Presence of the dinoflagellates Ceratium dens, C. fusus and C. furca (Gonyaulacales: Ceratiaceae) in Golfo de Nicoya, Costa Rica

Harmful Algae Blooms (HAB) are a frequent phenomenon in the Gulf of Nicoya, Costa Rica, as in other parts of the world. The morphology and physiology of these microalgae are important because HAB species have adaptive characteristics. The production of high concentrations of paralytic toxins by Ceratium dinoflagellates has only been documented at the experimental level. However, this genus has been associated with the mortality of aquatic organisms, including oyster and shrimp larva, and fish, and with decreased water quality. Recently, fishermen reported massive mortality of encaged fish near Tortuga Island (Gulf of Nicoya). Samples were taken from an algal bloom that had produced an orange coloration and had a strong foul-smelling odor. Ultrastructural details were examined with scanning electron microscopy. The dinoflagellates Ceratium dens, C. furca and C. fusus were found in samples taken at the surface. The cell count revealed four million cells of this genus per liter. The morphological variability of these species is high; therefore electron microscopy is an useful tool in the ultrastructural study of these organisms. This is the first time that three Ceratium species are reported concurrently producing harmful blooms in Costa Rica.     

PHENOTYPIC PLASTICITY INDUCED IN TRANSPLANT EXPERIMENTS IN A MUTUALISTIC ASSOCIATION BETWEEN THE RED ALGA JANIA ADHAERENS (RHODOPHYTA,CORALLINALES) AND THE SPONGE HALICLONA CAERULEA (PORIFERA: HAPLOSCLERIDA): MORPHOLOGICAL RESPONSES OF THE ALGA1

The association between the red macroalga Jania adhaerens J. V. Lamour. and the sponge Haliclona caerulea is the most successful life‐form between 2 and 4 m depth in Mazatlán Bay (Mexican Pacific). J. adhaerens colonizes the rocky intertidal area and penetrates into deeper areas only when it lives in association with H. caerulea. The aposymbiotic form of the sponge has not been reported in the bay. To understand the ecological success of this association, we examined the capacity of J. adhaerens to acclimate in Mazatlán Bay using transplant experiments. The transplanted aposymbiotic J. adhaerens did not survive the first 2 weeks; however, J. adhaerens when living in association with H. caerulea, acclimated easily to depth, showing no sign of mortality during the 103 d of the experiment. We conclude that the ability of J. adhaerens to colonize in deeper areas in this hydrodynamic environment may in part rely on the protection provided by the sponge to the algal canopy. Both species contribute to the shape of the associated form. Nevertheless, the morphological variation in the association appears to be dominated by the variation in J. adhaerens canopy to regulate pigment self‐shading under light‐limited conditions and/or tissue resistance under high hydrodynamics. Consequently, our results are consistent with light as the abiotic controlling factor, which regulates the lower depth distribution of the association in Mazatlán Bay, through limiting the growth rate of J. adhaerens. Hydrodynamics may determine the upper limit of the association by imposing high mass losses.     

Mass Stranding of Marine Birds Caused by a Surfactant-Producing Red Tide

In November-December 2007 a widespread seabird mortality event occurred in Monterey Bay, California, USA, coincident with a massive red tide caused by the dinoflagellate Akashiwo sanguinea. Affected birds had a slimy yellow-green material on their feathers, which were saturated with water, and they were severely hypothermic. We determined that foam containing surfactant-like proteins, derived from organic matter of the red tide, coated their feathers and neutralized natural water repellency and insulation. No evidence of exposure to petroleum or other oils or biotoxins were found. This is the first documented case of its kind, but previous similar events may have gone undetected. The frequency and amplitude of red tides have increased in Monterey Bay since 2004, suggesting that impacts on wintering marine birds may continue or increase.     

