Effects of induced triploidy on aggressive display in the fighting fish, Betta splendens Regan

Abstract. Triploidy was induced in the fighting fish, Betta splendens Regan, by varying all possible combinations of temperature (37-41°C), time after insemination (2-3min)and shock duration (2-4 min). Heat shock at 39°C for 3 min duration initiated 2-5 min after insemination gave high frequencies of triploids (86%) as assessed from chromosome number and red blood cell nuclear volume. There was no significant difference in the growth rate of triploid and diploid fish. Gonadal development in both sexes was retarded in triploids at 5 months of age. Eggs fertilized with milt from triploids developed to gastrulation. Beyond gastrulation there was increasing mortality associated with abnormalities and none of them hatched. The display frequencies of air gulping, erection of operculum and fins, striking and biting, and undulating movements were fewer in triploids compared to diploids. It appears that triploids are less aggressive than diploids. The aggressive behaviour of fighting fish may be related Io their reproductive activity.     

Freeze‐dried forms of mosquito larvae for feeding of Siamese fighting fish (Betta splendens Regan, 1910)

Live diets are preferably used for rearing Siamese fighting fish (Betta splendens Regan, 1910) since they provide superior growth over the dry feed. In the current study, three different preparations of mosquito larvae were prepared (frozen at ?20°C, F–20; freeze‐dried and kept at 4°C, FD4; freeze‐dried and kept at ambient temperature, FDAT) and were individually fed to 1‐month‐old red male fighting fish (1.18 ± 0.01 g initial body weight) over 6 weeks duration. At the end of experiment, there were no significant differences in growth performance and feed utilization across three dietary treatments (p ? 0.05). Specific activity of lipase was significantly lower in fish fed FD4 and FDAT than with the F–20 diet, while no differences in other enzymes were observed. The fish fed with FDAT diet significantly increased in viscerosomatic index relative to F–20 and FD4 treatments. Significant improvements in skin redness and flesh quality (RNA and RNA/protein ratio) were observed in the fish fed with FDAT diet relative to the other treatments. This preferred FDAT treatment also maintained the carcass composition. Analysis of digestive enzymes in FDAT mosquito larvae demonstrated the presence of protein‐, carbohydrate‐, and lipid‐digesting enzymes after 1 month of storage. The findings from our experiments indicate that the freeze‐dried form (FDAT) of mosquito larvae is suitable for rearing Siamese fighting fish. However, effective preparation protocol and appropriate storage times should be further studied.     

Pigment deposition and in vitro screening of natural pigment sources for enhancing pigmentation in male Siamese fighting fish (Betta splendens Regan, 1910)

Distribution in pigment deposition was studied in male Siamese fighting fish (Betta splendens Regan, 1910) of solid Blue and solid Red phenotypes. Growths were similar, and also indicated by similar activity ratio of trypsin to chymotrypsin. Growth strategy was different at maturing stage, whereas trypsin expression decreased in solid Blue while chymotrypsin expression decreased in solid Red (P ≤ 0.05). Ranking of total pigment contents was muscle > skin > caudal fin > anal fin > dorsal fin > pelvic fin > pectoral fin. Immature fish had similar total contents and concentrations of pigments, except solid Blue had higher total contents in dorsal fin than solid Red. At maturing stage, solid Blue showed higher total pigment contents than solid Red due to higher contents in the fins. Solid blue had higher concentration in pelvic fin, while solid Red had higher in pectoral and dorsal fins. Age, phenotype and their interaction influenced pigment deposition. Determinations of pigment levels, high temperature tolerant of pigments and in vitro digestibilities of pigments, protein and carbohydrate from twenty natural sources indicated that fermented red rice was the most suitable pigment source, following with Chlorococcum sp.     

