Loss of genetic variability in the captive stocks of tambaqui,Colossoma macropomum (Cuvier, 1818), at breeding centres in Brazil,and their divergence from wild populations

The loss of variability in farmed populations and the risks of interactions with wild populations support the need for the genetic monitoring of species farmed throughout the world. In Brazil, the tambaqui is the most widely farmed native fish species. Despite this, there are no data on the pedigree of the farmed stocks, and the potential for interactions with wild populations in the Amazon basin has raised concerns with regard to the genetic variability of these stocks. The present study analysed sequences of the mitochondrial Control Region and 12 microsatellites to characterize the genetic variability of seven historically important commercial tambaqui breeding centres located in four different regions of Brazil, and compared these sequences with those obtained from individuals collected from a wild population. High levels of genetic diversity were found in the wild population, whereas genetic diversity was reduced in both markers in most captive populations, except for the broodstock located near the Amazon River. High F_ST and D_EST indices were recorded between the wild population and most of the captive stocks analysed. The drastic reduction in genetic diversity found in most captive stocks and the difference between these stocks and the wild population may have been the result of the small size of the founding populations and the absence of breeding management. The renewal of the broodstocks and the application of breeding management techniques are recommended. In the Amazon region, in addition, the use of broodstocks that are genetically very different from local wild populations should be avoided.     

High immune diversity in farmed Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis> L.)

As with any other intensively farmed animal species, the Atlantic salmon has been selectively bred and cross-bred to maximize desirable traits. Selection tends to diminish genetic diversity in target populations, which among other negative effects is hypothesized to decrease their capacity to confront a broad variety of pathogens. We have analyzed mitochondrial (mtDNA) as well as major histocompatibility complex (MHC) DNA sequences from individuals collected from a single aquaculture cage in southern Chile. Interestingly, only two mtDNA haplotypes were obtained; however, several different MH alleles were detected, with divergence values that were compared with those of natural populations of salmonids. Thus, it seems evolutionary processes responsible of keeping MH diversity in the wild managed to retain MH variability in farmed Atlantic salmon, maintaining high immune diversity despite the generally lower levels of observed neutral genetic diversity.     

Reproductive strategies in small populations: using Atlantic salmon as a case study

Abstract –  Wild salmonid populations with only a few breeding adults may not exhibit a significant reduction in genetic variability compared with larger populations. Such an observation suggests that effective population sizes are larger than population size estimates based on direct adult counts and/or the mating strategy maximises outbreeding, contributing to increased heterozygosity. In the case of wild Atlantic salmon Salmo salar populations, stratification by age classes and sexes on the spawning grounds avoids inbreeding and increases genetic variability. We studied the breeding composition of four Spanish salmon populations. Over a 7-year period we concluded that the probability of within-cohort mating is very low: females generally reproduce after two sea-winters whereas males reproduce mostly as one sea-winter ( grilse ) and/or mature parr. Considering different levels of contribution of mature parr to spawning derived from field surveys, we developed a simple model for estimating effective population sizes and found that they doubled with 65% parr contribution expected for rivers at this latitude (43°N), and ranged from 100–800 individuals. The effect of between-cohort mating was modelled considering different ranges of differences in allele frequencies between cohorts and resulted in 28–50% increases in heterozygosity when considering a 65% parr contribution. The complex mating strategy of Atlantic salmon contributes to explain the high levels of genetic variability found for small populations of this species. This model can probably be extended to other animal species with mating strategies involving different cohorts.     

An assessment of genetic diversity in wild and captive populations of endangered Japanese bitterling <Emphasis Type="Italic">Tanakia tanago</Emphasis> (Cyprinidae) using amplified fragment length polymorphism (AFLP) markers

