Development of type I genetic markers from expressed sequence tags in highly polymorphic species

Expressed sequence tag (EST) databases provide a primary source of nuclear DNA sequences for genetic marker development in non-model organisms. To date, the process has been relatively inefficient for several reasons: - 1) priming site polymorphism in the template leads to inferior or erratic amplification; - 2) introns in the target amplicon are too large and/or numerous to allow effective amplification under standard screening conditions, and; - 3) at least occasionally, a PCR primer straddles an exon–intron junction and is unable to bind to genomic DNA template. The first is only a minor issue for species or strains with low heterozygosity but becomes a significant problem for species with high genomic variation, such as marine organisms with extremely large effective population sizes. Problems arising from unanticipated introns are unavoidable but are most pronounced in intron-rich species, such as vertebrates and lophotrochozoans. We present an approach to marker development in the Pacific oyster Crassostrea gigas, a highly polymorphic and intron-rich species, which minimizes these problems, and should be applicable to other non-model species for which EST databases are available. Placement of PCR primers in the 3′ end of coding sequence and 3′ UTR improved PCR success rate from 51% to 97%. Almost all (37 of 39) markers developed for the Pacific oyster were polymorphic in a small test panel of wild and domesticated oysters.

Improving the effectiveness of age-abundance indicators in the management of fisheries in Queensland, Australia

The development of fishery indicators is a crucial undertaking as it ultimately provides evidence to stakeholders about the status of fished species such as population size and survival rates. In Queensland, as in many other parts of the world, age-abundance indicators (e.g. fish catch rate and/or age composition data) are traditionally used as the evidence basis because they provide information on species life history traits as well as on changes in fishing pressures and population sizes. Often, however, the accuracy of the information from age-abundance indicators can be limited due to missing or biased data. Consequently, improved statistical methods are required to enhance the accuracy, precision and decision-support value of age-abundance indicators.

Causes and consequences of inbreeding in the ant Formica exsecta

Human actions cause destruction and fragmentation of natural habitats, predisposing populations to loss of genetic diversity and inbreeding, which may further decrease their fitness and survival. Understanding these processes is a main concern in conservation genetics. Yet data from natural populations is scarce, particularly on invertebrates, owing to difficulties in measuring both fitness and inbreeding in the wild. Ants are social insects, and a prime example of an ecologically important group for which the effects of inbreeding remain largely unstudied. Social insects serve key roles in all terrestrial ecosystems, and the division of labor between the females in the colonies queens reproduce, workers tend to the developing brood probably is central to their ecological success. Sociality also has important implications for the effects of inbreeding. Despite their relative abundance, the effective population sizes of social insects tend to be small, owing to the low numbers of reproductive individuals relative to the numbers of sterile workers. This may subject social insects to loss of genetic diversity and subsequent inbreeding depression. Moreover, both the workers and queens can be inbred, with different and possibly multiplicative consequences. The aim of this study was to investigate causes and consequences of inbreeding in a natural population of ants. I used a combination of long-term field and genetic data from colonies of the narrow-headed ant Formica exsecta to examine dispersal, mating behavior and the occurrence of inbreeding, and its consequences on individual and colony traits. Mating in this species takes place in nuptial flights that have been assumed to be population-wide and panmictic. My results, however, show that dispersal is local, with queens establishing new colonies as close as 60 meters from their natal colony. Even though actual sib-mating was rare, individuals from different but related colonies pair, which causes the population to be inbred. Furthermore, multiple mates of queens were related to each other, which also indicates localized mating flights. Hence, known mechanisms of inbreeding avoidance, dispersal and multiple mating, were not effective in this population, as neither reduced inbreeding level of the future colony. Inbreeding had negative consequences both at the individual and colony level. A queen that has mated with a related male produces inbred workers, which impairs the colony s reproductive success. The inbred colonies were less productive and, specifically, produced fewer new queens, possibly owing to effects of inbreeding on the caste determination of female larvae. A striking finding was that males raised in colonies with inbred workers were smaller, which reflects an effect of the social environment as males, being haploid, cannot be inbred themselves. The queens produced in the inbred colonies, in contrast, were not smaller, but their immune response was up-regulated. Inbreeding had no effect on queen dispersal, but inbred queens had a lower probability of successfully founding a new colony. Ultimately, queens that survived through the colony founding phase had a shorter lifespan. This supports the idea that inbreeding imposes a genetic stress, leading to inbreeding depression on both the queen and the colony level. My results show that inbreeding can have profound consequences on insects in the wild, and that in social species the effects of inbreeding may be multiplicative and mediated through the diversity of the social environment, as well as the genetic makeup of the individuals themselves. This emphasizes the need to take into account all levels of organization when assessing the effects of genetic diversity in social animals.

