk sac depletion. Thus, microplankton promoted early feeding but was not sufficient to alter life span and growth during the yolk sac phase. Given the importance of early feeding, field programs should place greater emphasis on the protozooplankton-ichthyoplankton link to better understand match-mismatch dynamics and bottom-up drivers of year class success in marine fish.  相似文献   

18.
The effects on newly fertilized eggs, embryos and larvae of zebrafish Danio rerio following exposure to sediment samples from the more heavily contaminated River Körsch, southern Germany, occurred earlier and were more prominent than in samples from the less contaminated Krähenbach. Dose- and time-related effects following exposure to Körsch sediment eluates and extracts included: (1) hatching failure and subsequent death of larvae exposed to undiluted aqueous sediment eluates and reduced hatching rates at sediment extract concentrations 0·0125%; (2) increased mortality after exposure to 25 and 50% dilutions of aqueous sediment eluates, and dilutions of 0·00625% sediment extracts; (3) reduction of heart beat frequency for 50% dilutions of sediment eluates and concentrations of 0·025% extracts; (4) increased frequency of heart and yolk sac oedema after exposure to 0·0125% sediment extracts. Since adverse effects of sediment extracts observed in zebrafish laboratory tests correlated with reproductive failure in natural populations of brown trout Salmo trutta f. fario in the severely polluted River Körsch, early life stages tests with zebrafish appear to be a suitable tool to assess the contamination rate of natural sediments.  相似文献   

19.
Trypan blue is a potent teratogen in vivo and in vitro in the rat. Many of the abnormalities produced by trypan blue--including swollen neural tube and pericardium, subectodermal blisters, hematomas, and generalized edema--may result from altered fluid balance in and around the embryo. The present study demonstrates relationships between changes in the fluid environment around the embryo and appearance of anomalies. Rat embryos were exposed in utero or in vitro to trypan blue during the early period of organogenesis. Both exposures resulted in defects that are typical of trypan blue treatment. Osmolality of exocoelomic fluid (ECF) was measured on gestation day 10 in vivo and day 12 in vitro, both after 48 hr of exposure to trypan blue. In both cases ECF osmolality was significantly lower than controls. This was correlated with the presence of edema-related anomalies in the embryo. On gestation day 11 in vivo, three days after maternal injection of trypan blue, ECF osmolalities were significantly higher than controls; however, there was tremendous variability in this parameter in day 11 treated embryos, and some had ECF osmolalities below the control range. Increased frequency of abnormalities was correlated with abnormal ECF osmolality, below and above the control range. Trypan blue probably exerts its teratogenic effects by disturbing the function of the visceral yolk sac. The movements of an amino acid and a monosaccharide across the visceral yolk sac were measured on gestation day 12 embryos in vitro. This aspect of yolk sac function was not altered by trypan blue exposure. Ultrastructure of the visceral yolk sac was observed after trypan blue exposure in vivo and in vitro. Endodermal cells in trypan blue-treated yolk sacs contained fewer large, electron dense lysosomes than controls. These were replaced by numerous small vacuoles, which may contain trypan blue. Trypan blue causes osmotic changes in the rat embryo in vivo and in vitro. These changes are correlated with embryonic malformations. Alterations in yolk sac ultrastructure indicate that trypan blue affects the function of this membrane.  相似文献   

20.
Ascorbic acid (AA) and α-tocopherol (α-TOH) levels in whole Atlantic halibut larvae were constant during the yolk sac stage at 170 and 131 ng individual−1, respectively. At hatching c . 80% of the AA and 97% of the α-TOH were contained within the yolk-sac compartment. With development, AA and α-TOH levels in the yolk decreased, at different rates. At first feeding (at 200 day degrees post hatch, D°PH)>95% of AA but <30% of α-TOH in the yolk at hatching had been transferred to the larval body. Transfer of α-TOH was completed at 360 D°PH, when the yolk was completely absorbed. The plankton offered to the larvae at first feeding (chiefly Temora longicornis ) contained 756 μg g−1 AA and 120 μg g−1α-TOH (dry weight). The AA content increased to 472 ng individual−1 within one week after first feeding, while it declined slightly in unfed larvae. In fed larvae the AA content reached c . 3500 ng individual−1 at 580 D)PH. The α-TOH content increased only slightly in the first week of feeding (206 to 431 D°PH), but then increased to > 800 ng individual−1 at 483 D°PH.  相似文献   

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