Spatial and Temporal Aspects of Mixotrophy In Chesapeake Bay Dinoflagellates

ABSTRACT. Gymnodinium sanguineum, Gyrodinium uncatenum , and Ceratium furca are large phototrophic dinoflagellates that commonly form red tides in the mesohaline portion of Chesapeake Bay during the summer. Examination of protargol-stained specimens revealed that these dinoflagellates also feed heterotrophically as indicated by the presence of food vacuoles containing partially digested prey. Ingested prey were generally identified as nanociliates (≥20 μm) belonging to the oligotrich genera Strobilidium and Strombidium ; occasionally other small ciliates (e.g. Balanion sp. and Mesodinium sp.), dinoflagellates, and diatoms were observed in early stages of digestion. the percentage of these mixotrophs that had ingested prey was usually less than 20%, but approached 30% in some samples. Occurrence of food vacuoles in Gymno. sanguineum was positively correlated with ≤20 μm oligotrichous ciliate density; limited data for Gyro. uncatenum suggests a similar relationship, but C. furca feeding was not related to nanociliate densities.     

Phased cell division in natural populations of marine dinoflagellates from shipboard cultures

In June 1975 samples were preserved every 2 h for 26 h from water collected from Santa Monica Bay, California and which was incubated on shipboard. The percentage of dividing dinoflagellate cells in the samples was determined by microscopic examination after applying a nuclear stain. The maximum proportion of dividing cells was at 03.30 h, 05.30 h, and 09.30 h for Ceratium dens Ostf. & Schmidt, C. furca (Ehrenb.) Claparède & Lachmann, and Dinophysis fortii Pavillard, respectively. Mean doubling times for these species, calculated from the percentage of dividing cells, were 3.3,4.6, and 1.4 days. Temporal stratification in the timing of cell division may reflect a selective process for minimizing interspecific competition.     

VARIATIONS OF PSP TOXIN PROFILES DURING DIFFERENT GROWTH PHASES IN GYMNODINIUM CATENATUM (DINOPHYCEAE) STRAINS ISOLATED FROM THREE LOCATIONS IN THE GULF OF CALIFORNIA,MEXICO1

In vitro experiments were performed with Gymnodinium catenatum Graham strains isolated from three locations in the Gulf of California to determine the variability in toxicity and toxin profiles. Strains were cultivated in GSe at 20°C±1°C, 150 μmol photons·m^?2·s^?1 (12:12 light:dark cycle), and harvested during different growth phases. Growth rates were higher than in previous studies, varying between 0.70 and 0.82 day^?1. The highest cell yields were reached at 16 and 19 days, with maximum densities between 1090 and 3393 cells·mL^?1. Bahía de La Paz (BAPAZ) and Bahía de Mazatlán (BAMAZ) were the most toxic (101 pg STXeq·cell^?1), whereas strains from Bahía Concepción (BACO) were the least toxic (13 pg STXeq·cell^?1). A strain isolated from cyst germination was one of the least toxic strains. No significant changes in toxin content with culture age were observed (0.2 and 0.6 pmol paralytic shellfish poisoning·cell^?1). All strains contained neosaxitoxin (NEOSTX), decarbamoyl‐saxitoxin (dcSTX), decarbamoyl‐gonyautoxin‐2,‐3, (dcGTX2‐3), N‐sulfo‐carbamoylsaxitoxin (B1), N‐sulfo‐carbamoylneosaxitoxin (B2), and N‐sulfo‐carbamoylgonyautoxin‐2,‐3 (C1‐2). Bahía Concepción strains had the highest content of C1; BAPAZ and BAMAZ strains had a higher percentage of NEOSTX. Differences in toxin composition with culture age were observed only in BAMAZ and BAPAZ strains. Cultures with a higher percentage of long chains had more NEOSTX, while those with a higher proportion of short chains had a lower content of NEOSTX. Gulf of California strains are characterized by a high proportion of NEOSTX, and seem to have evolved particular physiological responses to their environment that are reflected in the toxin profile, suggesting different populations.     

