Quantitative genetic properties of four measures of deformity in yellowtail kingfish Seriola lalandi Valenciennes, 1833.

The main aim of this study was to estimate the heritability for four measures of deformity and their genetic associations with growth (body weight and length), carcass (fillet weight and yield) and flesh-quality (fillet fat content) traits in yellowtail kingfish Seriola lalandi. The observed major deformities included lower jaw, nasal erosion, deformed operculum and skinny fish on 480 individuals from 22 families at Clean Seas Tuna Ltd. They were typically recorded as binary traits (presence or absence) and were analysed separately by both threshold generalized models and standard animal mixed models. Consistency of the models was evaluated by calculating simple Pearson correlation of breeding values of full-sib families for jaw deformity. Genetic and phenotypic correlations among traits were estimated using a multitrait linear mixed model in ASReml. Both threshold and linear mixed model analysis showed that there is additive genetic variation in the four measures of deformity, with the estimates of heritability obtained from the former (threshold) models on liability scale ranging from 0.14 to 0.66 (SE 0.32-0.56) and from the latter (linear animal and sire) models on original (observed) scale, 0.01-0.23 (SE 0.03-0.16). When the estimates on the underlying liability were transformed to the observed scale (0, 1), they were generally consistent between threshold and linear mixed models. Phenotypic correlations among deformity traits were weak (close to zero). The genetic correlations among deformity traits were not significantly different from zero. Body weight and fillet carcass showed significant positive genetic correlations with jaw deformity (0.75 and 0.95, respectively). Genetic correlation between body weight and operculum was negative (-0.51, P < 0.05). The genetic correlations' estimates of body and carcass traits with other deformity were not significant due to their relatively high standard errors. Our results showed that there are prospects for genetic selection to improve deformity in yellowtail kingfish and that measures of deformity should be included in the recording scheme, breeding objectives and selection index in practical selective breeding programmes due to the antagonistic genetic correlations of deformed jaws with body and carcass performance.

Strain variation in Mycobacterium marinum fish isolates.

A molecular characterization of two Mycobacterium marinum genes, 16S rRNA and hsp65, was carried out with a total of 21 isolates from various species of fish from both marine and freshwater environments of Israel, Europe, and the Far East. The nucleotide sequences of both genes revealed that all M. marinum isolates from fish in Israel belonged to two different strains, one infecting marine (cultured and wild) fish and the other infecting freshwater (cultured) fish. A restriction enzyme map based on the nucleotide sequences of both genes confirmed the divergence of the Israeli marine isolates from the freshwater isolates and differentiated the Israeli isolates from the foreign isolates, with the exception of one of three Greek isolates from marine fish which was identical to the Israeli marine isolates. The second isolate from Greece exhibited a single base alteration in the 16S rRNA sequence, whereas the third isolate was most likely a new Mycobacterium species. Isolates from Denmark and Thailand shared high sequence homology to complete identity with reference strain ATCC 927. Combined analysis of the two gene sequences increased the detection of intraspecific variations and was thus of importance in studying the taxonomy and epidemiology of this aquatic pathogen. Whether the Israeli M. marinum strain infecting marine fish is endemic to the Red Sea and found extremely susceptible hosts in the exotic species imported for aquaculture or rather was accidentally introduced with occasional imports of fingerlings from the Mediterranean Sea could not be determined.

Photobacterium damselae ssp. piscicida: detection by direct amplification of 16S rRNA gene sequences and genotypic variation as determined by amplified fragment length polymorphism (AFLP).

