鲤科鱼蓝黑鲮微卫星位点的种间扩增(英文)

本研究使用105对微卫星引物对7种鲤科鱼类进行跨越种间PCR扩增,共得到14个多态性微卫星位点.其中9个扩增效果较好的位点用于分析来自帕吉勒提河(Bhagirathi, n=20)和戈达瓦里河(Godavari, n=25)的蓝黑鲮(Labeo calbasu)样品的遗传多样性.结果显示,前者在每个位点的平均等位基因数为7.33,而后者为8 1,期望杂合度介于0.795(Bhagirathi)和0.801(Godavari)之间;4个位点MFW11* (Godavari)、R1*(Godavari)、R3* (Bhagirathi) 和 Lr38*(Bhagirathi和Godavari)都表现出明显的杂合子缺失和哈迪温伯格平衡偏离;而任意两位点间都未观测到连锁不平衡现象;位点R3*极可能存在无效等位基因.上述结果表明这些多态性微卫星位点作为共显性标记在蓝黑鲮群体遗传学研究中有着较好的应用前景.     

Establishment of a leukocyte cell line derived from peritoneal macrophages of fish,Labeo rohita (Hamilton, 1822)

A continuous leukocyte cell line with phagocytic activity was established from peritoneal macrophages of rohu, Labeo rohita (LRPM). LRPM was initiated from adherent mononuclear leukocytes isolated from peritoneal cavity of rohu, without use of any growth factors or feeder cells. These cells exhibited maximum growth at 30 °C in L-15 medium containing 20 % foetal bovine serum, and has been subcultured for more than 60 passages till date. The cells showed 85 % viability after 6 months of storage in liquid nitrogen. The species of origin of the LRPM was confirmed by the amplification and sequencing of 655 bp fragment of cytochrome oxidase subunit I of mitochondrial DNA. Functionally, LRPM showed phagocytic activity of yeast cells and fluorescent latex beads as evaluated by phase contrast and scanning electron microscopy, respectively. Immuno-modulators such as bacterial lipopolysaccharide and phorbol myristate acetate resulted in functional activation of LRPM; and enhanced their microbicidal activity through release of reactive oxygen species and nitric oxide. Culture supernatant from activated cells also revealed lysozyme activity. Cells of LRPM were positive for alpha-naphthyl acetate esterase enzyme indicating macrophage lineage. Our results indicate that this cell line can be a useful in vitro tool to study the role of macrophages in teleost immune system and to evaluate the effects of new aquaculture drugs. The LRPM cell line represents the first reported leukocyte cell line of peritoneal origin from any freshwater species of fish.     

Length‐weight relationship of two cichlid fish species,Etroplus suratensis (Bloch, 1790) and Etroplus maculatus (Bloch, 1795) from Lake Vembanad,Kerala, India

The present study describes the length–weight relationships (LWRs) of two cichlid fish species, Etroplus suratensis (Bloch, 1790) and Etroplus maculatus (Bloch, 1795) collected from Vembanad Lake, Kerala, India during the period June 2012 to May 2013 using gill nets with various mesh sizes (3.5–9 cm) and cast nets with a 1.5 cm mesh size. The values of parameters b in the LWR equation were estimated as 2.67 for E. suratensis and 2.72 for E. maculatus, respectively.     

A comparative study on the effects of selected pesticides on hemato-biochemistry and tissue histology of freshwater fish Cirrhinus mrigala (Hamilton, 1822)

The aim of the present study was to investigate the comparative effects of pesticides Chlorfenapyr, Dimethoate and Acetamiprid on the health of Cirrhinus mrigala under long term exposure. Eighty C. mrigala were divided in four equal groups; one control and three treated groups. The blood was collected from both control and treated groups at intervals of 10th, 20th and 30th days for hemato-biochemistry and histopathological alterations. The result indicates significant difference (P < 0.05) in RBCs, Hb, PCV and MCHC whereas elevation in WBCs and Platelets counts were recorded. In 10th day sampling, MCV value of Dimethoate and Acetamiprid treatment had no difference in comparison with the control group, however it is significantly increased (P < 0.05) in rest of sampling. The MCH value of exposed fish showed significant increased (P < 0.05) after 20th and 30th days for Chlorfenapyr and after 30th days for Acetamiprid exposure while insignificantly increased for rest of sampling. It was also found that these pesticides significantly decrease (p < 0.05) the T3 and T4 levels while increase in the TSH, cortical, ALP, AST, ALT and LDH levels in the serum of the treated fishes in contrast to control group. Similarly, histopathological analysis of gills and liver showed significant alterations in all the treated groups. Toxicity trends of these pesticides was ranked as Chlorfenapyr > Acetamiprid > Dimethoate. It is concluded that indiscriminate use of such pesticides poses a noxious threat to non-target organisms, harm the ecosystems and jeopardizes human health.     