Should I stay or should I go? : Reproductive and dispersal strategies of site-specific liverwort species

The area of intensively managed forests, in which required conditions for several liverwort species are seldom found, has expanded over the forest landscape during the last century. Liverworts are very sensitive to habitat changes, because they demand continuously moist microclimate. Consequently, about third of the forest liverworts have been classified as threatened or near threatened in Finland. The general objective of this thesis is to increase knowledge of the reproductive and dispersal strategies of the substrate-specific forest bryophytes. A further aim was to develop recommendations for conservation measures for species inhabiting unstable and stable habitats in forest landscape. Both population ecological and genetic methods have been applied in the research. Anastrophyllum hellerianum inhabits spatially and temporally limited substrate patches, decaying logs, which can be considered as unstable habitats. The results show that asexual reproduction by gemmae is the dominant mode of reproduction, whereas sexual reproduction is considerably infrequent. Unlike previously assumed, not only spores but also the asexual propagules may contribute to long-distance dispersal. The combination of occasional spore production and practically continuous, massive gemma production facilitates dispersal both on a local scale and over long distances, and it compensates for the great propagule losses that take place preceding successful establishment at suitable sites. However, establishment probability of spores may be restricted because of environmental and biological limitations linked to the low success of sexual reproduction. Long-lasting dry seasons are likely to result in a low success of sexual reproduction and decreased release rate of gemmae from the shoots, and consequent fluctuations in population sizes. In the long term, the substratum limitation is likely to restrict population sizes and cause local extinctions, especially in small-sized remnant populations. Contrastingly, larger forest fragments with more natural disturbance dynamics, to which the species is adapted, are pivotal to species survival. Trichocolea tomentella occupies stable spring and mesic habitats in woodland. The relatively small populations are increasingly fragmented with a high risk for extinction for extrinsic reasons. The results show that T. tomentella mainly invests in population persistence by effective clonal growth via forming independent ramets and in competitive ability, and considerably less in sexuality and dispersal potential. The populations possess relatively high levels of genetic diversity regardless of population size and of degree of isolation. Thus, the small-sized populations inhabiting stable habitats should not be neglected when establishing conservation strategies for the species and when considering the habitat protection of small spring sites. Restricted dispersal capacity, also on a relatively small spatial scale, is likely to prevent successful (re-)colonization in the potential habitat patches of recovering forest landscapes. By contrast, random short-range dispersal of detached vegetative fragments within populations at suitable habitat seems to be frequent. Thus, the restoration actions of spring and streamside habitats close to the populations of T. tomentella may contribute to population expansion. That, in turn, decreases the harmful effects of environmental stochasticity.

Pollinator limitation and the effect of breeding systems on plant reproduction in forest fragments

Reproduction of plants in fragmented habitats may be limited because of lower diversity or abundance of pollinators, and/or variation in local plant density. We assessed natural fruit set and pollinator limitation in ten species of woody plants in natural and restored fragments in the Pondicherry region of southern India, to see whether breeding system of plants (self-compatible and self-incompatible) affected fruit set. We tested whether the number of flowering individuals in the fragments affected the fruit set and further examined the adult and sapling densities of self-compatible (SC) and self-incompatible (SI) species. We measured the natural level of fruit set and pollinator limitation (calculated as the difference in fruit set between hand cross-pollinated and naturally pollinated flowers). Our results demonstrate that there was a higher level of pollinator limitation and hence lower levels of natural fruit set in self-incompatible species as compared to self-compatible species. However, the hand cross-pollinated flowers in SC and SI species produced similar levels of fruit set,further indicating that lower fruit set was due to pollinator limitation and not due to lack of cross-compatible individuals in the fragments. There was no significant relation between number of flowering individuals and the levels of natural fruit set, except for two species Derris ovalifolia, Ixora pavetta. In these species the natural fruit set decreased with increasing population size, again indicating pollinator limitation. The adult and sapling densities in self-compatible species were significantly higher than inself-incompatible species. These findings indicate that the low reproductive output in self-incompatible species may eventually lead to lower population sizes. Restoration of pollinator services along with plant species in fragmented habitats is important for the long-term conservation of biodiversity. (C) 2009 Elsevier Masson SAS. All rights reserved.