Effect of substituting live feed with formulated feed on the reproductive performance and fry survival of Siamese fighting fish, Betta splendens (Regan, 1910)

This study aimed to elucidate the effect of partial or complete replacement of live feed (LF) (Tubifex) with formulated feed (FF) on the reproductive performance of Betta splendens. Three hundred B. splendens fry (average weight 0.19 ± 0.01 g) were equally distributed into five different groups, each with three replicates. They were fed for 105 days with following different diets: control (C)—100% LF; T1—75% LF, 25% FF; T2—50% LF, 50% FF; T3—25% LF, 75% FF, and T4—100% FF. Results showed that the average number of hatched larvae (654 ± 101) and fry survival after 2 weeks of rearing (428 ± 70), after completion of three spawning, were recorded highest in the control group, which was, however, not significantly different from the T1, T2, and T3 groups. At the end of the feeding trial, the highest hatching percentage (90.3%) was registered in the T2 group, which was not significantly different from the control and T1 groups. The T2 group also recorded highest fry survival (65.54%) after completion of three spawning, which was not significantly (P < 0.05) different from the T1 and T3 groups. Control diet contained higher saturated fatty acid (63.23%) than formulated diet (29.80%). In the whole-body tissue, highest level of EPA (0.42%) and DHA (3.13%) were found in the T4 group followed by T3 group. The DHA/EPA ration was recorded highest in the T2 group (10.96%), which did not differ significantly from the T1 and T3 groups. Significant positive correlation was observed between saturated fatty acid levels in fish whole-body tissue and number of hatched larvae (Y = 30.81 × −825.3, R ² = 0.968) and fry survival after 2 weeks of rearing (Y = 21.38 × −580.9, R ² = 0.967). Considering all these factors, it can be concluded that the live feed Tubifex can be replaced up to 50% without any adverse effect on the reproductive performance and fry survival of B. splendens.     

Replacement of live feed by formulated feed: effect on the growth and spawning performance of Siamese fighting fish (Betta splendens,Regan, 1910)

Partial replacement of live feed (LF) with formulated feed (FF) was performed in Betta splendens to evaluate their growth, survival and reproductive performance. Three hundred B. splendens fry of uniform size (mean weight 0.19 g±0.01) were equally distributed in five treatment groups with each of three replicates in glass aquaria of 35 L capacity. Fish were offered diets of different ratio of LF and FF like T1 (control) – 100% LF; T2 – 75% LF, 25% FF; T3 – 50% LF, 50% FF; T4 – 25% LF, 75% FF and T5 – 100% FF. Highest (P<0.05) body weight gain (% BWG) (67.1±1.03) and specific growth rate (2.34±0.02) were observed in the T2 group. Best feed conversion ratio was also found in T2 (2.40±0.11), which was similar to T1 and T3. The highest survival rate (%) was recorded in T1, T2 and T3 groups (97±1.7), which was similar to T4 and the lowest in T5 (49±2.0). The highest gonadal weight (0.12 g±0.01) and gonadosomatic index (15.17±0.50) was observed in the T3 group during the first sampling after 63 days. No significant changes in spawning performance were observed in the second sampling after 21 days after first sampling. However, T4 group registered the highest fecundity and fertilized eggs at the end of 105 days of experiment. From the present study, it concludes that LF can be successfully replaced to the extent of 25% by FF without any detrimental effect on the growth, survival and spawning performance of B. splendens.     

Induction of heterozygous and homozygous diploid gynogenesis in Betta splendens (Regan) using hydrostatic pressure

Abstract Optimum conditions for hydrostatic pressure treatment for duplication of chromosome set in gynogenetically activated fighting fish, Betta splendens (Regan), eggs were identified. Maximum survival of heterozygous gynogens was 50%, when 2·5-min-old eggs, after insemination with UV irradiated tilapia sperm, were pressure shocked at 7000psi for 6 min. The frequency (21%) of homozygous gynogenetic fry was high, when the 34min (post-insemination) old eggs, which were inseminated with tilapia sperm, were pressure shocked for 5 min. Sex ratio of gynogenetic progeny suggested that the mechanism of sex determination in this fish is homogametic female and heterogametic male.     

