Bruce W. Frost

Closing the microbial loop: dissolved carbon pathway to heterotrophic bacteria from incomplete ingestion, digestion and absorption in animals

Abstract A new extension of digestion theory and re-interpretation of published empirical evidence suggest that the principal pathway of dissolved organic carbon (DOC) from phytoplankton to bacteria is through the byproducts of animal ingestion and digestion rather than via excretion of DOC directly from intact phytoplankton. Simple model calculations reveal that for a substance with diffusion coefficient equalling 10−5 cm2 s−1, excess (over ambient) concentrations of solute in a fecal pellet of typical size (diam. ⩽ 1 mm) are lost rapidly; ⩾ 50% of any excess is diffused out of the pellet within 5 min—even in a stagnant water column and without particle sinking. Reasons for rapid loss and its insensitivity to fluid dynamic conditions are small size of the pelletal reservoir and the sharp concentration gradient between pelletal and ambient concentrations upon pellet release. As a consequence, most solutes initially contained in fecal pellets of zooplankton generally will remain in the 10–100 m thick water layer within which the pellets initially are deposited. Focus on animal-caused organic release over these very short time scales may help to resolve some of the growing paradoxes of DOC standing stocks and fluxes in the upper ocean.

Life histories of large, grazing copepods in a subarctic ocean gyre: Neocalanus plumchrus, Neocalanus cristatus, and Eucalanus bungii in the Northeast Pacific

Life histories for the dominant, larger copepods of the subartic Pacific have been constructed by sampling from weatherships patrolling Ocean Station P (50°N, 145°W) during 1980 and 1981. Neocalanus plumchrus reproduced at depths below 250 m from July through February. Copepodite stages were present in surface layers from October through August with a large peak in numbers and biomass in spring. Fifth copepodites prepared for diapuse in 38 days during spring and descended to depths below 250 m. They commenced immediately to mature, and the females reproduced without renewed feeding. This schedule contrasts with that of the population in the Strait of Georgia, which remains in diapause from July to January and matures exclusively in January and February. There appears to be a difference between the coastal and oceanic habitats in preparing the diapausing individuals for maturation. Maturation of the diapausing stock of N. plumchrus maintained constant adult populations, averaging 714 males m−2 from June through October and 1,434 females m−2 from August through January. This constancy, together with the exponential pattern of decline in the diapause stock from September through February, suggests that density of adults may regulate maturation of fifth copepodites. Offspring of individuals delaying maturation and, thus, reproduction would benefit from the resulting moderation of intraspecific competition, probably that among copepodites. Reproduction of Neocalanus cristatus also occurred below 250 m, and, while spawning was continuous through the year, there was a substantial peak in November. That resulted in a peak of abundance for early copepodite stages in mid-winter, and a peak for the fifth copepodite stage in June. Stocking of the population of fifth copepodites in diapause below 250 m occurred from July through October. Some fifth copepodites were present in surface layers through the entire summer, and some younger copepodites persisted through the summer in progressively declining abundance just below the mixed layer. In autumn 1980 resurgence of early copepodite populations was rapid, occurring during the course of a prolonged October storm. The storm may have improved the habitat either by cooling the mixed layer or by resupplying nutrients to the euphotic zone. Eucalanus bungii reproduced in the mixed layer in early May and in early July. The first event was a spawning by females that had previously spawned in 1979 and then had returned to diapause. The second, heavier spawning (more females, more eggs per female) was by newly matured females from stocks that had overwintered as fifth copepodites. Nauplii peaked sharply in abundance on 19 July, one week after the peak in spawning. First and second copepodites peaked on 1 August, and all had advanced to the third copepodite stage by September. The diapause stock was established by September, principally between 250 and 500 m, and consisted of copepodite stages from third to sixth. Duration of the E. bungii life cycle appears to be typically two years. New nauplii develop as far as the third or fourth copepodite stage during their first summer, then enter diapause. The second summer they advance to the fifth copepodite stage and reenter diapause. Fifth copepodites mature in their third summer at two years of age. The males remain at depth and mate without subsequent feeding. Females migrate at night to the mixed layer where spawning occurs. About 20% of females that had already spawned in 1980 reentered diapause. They would reproduce again in their fourth summer at three years of age. All aspects of the life cycle suggest low mortality rates for copepodite stages, particularly at depth in the habitat occupied during diapuse. There can be no premium on rapid reproduction for E. bungii in the subartic Pacific, and there must even be benefit from spreading reproduction between years. This iteroparity may amount to a “bet-hedging” tactic, the young from a given mother having more than one chance to find sustaining conditions. It also produces gene flow between the year classes of the biennial life cycle.

Reverse Diel Vertical Migration: An Escape from Invertebrate Predators

The marine copepod Pseudocalanus sp. exhibits an unusual reverse diel vertical migration in Dabob Bay, Washington, concurrently with a normal vertical migration by nocturnal invertebrate predators. Reduced spatial overlap with predators appears to decrease mortality rate of adult female Pseudocalanus. A life table analysis suggests that the demographic disadvantage associated with daily migration across a thermal gradient can be overcome if mortality is reduced by as little as 16 percent.

