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1.
In this article, an in vitro investigation was carried out to ascertain the roles of hormones and growth factor in the inductions of oocyte maturation and steroidogenesis of the postvitellogenic follicles in an Indian estuarine grey mullet, Mugil cephalus L. Oocyte maturation was evaluated by scoring the germinal vesicle breakdown (GVBD) percent of the postvitellogenic follicles. All the sex [17α,20β-dihydroxy-4-pregnane-3-one (DHP), estradiol 17β (E2), progesterone (P), 17α-OH progesterone (17-OH-P) and testosterone] and other [bovine-insulin and salmon-calcitonin, human chorionic gonadotropin (hCG), luteinizing hormone (LH) or hCG+DHP] hormones and insulin-like growth factor-I (IGF-I) significantly increased GVBD% in 9 h culture. DHP had a maximum effect (75 %) compared to other effectors. Some effectors (hCG: 82.14 %, LH: 78.94 %, hCG plus DHP: 81.81 %, E2: 80 % and IGF-I: 74.19 %) including DHP (79 %) further increased GVBD% in 15-h culture. All the hormones (except DHP) and IGF-I increased DHP, E2 and testosterone productions by the postvitellogenic ovarian follicles in vitro. DHP and testosterone productions were increased with the increase of incubation time from 9 h through 15 h. E2 production was not further increased beyond 12 h. DHP production was highest by hCG compared to other effectors. The hCG of all the test compounds was most effective in both the induction of GVBD% and steroid production. DHP is the most potent inducer of oocyte maturation in Indian estuarine flat head grey mullet. Involvement of estrogen in mullet oocyte maturation is indicated. hCG, like DHP, is equally potent and induces oocyte maturation via DHP production in vitro. hCG with DHP has synergistic action on oocyte maturation in mullet ovary. Interplay of several hormones (hCG, LH, and probably E2 and testosterone) and IGF-I on oocyte maturation is suggested in the mullet.  相似文献   

2.
Repeated injections of salmon pituitary extract (20 mg per fish per week) induced vitellogenesis in feminized, cultivated Japanese eels (Anguilla japonica). Oocytes were attained at the migratory nucleus stage after 11 or 12 injections. Addition of 17,20-dihydroxy-4-pregnen-3-one (DHP) into the incubation medium induced germinal vesicle breakdown (GVBD) in the oocytes at the migratory nucleus stage. An injection of DHP (2 µg g-1 BW), given 24h after an injection of salmon pituitary extract (20 mg fish-1), succeeded in inducing maturation and ovulation in females which contained occytes at the migratory nucleus stage. Most fish ovulated 15–18h following the DHP injection. Eggs that were ovulated within 15h after the DHP injection showed high fertility and hatchability, but eggs ovulated 18 or 21h after the DHP injection, showed considerably lower fertility and hatchability. A delay between ovulation and stripping of the eggs rapidly decreased both the fertility and hatchability within 6–9h after ovulation, indicating that artificial fertilization must be carried out immediately after ovulation. Repeated injections of human chorionic gonadotropin (hCG) at a concentration of 1 IU g-1 BW week-1 induced spermatogenesis, spermiation, and the acquisition of potential for sperm motility in cultivated males. Most males spermiated after the fifth or sixth injection of hCG, and the milt weight gradually increased and remained constant (1–2 g) from the 11th to 31th injection. Sperm motility peaked 24h after each weekly injection, and decreased from the 3rd day after the injection. Potassium ions are an essential constituent for the maintenance of motility in the eel spermatozoa. Artificial seminal plasma containing 15.2 mM KCl is applicable as a milt diluent. Using these techniques developed for female and male eels, we have succeeded in obtaining many fertilized eggs from cultivated eels.  相似文献   