ABSTRACT:   The Japanese bitterling Tanakia tanago (Cyprinidae) is on the verge of extinction in the wild, placing great importance on captive breeding programs for current conservation of the species. However, the loss of genetic diversity during captive breeding is an ongoing matter of concern. Since some captive populations have been almost monomorphic in mitochondrial DNA (mtDNA), this hampers assessments of their genetic diversity during captive breeding. To more accurately assess their genetic diversity, one wild and three captive populations were examined using amplified fragment length polymorphism (AFLP) markers. Estimates of average heterozygosity and nucleotide diversity ranged 0.0479–0.1920 and 0.0023–0.0088, respectively, enabling comparison of genetic diversity among the wild and captive populations, and among year-classes of captive populations. Significant differences in numbers of amplified fragments and proportions of polymorphic fragments were observed among year-classes of all populations. The indices of genetic diversity calculated from AFLP seemed to be, however, less sensitive to weak bottlenecks. No continuous decrease in genetic diversity in nuclear DNA was detected in presently captive populations. This supports the possibility of re-introduction of the captive populations into the original habitats, although survival and reproductive ability in the wild must be taken into consideration.     

Genetic variation in farmed populations of the gilthead sea bream Sparus aurata in Greece using microsatellite DNA markers

Genetic variation in seven reared stocks of gilthead sea bream Sparus aurata, originating from Greek commercial farms, was assessed using five polymorphic microsatellite markers and was compared with that of two natural populations from the Ionian and the Adriatic Seas. The total number of alleles per marker ranged from 11 to 19 alleles, and hatchery samples showed the same levels of observed heterozygosity with samples from the wild but substantially smaller allelic diversity and expected heterozygosity. The global genetic differentiation for the cultivated samples was significant as indicated by F_st analysis, which might indicate random genetic drift and inbreeding events operating in the hatcheries. On the contrary, no significant difference was found between the two wild populations. Population pairwise tests between farmed and wild stocks were also significant, with the exception of one hatchery sample, the Central Greece 1, which was not significantly different from the two wild samples perhaps due to its recent use in aquaculture from wild‐caught animals. The UPGMA tree topology grouped the wild samples together with the Central Greece 1 stock, and showed a clear division between wild and farmed sample sets for the six remaining hatchery samples. Knowledge of the genetic variation in S. aurata cultured populations compared with that in the wild ones is essential for setting up appropriate guidelines for the proper monitoring and management of the stocks either under traditional practices or for the implementation of selective breeding programmes.     

团头鲂3个选育群体遗传潜力的微卫星分析

为从遗传多样性的角度了解团头鲂(Megalobrama amblycephala)3个选育群体的遗传潜力,该研究以团头鲂"浦江1号"选育奠基群体(F_0)为对照组,采用14个多态性转录组微卫星标记评估了团头鲂3个选育群体的遗传多样性,分析其遗传潜力。结果显示,3个选育群体平均每个位点的等位基因数(A)为7.928 6~8.785 7,有效等位基因数(A_E)为4.409 4~4.878 4,观察杂合度(H_O)为0.491 1~0.574 4,期望杂合度(HE)为0.741 3~0.751 8,多态信息含量(PIC)为0.691 2~0.705 2,近交系数(FIS)为0.229~0.352。3个选育群体的遗传多样性水平(AE、HE)均高于F0群体,但不存在显著差异(P0.05)。3个选育群体的有效群体大小(N_e)为11.0~29.3,在近期可能经历过遗传瓶颈。3个选育群体间D_A、D_(SW)遗传距离分别为0.175 4~0.358 8、0.804 7~1.054 4。该结果表明,3个选育群体的遗传多样性较高,遗传潜力较大,但因有效群体数量较少和瓶颈效应的影响,存在杂合度下降和近交衰退的风险,今后需采取科学措施来保护选育群体的遗传潜力。     

A comparison of the swimming and cardiac performance of farmed and wild Atlantic salmon, Salmo salar, before and after gamete stripping

Farmed Atlantic salmon, Salmo salar, frequently escape from the aquaculture industry and interact with wild populations. The impact of these interactions on the wild populations will depend, in part, on differences in their performances. This study compared the swimming and cardiac performance of farmed salmon (Aquagen) with their founder population from the River Namsen both before and after gamete stripping. Cardiac output (CO), heart rate (HR), and stroke volume (SV), which were measured by placing Doppler flow probes around the ventral aorta of the fish, increased with exercise, but the response did not significantly differ between farmed and wild salmon. Similarly, the swimming performance of wild salmon never significantly differed from the farmed salmon. The overall similarity in swimming and cardiac performance between farmed and wild Atlantic salmon observed in the present study suggests that cultured salmon may have the ability to be competitive with the wild salmon in native waters.     