Morphological stasis in the Eurytemora affinis species complex (Copepoda: Temoridae)

Morphological stasis has long been regarded as one of the most challenging problems in evolutionary biology. This study focused on the copepod species complex, Eurytemora affinis, as a model system to determine pattern and degree of morphological stasis. This study revealed discordant rates of morphological differentiation, molecular evolution, and reproductive isolation, where speciation was accompanied by lack of morphological differentiation in secondary sex characters. Comparisons were made among phylogenies based on morphometrics, nuclear (allozyme) loci, and mitochondrial DNA (mtDNA) sequences from cytochrome oxidase I, for a total of 43 populations within the complex. These systematic relationships were also compared to patterns of reproductive isolation. In addition, genetic subdivision of nuclear molecular (allozyme) markers (GST) and quantitative (morphological) characters (QST) were determined to infer evolutionary forces driving morphological differentiation. The morphometric phylogeny revealed that all clades, excluding the European clade, were morphologically undifferentiated and formed a polytomy (multifurcation). Morphometric distances were not correlated with mtDNA distances, or with patterns of reproductive isolation. In contrast, nuclear and mtDNA phylogenies were mostly congruent. Reproductive isolation proved to be the most sensitive indicator of speciation, given that two genetically and morphologically proximate populations showed evidence of hybrid breakdown. Quantitative genetic (morphological) subdivision (QST = 0.162) was lower than nuclear genetic subdivision (GST = 0.617) for four laboratory-reared North American populations, indicating retarded evolution of morphological characters. This result contrasts with most other species, where QST typically exceeds GST as a result of directional selection. Thus, in all but the European populations, evolution of the secondary sex characters was marked by morphological stasis, even between reproductively-isolated populations.

Measuring feeding rates of pelagic herbivores: analysis of experimental design and methods

We have re-evaluated the experimental methods and statistical procedures used to determine the relationship between feeding rates of pelagic herbivores and food concentration. Analysis of our own experiments, on Calanus pacificus feeding on Gyrodinium resplendens, and of other published research on this subject suggests the need for improvements in experimental design and methodology. We show that the use of “mean concentration” is statistically erroneous. First, it produces an artificial increase in the degrees of freedom that may result in the acceptance of nonsignificant regression lines. Second, it negates the value of replication, which is required to estimate sources of error. We present an example of how replication may be used to improve control over sources of error. Furthermore, we recommend the use of initial concentration rather than mean concentration. Finally, we introduce alternative methods to determine clearance and ingestion rates that enable the investigator to use replication and thus to estimate experimental errors.

Calanus marshallae, a new species of calanoid copepod closely allied to the sibling species C. finmarchicus and C. glacialis

Species of the copepod genus Calanus frequently dominate the marine zooplankton in boreal and arctic waters. Up to now there have been no operational means of identifying several species closely related to Calanus finmarchicus. Reanalysis of these taxa, using material from plankton samples collected throughout Northern Hemisphere polar and coreal waters, shows that there are 3 sibling species which have been previously combined under the names C. finmarchicus and C. glacialis. Several new taxonomic characters permit unequivocal identification of C. finmarchicus, C. glacialis, and a new species, C. marshallae. Claims that C. finmarchicus and C. glacialis are subspecies are refuted; there is no evidence that the two species continuously intergrade either where they co-occur or where they are allopatric, nor is there evidence that the two species hybridize. C. finmarchicus is basically restricted to the North Atlantic Ocean and C. marshallae to the North Pacific Ocean and Bering Sea; C. glacialis is primarily an Arctic species, but its geographical distribution slightly overlaps those of the other two species. Taxa closely related to C. finmarchicus and C. helgolandicus probably represent two separate, but closely linked evolutionary lineage; species of these two lineages are placed in one of two species groups, the finmarchicus group and the helgolandicus group.

Molecular systematics of six Calanus and three Metridia species (Calanoida: Copepoda)

The discrimination of species of the copepod genus, Calanus (Copepoda; Calanoida), is problematical-especially in regions of sympatry. Although the species of Calanus exhibit exceptional morphological similarity, they are quite distinct in genetic character. The DNA base sequences of the mitochondrial large subunit (16S) ribosomal RNA (rRNA) gene unambiguously discriminated C. finmarchicus (Gunnerus 1765), C. glacialis (Jaschnov 1955), C. marshallae (Frost 1974), C. helgolandicus (Claus 1863), C. pacificus (Brodsky 1948), C. sinicus (Brodsky 1965), and C. hyperboreus (Kroyer 1838). Sequence differences among Calanus species for this gene portion range from 7.3% (between C. glacialis and C. marshallae) to 23.9% (between C. glacialis and C. sinicus). Differences among conspecific individuals were approximately 1 to 2%. [These sequence data were determined between April and November 1993; the sequenced domain is similar to that published previously in Bucklin et al. (1992) but are derived from analysis of additional individuals.] Statistical analysis of the sequence data using a variety of tree-building algorithms separated the taxa into one group of species corresponding to the C. finmarchicus group (C. finmarchicus, C. marshallae, and C. glacialis) and another ungrouped set of species corresponding to the C. helgolandicus group (C. helgolandicus, C. pacificus, and C. sinicus). The C. helgolandicus group may be older than the C. finmarchicus group, making the tree topology less reliable in this area. Calanus hyperboreus was an outlier; Nannocalanus minor (Claus 1863) was the outgroup. Similar analysis of Metridia species confirmed that M. lucens (Boeck 1864) and M. pacifica (Brodsky 1948) are distinct species; M. longa (Lubbock 1854) was still more divergent. These sequence data will allow the design of simple, molecular tools for taxonomic identifications. Diagnostic characters, assayed by rapid molecular protocols, will enable biological oceanographers to answer important questions about the distribution and abundance of all life stages (as well as patterns of reproduction) of morphologically similar species, such as those of Calanus.