3.
Vitamin A (VA) is an essential micronutrient in fish, vital for reproduction, vision and immunity. Freshwater fish species store and use VA in the form of retinol (ROL), as well as uncommon forms such as 3,4‐didehydroretinol (DROL) and 3‐hydroxyretinol (3ROL). We developed and validated an HPLC method capable of measuring ROL, DROL and 3ROL in the feed and tissues of North American freshwater fish, for which little information on retinoid concentrations is available. DROL and ROL were higher than expected in both fish feed and fish liver (at 9.5–29.6 mg kg?1 DROL and 26.9–79.2 mg kg?1 ROL for feed; and 19.8–548.8 μg g?1 DROL and 10.4–128.8 μg g?1 ROL for liver). Interestingly, DROL concentrations were lower than ROL in fish feed, but often substantially higher in fish liver. Concentrations of ROL and DROL were similar to each other in muscle. 3ROL was detected in fish liver only. To our knowledge, this is the first publication to measure DROL concentrations in North American farm‐raised fish and commercial fish feed. Monitoring of not only ROL, but DROL and 3ROL as well, may be advisable to better assess the VA content of fish feed and tissue.  相似文献   

4.
In this study, we examined the effects of 500 IU mL?1 penicillin + 500 μg mL?1 streptomycin sulphate on semen quality indices of endangered caspian brown during 12 days short‐term storage at 4°C. Twenty‐four millilitre semen samples with good quality were considered for the experiment. The semen samples were then stored in the presence and absence of 500 IU mL?1 penicillin + 500 μg mL?1 streptomycin sulphate. The semen quality parameters including percentage and duration of sperm motility were measured 0, 3, 6, 9 and 12 days after storage. In the antibiotic receiving group, the values of percentage and duration of sperm motility reduced 3 and 6 days after storage respectively and reduced to lowest levels at day 12. In the antibiotic‐free group, the duration and percentage of sperm motility decreased significantly after 3 days of storage and reached to lowest values at day 12. Also, percentage and duration of sperm motility in each storage time were significantly higher in the antibiotic receiving group than in the antibiotic‐free group. The overall values of percentage and duration of sperm motility for all storage periods were higher in the antibiotic receiving group than in the antibiotic‐free group. In conclusion, our results demonstrated that 500 IU mL?1 penicillin + 500 μg mL?1 streptomycin sulphate improves the viability of caspian brown trout during short‐term storage.  相似文献   

5.
Recently, we developed an intraspecies spermatogonial transplantation technique in a pelagic egg spawning marine teleost, nibe croaker Nibea mitsukurii. Nibe croaker is an ideal candidate recipient for spermatogonial transplantation since it has a short generation time and small body size. In the present study, yellowtail Seriola quinqueradiata spermatogonia were transplanted into nibe croaker larvae, and the behavior of transplanted spermatogonia in recipient gonads was observed. Three weeks post-transplantation, yellowtail spermatogonia were incorporated into the gonads of 72 out of 88 recipients. An antiproliferating cell nuclear antigen was detected in incorporated yellowtail spermatogonia, suggesting that the xenogenic germ cells were proliferating in recipient gonads. Yellowtail vasa-positive spermatogonia survived for 11 months after transplantation in the gonads of recipient fish. Thus, we showed that the microenvironment in nibe croaker gonads can support the colonization, proliferation, and survival of germ cells derived from a different taxonomic family.  相似文献   

6.
Partial characterization of digestive proteases in the three‐spot cichlid Cichlasoma trimaculatum juveniles was conducted. It was determined that there is higher alkaline proteases activity (3.95 ± 0.32 IU mg?1 protein) compared to acidic proteases (2.01 ± 0.57 IU mg?1 protein). Optimal temperature for alkaline proteases is 60 °C which resulted in more thermostability to temperature changes. On the other hand, optimal temperature for acidic proteases is 50 °C. Optimal pH for acidic proteases was pH 2, while for alkaline proteases, it was pH 10, which resulted in more stability in relation to pH changes than acidic protease. The use of specific inhibitors and the SDS‐PAGE electrophoresis analysis revealed seven types of bands for alkaline proteases, which make evident the main presence of serine proteases. In acidic proteases, more than 98 g kg?1 of the activity was inhibited with pepstatin A inhibitor. Therefore, it is evident that C. trimaculatum digestion is composed by acidic and alkaline proteases; thus, it should be considered an omnivorous fish.  相似文献   