Stochastic Simulations Suggest that HIV-1 Survives Close to Its Error Threshold

The use of mutagenic drugs to drive HIV-1 past its error threshold presents a novel intervention strategy, as suggested by the quasispecies theory, that may be less susceptible to failure via viral mutation-induced emergence of drug resistance than current strategies. The error threshold of HIV-1, mu(c), however, is not known. Application of the quasispecies theory to determine mu(c) poses significant challenges: Whereas the quasispecies theory considers the asexual reproduction of an infinitely large population of haploid individuals, HIV-1 is diploid, undergoes recombination, and is estimated to have a small effective population size in vivo. We performed population genetics-based stochastic simulations of the within-host evolution of HIV-1 and estimated the structure of the HIV-1 quasispecies and mu(c). We found that with small mutation rates, the quasispecies was dominated by genomes with few mutations. Upon increasing the mutation rate, a sharp error catastrophe occurred where the quasispecies became delocalized in sequence space. Using parameter values that quantitatively captured data of viral diversification in HIV-1 patients, we estimated mu(c) to be 7 x 10(-5) -1 x 10(-4) substitutions/site/replication, similar to 2-6 fold higher than the natural mutation rate of HIV-1, suggesting that HIV-1 survives close to its error threshold and may be readily susceptible to mutagenic drugs. The latter estimate was weakly dependent on the within-host effective population size of HIV-1. With large population sizes and in the absence of recombination, our simulations converged to the quasispecies theory, bridging the gap between quasispecies theory and population genetics-based approaches to describing HIV-1 evolution. Further, mu(c) increased with the recombination rate, rendering HIV-1 less susceptible to error catastrophe, thus elucidating an added benefit of recombination to HIV-1. Our estimate of mu(c) may serve as a quantitative guideline for the use of mutagenic drugs against HIV-1.

Blue Crab target setting: final report

We have developed a hierarchy of target levels, designated to address sustainability, efficiency, and recovery scenarios. Targets were derived from: 1) reported catches and effort in the commercial fishery, 2) statistics from fishery-independent surveys, and 3) knowledge of the biology of blue crab. Targets that are recommended include population sizes, catches, and effort levels, as well as reference fishing mortality rates. They are intended to be conservative and risk-averse. (PDF contains 182 pages)

Densidade, genética e saúde populacional como ferramentas para propor um plano de controle e erradicação de invasão biológica: o caso de Callithrix aurita (Primates) no Parque Nacional da Serra dos Órgãos, RJ, Brasil

A introdução de espécies em locais fora de sua distribuição natural é uma preocupação importante na conservação da biodiversidade. A espécie Callithrix aurita é endêmica das regiões de floresta de altitude da Mata Atlântica do Sudeste do Brasil. Os critérios mais relevantes que a enquadram como espécie ameaçada de extinção são: destruição do habitat, incapacidade de adaptação a florestas secundárias degradadas, declínio populacional, distribuição restrita e introdução de espécies exóticas invasoras. Estes critérios, aliados à evidente raridade, explicam a sua inclusão na Lista Oficial de Espécies da Fauna Brasileira Ameaçadas de Extinção. Os objetivos do trabalho são: estimar o tamanho populacional de C. aurita, C. penicillata e seus híbridos no Parque Nacional da Serra dos Órgãos, avaliar a hibridação entre as espécies por caracteres morfológicos e laboratoriais, verificar o estado de saúde e confirmar a participação de C. aurita na paternidade dos animais capturados, propor um plano de erradicação e de controle de invasão de C. penicillata no Parque. Os tamanhos populacionais das duas espécies de primatas foram estimados através do método Distance Sampling. Um total de sete sagüis foi capturado com armadilhas de captura viva para a contenção física e química e posterior realização dos procedimentos. Para o hemograma, as dosagens bioquímicas e as análises genéticas, o sangue foi recolhido em um tubo de ensaio contendo anticoagulante e mantido em temperatura de refrigeração até o momento da manipulação / processamento das amostras. Callithrix aurita parece estar bem preservada apenas na área do Parque correspondente ao trecho situado no município de Petrópolis. As análises citogenéticas e moleculares dos híbridos são uma ferramenta útil para confirmar se há ou não hibridação, identificando as espécies envolvidas e verificando se há tendência nos retrocruzamentos. Pode-se sugerir que existe uma tendência à diferenciação das espécies e identificação de indivíduos híbridos pelo padrão hematológico e bioquímico, a ser confirmada com uma amostragem maior de animais da espécie C. aurita, preferencialmente da mesma localidade e nas mesmas condições. No caso de C. aurita, as principais recomendações para sua conservação incluem pesquisas para o registro de outras populações em áreas de distribuição livres de invasão, para que se possa avaliar as chances de recuperação populacional e sobrevivência da espécie. A criação de novas Unidades de Conservação deve ser estimulada, assim como estudos mais aprofundados sobre a espécie nos locais já conhecidos de ocorrência, além de um programa seguro de criação em cativeiro.