A PCR protocol for the rapid diagnosis of fish 'pasteurellosis' based on 16S rRNA gene sequences was developed. The procedure combines low annealing temperature that detects low titers of Photobacterium damselae but also related species, and high annealing temperature for the specific identification of P. damselae directly from infected fish. The PCR protocol was validated on 19 piscine isolates of P. damselae ssp. piscicida from different geographic regions (Japan, Italy, Spain, Greece and Israel), on spontaneously infected sea bream Sparus aurata and sea bass Dicentrarchus labrax, and on closely related American Type Culture Collection (ATCC) reference strains. PCR using high annealing temperature (64 degrees C) discriminated between P. damselae and closely related reference strains, including P. histaminum. Sixteen isolates of P. damselae ssp. piscicida, 2 P. damselae ssp. piscicida reference strains and 1 P. damselae ssp. damselae reference strain were subjected to Amplified Fragment Length Polymorphism (AFLP) analysis, and a similarity matrix was produced. Accordingly, the Japanese isolates of P. damselae ssp. piscicida were distinguished from the Mediterranean/European isolates at a cut-off value of 83% similarity. A further subclustering at a cut-off value of 97% allowed discrimination between the Israeli P. damselae ssp. piscicida isolates and the other Mediterranean/European isolates. The combination of PCR direct amplification and AFLP provides a 2-step procedure, where P. damselae is rapidly identified at genus level on the basis of its 16S rRNA gene sequence and then grouped into distinct clusters on the basis of AFLP polymorphisms. The first step of direct amplification is highly sensitive and has immediate practical consequences, offering fish farmers a rapid diagnosis, while the AFLP is more specific and detects intraspecific variation which, in our study, also reflected geographic correspondence. Because of its superior discriminative properties, AFLP can be an important tool for epidemiological and taxonomic studies of this highly homogeneous genus.

Mycobacteriosis in wild rabbitfish Siganus rivulatus associated with cage farming in the Gulf of Eilat, Red Sea.

Infection patterns of Mycobacterium marinum were studied over a period of 3 yr in wild rabbitfish Siganus nivulatus populations associated with commercial mariculture cages and inhabiting various sites along the Israeli Red Sea coastline. Mycobacteriosis was first recorded from the Red Sea in 1990 in farmed sea bass Dicentrarchus labrax and is absent from records of studies on parasites and diseases of wild rabbitfish carried out in the 1970s and 1980s. A sharp increase in the prevalence of the disease in cultured and wild fish in the region has occurred since. A total of 1142 rabbitfish were examined over a 3 yr period from inside mariculture net cages, from the cage surroundings and from several sites along the coast. Histological sections of spleens were examined for presence of granulomatous lesions. Overall prevalence levels of 50% were recorded in the rabbitfish sampled inside the net cages and 39% at the cages' close surroundings, 21% at a sandy beach site 1.2 km westwards, 35% at Eilat harbour 3 km to the south and 42% at a coral reef site about 10 km south of the cages. In addition, 147 fish belonging to 18 native Red Sea species were sampled from 2 sites, the net cage farm perimeter and the coral reef area, and examined for similar lesions. None of those from the coral reef were infected with Mycobacterium; however, 9 of 14 species collected from the cage surroundings were infected. An increase in prevalence of mycobacteriosis in the mariculture farm area was noted from 1995 to 1997. At the same time, a significant increase in prevalence was also apparent at the coral reef sampling site. Two M. marinum isolates from rabbitfish captured at Eilat harbour and the coral reef site were shown by 16S rDNA sequencing analysis to be identical to isolates from rabbitfish trapped inside the mariculture cages as well as isolates from locally cultured sea bass D. labrax. The implications of spreading of M. marinum infection in wild fish populations in the Gulf of Eilat are discussed.

Association of Foreign DNA with Sperm of Gilthead Seabream, Sparus aurata, After Sonication, Freezing, and Dimethyl Sulfoxide Treatments.

: The association of foreign DNA with gilthead seabream (Sparus aurata) sperm was enhanced relative to simple coincubation by sonication, freezing, dimethyl sulfoxide and polyethylene glycol treatments. Sonication yielded the strongest dot blot signals, equivalent to 250 to 380 foreign DNA copies per spermatozoa. We are unaware of previous reports attempting to associate DNA with ultrasound for fish or elsewhere. However, no or negligible foreign DNA was evident in 1- and 2-day-old fish larvae resulting from eggs fertilized with sonicated or frozen sperm.

Cloning and sequencing of dolphinfish (Coryphaena hippurus, Coryphaenidae) growth hormone-encoding cDNA.

The cDNA encoding the preprotein growth hormone from the dolphinfish (Coryphaena hippurus) has been cloned and sequenced. The cDNA was derived by reverse transcription of RNA from the pituitary of a young fish using the method known as Rapid Amplification of cDNA Ends (RACE). An oligonucleotide primer corresponding to the 5' region of Pagrus major and the universal RACE primer enabled amplification using the Polymerase Chain Reaction (PCR). The dolphinfish and yellow-tail, Seriola quineqeradiata, are both members of the sub-order Percoidei (Perciforme) and their GH sequences show a high level of homology.