Identification of Mycobacterium spp. isolated from snakehead, Channa striata (Fowler), and Siamese fighting fish, Betta splendens (Regan), using polymerase chain reaction–reverse cross blot hybridization (PCR–RCBH)

A variety of methods have been used to identify Mycobacterium spp. isolated from snakehead and Siamese fighting fish, including biochemistry, mycolic acid profiles and antibody-based methods. However, these methods are unable to differentiate between different species of Mycobacterium . Polymerase chain reaction (PCR) followed by reverse cross blot hybridization (RCBH) was adapted in this study to speciate aquatic mycobacteria. The method was highly specific for Mycobacterium spp. and identified the bacteria to species level with a detection limit of 100 fg DNA, equivalent to 20 mycobacteria. Twenty-nine isolates previously collected and cultured from Siamese fighting fish (10 isolates) and snakehead (19 isolates) during outbreaks of mycobacteriosis were analysed using PCR–RCBH. Six of the Siamese fighting fish isolates and nine of the snakehead isolates were identified as Mycobacterium fortuitum , while the remainder were classified as M. marinum . Notably, two isolates recovered from snakehead and Siamese fighting fish, previously identified as M. poriferae and M. piscicida , respectively, were confirmed to be M. fortuitum .     

Minimal water volume for intensively producing male Siamese fighting fish (<Emphasis Type="Italic">Betta splendens</Emphasis> Regan, 1910)

Water volume is a key parameter affecting the individual rearing of male Siamese fighting fish (Betta splendens Regan, 1910). In this study, minimization of water volume was pursued by assessing growth, feed utilization, digestive enzyme activities, color coordinates, muscle quality, and carcass composition. One-month-old solid-red male fish (0.97?±?0.01 g initial body weight) were distributed individually into glass aquaria with five alternative water volumes (100, 150, 200, 250, and 300 mL), comprising 15 fish per treatment (n?=?15), over 8 weeks duration. No mortality of the reared fish was found during the study. Growth performance and feed utilization of the fish reared in 150 mL water were superior to the other treatments. The water volume significantly affected specific activities of the digestive enzymes (P ? 0.05), except for amylase, and no differences in enzyme activities were observed between fish reared in 150 and in 300 mL water. The preferred treatment maintained skin lightness (L*) and had the highest redness (a* and a*/b*) among the treatments. Protein synthesis (RNA concentration) and its turnover rate (RNA/protein ratio) and myosin and actin in muscle also benefited from this treatment. Carcass composition, in terms of moisture, crude protein, and crude ash, was maintained, but the amount of crude lipid fluctuated with water volume. Based on our experiments, the preferred minimal water volume for individual rearing of male Siamese fighting fish should be about 150 mL.     

暹罗斗鱼的饲养与繁殖

1名称与原产地暹罗斗鱼(Betta splendensRegan)又名五彩搏鱼,隶属于鲈形目(Perciformes)、攀鲈亚目(Anabantoidei)、斗鱼科(Belontiidae)。暹罗斗鱼原生长在热带地区的湖沼、稻田等水流缓慢水域,个体小,没有绚丽缤纷的色彩。它主产于东南亚的暹罗(即如今的泰国),让人啧啧称奇的     

Effects of live feed containing Panagrellus redivivus and water depth on growth of Betta splendens larvae

Panagrellus redivivus is a nematode with a high protein content and low‐cost production. It is successfully used in the larviculture of shrimp, however, it has not yet been evaluated as feed for Betta splendens larvae. Thus, the objective of this study was to evaluate P. redivivus as feed for betta larvae maintained at different depths of water. The experiment was based on a completely randomized 3 × 2 factorial design, represented by two feed types and one alternating feeding regime plus two water depths (2.5 and 5.0 cm). The Artemia feed promoted the highest weight gain, followed by the alternating feeding regime. The single use of the nematode as live feed produced the poorest weight gain. The water depth and the feeding method influenced the specific growth rate (SGR). At a depth of 2.5 cm, the alternating feeding promoted similar SGR compared to fish fed with Artemia only. Furthermore, at the greater water depth an improvement in SGR was observed in fish fed Artemia only, but the similarity to alternating feeding regime remains. Thus, the inclusion of nematode is an appropriate and lower cost feed strategy for the betta larviculture independent of water depths.     