Age and length composition of Columbia Basin chinook, sockeye, and coho salmon at Bonneville Dam in 2001

In 2001, representative samples of adult Columbia Basin chinook (Oncorhynchus tshawytscha), sockeye (O. nerka), and coho salmon (O. kisutch) populations at Bonneville Dam were collected. Fish were trapped, anesthetized, sampled for scales and biological data, revived, and then released adult migrating salmonids. Scales were examined to estimate age composition; the results contributed to an ongoing database for age class structure of Columbia Basin salmon populations. Based on scale analysis of chinook salmon, four-year-old fish (from brood year [BY] 1997) comprised 88% of the spring chinook, 67% of the summer chinook, and 42% of the Bright fall chinook salmon population. Five-year-old fish (BY 1996) comprised 9% of the spring chinook, 14% of the summer chinook, and 9% of the fall chinook salmon population. The sockeye salmon population at Bonneville was predominantly four-year-old fish (81%), with 18% returning as five-year-olds in 2001. The coho salmon population was 96% three-year-old fish (Age 1.1). Length analysis of the 2001 returns indicated that chinook salmon with a stream-type life history are larger (mean length) than the chinook salmon with an ocean-type life history. Trends in mean length over the sampling period for returning 2001 chinook salmon were analyzed. Chinook salmon of age classes 0.2 and 1.3 show a significant increase in mean length over time. Age classes 0.1, 0.3, 0.4, 1.1, 1.2, and 1.4 show no significant change over time. A year class regression over the past 12 years of data was used to predict spring, summer, and Bright fall chinook salmon population sizes for 2002. Based on three-year-old returns, the relationship predicts four-year-old returns of 132,600 (± 46,300, 90% predictive interval [PI]) spring chinook and 44,200 (± 11,700, 90% PI) summer chinook salmon for the 2002 runs. Based on four-year-old returns, the relationship predicts five-year-old returns of 87,800 (± 54,500, 90% PI) spring, 33,500 (± 11,500, 90% PI) summer, and 77,100 (± 25,800, 90% PI) Bright fall chinook salmon for the 2002 runs. The 2002 run size predictions should be used with caution; some of these predictions are well beyond the range of previously observed data.

Age and length composition of Columbia Basin chinook, sockeye, and coho salmon at Bonneville Dam in 2000

In 2000, representative samples of adult Columbia Basin chinook (Oncorhynchus tshawytscha), sockeye (O. nerka), and coho salmon (O. kisutch), populations were collected at Bonneville Dam. Fish were trapped, anesthetized, sampled for scales and biological data, allowed to revive, and then released. Scales were examined to estimate age composition and the results contribute to an ongoing database for age class structure of Columbia Basin salmon populations. Based on scale analysis, four-year-old fish (from brood year (BY) 1996) were estimated to comprise 83% of the spring chinook, 31% of the summer chinook, and 32% of the upriver bright fall chinook salmon population. Five-year-old fish (BY 1995) were estimated to comprise 2% of the spring chinook, 26% of the summer chinook, and 40% of the fall chinook salmon population. Three-year-old fish (BY 1997) were estimated to comprise 14% of the spring chinook, 42% of the summer chinook, and 17% of the fall chinook salmon population. Two-year-olds accounted for approximately 11% of the fall chinook population. The sockeye salmon population sampled at Bonneville was predominantly four-year-old fish (95%), and the coho salmon population was 99.9% three-year-old fish (Age 1.1). Length analysis of the 2000 returns indicated that chinook salmon with a stream-type life history are larger (mean length) than the chinook salmon with an ocean-type life history. Trends in mean length over the sampling period were also analysis for returning 2000 chinook salmon. Fish of age classes 0.2, 1.1, 1.2, and 1.3 have a significant increase in mean length over time. Age classes 0.3 and 0.4 have no significant change over time and age 0.1 chinook salmon had a significant decrease in mean length over time. A year class regression over the past 11 years of data was used to predict spring and summer chinook salmon population sizes for 2001. Based on three-year-old returns, the relationship predicts four-year-old returns of 325,000 (± 111,600, 90% Predictive Interval [PI]) spring chinook and 27,800 (± 29,750, 90% PI) summer chinook salmon. Based on four-year-old returns, the relationship predicts five-year-old returns of 54,300 (± 40,600, 90% PI) spring chinook and 11,000 (± 3,250, 90% PI) summer chinook salmon. The 2001 run size predictions used in this report should be used with caution, these predictions are well beyond the range of previously observed data.