Detection and identification of a pathogenic marine Mycobacterium from the European seabass Dicentrarchus labrax using polymerase chain reaction and direct sequencing of 16S rDNA sequences.

Mycobacteriosis has become a major concern for the commercial mariculture of the European sea bass Dicentrachus labrax in Israel. The disease remains asymptomatic for a long time, is virtually impossible to eradicate with antibiotics, stunts the growth of the fish and renders the fish unmarketable. The pathogen was identified as Mycobacterium marinum by direct sequencing and analysis of approximately 600 bp of the pathogen ribosomal encoding DNA (rDNA). The polymerase chain reaction technique was evaluated as a diagnostic tool for detecting the infection in D. labrax and found to be highly specific and sensitive.

Genetics and linkage mapping of Drosophila buzzatii.

Of 51 visible mutants isolated from natural or laboratory populations of Drosophila buzzatii, or X-ray induced, 42 have been assigned to chromosomes, and linkage maps have been constructed. About half of the autosomal mutants map to chromosome 2, with only two on chromosome 3 and none on chromosome 4. For the whole repleta group, chromosome 2 also exhibits much greater inversion variability than other chromosomes, which suggests variation among chromosomes in apparent mutability. The chromosomes of D. buzzatii are homologized to those of D. melanogaster and to the standard chromosomal elements of Drosophila. Sequence comparisons for six X chromosome mutant genes, whose homology is reasonably certain, in 13 Drosophila species confirm linkage group conservation but great variation among species in gene order. The linkage group conservation of single-copy genes stands in contrast to observed transpositions between elements for tandem repeat genes.

Genetic analysis of chromosomal region 97D2-9 of Drosophila melanogaster.

The rough homeobox gene of D. melanogaster is required for the correct patterning of the developing eye. The locus maps to cytological location 97D2-5, a region which has not been extensively characterised. As part of our genetic and molecular characterization of rough we carried out an EMS mutagenesis to generate mutants that map to the surrounding region, 97D2-9 which is deleted in Df(3R)ro-XB3. We have generated 1 visible and 13 lethal mutations which, together with the previously described Toll and ms(3)K10 loci, and other unpublished lethals, define nine complementation groups--four lethal, three semi-lethal, one visible and one male-sterile. In addition to rough, one other locus within this region, 1(3)97De, was shown to be required for formation of the normal pattern of photoreceptor cells in the compound eye.

Molecular cloning and sequencing of Australian black bream Acanthopagrus butcheri and barramundi Lates calcarifer fish growth hormone cDNA using polymerase chain reaction.

The complementary DNA (cDNA) encoding the preprotein growth hormone (pre-GH) from two Australian marine fish species, namely Acanthopagrus butcheri and Lates calcarifer, have been isolated, cloned and sequenced. The sequences were amplified from reverse transcribed total RNA of whole brains using Polymerase Chain Reaction (PCR) and oligonucleotide primers corresponding to the 5' and 3' regions of Pagrus major. Use of PCR offers a rapid method of isolating fish GH cDNA sequences for commercial and taxonomic applications. Sequence comparison indicates a high degree of conservation for GH cDNAs within the family Sparidae.

The genetics of abnormal abdomen, incomplete abdomen, and bobbed in Drosophila buzzatii.

Five stocks of Drosophila buzzatii with superficially similar abdominal disruptions including partial tergite and sternite loss were isolated by inbreeding. Three of the stocks have indistinguishable phenotypes, the inheritance of which is maternally influenced. This phenotype and its mode of inheritance bear similarities with those of Abnormal abdomen in D. melanogaster. The phenotype in the fourth stock is slightly different and is due to a single autosomal recessive gene, which we denote incomplete abdomen. In the fifth stock the trait is limited to females, and in appearance and mode of inheritance resembles bobbed in D. melanogaster. Furthermore, only in this stock are rDNA deletions evident. The combined frequencies of the three types of abdominal aberration were found to be around 1% in several samples from wild and laboratory populations of D. buzzatii.

Polymorphic inversion and esterase loci complex on chromosome 2 of Drosophila buzzatii. II. Spatial variation.