7.
Aquaflor® [50% w w?1 florfenicol (FFC)], is approved for use in freshwater‐reared warmwater finfish which include tilapia Oreochromis spp. in the United States to control mortality from Streptococcus iniae. The depletion of florfenicol amine (FFA), the marker residue of FFC, was evaluated after feeding FFC‐medicated feed to deliver a nominal 20 mg FFC kg?1 BW d?1 dose (1.33× the label use of 15 mg FFC kg?1 BW d?1) to Nile tilapia O. niloticus and hybrid tilapia O. niloticus × O. aureus held in a recirculating aquaculture system (RAS) at production‐scale holding densities. Florfenicol amine concentrations were determined in fillets taken from 10 fish before dosing and from 20 fish at nine time points after dosing (from 1 to 240 h post‐dosing). Water samples were assayed for FFC before, during and after the dosing period. Parameters monitored included daily feed consumption and biofilter function (levels of ammonia, nitrite and nitrate). Mean fillet FFA concentration decreased from 13.77 μg g?1 at 1‐h post dosing to 0.39 μg g?1 at 240‐h post dosing. Water FFC concentration decreased from a maximum of 1400 ng mL?1 at 1 day post‐dosing to 847 ng mL?1 at 240 h post‐dosing. There were no adverse effects noted on fish, feed consumption or biofilter function associated with FFC‐medicated feed administration to tilapia.  相似文献   

8.
This study investigated the effect of the replacement of fish oil (FO) with DHA‐Gold (DHA‐G)‐supplemented plant oils (PO) in rainbow trout fed plant‐protein‐based diets. Five diets (450 mg g?1 digestible protein and 150 mg g?1 crude lipid) were fed to rainbow trout (initial weight 37 ± 0.5 g) for 12 weeks in a 15 °C recirculating water system. The lipid inclusion types and levels were FO, PO and PO with DHA‐G supplemented at 30 mg g?1, 60 mg g?1 or 90 mg g?1 of the diet replacement for corn oil. Fish fed 90 mg g?1 DHA‐G were significantly larger and consumed more feed than fish‐fed PO or FO (218 g and 2.6% bwd?1 versus 181 g and 2.4% and 190 g and 2.3%, respectively). Feed conversion ratio was significantly increased in fish fed 90 mg g?1 DHA‐G (0.99) as compared to fish‐fed FO (0.90) and 30 mg g?1 DHA‐G (0.91). Panellists found trout fillets from fish fed the 90 mg g?1 DHA‐G diet to have significantly fishier aroma and flavour than fish fed the FO diet. Fatty acid analysis demonstrated that 60 mg g?1 or 90 mg g?1 DHA‐G supplementation increased PO fed fish fillet DHA to fatty acid levels equivalent or higher than those fish fed a FO diet.  相似文献   