Assessment of Epinephelus coioides (Hamilton, 1822) aquaculture systems in the Gulf of Tonkin,Vietnam, by using fish parasites

The parasite fauna of Epinephelus coioides can be used as an indicator to assess different aquaculture systems. A total of 133 E. coioides were collected from fishermen (wild fish) and marine fish farms (net cages, pond farms) in the Gulf of Tonkin (Ha Long, Cat Ba and Nghia Hung), Vietnam in 2013 and 2014, and examined for the presence of protozoan and metazoan parasites. Different patterns of parasite infections were found. The monogenean Pseudorhabdosynochus spp. was predominant in the net cage system; and the protozoans Trichodina spp. and Cryptocaryon irritans were found exclusively in pond and net cage cultivation sites in Nghia Hung and Cat Ba. These three species were assessed in addition to seven further indicator species and two dominance indices, to visualize the potential of assessing different aquaculture systems in Vietnam. This suggests the use of former shrimp ponds for grouper cultivation, reducing the pressure to further extend net cage cultivation in already highly influenced mariculture regions in Vietnam.     

Differential C-heterochromatin distribution in two species of freshwater fish, Anabas testudineus (Bloch.) and Puntius sarana (Hamilton.)

Anabas testudineus (2n = 46) had the more conserved pattern of its C-heterochromatin distributed mainly in the centromeric region, whereas Puntius sarana (2n = 50) exhibited a rather unorthodox pattern, many chromosomes showing interstitial, some telomeric and a few chromosomes showing centromeric C-band localization. Further, lateral asymmetry in distribution of heterochromatin was also noted in two pairs of chromosomes in P. sarana. The possible implications of the differential distribution noted in these two species has been discussed.     

Cryopreservation of sperm of an indigenous endangered fish species Nandus nandus (Hamilton, 1822) for ex-situ conservation

This study dealt with the development of cryopreservation protocol for Nandus nandus, which entailed a number of experiments. Sperm was collected by sacrificing males. The collected sperm was suspended in extenders. Activation of sperm motility was evaluated in different osmolalities of NaCl. Motility of sperm decreased as the osmolality of the extender increased and was completely inhibited at almost 319 mOsmol/kg. To evaluate the toxicity of cryoprotectant, sperm was incubated with DMSO, methanol and ethanol at 5%, 10% and 15% concentrations, respectively, for 5–35 min. Five and ten percent of cryoprotectants produced better motility during 5 and 10 min incubation. Sperm incubated with 15% cryoprotectant seemed to be toxic and this concentration was excluded in the subsequent trials. Three extenders, namely, Alsever’s solution, egg-yolk citrate and urea egg-yolk and three cryoprotectants, DMSO, methanol and ethanol were employed to preserve the sperm. Alsever’s solution with 10% DMSO showed best performance producing 90.0 ± 1.8% and 75.0 ± 2.5% equilibration and post-thaw motility followed by that of 82.5 ± 4.2% and 62.5 ± 5.5% with Alsever’s solution plus methanol, respectively. Between two diluents, sperm preserved with Alsever’s solution plus DMSO produced highest fertilization (76.7 ± 3.3%) and hatching (43.8 ± 7.9%) while fresh sperm yielded 83.3 ± 6.7% and 64.0 ± 10.4% fertilization and hatching, respectively. The protocol developed through the study can be applied for long-term conservation of genetic materials of the endangered fish N. nandus and the cryopreserved sperm can be used in artificial breeding for generating new individuals.     