Modulation of monoamine neurotransmitters in fighting fish <Emphasis Type="Italic">Betta splendens</Emphasis> exposed to waterborne phytoestrogens

Endogenous estrogens are known to affect the activity of monoamine neurotransmitters in vertebrate animals, but the effects of exogenous estrogens on neurotransmitters are relatively poorly understood. We exposed sexually mature male fighting fish Betta splendens to environmentally relevant and pharmacological doses of three phytoestrogens that are potential endocrine disruptors in wild fish populations: genistein, equol, and β-sitosterol. We also exposed fish to two doses of the endogenous estrogen 17β-estradiol, which we selected as a positive control because phytoestrogens are putative estrogen mimics. Our results were variable, but the effects were generally modest. Genistein increased dopamine levels in the forebrains of B. splendens at both environmentally relevant and pharmacological doses. The environmentally relevant dose of equol increased dopamine levels in B. splendens forebrains, and the pharmacological dose decreased norepinephrine (forebrain), dopamine (hindbrain), and serotonin (forebrain) levels. The environmentally relevant dose of β-sitosterol decreased norepinephrine and dopamine in the forebrain and hindbrain, respectively. Our results suggest that sources of environmental phytoestrogens, such as runoff or effluent from agricultural fields, wood pulp mills, and sewage treatment plants, have the potential to modulate neurotransmitter activity in free-living fishes in a way that could interfere with normal behavioral processes.     

Genetic variation in different varieties of Siamese fighting fish using isoelectric focusing of sarcoplasmic proteins

Sarcoplasmic proteins of four colour varieties of Siamese fighting fish, Betta splendens Regan, were analysed by isoelectric focusing (IEF). Genetic polymorphisms were detected in six loci having isoelectric points: 4.92-5.00 (Sp-1), 5.33-5.48 (Sp-2), 5.90 (Sp-3), 6.31-6.35 (Sp-4), 6.58-6.70 (Sp-5) and 6.90 (Sp-6). For the Sp-1, Sp-2, Sp-4 and Sp-5 loci, there were two co-dominant alleles. At Sp-3 and Sp-6, one allele was dominant and the other null. Individuals within each locus were classified into phenotypes A, AB and B. Only homozygotes were found at Sp-2. Allele frequencies were calculated for each locus and tested for accordance with the Hardy-Weinberg equilibrium. Observed values for each phenotype at Sp-1, Sp-3, Sp-4, Sp-5 and Sp-6 agreed well with the expected ones. At Sp-2, genotypic frequencies for each variety deviated markedly (p < 0.01) from the equilibrium. When tested for homogeneity, no significant differences (p 0.95) in gene frequencies were found among the varieties except at Sp-2. Band sharing index (BSI), computed using the BIO-GENE v5.04 program, revealed high intravariety genetic similarities (0.701-0.740) and slightly above-average intervariety values (0.550-0.583). In this study, we estimated the levels of genetic variability in B. splendens, and proposed a genetic model for each polymorphic locus.     

Size,growth, temperature and the natural mortality of marine fish

The natural mortality of exploited fish populations is often assumed to be a species‐specific constant independent of body size. This assumption has important implications for size‐based fish population models and for predicting the outcome of size‐dependent fisheries management measures such as mesh‐size regulations. To test the assumption, we critically review the empirical estimates of the natural mortality, M (year^?1), of marine and brackish water fish stocks and model them as a function of von Bertalanffy growth parameters, L (cm) and K (year^?1), temperature (Kelvin) and length, L (cm). Using the Arrhenius equation to describe the relationship between M and temperature, we find M to be significantly related to length, L and K, but not to temperature (R² = 0.62, P < 0.0001, n = 168). Temperature and K are significantly correlated and when K is removed from the model the temperature term becomes significant, but the resulting model explains less of the total variance (R² = 0.42, P < 0.0001, n = 168). The relationships between M, L, L, K and temperature are shown to be in general accordance with previous theoretical and empirical investigations. We conclude that natural mortality is significantly related to length and growth characteristics and recommend to use the empirical formula: ln(M) = 0.55 ? 1.61ln(L) + 1.44ln(L) + ln(K), for estimating the natural mortality of marine and brackish water fish.     