Age and length composition of Columbia Basin chinook, sockeye, and coho salmon at Bonneville Dam in 2002

In 2002, representative samples of migrating Columbia Basin chinook (Oncorhynchus tshawytscha), sockeye (O. nerka), and coho salmon (O. kisutch) adult populations were collected at Bonneville Dam. Fish were trapped, anesthetized, sampled for scales and biological data, revived, and then released. Scales were examined to estimate age composition; the results contributed to an ongoing database for age class structure of Columbia Basin salmon populations. Based on scale analysis of chinook salmon, four-year-old fish (from brood year [BY] 1998) comprised 86% of the spring chinook, 51% of the summer chinook, and 51% of the bright fall chinook salmon population. Five-year-old fish (BY 1997) comprised 13% of the spring chinook, 43% of the summer chinook, and 11% of the bright fall chinook salmon population. The sockeye salmon population at Bonneville was predominantly five-year-old fish (55%), with 40% returning as four-year-olds in 2002. For the coho salmon population, 88% of the population was three-year-old fish of age class 1.1, while 12% were age class 1.0. Length analysis of the 2002 returns indicated that chinook salmon with a stream-type life history are larger (mean length) at age than the chinook salmon with an ocean-type life history. Trends in mean length over the sampling period for returning 2002 chinook salmon were analyzed. Chinook salmon of age classes 1.2 and 1.3 show a significant increase in mean length over the duration of the migration. A year class regression over the past 14 years of data was used to predict spring, summer, and bright fall chinook salmon population sizes for 2003. Based on three-year-old returns, the relationship predicts four-year-old returns of 54,200 (± 66,600, 90% predictive interval [PI]) spring chinook, 23,800 (± 19,100, 90% PI) summer, and 169,100 (± 139,500, 90% PI) bright fall chinook salmon for the 2003 runs. Based on four-year-old returns, the relationship predicts five-year-old returns of 36,300 (± 35,400, 90% PI) spring, 63,800 (± 10,300, 90% PI) summer, and 91,100 (± 69,400, 90% PI) bright fall chinook salmon for the 2003 runs. The 2003 run size predictions should be used with caution; some of these predictions are well beyond the range of previously observed data.

The relative value of different estuarine nursery areas in North Carolina for transient juvenile marine fishes

Offshore winter-spawned fishes dominate the nekton of south-eastern United States estuaries. Their juveniles reside for several months in shallow, soft bottom estuarine creeks and bays called primary nursery areas. Despite similarity in many nursery characteristics, there is, between and within species, variability in the occupation of these habitats. Whether all occupied habitats are equally valuable to individuals of the same species or whether most recruiting juveniles end up in the best habitats is not known. If nursery quality varies, then factors controlling variation in pre-settlement fish distribution are important to year-class success. If nursery areas have similar values, interannual variation in distribution across nursery creeks should have less effect on population sizes or production. I used early nursery period age-specific growth and mortality rates of spot (Leiostomus xanthurus) and Atlantic croaker (Micropogonias undulatus)—two dominant estuarine fishes—to assess relative habitat quality across a wide variety of nursery conditions, assuming that fish growth and mortality rates were direct reflections of overall physical and biological conditions in the nurseries. I tested the hypothesis that habitat quality varies for these fishes by comparing growth and mortality rates and distribution patterns across a wide range of typical nursery habitats at extreme ends of two systems. Juvenile spot and Atlantic croaker were collected from 10 creeks in the Cape Fear River estuary and from 18 creeks in the Pamlico Sound system, North Carolina, during the 1987 recruitment season (mid-March–mid-June). Sampled creeks were similar in size, depth, and substrates but varied in salinities, tidal regimes, and distances from inlets. Spot was widely distributed among all the estuarine creeks, but was least abundant in the creeks in middle reaches of both systems. Atlantic croaker occurred in the greatest abundance in oligohaline creeks of both systems. Instantaneous growth rates derived from daily otolith ages were generally similar for all creeks and for both species, except that spot exhibited a short-term growth depression in the upriver Pamlico system creeks—perhaps the result of the long migration distance of this species to this area. Spot and Atlantic croaker from upriver oligohaline creeks exhibited lower mortality rates than fish from downstream polyhaline creeks. These results indicated that even though growth was similar at the ends of the estuaries, the upstream habitats provided conditions that may optimize fitness through improved survival.

