Inbreeding depression in the Zebra fish Brachydanio rerio (Hamilton Buchanan)

Inbreeding depression was observed in Half and Full sib matings (Inbreeding Coefficient 0.125 and 0.25) of Zebra Danios ( Brachydanio rerio ). Inbreeding at these levels reduced fertility, survival to 30 days, and length at 30 days. There was an increase in the incidence of crippled fry. No effect of inbreeding on either number of eggs per spawn or hatchability were found.     

Effects of group size on the responsiveness of zebrafish, Brachydanio rerio (Hamilton Buchanan), to alanine, a chemical attractant

Previous studies have examined the effects of grouping on the locating (search) phase of foraging and feeding behaviour in fishes. Few studies have examined whether schooling in fishes may facilitate individual foraging by enhancing a group's responsiveness to food odours. The purpose of the current study was to assess the effect of increasing group size on the responsiveness of zebrafish, Brachydanio rerio (Hamilton Buchanan), to L-alanine, an amino acid which is an important constituent of prey odours for many fishes. Based on the results of previous studies, either an increasing or decreasing linear relationship or a unimodal (convex or concave) relationship between responsiveness and group size was expected; the results, however, were bimodal. Groups of four fish were most responsive to alanine, as determined by the mean percentage of occurrences of fish in the area of a behavioural arena (an octagonal fluviarium) into which alanine was infused (at 10⁻³, 10⁻⁴, or 10⁻⁵ M). Groups of two, six and eight fish were significantly less responsive ( P < 0.05) than either groups of four fish or individual fish. The responses of groups of two, six and eight fish were not significantly different from each other.     

Gill morphology in the zebrafish, Brachydanio rerio (Hamilton- Buchanan)

A light and electron microscopic study of the gills of the zebrafish, Brachydanio rerio , were made to serve as a morphological basis for future investigations. It was found that for fixation of B. rerio gills, a mixture of 1·5% gluturaldehyde and 1·5% paraformaldehyde gives a mucus-free surface. Morphometric measurements of structural components of the gill secondary lamellae were made. Observations at SEM were correlated with those made at TEM. The different cell types in the branchial epithelium were characterized. Chloride cells were mainly located in the interlamellar regions and on the afferent side of the primary lamellae. Two morphologically different chloride cells were seen. The first type communicates with the external environment through a reservoir-like lumen, which is normally absent in freshwater fishes. The second type of chloride cell has more direct contact with the ambient water, resembling chloride cells from other freshwater fishes. Another cell type with features similar to those of the rodlet cell was frequently observed. This cell is interposed between other types of cells in the epithelium, and sometimes junctional complexes were present between the rodlet cell and surrounding cells.     

Inheritance and longevity of infectious pancreatic necrosis virus in the zebra fish, Brachydanio rerio (Hamilton-Buchanan).

Zebra fish (Brachydanio rerio) were injected with infectious pancreatic necrosis virus (IPNV) and then spawned to determine whether the virus was passed on to the eggs, and if it was, how long it remained in the free-swimming F1. The mating variations included parents receiving one or two injections of virus, and within these categories, matings in which both parents were treated or only one parent was treated. The results showed that transmission of IPNV to the egg did occur, and that this transmission was via the female alone. However, if the female was allowed to produce antibodies to the virus, as when she received two injections of IPNV, she transmitted the virus to the eggs for only a short period of time. In addition, when the virus was transmitted to the egg, it remained in the free-swimming F1 for a period of at least 5 months.     

Inheritance and longevity of infectious pancreatic necrosis virus in the zebra fish, Brachydanio rerio (Hamilton-Buchanan).

Zebra fish (Brachydanio rerio) were injected with infectious pancreatic necrosis virus (IPNV) and then spawned to determine whether the virus was passed on to the eggs, and if it was, how long it remained in the free-swimming F1. The mating variations included parents receiving one or two injections of virus, and within these categories, matings in which both parents were treated or only one parent was treated. The results showed that transmission of IPNV to the egg did occur, and that this transmission was via the female alone. However, if the female was allowed to produce antibodies to the virus, as when she received two injections of IPNV, she transmitted the virus to the eggs for only a short period of time. In addition, when the virus was transmitted to the egg, it remained in the free-swimming F1 for a period of at least 5 months.     