9.
The acceleration of final oocyte maturation and ovulation is recognized as a principal process in Caspian kutum Rutilus frisii kutum, a commercial and valuable species in Caspian Sea. The aim of this study was to evaluate the effects of catecholaminergic pharmaceutics which include Salbutamol (β2‐adrenergic receptor agonist), Metoprolol (β1‐adrenergic receptor antagonist), Clozapine and Olanzapine (the third generation of D4 and D2 dopamine receptor antagonists) which can be more efficient than the first and second generations, in combination with Buserelin ([D‐Ser(tBu)6,Pro9‐NEt]‐GnRHa) and Ovaprim (D‐Ala6,Pro9‐Net)‐sGnRH+Domperidone on ovulation and spawning induction in Caspian kutum. One hundred eight adult females were injected once with Buserelin Acetate (BUS) 5 μg kg?1 BW, Ovaprim 0.5 mL kg?1 BW, Salbutamol Sulphate (SLB) 4 mg kg?1 BW, Olanzapine (OLZ) 5 mg kg?1 BW, Clozapine (CZ) 12 mg kg?1 BW and Metoprolol Tartrate (MTP) 5 mg kg?1 BW, being divided into 12 groups: group 1, intact (negative control); group 2, Ovaprim (positive control); group 3, OLZ+SLB; group 4, OLZ+MTP; group 5, OLZ+BUS; group 6, CZ+SLB; group 7, CZ+MTP; group 8, CZ+BUS; group 9, OLZ+BUS+SLB; group 10, OLZ+BUS+MTP; group 11, CZ+BUS+SLB and group 12, CZ+BUS+MTP (N = 9). The results showed that the highest mean value in ovulation success, ovulation index, fertilization success, relative fecundity and the number of eggs belonged to Ovaprim treatment. On the other hand, spawning was successful in OLZ+BUS+SLB and CZ+BUS treatments (P < 0.05), whereas it was lower in CZ+MTP, CZ+BUS+MTP and OLZ+BUS than the other treatments. Therefore, it could be concluded that Clozapine and Olanzapine potentiated the effect of Buserelin treatment in ovulation and spawning induction, while Metoprolol blocked the stimulatory effects of GnRHa, Clozapine and Olanzapine. Salbutamol also can amplify stimulatory effect of all mentioned pharmaceutics.  相似文献   

10.
A 9‐week feeding experiment was conducted to determine the effect of dietary biotin levels on growth performance and non‐specific immune response of large yellow croaker. Fish (6.16 ± 0.09 g) were fed twice daily to apparent satiation with diets containing 0.00 (as the basal diet), 0.01, 0.05, 0.25, 1.24 and 6.22 mg biotin kg?1 diet. Results showed that fish fed the basal diet had the lowest survival rate, and fish fed 0.05 mg kg?1 dietary biotin achieved significantly higher final weight and weight gain. Dietary biotin levels had no significant influence on carcass crude lipid, moisture and ash content, but significantly influenced the carcass crude protein. Liver biotin concentration significantly increased with the supplementation of biotin, but no tissue saturation was found within the supplementation scope of biotin. Broken‐line regression analysis of weight gain showed that juvenile large yellow croaker requires a minimum dietary biotin of 0.039 mg kg?1 for maximal growth. The analyses of serum parameters showed that the moderate‐ (0.05 mg kg?1) and high‐dose (6.22 mg kg?1) dietary biotin significantly improved both lysozyme and alternative complement pathway activities, indicating dietary biotin within a certain range could improve the non‐specific immune response of large yellow croaker.  相似文献   

11.
An attempt was made to increase free amino acids (FAA) concentrations in eggs of red snapper Lutjanus campechanus as part of an induced spawning protocol. Mature female red snapper were given intramuscular injections of human chorionic gonadotropin (HCG) at a 1100 IU kg?1 of female body weight. Immediately after the HCG injection, one set of females received an intramuscular injection of FAA in a buffered saline solution at a dose of 20 mg FAA cocktail kg?1 of body weight. The FAA cocktail was 25% isoleucine, 25% leucine, 25% lysine and 25% valine on a molecular weight basis. A second set of fish did not receive the FAA supplement. Both sets of fish were equally successful in spawning with ovulation occurring 21–32.5 h post injection. Injection of FAA into brooders as eggs began final maturation and hydration resulted in an increase in leucine (nmoles mg egg?1) and a slight increase in isoleucine (nmoles egg?1) in recently ovulated eggs. FAA supplementation via brood injection altered FAA utilization rates by embryos and during larval development, resulted in a greater yolk sac and oil globule reserve in larvae as they approached the time of first feeding.  相似文献   