Cryopreservation of sperm of an indigenous endangered fish species Nandus nandus (Hamilton, 1822) for ex-situ conservation

This study dealt with the development of cryopreservation protocol for Nandus nandus, which entailed a number of experiments. Sperm was collected by sacrificing males. The collected sperm was suspended in extenders. Activation of sperm motility was evaluated in different osmolalities of NaCl. Motility of sperm decreased as the osmolality of the extender increased and was completely inhibited at almost 319 mOsmol/kg. To evaluate the toxicity of cryoprotectant, sperm was incubated with DMSO, methanol and ethanol at 5%, 10% and 15% concentrations, respectively, for 5–35 min. Five and ten percent of cryoprotectants produced better motility during 5 and 10 min incubation. Sperm incubated with 15% cryoprotectant seemed to be toxic and this concentration was excluded in the subsequent trials. Three extenders, namely, Alsever’s solution, egg-yolk citrate and urea egg-yolk and three cryoprotectants, DMSO, methanol and ethanol were employed to preserve the sperm. Alsever’s solution with 10% DMSO showed best performance producing 90.0 ± 1.8% and 75.0 ± 2.5% equilibration and post-thaw motility followed by that of 82.5 ± 4.2% and 62.5 ± 5.5% with Alsever’s solution plus methanol, respectively. Between two diluents, sperm preserved with Alsever’s solution plus DMSO produced highest fertilization (76.7 ± 3.3%) and hatching (43.8 ± 7.9%) while fresh sperm yielded 83.3 ± 6.7% and 64.0 ± 10.4% fertilization and hatching, respectively. The protocol developed through the study can be applied for long-term conservation of genetic materials of the endangered fish N. nandus and the cryopreserved sperm can be used in artificial breeding for generating new individuals.     

Length–weight relationships of two cyprinid fish species,Crossocheilus latius (Hamilton, 1822) and Garra gotyla gotyla (Gray, 1830) from the Ganga River Basin,India

This study provides length–weight relationship (LWR) information for two fish species (family Cyprinidae), Crossocheilus latius (Hamilton, 1822) and Garra gotyla gotyla (Gray, 1830), from a tributary of the Ganga River Basin, India. Both species had no previous LWR estimates as per Fishbase 2014.     

Responses of Corpuscles of Stannius to intra-peritoneal vitamin-D3 administration in teleost Labeo rohita (Hamilton, 1822) reared in water with two different levels of calcium concentration

This study assessed the responses of vitamin-D₃ intraperitoneally injected to Rohu, Labeo rohita @ of 0 IU/kg bw (only solvent), 100 IU/kg bw and 500 IU/kg bw reared in 20 and 40 ppm of calcium (Ca) enriched water. The cellular changes in Corpuscles of Stannius (CS) gland, serum Ca, and inorganic phosphate (P_i) level were analysed up to the 60th day. Rohu administered with 100 IU/kg bw D₃ and exposed to 40 ppm Ca-rich water exhibited notable hyperplasia of CS compared with their control groups. Notable changes with high serum Ca level (13.87 ± 0.3 mg/dl) was detected on the 5th day in fish exposed to 40 ppm Ca-rich water, while related values attained (13.74 ± 0.1 mg/dl) only after 7 days in 20 ppm Ca-rich water of 500 IU/kg bw vitamin D₃ injection. Similarly, high serum P_i level (7.66 ± 0.2 mg/dl) in 40 ppm Ca injected with D₃ at 500 IU/kg bw. The results demonstrated that the Ca homeostasis of Labeo rohita is influenced by intra-peritoneal vitamin D₃. Progressive studies should be conducted by increasing the dose of vitamin D₃ to investigate optimum dose/supplement in feed for commercially important aquaculture teleost Labeo rohita for maximum and sustainable absorption of Ca from the variable water Calcium levels to maintain Ca²⁺ homeostasis.     