Genetic relationship between broodstocks of olive flounder, Paralichthys olivaceus (Temminck and Schlegel) using microsatellite markers

For the first generation of a selective breeding programme, it is important to minimize the possibility of inbreeding. This mostly occurs by mating between closely related individuals, while proper mating can provide an opportunity to establish the base families with wide genetic variation from which selection for subsequent generations can be more effective. Genotyping with microsatellite‐based DNA markers can help us determine the genetic distances between the base populations. The genetic markers further facilitate the identification of the correct parents of the offspring (parentage assignments) reared together with many other families after hatching. We established a genetic analysis system with microsatellite DNA markers and analysed the genetic distances of three farmed stocks and a group of fish collected from wild populations using eight microsatellite markers. The averaged heterozygosity of the farming stocks was 0.826 and that of the wild population was 0.868. The hatchery strains had an average of 8.6 alleles per marker, which was less than a wild population that carried an average of 14.3 alleles per marker. Significant Hardy–Weinberg disequilibrium (HWDE) was observed in two farming stocks (P<0.05). Despite relatively low inbreeding coefficiency of the hatchery populations, the frequency of a few alleles was highly represented over others. It suggests that the hatchery stocks to some extent have experienced inbreeding or they originated from closely related individuals. We will develop a selective program using the DNA markers and will widen the usage of the DNA‐based genetic analysis system to other fish species.     

Genetic comparison of experimental farmed strains and wild Icelandic populations of Atlantic cod (Gadus morhua L.)

Microsatellite DNA loci and the Pantophysin locus (Pan I) were used to investigate levels of genetic diversity within farmed strains of Atlantic cod Gadus morhua and to compare them with the wild source population. A total of 282 farmed samples originating from a spawning ground off the south-west coast of Iceland were sampled in the years 2002 and 2003, and 258 wild cod were collected at the same spawning ground in the same years. The farmed strains exhibited a lower mean number of alleles and allelic diversity than the wild samples at the microsatellite loci. Significant differences were observed between wild and farmed samples both in allele and genotype frequencies at the Pan I locus. We argue that the genetic divergence of wild and farmed samples of Atlantic cod may be due to a small number of effective founding breeders contributing to the genetic variation of the farmed strains, inducing a reduction in allelic diversity. We discuss the potential effect of breeding practices on the genetic diversity of Atlantic cod.     

Microsatellite analysis in domesticated and wild Atlantic salmon (Salmo salar L.): allelic diversity and identification of individuals

Samples of wild and domesticated salmon in Norway were genotyped at 12 microsatellite loci to compare allelic variability and investigate the potential of microsatellite markers for identification of individuals. The following loci were amplified: Ssa20, Ssa62NVH, Ssa71NVH, Ssa90NVH, Ssa103NVH, Ssa105NVH, SsaF43; Ssa20.19; Ssa13.37; SsOSL85; Ssa197; Ssa28. All domesticated strain samples displayed reduced variability compared to wild salmon. On average 58% of the allelic richness observed within the four wild stocks were present in the samples taken from domesticated strains. No systematic differences in heterozygosity were observed between samples representing the two groups.

Pairwise genetic distances, as estimated by Fst values and Nei [1978] was 2–8 times higher among domesticated strains than among wild strains. Among the wild stocks, the highest genetic distances were observed between the river Neiden, located in northern Norway, and the other wild stocks located in the southwest of Norway.

Assignment tests indicated that the wild and domesticated salmon could be distinguished with high precision. Less than 4% of domesticated salmon were misassigned as wild salmon, and less than 3% of wild fish were misassigned as domesticated salmon. Fish from individual domesticated strains were identified with similarly high precision. Assignment to wild salmon stocks was less accurate, with the exception of the sample taken from the river Neiden, for which 93% of the individuals were correctly assigned.     