12.
An optimal chemical shell marking protocol was developed for the New Zealand green‐lipped mussel, Perna canaliculus with a view to its future use in larval tracking experiments. Larval P. canaliculus aged either 10, 15 or 19 days post fertilization were immersed in treatments of 50, 100 and 200 mg L?1 of calcein for a period of 24 h before measurements of shell mark brightness were taken. There was 100% marking success in all calcein treatments for all age classes, with 19‐day larvae immersed in 200 mg L?1 calcein producing the brightest mark. Growth was not affected by calcein immersion; however, 10‐day larvae exhibited significantly higher levels of mortality compared with 15‐ and 19‐day larvae suggesting a reduced resilience to the marking protocols in younger larvae. In a mass staining experiment, a solution of 100 mg L?1 calcein was used to successfully stain15.6 million hatchery reared P. canaliculus larvae. Calcein, therefore, offers a low impact method with which to stain the sensitive early life stages of this species thus providing a rapid method for identifying individuals of interest, i.e. individuals released in the wild or specific family lines within a hatchery environment.  相似文献   

13.
An 8 weeks feeding trial was conducted to determine the dietary methionine requirement of fingerling Indian catfish, Heteropneustes fossilis (6.08 ± 0.95 cm; 4.33 ± 0.52 g). Six isonitrogenous (40%) and isoenergetic (17.90 kJ g?1 GE) amino acid test diets were formulated with gradation of 0.25 g 100 g?1containing graded levels of L‐methionine (0.30, 0.55, 0.80, 1.05, 1.30 and 1.55 g 100 g?1, dry diet) with 0.40 g 100 g?1 constant level of cystine. Twenty fish were stocked in triplicate groups, in 75‐L circular trough with continuous flow‐through system and fed experimental diets at 4% BW/day twice daily, at 08:00 and 18:00 hours. Maximum live weight gain (296%), best feed conversion ratio (1.56) and protein efficiency ratio (1.60) were occurred at 1.05 g 100 g?1 methionine, beyond which they showed declining tendency. However, quadratic regression analysis of weight gain, feed conversion ratio (FCR), protein efficiency ratio (PER) and body protein deposition (BPD) data indicated requirement for methionine at 1.15, 1.08, 1.06 and 1.05 g 100 g?1 of dry diet respectively. Significantly (< 0.05), higher whole body protein content, minimum moisture and intermediate fat contents were recorded at 1.05 g 100 g?1 dietary methionine level. Ash content remained insignificantly (> 0.05) low among all the treatments, excepting at diet I and diet II. Body protein deposition was also found to be significantly (< 0.05) higher at 1.05 g 100 g?1 methionine level. Best somatic and haematological indices values were also obtained at the requirement level. Based on above results, it is recommended that the diet for young H. fossilis should contain methionine at 1.09 g 100 g?1 dry diet, corresponding to 2.73 g 100 g?1 dietary protein with 0.40 g 100 g1 cystine concentration for optimum growth and efficient feed utilization. Thus, the total sulphur amino acid requirement of H. fossilis would be (1.09 + 0.40) 1.49 g 100 g?1 of dry diet, corresponding to 3.73 g 100 g?1 of dietary protein.  相似文献   

14.
The effect of docosahexaenoic acid (DHA) on the growth performance, survival and swim bladder inflation of larval Seriola dumerili during the rotifer feeding period was investigated in two feeding experiments. Amberjack larvae at 3 day post hatching were fed rotifers enriched with (1) freshwater C hlorella (Chlo), (2) a mixture (2:1, v/v) of Chlo and DHA‐enriched C hlorella (DHA‐Chlo), (3) DHA‐Chlo and (4) DHA‐Chlo and commercial DHA emulsion, in triplicate for 7 days. The average DHA contents of the rotifers were 0.0, 0.4, 1.0 and 1.9 mg g?1 DM respectively. The survival rate was improved by the enrichment of rotifers with DHA‐Chlo alone, and DHA‐Chlo and emulsion. Growth and swim bladder inflation of fish fed rotifers enriched with DHA‐Chlo were significantly (< 0.05) improved, however, with increased levels of DHA further improvement was not found. DHA content in the larval whole body proportionally increased with the DHA level in the rotifers. These results suggest that DHA enrichment of rotifers is effective to improve the growth, survival rate and swim bladder inflation of amberjack larvae. The DHA requirement of amberjack larvae is estimated to be 1.5 mg g?1 on a dry matter basis of rotifers.  相似文献   