The potential influence of linked inversions on allele frequency variation at the Est-1 and Est-2 loci among Australian populations of D. buzzatii was determined by statistical analyses allele and inversion gametic frequencies. Most of the significant spatial and climatic associations found for all Est-1 allele frequencies, and for one allele only of Est-2 (Est-2c+), were accounted for by their linkage disequilibria with the inversions, which covaried with environmental variables. Consistent with this result, the spatial and climatic associations for conditional Est-1 and Est-2 allele frequencies tended to be different from those for the respective unadjusted allele frequencies. In one important respect, the results for Est-1 and Est-2 were not altered by inversions. For both unadjusted and conditional Est-1 allele frequencies, few climatic associations remain after correcting for geographic location, whereas for both unadjusted and conditional Est-2 allele frequencies, climatic associations remain after correcting for geographic location. Thus, apparent selection affecting allele frequencies at the Est-2 locus is not accounted for by linked inversions.

Selection affecting enzyme polymorphisms in enclosed Drosophila populations maintained in a natural environment.

Allele frequencies for the Adh, Gpdh, and Est6 enzyme polymorphisms of Drosophila melanogaster show large-scale latitudinal clines, whereas those for Pgm do not vary systematically with latitude. To elucidate possible mechanisms of selection underlying these distributions, large collections of the species were made from five Australasian localities spanning 24 degrees of latitude. Two replicate experimental populations were established from each collection, and each replicate was then released into an enclosure surrounding a natural habitat at a central-latitude locality. Genotype frequencies at the four loci were monitored for 15 months, covering 12 discrete generations, and selection coefficients on each polymorphism were then estimated by maximum likelihood procedures. For Est6 no coefficients were found to be significantly different from zero. For Pgm some nonzero coefficients were estimated, but these were heterogeneous across experimental populations of different geographic origins. For both Adh and Gpdh, nonzero selection coefficients were estimated that were homogeneous across populations and indicated heterozygote advantage. Predicted Adh and Gpdh equilibrium allele frequencies were consistent with those found in adjacent free-living populations. It is concluded that, at such intermediate latitudes at least, selection operates on the Adh and Gpdh polymorphisms to the advantage of heterozygotes.

Observations on the extent and temporal stability of latitudinal clines for alcohol dehydrogenase allozymes and four chromosome inversions in Drosophila melanogaster.

Previously we have presented evidence of large-scale latitudinal clines in the frequencies of four chromosome inversions and alleles at six enzyme loci in populations of D. melanogaster in Australasia, Asia and North America. Subsequent sampling by others in Japan and western U.S.A. has failed to repeat this observation for the steepest of the clines (alcohol dehydrogenase and the four chromosome inversions). We argue that this failure reflects the few populations and small latitudinal range sampled in these later studies. From extensive sampling over a long latitudinal transect in Australasia we here document Adh and inversion clines which are virtually identical to those originally obtained in different Australian populations four years earlier. We also repeat our observation that the Adh cline is largely independent of the cline in the linked inversion In(2L)t. We therefore retain our original conclusion that these polymorphisms are subject to natural selection. However the new Australasian data do not indicate an association between Adh and maximum rainfall which had been evident in the earlier data for Australasia, Asia and North America. We therefore retract our claim that the selective agent on Adh is related to rainfall.

Chromosome Inversion Polymorphisms in DROSOPHILA MELANOGASTER. I. Latitudinal Clines and Associations between Inversions in Australasian Populations.

Nineteen Australasian populations of Drosophila melanogaster have been screened for chromosome inversion polymorphisms. All 15 of the inversion types found are paracentric and autosomal, but only four of these, one on each of the major autosome arms, are common and cosmopolitan. North-south clines occur, with the frequencies of all four of the common cosmopolitan inversions increasing toward the equator. These clines in the Southern Hemisphere mirror north-south clines in the Northern Hemisphere, where the frequencies of all four of the common cosmopolitan inversions again increase towards the equator.-While few of the Australasian populations show significant disequilibrium between linked common cosmopolitan inversions, those that do invariably have excesses of coupling gametes, which is consistent with other reports. We also find nonrandom associations between the two major autosomes, with the northern populations in Australasia (those with high inversion frequencies) tending to be deficient in gametes with common cosmopolitan inversions on both major autosomes, while the southern populations in Australasia (low inversion frequencies) tend to have an excess of this class of gametes.-The clines and the nonrandom associations between the two major autosomes are best interpreted in terms of selection operating to maintain the common cosmopolitan inversion polymorphisms in natural populations of D. melanogaster.