Relationships between dynamics of red tide-causing raphidophycean flagellates and algicidal micro-organisms in the coastal sea of Japan

Temporal fluctuations of algicidal micro-organisms against the red tide causing raphidophycean flagellates Chattonella antiqua (Hada) Ono and Heterosigma akashiwo (Hada) Hada ex Hara et Chihara were investigated using the microplate most probable number (MPN) method in northern Hiroshima Bay and Harima-Nada, the Seto Inland Sea, in 1992 and 1993. In Har-ima-Nada, both flagellates appeared at low levels (< 1 cell mL^?1), and killer micro-organisms against the two flagellates (C-killer for C. antiqua and H-killer for H. akashiwo) also appeared at low densities (< 2 mL^?1). In northern Hiroshima Bay, C. antiqua cells were scarce (< 1 cell mL^?1), and C-killers occurred at a low level (≤ 3.4 mL^?1). Conversely, red tides of H. akashiwo occurred there in June of both years. The dynamics of H-killers revealed a close relationship with that of H. akashiwo populations. H-killers followed the increase of H. akashiwo cells, reached a maximum level after the beginning of decline of H. akashiwo, maintained a high level for at least 1 week after the crash of bloom, and then decreased. C-killers consistently remained at low densities during the period of H. akashiwo red tides in both 1992 and 1993. Hence, algicidal micro-organisms specifically associated with the occurrence and crash of H. akashiwo red tides, and presumably contributed to the rapid termination of the red tides in the coastal seas such as northern Hiroshima Bay.     

Blooms of Cochlodinium polykrikoides (Gymnodiniaceae) in the Gulf of California, Mexico

Cochlodinium polykrikoides was the species responsible for the discoloration that occurred between September 15th and 27th, 2000 in a shallow coastal lagoon located in the southern part of the Bahia de La Paz, on the west side of the Gulf of California. Blooms of C. polykrikoides were observed four days after two rainy days with a seawater temperature of 29 to 31 degrees C. Nutrient concentration ranges during the bloom were 0.165-0.897 microM NO2+NO3, 0.16-3.25 microM PO4, and 1.0-35.36 microM SiO4. Abundance of C. polykrikoides ranged from 360 x 10(3) to 7.05 x 10(6)/cells l(-1). Biomass expressed in terms of chlorophyll a was high, ranging from 2.7 to 56.8 mg/m3. A typical dinoflagellate pigment profile (chlorophyll a and c, peridinin, diadinoxantin, and beta-carotene) was recorded. In this study, the red tide occurred in front of several fish and shrimp-culture ponds. No PST toxins were found in the samples. However, 180 fish were found dead in the infected fish-pond; the gills were the most affected part. C. polykrikoides is a cyst-forming species that recurs in this area. New blooms were observed in November 2000 and September-November 2001 in the same area. Anthropogenic activities, such as eutrophication caused by water discharge in this shallow lagoon, and nutrient enrichment in the culture ponds, as well as effects from precipitation and wind stress, could have favored the outbreak of this dinoflagellate.     

Ecology of the red-tide dinoflagellate Ceratium furca: distribution, mixotrophy, and grazing impact on ciliate populations of Chesapeake Bay

Ceratium furca is a primarily photosynthetic dinoflagellate also capable of ingesting other protists. During 1995 and 1996, we documented the abundance of C. furca in Chesapeake Bay and determined grazing rates on prey labeled with fluorescent microspheres. Abundance usually remained below 20 cells ml(-1), although the species was capable of localized late-summer blooms (< or = 478 cells ml(-1)) in the more saline lower to mid-Bay region. Feeding rates ranged from 0 to 0.11 prey dinoflagellate(-1) h(-1) or from 0 to 37 pg C dinoflagellate(-1) h(-1) and were highest at lower salinities. Clearance rates averaged 2.5 +/- 0.35 microl dinoflagellate(-1) h(-1). Impact of C. furca feeding on prey populations was higher in the lower Bay, averaging 67% of Strobilidium spp. removed d(-1). Ingestion rates were positively correlated with prey abundance and dissolved inorganic nitrogen, but negatively with salinity, depth, dissolved inorganic phosphorus, and inorganic P:N ratio. Daily consumption of prey biomass by C. furca averaged 4.6% of body carbon, 6.5% of body nitrogen, and 4.0% of body phosphorus. with maximal values of 36, 51, and 32%, respectively. Thus, the ability to exploit an organic nutrient source when inorganic nutrients are limiting may give C. furca a competitive advantage over purely photosynthetic species.     

Does damming of the Colorado River affect the nursery area of blue shrimp Litopenaeus stylirostris (Decapoda: Penaeidae) in the Upper Gulf of California?