Population genetic structure and phylogeography of cyprinid fish, <Emphasis Type="Italic">Labeo dero</Emphasis> (Hamilton, 1822) inferred from allozyme and microsatellite DNA marker analysis

We examined population structure of Labeo dero (Hamilton, 1822) from different riverine locations in India using 10 polymorphic allozyme and eight microsatellite loci. For analysis, 591 different tissue samples were obtained from commercial catches covering a wide geographic range. Allozyme variability (An = 1.28–1.43, Ho = 0.029–0.071) was much lower than for microsatellites (An = 4.625–6.125, Ho = 0.538–0.633). Existence of rare alleles was found at three allozyme (MDH-2*, GPI* and PGDH*) and at two microsatellite loci (R-3* and MFW-15*). Deviation from Hardy–Weinberg equilibrium (P < 0.05, after the critical probability levels were adjusted for sequential Bonferroni adjustment) could be detected at three loci (EST-1*, -2* and XDH*) whereas, after correction for null alleles, two microsatellite loci (MFW-1*,-15*) deviated from HWE in the river Yamuna. Fst for all the samples combined over all allozyme loci was found to be 0.059 suggesting that 5.9% of the total variation was due to genetic differentiation while microsatellite analysis yielded 0.019 which was concordant to mean Rst (0.02). Hierarchical partition of genetic diversity (AMOVA) showed that greater variability (approx. 95%) was due to within population component than between geographical regions. Based on distribution of genetic differentiation detected by both markers, at least five different genetic stocks of L. dero across its natural distribution could be identified. These results are useful for the evaluation and conservation of L. dero in natural water bodies.     

Reproductive biology,growth, and age composition of non‐native Indian glassy fish Parambassis ranga (Hamilton, 1822) in Haebaru Reservoir,Okinawa‐jima Island,southern Japan

Age composition, growth, and reproductive biology of the non‐native Indian glassy fish Parambassis ranga (Hamilton, 1822) were surveyed in the Haebaru Reservoir on Okinawa‐jima Island, southern Japan. Standard lengths (SLs) of males and females ranged from 19.7 to 44.0 mm and from 19.2 to 52.4 mm, respectively. The overall sex ratio was significantly female‐biased, with the monthly percentage of females ranging from 71.4 to 100.0. Marginal growth analysis of sectioned otoliths revealed opaque zones formed annually from November to May. Observed age ranged from 0 to 3 years for both sexes, although the 1‐year age class comprised the majority of the sampled population. Von Bertalanffy growth parameters were L_∞ = 48.8 mm (SL), K = 0.43 year^?1, and t₀ = ?1.47 years for males, and L_∞ = 43.7 mm, K =0.72 year^?1, and t₀ = ?1.29 years for females. Length at 50% maturity was estimated to be 25.8 mm SL, and maturation was within 1 year after hatching. The main spawning season of P. ranga was estimated to occur from February to October, peaking in April.     

Length‐weight and length‐length relationships of two gobiid fishes Eleotris fusca (Forster, 1801) and Odontamblyopus rubicundus (Hamilton, 1822) from the Payra River,southern Bangladesh

The present study provides the estimates of length‐weight relationships (LWRs) and length‐length relationships (LLRs) of two gobiid fishes Eleotris fusca and Odontamblyopus rubicundus from the Payra River, southern Bangladesh. Fish samples were collected from the set bagnet (mesh size <2 mm) fishers of the Payra River in July 2016. Allometric coefficient (b) values were 2.95 for E. fusca and 2.76 for O. rubicundus. Additionally, the LLRs were highly significant with r² ≥ .955. While the estimate of the LWR for O. rubicundus includes a substantial larger sample size and thus provides a more reliable LWR than the one published so far, the LWR for E. fusca must be considered as tentative because the sample includes only juveniles for a fish that reaches a maximum standard length more than three times as much. Additional studies are needed to further improve our LWR knowledge base for these fish species.     

Competitive interactions between two pest species of earth mites, Halotydeus destructor and Penthaleus major (Acarina: Penthaleidae)

Competition was investigated between two pest earth mite species, Penthaleus major Dugés and Halotydeus destructor Tucker, which occur sympatrically and are major pests of southern Australian pastures and crops. Three geographically separate pasture sites within Victoria were sampled monthly. Abundance patterns were similar across sites in that there was a marked increase in number of H. destructor in spring. Field competition plots were set up on pasture. In the first season, H. destructor had a negative effect on P. major but not vice versa. During the summer, diapause and ensuing generation, density-dependent (intraspecific) competition was evident in P. major but not in H. destructor. The interspecific asymmetry was switched in the following season, with P. major dominating most plots and having a negative effect on the reproductive output of H. destructor. These findings indicate that suppression or eradication of one species of earth mite from pastures may result in another species increasing in relative abundance.     