15.
Abstract:   The question of whether the ovulation and spawning time in chub mackerel Scomber japonicus is entrained by a circadian rhythm was raised by our previous experiments. Further questions were also raised about whether the time course of human chorionic gonadotropin (hCG)-induced final oocyte maturation (FOM) and ovulation reflected the natural time course induced by endogeneous pituitary gonadotropin (GtH). To address these questions, hCG and gonadotropin-releasing hormone analog (GnRHa) were administered at two 'opposite' times, 14:00 and 02:00 hours, and the time courses of FOM and ovulation were compared. When hCG was injected, ovulation occurred 33 h post-injection in both groups, regardless of the timing of the hCG injection. The timing of ovulation in chub mackerel depends on the timing of hCG injection, but apparently not on circadian rhythms. When GnRHa was injected, ovulation began at 36 h post-injection of GnRHa, regardless of the timing of injection. These results indicate that the time course of FOM and ovulation in the chub mackerel followed a similar pattern whether stimulated by hCG injection or spontaneous luteinizing hormone (LH) surge because GnRHa induces the secretion of endogenous GtH (primarily LH) from the fish pituitary. Thus, it is concluded that the time course of hCG-induced FOM and ovulation in chub mackerel follows the natural time course induced by endogenous pituitary LH.  相似文献   

16.
Female northern whitefish were treated with salmon [D‐Arg6Pro9Net]‐sGnRHa emulsified in Freund′s incomplete adjuvant as a sustained release treatment (sGnRHa‐FIA) or with [D‐Arg6Pro9Net]‐sGnRHa dissolved in physiological saline as acute release treatment (sGnRHa‐PS). Females in four experimental groups (A, B, C, D) and one control group (E) were injected intraperitoneally as follows: A and B – sGnRHa‐FIA at doses of 50 and 25 μg kg?1, respectively; C – double injection (DI) of sGnRHa‐PS at 25 μg kg?1 administered 3 days apart; D – single injection (SI) of sGnRHa‐PS at 25 μg kg?1; and E – control group receiving a 1:1 mixture of FIA and physiological saline. All treatments induced and synchronized ovulation. Mean time to ovulation was significantly reduced (< 0.05) in GnRHa‐treated groups compared with control. Hormone treatments did not affect the relative fecundity. Slight differences were found in ovarian fluid pH between group A and D (< 0.05). Except for group C, egg size was significantly reduced (P < 0.01) in hormonally synchronized groups compared with the control. Survival to the eyed stage of eggs from females in groups A, B and C was significantly lower (< 0.01) than in groups D and E. Per cent hatched alevins was lower (P < 0.01) in groups A, B and C than in groups D and E. We conclude that synchronization of ovulation in northern whitefish can be induced by a single acute injection of [D‐Arg6Pro9NEt]‐sGnRHa at 25 μg kg?1 BW if given near the natural spawning time determined by water temperature below 2°C.  相似文献   

17.
To induce synchronized ovulation, migrating wild Caspian brown trout (Salmo trutta caspius) females were treated with two interperitoneal injections of Des‐Gly10, d ‐Ala6 LHRH (LHRHa), given 3 days apart. Two injections of 100 μg kg?1 body weight of this hormone effectively induced ovulation. Within 27 days from the second injection, all fish injected with 100 μg kg?1 LHRHa had ovulated compared with 54.5% of the controls. The mean time to ovulation was reduced significantly (P<0.05) from 31.67±4.84 days in control fish and 28.83±7.31 days in sham‐treated fish to 16.36±1.61 days in fish injected with 100 μg kg?1 LHRHa. The fertilization rate in 50 and 100 μg kg?1 LHRHa‐injected fish was significantly lower than that in the control fish (P<0.05). In fish injected with 50 and 100 μg kg?1 LHRHa, significant (P<0.05) changes in testosterone (T) and 17α‐hydroxyprogestrone (OHP) levels were observed. After the second LHRHa injection, the fish injected with 100 μg kg?1 showed the highest serum levels of testosterone and OHP. These results demonstrate that the use of LHRHa can effectively reduce the mean time to ovulation and induce synchronized ovulation in Caspian brown trout.  相似文献   