人工雌核发育鲢近交F_2微卫星DNA变异分析

采用17对鲢微卫星引物,以野生鲢、养殖鲢、雄鲤作对照对人工雌核发育鲢近交F2及其亲本进行了微卫星分析。结果表明:人工雌核发育鲢近交F2、养殖鲢和野生鲢群体的平均等位基因数范围为2.1～4.0;平均观测杂合度范围为0.2762～0.9588;期望杂合度范围为0.2774～0.7360;遗传多样性指数范围为0.4500～1.2258,人工雌核鲢近交F2为0.4500,显著低于养殖鲢(1.0273)和野生鲢(1.2258),揭示人工雌核发育鲢近交F2遗传多样性水平较低,纯合度较高。从遗传距离来看,人工雌核发育鲢近交F2与对照群体之间的遗传距离都要大于野生鲢与养殖鲢群体之间遗传距离,表明人工雌核发育鲢近交F2发生了一定程度的遗传分化。     

Potential disease interaction reinforced: double-virus-infected escaped farmed Atlantic salmon,Salmo salar L., recaptured in a nearby river

The role of escaped farmed salmon in spreading infectious agents from aquaculture to wild salmonid populations is largely unknown. This is a case study of potential disease interaction between escaped farmed and wild fish populations. In summer 2012, significant numbers of farmed Atlantic salmon were captured in the Hardangerfjord and in a local river. Genetic analyses of 59 of the escaped salmon and samples collected from six local salmon farms pointed out the most likely source farm, but two other farms had an overlapping genetic profile. The escapees were also analysed for three viruses that are prevalent in fish farming in Norway. Almost all the escaped salmon were infected with salmon alphavirus (SAV) and piscine reovirus (PRV). To use the infection profile to assist genetic methods in identifying the likely farm of origin, samples from the farms were also tested for these viruses. However, in the current case, all the three farms had an infection profile that was similar to that of the escapees. We have shown that double-virus-infected escaped salmon ascend a river close to the likely source farms, reinforcing the potential for spread of viruses to wild salmonids.     

Migratory behaviour of adult wild and escaped farmed Atlantic salmon, Salmo salar L., before, during and after spawning in a Norwegian river

The migratory behaviour of adult wild and escaped farmed Atlantic salmon, Salmo salar L., before, during after spawning in the River Namsen, Norway, was analysed using radio telemetry. The fish were caught, radio tagged and released into the fjord between 7 and 25 km from the river mouth. A significantly higher proportion of wild (74%) than farmed (43%) salmon was subsequently recorded in the river. Wild salmon (33%) were more frequently captured in the sea and in rivers than farmed salmon (14%). The migration speed from release to passing a data logger 11 km upstream from the river mouth was not significantly different between wild (20.6 km day^?1) and farmed (19.8 km day^?1) salmon. Wild salmon tagged when water flow in the river was increasing had a significantly higher migration speed than wild salmon tagged when water flow was decreasing. This was not true for farmed salmon. Farmed salmon were distributed significantly higher up the river than wild salmon during spawning, although both types of fish were found together in spawning areas. Thus, there was no geographical isolation to prevent spawning between wild and escaped farmed salmon. Farmed salmon had significantly more and longer up- and downstream movements than wild salmon during the spawning period. Unlike farmed salmon, the number of riverine movements by wild salmon increased significantly when variation in water flow increased. A smaller proportion of wild (9%) than farmed (77%) salmon survived through the winter after spawning.     