New maximum size records and length–weight relationships for two species,Corica soborna (Hamilton, 1822) and Mystus bleekeri (Day, 1877), from the Ganges River (NW Bangladesh)

This study records new maximum lengths and provides length–weight relationships (LWRs) for two small indigenous fishes, Corica soborna (Hamilton, 1822) and Mystus bleekeri (Day, 1877), from the Ganges River in northwestern Bangladesh. Fishes were sampled sporadically using gill nets and cast nets from August 2015 to July 2016. New maximum recorded lengths weere 5.3 cm total length (TL) for C. soborna and 17.7 cm TL for M. bleekeri. The LWRs were highly significant (p < .001) with r² values >.958. These findings will play a significant role in the fisheries resource management.     

Assessment of growth characteristics,the survival rate and body composition of Asian sea bass Lates calcarifer (Bloch, 1790) under different feeding rates in closed aquaculture system

One of the essential factors to be addressed in the development of aquaculture is the feeding regime. This study was investigated to assess the effects of feeding rate on growth performance, feed utilization, chemical body composition survival rate, cannibalism and morphological indices of Asian Seabass, Lates calcarifer. Intended for the trial, one hundred forty sea bass individuals with an average weight 5.47 ± 0.11 g were randomly distributed in 4 concrete tanks (914 cm × 183 cm 122 cm) each; length × width × depth) and volume 18,399 L, for total 68 days. The fishes were fed with a pelleted diet containing 46% crude protein for different feeding groups designated as (T1, T2, T3 and T4). The feed was supplied with a rate of (T1) 3%, (T2) 4%, (T3) 6%, and (T4) 9% of fish biomass per day and feeding frequency were maintained three times per day to all the groups. At the end of the trial water physicochemical parameters was in acceptable range for Asian sea bass growth. The average daily weight gain (g), weight gain (g) and specific growth rate (%) was significantly higher (p < 0.05) in T3 and T4 as compared to T1 and T2 group. The poorest feed conversion ratio was recorded in T1 group with 3% biomass per day. The cannibalism rate was significantly (p < 0.05) higher in T1 (3%) compared to T3 and T4 treatment. The morphological indices, condition factors (CF), viscerosomatic index and hepatosomatic index (HSI) was significantly higher in T4 group as compared to other treatments. The protein, moisture and ash contents of the whole biomass of the Asian sea bass were not significantly influenced by feeding rate. The fat levels in the fish bodies increased significantly (p < 0.05) with increasing feeding ratio. The phenomenal regression indicates that 6.5% feeding rate per day is optimum for best growth performance, survival and minimum cannibalism rate for Asian sea bass in captivity. The outcome of the finding will help in promotion for not only the coastal aquaculture in Pakistan abut also elsewhere.     

A comparative study of neuropeptides in the intestine of two stomachless teleosts (Poecilia reticulata,Leuciscus idus melanotus) under conditions of feeding and starvation

Summary Endocrine cells containing bombesin-, enkephalin-, gastrin/CCK-, 5-HT-, and substance P-like material were demonstrated in the alimentary tract of Poecilia reticulata and Leuciscus idus melanotus. Endocrine cells with neuropeptide-Y-like immunoreactivity were found only in P. reticulata, those with VIP-like immunoreactivity only in L. idus melanotus. Gut nerves showing bombesin-, G/CCK-5-HT-, neurotensin-, substance P-and VIP-like immunoreactivity were observed in both species investigated, enkephalin- and neuropeptide Y-like immunoreactivity in P. reticulata alone. The distribution and amount of endocrine cells and nerves along the gut as visualized with the appropriate antisera varied in both teleosts. Histologically, the intestinal tract of these stomachless fish can be divided into three regions. A large number of endocrine cells with VIP-like immunoreactivity was noted in the rectum of L. idus melanotus. Endocrine cells containing bombesin-, enkepha-lin- and substance P-like material were found only in intestinal parts I and II in L. idus melanotus. Neuropeptide Y-like immunoreactivity was absent from intestinal part I of P. reticulata. The influence of starvation on the immunoreactivity of nerves and enteroendocrine cells in the teleost intestine was examined. After a starvation period of more than 6 weeks, no alterations were observed either in the appearance or amount of nerve and endocrine cell immunoreactivity.