Cadmium-induced changes in gill morphology of zebrafish, Brachydanio rerio (Hamilton–Buchanan), and rainbow trout, Salmo gairdneri Richardson

Freshwater zebrafish and brackish water rainbow trout were exposed to different concentrations of cadmium for up to 6 weeks. The gill morphology was examined by light and electron microscopy, and a morphometric analysis was performed. The morphometric study of the secondary lamellae revealed an increase in the portion exterior to the basal lamina, resulting in an increased diffusion distance, after exposure to cadmium concentrations of 10μg l⁻¹ and above. In both species this was due to an increase in volume of the non-tissue spaces of the secondary lamellar epithelium. Furthermore, the water space between neighbouring secondary lamellae was clearly reduced. Morphological examination revealed some gross alterations compared with control fish. These initially consisted in curling of the secondary lamellae and finally resulted in local teleangiectasia. Partial lifting of the secondary lamellar epithelium from the pillar cells resulted in large non-tissue spaces which were invaded by leucocytes. The first sign of degeneration was observed in the chloride cells, which were characterized by a dispersed cytoplasm and a smooth apical plasma membrane.     

High mortality in a large-scale zebrafish colony (Brachydanio rerio Hamilton & Buchanan, 1822) associated with Lecythophora mutabilis (van Beyma) W. Gams & McGinnis

Zebrafish (Brachydanio rerio) have become an important model system for studying vertebrate embryonic development and gene function through manipulation of genotype and characterization of resultant phenotypes. An established research zebrafish colony without substantial disease problems for more than 7 years of operation began experiencing appreciable mortalities in November of 1997. Young fish (fry), from five to 24 days after hatching, spontaneously developed elongate strands of organic material protruding from the mouth, operculum, and anal pore, leading workers in the laboratory to describe the infected fish as "bearded." Unlike typical freshwater fish fungal infections, the skin surface did not have evidence of fungal colonization. The disease was associated with progressive lethargy, reduced feeding, and subsequent mortality. From 10 to 100% of the fry in a given tank were affected. Initial examination indicated that the biofilm around the head of affected fry consisted of bundles of septate fungal hyphae, large numbers of mixed bacterial populations, and protozoans. Environmental samples of air and water in the laboratory were obtained to ascertain the source of the infective agent and to isolate and identify the fungus. A fungus identified as Lecythophora mutabilis was isolated repeatedly from infected fish and water samples from infected fish tanks, and from the main laboratory water supply tanks, but not from laboratory air. Some biofilm beards on fish were found to consist of relatively pure bacterial populations, and beards on occasional fish examined in the later part of the study consisted of hyphae and spores of the oomycete genus Aphanomyces. Lecythophora mutabilis did not invade tissues; however, elimination of the epizootic correlated with reduction in the number of L. mutabilis conidia in the water following modification of the laboratory water system by use of new filtration and sterilization systems. We conclude that the dense hyphal strands of L. mutabilis composing the predominant biofilm type, along with mixed bacteria and protozoa, contributed to the die-off in young fry by occluding the oral cavity and/or gills, leading to starvation and/or asphyxiation.     

The influence of alarm substance and shoal size on the behaviour of zebra danios, Brachydanio rerio (Cyprinidae)

Timidity, vigilance and response to alarm substance were examined in shoals consisting of one to 12 zebra danios, Brachydanio rerio . Measures of timidity were not clearly related to shoal size. Fish in larger shoals spent more time in central areas of the tank away from cover than fish in smaller shoals. However, fish in small shoals appeared to be as aggressive as those in larger shoals. Vigilance and foraging rates did not appear to be related to shoal size. Per capita foraging rates and shoal size were not correlated. After being frightened by alarm substance, danios in larger shoals did not return to foraging sooner than those in smaller groups. Zebra danios in all shoal sizes responded behaviourally to alarm substance. It appears that the presence of conspecifics is unnecessary for alarm behaviours to occur and that the nature of the behaviours are independent of shoal size.     

Different methods of tissue preparation for X-ray microanalysis combined with scanning electron microscopy based on zebra fish, Brachydanio rerio muscle fibres

Four preparation methods for electron probe X-ray microanalysis (EPXMA) of muscle fibres were compared: (1) dissection, freezing in LN2 (liquid nitrogen), cutting in cryostat, freeze-drying and analysis; (2) dissection, immersing in Tissue-Tek, freezing in LN2, cutting in cryostat, the following steps as in method 1; (3) dissection, freezing in LN2, freeze-drying, separation of fibres into groups; (4) dissection, cutting into 2 mm thick slices by razor blade, freezing in LN2, following steps as in method 1. The contents of Na, Cl, and K as well as K/Na, Cl/K, ratios were taken as criteria of good preservation of muscle fibres. The best results were obtained by method 1. Good morphological preservation can be routinely observed in sections prepared by methods 3 and 4. However, the diffusible elements and K/Na, Cl/K ratios were not retained at relatively constant level among the individual samples. Fibres prepared by methods 1 and 3 showed, despite of freezing artefacts, high K/Na, and low Cl/K ratio. After method 1, high level of the elemental contents was retained, and it did not differ significantly among the samples, showing relatively low standard deviation values.     