Turtle excluder devices -- Are the escape openings large enough?

All five species of sea turtles in continental U.S. waters are protected under the Endangered Species Act of 1973 and the population sizes of all species remain well below historic levels. Shrimp trawling was determined to be the largest source of anthropogenic mortality of many of the species. As a mechanism to reduce the incidental catch of turtles in trawl nets, turtle excluder devices have been required intermittently in the shrimp fishery since 1987, and at all times since 1994. The expanded turtle excluder device (TED) regulations, implemented in 1994, were expected to reduce shrimp trawl capture of sea turtles by 97%. Recent evidence has indicated that the sizes of turtles stranding were not representative of the animals subjected to being captured by the shrimp trawlers. The purpose of our study was to compare the sizes of stranded sea turtles with the size of the TED openings. We compared the sizes of stranded loggerhead (Caretta caretta), green (Chelonia mydas), and Kemp’s ridley (Lepidochelys kempii) sea turtles, the three species most commonly found stranded, to the minimum widths and heights of TED openings. We found that annually a large proportion of stranded loggerhead turtles (33–47%) and a small proportion of stranded green turtles (1–7%) are too large to fit through the required minimum-size TED openings. The continued high mortality of sea turtles caused by bottom trawling is reason for concern, especially for the northern subpopulation of loggerhead turtles, which currently is not projected to achieve the federal recovery goal of reaching and maintaining prelisting levels of nesting.

Microsatellite polymorphisms of Sichuan golden monkeys

Previous study using protein electrophoresis shows no polymorphism in 44 nuclear loci of Sichuan golden monkey (Rhinopithecus roxellana), which limits our understandings of its population genetic patterns in the nuclear genome. In order to obtain sufficient information, we scanned 14 microsatellite loci in a sample of 32 individuals from its three major habitats (Minshan, Qinling and Shennongiia). A considerable amount of polymorphisms were detected. The average heterozygosities in the local populations were all above 0.5. The differentiations among local populations were significant. There was evidence of geneflow among subpopullations, but geneflow between Qinling and Shennongjia local populations was the weakest. Minshan and Qinling populations might have gone through recent bottlenecks. The estimation of the ratio of the effective population sizes among local populations was close to that from census sizes. Comparisons to available mitochondria data suggested that R. roxellana's social structures played an important role in shaping its population genetic patterns. Our study showed that the polymorphism level of R. roxellana was no higher than other endangered species; therefore, measures should be taken to preserve genetic diversity of this species.

The picophytoplankton in three Chinese lakes of different trophic status and its relationship to fish populations

A year-round comparison was made of the picophytoplankton populations in three lakes of different trophic status (oligotrophic, mesotrophic, and hypereutrophic), as well as in experimental enclosures stocked with various densities of fish. Picophytoplankton abundance was significantly different between the hypereutrophic lake and the oligotrophic lake (P<0.018) and between the hypereutrophic lake and the mesotrophic lake (P<0.021), whereas the difference between the mesotrophic and oligotrophic lakes was not significant (P<0.745). The effect of total nitrogen:total phosphorus ratio on the abundance of picophytoplankton was demonstrated in the oligotrophic lake, but such effect did not manifest itself in the other lakes or in the experimental enclosures. The average annual picophytoplankton population sizes in the three lakes in 1995-1998 were correlated with estimated fish abundance (r=0.824, n=9, P=0.006). The results of enclosure experiments demonstrated that the abundance of picophytoplankton increased with the stocking density of fish (r=0.619, n=8, P=0.100).