(鱼康)(鱼良)白鱼的人工采卵孵化和苗种培育

(鱼康)(鱼良)白鱼是云南特有的名贵经济鱼类,正面临濒危.(鱼康)(鱼良)白鱼的人工采卵孵化和苗种培育,具有十分重要的意义.它不仅是人工增殖技术和种群恢复研究的重要环节,而且是对其进行人工驯养繁殖研究的基础.杂食性鱼类在稚鱼的生长发育过程中,都存在着食性转化现象,(鱼康)(鱼良)白鱼亦不例外.为此,我们在苗种培育中进行了(鱼康)(鱼良)白鱼饵料的人工培育、人工替代饵料的筛选;在人工孵化中进行了(鱼康)(鱼良)白鱼的人工受精孵化;上述尝试均取得了初步成功,现小结如下.     

Ultrasound Enhanced Immersion Protocols for Masculinization of Nile Tilapia, Oreochromis niloticus

Androgen immersion protocols have been unsuccessful in consistently producing all-male tilapia at a high enough ratio for them to be commercially viable. This study explored the use of ultrasound to improve on the results of previous immersion studies. Variables tested include two hormones (trenbolone acetate-TBA and 17α-methyldihydrotestoterone-MDHT) at two concentrations (100 and 250 μg/L) and with or without ultrasound (cavitation level). All hormone treatments with ultrasound and non-ultrasound resulted in significantly higher masculinization than the appropriate controls ( P < 0.05). Among pairs of treatments of the same hormone at the same dose, all ultrasound treatments resulted in significantly higher number of males compared with non-ultrasound treatments with the exception of MDHT 250 μg/L ( P 0.05). Comparing across all ultrasound treatments, TBA 250 μg/L with ultrasound had higher masculinization than all the other ultrasound treatments ( P 0.05). Comparing across all non-ultrasound treatments, TBA 250 μg/L had higher ( P 0.05) masculinization than MDHT 100 μg/L and TBA 100 μg/L with non-ultrasound. Two of the three replicates of TBA 250 μg/L ultrasound treatment resulted in 100% males and the highest mean percentage (98%) of males. This study thus demonstrated the potential of a short-term immersion protocol using ultrasound to more predictably produce all-male, commercially viable tilapia seed.     

坑塘流水养鱼技术

开化地处浙江西部,和江西、安徽两省交界,是个“九山半水半分田”的山区县。开化农民从明朝就开始利用山溪流水在房前屋后、溪边沟旁挖塘养鱼,水流常年不断,品种以草鱼为主,完全投喂青饲料,后把这种方式养出的鱼称作“清水鱼”。独具的生态优势和特有的养殖方式,使流水坑塘养出的清水鱼背部溜黑、肚皮亮白、肉细腻鲜嫩而有弹性。开化清水鱼以其优异的品质,深受杭州、嘉兴、上海等大中城市消费者喜爱,市场上供不应求。     

Detection of the bacterium Flavobacterium psychrophilum from a natural infection in rainbow trout, Oncorhynchus mykiss (Walbaum), using formalin-fixed, wax-embedded fish tissues

The bacterial pathogen Flavobacterium psychrophilum was successfully identified from formalin-fixed, wax-embedded tissue blocks of infected rainbow trout heart and spleen tissues, using a polymerase chain reaction (PCR)-based assay. Filamentous bacteria were observed in haematoxylin and eosin and Giemsa-stained sections but no bacteria were recovered from the diseased fish using standard bacteriology isolation techniques. All infected fish had histopathological evidence of myocarditis or rainbow trout fry syndrome. Immunohistochemistry was attempted using three different anti-F. psychrophilum sera but the results were inconclusive, and an alternative molecular approach was therefore attempted. This paper describes the use of a PCR-based assay to help identify bacteria present in formalin-fixed, wax-embedded tissue samples. This is the first time that this technique has been used for the detection of fish bacteria from diagnostic samples.     

河流型鱼类自然保护区管理思考

分析了河流型鱼类自然保护区上游来流主导、水体及其环境依赖、线性开放系统的主要特征。阐述了河流型鱼类自然保护区面临的保护与开发矛盾突出,封闭性管理弊端凸显,保护区本底调查缺乏及相关基础科研的投入不足严重制约保护提升的困难与挑战。针对河流型自然保护区的实际,在共性管理措施基础上,提出以下保护管理建议：（1）关注上游影响以从河流生态系统角度开展保护;（2）深入开展保护区基础调查以夯实保护基础;（3）实施适应性管理。