微囊藻毒素对斑马鱼重要细胞因子表达的影响

微囊藻毒素是一种肝毒素,它攻击的靶位是肝细胞,能导致鱼的肝脏发生病理变化。该研究以斑马鱼为研究对象,采用RT-PCR方法研究了微囊藻毒素对斑马鱼白细胞介素1β(Interleukin 1β,IL-1β),白细胞介素8(Interleukin 8,IL-8),白细胞介素10(Interleukin 10,IL-10),干扰素(Interferon,IFN),肿瘤坏死因子(Tumor Necrosis Factor,TNF),CXC,CXCa等几个重要细胞因子表达的影响。实验表明,25d时,仅毒素组的斑马鱼肝中有IFN和TNF表达。35d时,毒素组斑马鱼鳃中表达了IL-10,肝中表达了CXCa;毒素组和对照组的斑马鱼肝中都表达了IL-1β,TNF,IL-10,CXC,IFN,且其IL-1β,TNF,IL-10的条带亮于对照组。45d时,IFN仅在毒素组鱼鳃和肝中检测到表达;CXC仅在毒素组的鳃中检测到表达;IL-10在对照组的鳃、肠、肾和肌肉中有表达,在毒素组的肝、卵、肾、肌肉中有表达;IL-1β在对照组和毒素组的肝、卵、肾和肌肉中均有表达,且其条带亮于对照组。结果表明:毒素组斑马鱼的肝和鳃组织发生了炎症反应。这为进一步研究微囊藻毒素对鱼类免疫系统的毒害作用奠定了基础。     

The escape response of the zebra danio (Brachydanio rerio) II. The effect of experience

Naïve zebra danios (Brachydanio rerio) flee from an approaching predator when the angle subtended at the danio's eye by the predator changes at a rate greater than some characteristic threshold value. The hypothesis was tested that this value declines, and hence reactive distance increases, as a function of the number of past experiences with the predator. Artificial predators of two types, a model and a film of an approaching object, were presented to the danios daily and changes in behaviour recorded. The rate of change of visual angle at the time of response to the predator (dα/dt threshold) declined asymptotically with increasing experience. Consequently, flight distance increased asymptotically. The change was not due to growth, maturation, or increased familiarity with the testing situation. The danios apparently associated the visual characteristics of the predator with its subsequent high rate of angular growth and began to respond to these visual characteristics prior to dα/dt exceeding the threshold. The angle of escape was not affected by experience. Escape velocity increased with experience of the film predator, but not with experience of the model predator. No decrement of the learned change in reactive distance was evident after 10 days without experience with the film predator.     

The escape response of the zebra danio (Brachydanio rerio) I. The stimulus for escape

The hypothesis was tested that zebra danios (Brachydanio rerio) react to an approaching predator when the rate of change of the angle subtended by the predator at the prey's eye (dα/dt) exceeds some threshold level. Danios were presented with real predators (largemouth bass, Micropterus salmoides) and artificial predators of two types: a model predator, whose size and velocity could be varied, and a film of an approaching object. The reactive distance for flight (RD) could be predicted from the relationship: RD=[(VS/k)?(S²/4)]^1/2 where V is the predator approach velocity (cm/s), S is the predator front diameter (cm), and k is the threshold rate of change of visual angle (radians/s). The best overall estimate of k was 0·43 rad/s. The reactive field was shown to be circular, demonstrating that the mechanism is not solely a binocular one. Escape velocity, though higher in response to real than to artificial predators, was not correlated with reactive distance.     