福瑞鲤与豫选黄河鲤选育群体的遗传结构及亲本间遗传距离分布

用微卫星标记分析了鲤鱼(Cyprinus carpio L.)的2个品种福瑞鲤和豫选黄河鲤选育群体的遗传结构,并揭示了雌雄个体间遗传距离的分布规律。结果表明,23个微卫星标记在福瑞鲤(FR,n=192)和豫选黄河鲤(YX,n=96)中各检测到160个和131个等位基因。福瑞鲤的平均有效等位基因数(N_e)、观测杂合度(H_o)、期望杂合度(H_e)和多态信息含量(PIC)分别为4.559、0.695、0.741和0.702,群体处于高度多态水平(PIC≥0.5);豫选黄河鲤的4项遗传多样性参数分别为3.620、0.665、0.642和0.600。虽然豫选黄河鲤同样处于高度多态水平(PIC≥0.5),但是N_e、H_e和PIC均极显著低于福瑞鲤(P0.01),说明福瑞鲤的杂交选育背景决定了其较系统选育的豫选黄河鲤具有较多的来源于不同亲本的等位基因;而两者H_o差异不显著(P0.05),说明豫选黄河鲤种内也保持了较高的遗传杂合度。分别统计福瑞鲤与豫选黄河鲤雌雄个体间的遗传距离,结果表明两两雌雄个体间遗传距离呈正态分布。福瑞鲤个体间遗传距离的中间值位于0.8~1.0,占37.39%;而豫选黄河鲤个体间遗传距离中间值位于0.5~0.7,占49.33%。建议福瑞鲤和豫选黄河鲤在家系配组时,选择亲本间遗传距离阈值范围在0.8~1.0和0.5~0.7为宜。     

Genetic implications of translocation and stocking of fish species, with particular reference to Western Australia

Species or strains of fish may be translocated for farming, where the only access to the wild is via inadvertent escapes, or for stocking, where deliberate releases are undertaken. In either case, it is important that the translocated animals are representative of the donor population(s) in terms of genetic composition and level of variability. Many studies have shown that this ideal is difficult to achieve, the major reason being the use of inadequate numbers or composition of broodstock as founders of a strain. Also, where more than one conspecific population is involved, there may be outbreeding depression problems. In the case of farming, measures to improve the introduced strain genetically are likely to be undertaken, e.g. breeding programmes, manipulation of sex and ploidy, transgenic techniques. Such approaches are necessary economically, but can alter genetic make‐up. Thus, stringent attempts must be made to minimize escapes or reduce their impact should they occur. With stocking, genetic change during captive rearing should be avoided. No strain manipulation should be undertaken, and other agents of change should be minimized. Stocking may result in hybridization with related species or with endemic populations of the same species. In either case, there can be detrimental genetic effects on the native forms. To be able to identify subsequently any genetic changes in reared strains, whether intended for farming or stocking, wild population composition should be determined, using appropriate molecular techniques. Such molecular methods will demonstrate the degree of interpopulation differentiation and, thus, reproductive isolation. The same markers should then be used in each subsequent generation (in the hatchery and after escape or reintroduction to the wild) to monitor any changes in genetic composition or variability. Markers should include microsatellite DNA loci, but the inclusion of more than one type of marker is recommended. However, as the aforementioned markers are not considered to be influenced by natural selection, they give no information on the adaptive nature of such differences. For this reason, it is suggested that markers influenced by selection should be investigated. Monitoring a strain subsequent to deliberate or inadvertent release can be undertaken using genetic markers, either deliberately enhanced by breeding or occurring naturally. Highly variable minisatellite DNA loci have been used as family markers in farmed escape studies with Atlantic salmon. These investigations have demonstrated significantly superior survival of native strains compared with farmed salmon in natural stream conditions. These latter results, demonstrating fitness differences, were strongly indicative of local adaptation. Thus, methods exist to monitor the genetic effects of translocation and stocking. However, a holistic approach should be taken to such exercises, where genetics forms part of a wider suite of considerations.     

Identification of differential broodstock contribution affecting genetic variability in hatchery stocks of Atlantic salmon (Salmo salar)

Supportive breeding of Atlantic salmon (Salmo salar) is commonly employed to maintain numbers of fish where the species has become locally endangered. Increasingly, one of the main aims of population management is the preservation of natural genetic diversity. If the stocks employed in supportive breeding exhibit reduced variation they can alter the natural pattern of genetic variation observed in wild populations. In northern Spain, wild adult salmon are caught every year from local rivers and artificially crossed in order to create supportive stocks. The offspring are hatchery reared until the juvenile stage, then released into the same river where their parents were caught. In the current study, our findings demonstrate that although adult broodstock exhibit a pattern of variation similar to the wild populations, variability at microsatellite loci was drastically reduced in the juveniles released into one of three rivers analyzed. The contribution of broodstock to this juvenile stock was examined by pedigree analysis. A restricted number of females contributing to the hatchery stock was identified as the main cause of loss in genetic variation, possibly due to overmaturity of some multi-sea-winter females. We suggest that better monitoring and control of parental contribution will help in solving the problem of loss of genetic diversity in hatchery populations.     