A modelling study of processes regulating plankton standing stock and production in the open subarctic Pacific Ocean

Abstract At Ocean Station P (50°N 145°W) in the open subarctic Pacific Ocean recent estimates of phytoplankton production rate were consistently much higher than those obtained during previous decades of weathership operations. Moreover, the production was supported to a major extent by regenerated forms of nitrogen despite high concentrations of NO 3 . An ecosystem process model, simulating the interactions among phytoplankton, herbivorous microzooplankton and nutrients, was used to evaluate the possible mechanisms underlying both the high production rates and the patterns of utilization of dissolved nitrogenous nutrients. The model results suggest that at Station P the phytoplankton have specific growth rates expected for the ambient light and temperature regime, and furthermore that two processes, grazing control of phytoplankton stock and preferential utilization of NH 4 by the phytoplankton, can account for the continuously low standing stock of phytoplankton and the underutilization of NO 3 in the surface layer in summer. However, moderate limitation of phytoplankton specific growth rate by a trace nutrient such as dissolved iron cannot be ruled out. Winter observations of plankton dynamics at Station P would afford a strong test of the model predictions. In addition, the model highlights several properties of the subarctic Pacific epipelagic ecosystem in need of further investigation. These include possible trace nutrient limitation of maximum specific growth rate of the phytoplankton assemblage; the abundance, types and physiological characteristics of microzooplankton grazers; the sources and intensity of predatory mortality of grazers; effects of daily variability of physical processes on plankton dynamics; and control of growth rates of individual phytoplankton species, especially microphytoplankters whose specific growth rates may be limited by availability of trace nutrients.

Chemical Communication in the Marine Planktonic Copepods Calanus Pacificus and Pseudocalanus Sp

[Es wurde experimentell untersucht, ob intraspezifische Paarungsbegegnungen bei den marinen planktonischen Copepoden Calanus parificus und Pseudocalanus sp. durch Pheromone kontrolliert werden, die durch das adulte Weibchen erzeugt werden. Nach Uberfuhrung in Wasser, das vorher frisch gehautete adulte Weibchen enthielt, zeigen adulte Mannchen ein fur sie charakteristisches Schwimmverhalten. Durch direkte Messungen und autoradiographische Methoden wird nachgewiesen, das Mannchen 14C- markierte, geloste organische Substanzen aufnehmen, vermutlich Pheromone von kurzlich gehauteten Weibchen. Die Hauptrezeptoren fur Pheromone befinden sich auf den mannlichen Antennulen und scheinen Aestheten zu sein., Es wurde experimentell untersucht, ob intraspezifische Paarungsbegegnungen bei den marinen planktonischen Copepoden Calanus parificus und Pseudocalanus sp. durch Pheromone kontrolliert werden, die durch das adulte Weibchen erzeugt werden. Nach Uberfuhrung in Wasser, das vorher frisch gehautete adulte Weibchen enthielt, zeigen adulte Mannchen ein fur sie charakteristisches Schwimmverhalten. Durch direkte Messungen und autoradiographische Methoden wird nachgewiesen, das Mannchen 14C- markierte, geloste organische Substanzen aufnehmen, vermutlich Pheromone von kurzlich gehauteten Weibchen. Die Hauptrezeptoren fur Pheromone befinden sich auf den mannlichen Antennulen und scheinen Aestheten zu sein.]

Diel vertical migration in zooplankton: field evidence in support of the predator avoidance hypothesis

We report results of a field test of the predator avoidance hypothesis as an explanation of the adaptive significance of diel vertical migration in zooplankton. We determined the vertical distribution and diel migration of the planktonic copepod Acartia hudsonica, concurrently with the abundance of pelagic fish, transparency and thermal stratification of the water column, on six cruises over a one year period in a temperate marine lagoon (Jakles Lagoon, San Juan Island, Washington, USA). Striking seasonal variability was observed in all biological and environmental variables. Linear regressions of the strength of diel vertical migration in A. hudsonica on these environmental variables resulted in only one statistically significant relationship, that between copepod diel vertical migration and predator abundance. These results, together with those of previous studies, point to diel vertical migration as a widespread behavioral response of planktonic prey to the presence of their predators.