Developmental variability during early embryonic development of zebra fish, Danio rerio

Early vertebrate embryos pass through a period of remarkable morphological similarity. Possible causes for such similarity of early embryos include modularity, developmental constraints, stabilizing selection, canalization, and exhausted genetic variability. Supposedly, each process creates different patterns of variation and covariation of embryonic traits. We study the patterns of variation of the embryonic phenotype to test ideas about possible evolutionary mechanisms shaping the early embryonic development. We use the zebra fish, Danio rerio, as a model organism and apply repeated measures of individual embryos to study temporal changes of phenotypic variability during development. In particular, we are looking at the embryonic development from 12 hours post fertilization until 27 hours post fertilization. During this time period, the development of individual embryos is documented at hourly intervals. We measured maximum diameter of the eye, length of embryo, number of somites, inclination of somites, and the yolk size (as a maternal effect). The coefficient of variation (CV) was used as a measure of variability that was independent of size. We used a principal component analysis for analysis of morphological integration. The experimental setup kept environment x genotype interactions constant. Nongenetic parental contributions had no significant effects on interindividual variability. Thus all observed phenotypic variation was based on additive genetic variance and error variance. The average CV declined from 14% to 7.7%. The decline of the CV was in particular expressed during 15-19 h post fertilization and occurred in association with multiple correlations among embryonic traits and a relatively high degree of morphological integration. We suggest that internal constraints determine the patterns of variability during early embryonic development of zebra fish.     

Reproduction of the annual fish Austrolebias nigrofasciatus (Rivulidae) maintained at different temperatures

Temperature is considered an important factor that influences the life cycle of annual fishes, however the thermal preferendum of Austrolebias nigrofasciatus reproduction has not been defined. The hypothesis was that the fecundity of the species would respond differently within the temperature range analyzed, presenting an optimum thermal. Thus, this study investigated the effect of temperature on the fecundity of A. nigrofasciatus for the first time under laboratory conditions. Pairs of A. nigrofasciatus were placed in aquaria with temperatures of 17, 21 or 25°C, with four pairs (replicates) for each treatment, for five weeks. Contrary to expected, there were no significant differences in fecundity parameters among the tested temperatures, but the weekly fecundity at 17 and 21°C was more homogeneous than at 25°C. During the experimental period, the temperature did not affect male growth; however, at 25°C there was a negative effect on female body growth. The condition factor was also influenced, suggesting a tendency towards a negative energy balance with increasing temperature. The results indicate that temperatures between 17 and 21°C are suitable for broodstock maintenance.     

Growth and development of Hymenolepis nana in mice maintained at different environmental temperatures

Growth and development of Hymenolepis nana in mice maintained at different environmental temperatures. International Journal for Parasitology16: 13–17. On days 3 and 4 post infection (p.i.) the number of Hymenolepis nana cysticercoids in the villi of male mice kept at 5°C did not differ from those in controls (21°C), but fewer larvae were observed in hosts at 35°C. However, on day 10 and 14 p.i. in cysticercoid and egg-induced infections respectively, the incidence of infection was higher and significantly more worms per host were found in mice at 5°C than in those kept at 21 or 35°C. Also, worms from mice maintained at 5°C were significantly heavier and became patent 1 day earlier than those from 21°C, which, in turn, were significantly heavier than those grown in hosts at 35°C.     

The Use of the Zebra fish,Brachydanio rerio,for the Study of Embryological Development in Schools

Courtship behaviour and mating patterns are easily observed in millipedes and experiments can be performed to demonstrate the functional significance of behaviours that have evolved through male-male competition or female choice. We describe simple methodologies and give examples of likely results together with a brief theoretical interpretation for two series of laboratory experiments. These experiments provide a useful framework for tackling the abstract concepts of sexual selection in practical classes at undergraduate level.     

Identification of two families of satellite-like repetitive DNA sequences from the zebrafish (Brachydanio rerio).

To further our understanding of the structure and organization of the zebrafish genome, we have undertaken the analysis of highly and middle-repetitive DNA sequences. We have cloned and sequenced two families of tandemly repeated DNA fragments. The monomer units of the Type I satellite-like sequence are 186 bp long, A+T-rich (65%), and exhibit a high degree of sequence conservation. The Type I satellite-like sequence constitutes 8% of the zebrafish genome, or approximately 8 x 10(5) copies per haploid genome. Southern analysis of genomic DNA, digested with several restriction endonucleases, shows a ladder of hybridizing bands, consistent with a tandem array, and suggests longer range periodic variations in the sequence of the tandem repeats. The Type II satellite has a monomer length of 165 bp, is also A+T-rich (68%), and constitutes 0.2% of the zebrafish genome (22,000 copies per haploid genome). Southern analysis reveals a complex pattern rather than a ladder of regularly spaced hybridizing bands.