18.
Development of alligator gar (Atractosteus spatula) and spotted gar (Lepisosteus oculatus) larvae was evaluated after exposure of embryos to thyroid hormones (TH) by two different methods of administration. For the first method, alligator gar embryos were placed in a water bath with triiodothyronine (T3; 1 ppm for 2 h), and control treatment embryos were placed in a water bath without T3. For the second method, spotted gar adult males and females were injected with different TH during induced spawning: a group was injected with T3 [20 mg kg?1 body weight (BW)], a second group with thyroxine (T4; 20 mg kg?1 BW), a third group with thyroid‐stimulating hormone (TSH; 4 IU kg?1BW) and a control group with dimethyl sulfoxide. Both methods resulted in increases in the concentrations of T3 and T4 in embryos of both species. An increase in the alkaline enzymatic activity of TH‐treated larvae was observed as well as an increase in the RNA:DNA, RNA:Dry weight and DNA:Dry weight ratios at hatching. Some positive effects of TH application were: a significant increase in the hatching rate of spotted gar larvae from TH‐injected parents and a higher percentage of successful spawns. Evenness of larval sizes and a higher survival rate were observed in alligator gar larvae. Snout development was accelerated by TH treatment in both species.  相似文献   

19.
20.
To assess the effects of A3α‐peptidoglycan (A3α‐PG) extracted from Bifidobacterium sp. on the immune response and disease resistance of sea cucumber, different concentrations (0, 0.5, 5 and 50 mg mL?1) of A3α‐PG suspensions were used to perform hypodermic injection on Apostichopus japonicus, followed by a Vibrio splendidus challenge. Total coelomocyte count (TCC), phagocytosis activity and activities of four immunological enzymes in both cell‐free coelomic fluid (extra‐cellular, EC) and coelomocyte lysate supernatant (intracellular, IC), including acid phosphatase (ACP), alkaline phosphatase (ALP), superoxide dismutase (SOD) and peroxidase (POD), were measured at 2, 6, 14 and 24 h post injection (hpi). The TCC was not significantly affected (> 0.05) by A3α‐PG, ranging from 1.84 × 106 to 3.53 × 106 cells mL?1. The coelomocyte phagocytosis activity was significantly activated (< 0.05) in all the A3α‐PG treatments, whereas no significant difference was observed between them except 24 hpi (> 0.05). The EC‐ACP activity in the 5.0 mg mL?1 treatment increased significantly (< 0.05) at all sampling times, while the IC‐ACP activity in the 50 mg mL?1 treatment increased significantly (< 0.05) at 2 hpi. Also, the 5.0 mg mL?1 treatment had significant (< 0.05) increase in the EC‐ALP activity within 14 hpi and the EC‐POD activity at 2 hpi, respectively, while significantly (< 0.05) enhanced IC‐ALP and IC‐POD activities were observed in the 50 mg mL?1 treatment within 6 hpi and at 2 hpi, respectively. Only the 5.0 mg mL?1 treatment showed significant (< 0.05) increase in the EC‐SOD activity at 2 hpi and IC‐SOD activity within 14 hpi, respectively. The challenge test showed that the animals treated with 50 mg mL?1 of A3α‐PG had notably lower cumulative mortality after 14 days following V. splendidus exposure. All together, these results suggest that A3α‐PG could positively enhance immune response that effectively promotes the health status of A. japonicus against V. splendidus infection.  相似文献   

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