Impact of stocked Atlantic salmon (Salmo salar L.) on habitat use by the wild population

Laffaille P. Impact of stocked Atlantic salmon (Salmo salar L.) on habitat use by the wild population.
Ecology of Freshwater Fish 2011: 20: 67–73. © 2010 John Wiley & Sons A/S Abstract – We investigated the summer habitat occupied by populations of young‐of‐the‐year wild and stocked (farmed populations released into the native range) Atlantic salmon under allopatric and sympatric conditions. Under allopatric conditions, farmed and wild salmon occupied habitats with the same characteristics. The salmon preferentially occupied the riffle areas. However, under sympatric conditions, the fish occupied meso‐ and micro‐habitats with different characteristics. Wild salmon avoided habitats used by farmed salmon and preferred glide areas with considerable vegetation cover. This study suggests that differences in the pattern of habitats used by young Atlantic salmon were both size‐ and origin‐dependent and may result from intra‐species competition between farmed and wild populations. Given that stocking with farmed Atlantic salmon is carried out intensively to enhance recreational angling or to conserve salmon populations, this study warns that this can have a negative impact on the extant wild Atlantic salmon population.     

Genestic differences between successive year classes of two strains of reared rainbow trout, Oncorhynchus mykiss (Walbaum)

The genetic composition of consecutive year classes of two farmed rainbow trout, Oncorhynchus mykiss (Walbaum), strains was assessed, using starch gel electrophoresis of 11 enzymes encoded by a minimum of 23 loci, many of which have been shown to be polymorphic in previous studies. Angle frequencies at the majority of polymorphic loci varied significantly between year classes of each strain. Several alleles which were present at low frequency in the 1900 year classes, were absent in the samples from the 1991 cohorts. However, mean heterozygosity per locus (H) did not differ significantly between year classes of either strain, illustrating that allelic diversity is a more sensitive indicator of loss of genetic variability than mean heterozygosity. This heterogeneity between cohorts is probably due either to broodstock maintenance practices such as the use of insufficient numbers of spawners, or, in the case of one strain, to bottlenecking caused by selection for late maturation and increased growth rate. Genetic monitoring of all year classes of reared strains is suggested, if insufficient breeding and distribution records are available from egg producers. Such records are often unavailable in commercial situations.     

Genetic variation of wild and cultured populations of the Kuruma prawn Marsupenaeus japonicus (Bate 1888) using microsatellites

The microsatellite DNA technique was used to detect the genetic variations between wild and cultured populations of Kuruma prawn Marsupenaeus japonicus Bate 1888. All the six microsatellite loci screened in this study showed high polymorphism for their PIC (0.6701–0.8989), which was much more than the standard value of 0.5. A total of 73 alleles were observed over six loci from 93 shrimps. The mean number of allele locus ranged from 9.83 (cultured) to 11.83 (wild). The number of effective alleles varied from 6.86 (cultured) to 8.58 (wild). The average of observed heterozygosity (H_o) of populations varied from 0.6935 (cultured) to 0.7370 (wild), and that of expected heterozygosity (He) was 0.8169 (wild) and 0.8209 (cultured). Tests of Hardy–Weinberg showed that these loci deviated significantly or highly significantly in one or both populations. Compared with the wild population, the cultured population showed little reduction in genetic variation. The total number of alleles (71, 59) was not significantly (P=0.296) different between wild and cultured populations. The paired‐samples t test of observed heterozygosity and expected heterozygosity implied that there was no significant difference (P=0.572 and 0.891 respectively) between wild and cultured populations. However, some rare allele loss might have occurred in the cultured population. A total of 14 unique alleles were found in the wild population, but only two unique alleles were observed in the cultured population. Therefore, there is a need to monitor genetic variability of cultured population, and to improve the hatchery program for the conservation of wild Kuruma prawn resources.