After damming the Colorado River the freshwater flow was reduced to 1% of its virgin flow to the Upper Gulf of California (UGC). The ecological effects need to be properly documented. The UGC is the nursery area for Litopenaeus stylirostris, the most profitable fishery in the zone. In order to know the relative abundance of L. stylirostris postlarval stage we conducted a sampled survey every 14 days in 1993, 1994 and 1997, plus an intensive sampling during a complete tide cycle in July 1995 and 1996. We did 10 min trawls each hour during the flood tide. Relative abundance of postlarvae was higher (p < 0.05) in those years when freshwater flow reached the UGC.     

Observations on Alexandrium tamarense (Lebour) Balech and Other Dinoflagellate Populations in Golfo Nuevo, Patagonia (Argentina)

The toxic dinoflagellate Alexandrium tamarense and other dinoflagellatespecies were studied, along with water temperature and nutrientconcentrations, from September 1995 to December 1998 in theGolfo Nuevo, Chubut, Argentina. Nutrient concentrations werelow, showing a peak of high concentration in winter and a phaseof depletion in late spring and summer. Dinoflagellates tendedto be abundant during spring and summer, when Prorocentrum micanswas the most important species. Other dinoflagellates were Pyrophacushorologium and Dinophysis acuminata. Ceratium tripos, C. fususand C. horridum were present during the autumn, and a C. tripospeak up to 5.9 x 10³ cell l^–1 was observed in May 1997.Alexandrium tamarense showed strong interannual variation, thehighest concentration being found in spring (September–October)1995, with densities up to 15 x 10³ cells l^–1. The secondA.tamarense peak was observed during October–November1998 with maximal densities up to 5 x 10³ cells l^–1. Moderatelyhigh A. tamarense cyst densities, up to 300 cysts cm^–3of sediment, were found in the deep zone of the Golfo Nuevobasin. Among meteorolog-ical variables, increased late winterrain and higher solar radiation during spring may have influencedA. tamarense blooms.     

The emergence of Cochlodinium along the California Coast (USA)

A sudden and nearly synchronous emergence of the red tide forming dinoflagellate Cochlodinium along more than 800 km of California coastline was initially observed in late summer 2004. Thereafter high cell concentrations have been detected on an annual basis. Here, we present quantitative and semi-quantitative data indicating that Cochlodinium was uncommon in the phytoplankton community in California prior to 2004 and is now persisting as a more regular component and one that seasonally can cause red tides. The quantitative portion of this study was primarily conducted in Monterey Bay, where cell densities reached at least 6 × 10⁴ cells L⁻¹ during the initial outbreak. A semi-quantitative comparison of California coastal counties by the California Department of Health Services (CDHS) was also made: of the 15 counties surveyed (most with multiple sites per county), cells were detected only from Los Angeles County in the south to San Mateo County in the central region (seven counties), but not in the northern part of the state (six counties). Two counties in the central region of the state, San Luis Obispo and Santa Cruz, displayed intense and frequent periods of elevated Cochlodinium cell abundances. Although not observed in the state-wide CDHS survey, we occasionally found cells in San Diego County with densities up to 2.7 × 10⁴ cells L⁻¹. Though these colonial dinoflagellates have been recognized in California for over 80 years, with several “blooms” recorded prior to 2004, the species’ geographic range and abundance in recent years suggest significant shifts in the nearshore phytoplankton community of this region of the eastern Pacific.     

Heterosigma akashiwo in central California waters

Heterosigma akashiwo (Hada) gives rise to red tides along the Atlantic and Pacific coasts and is known to produce brevetoxins. This investigation establishes baseline information showing the presence of H. akashiwo along the central California coast based on water samples collected from the Santa Cruz pier in Monterey Bay (on the open coast) and the Berkeley pier in San Francisco Bay. Light and electron microscopy as well as two species-specific DNA probe methods based on cell homogenates preparations were employed to detect H. akashiwo during the 2001–2002 field study. The DNA probe methods consisted of a sandwich hybridization assay (SHA), which targets ribosomal RNA (rRNA), and an end-point polymerase chain reaction (PCR) assay, which targets internal transcribed spacer (ITS) sequences of rRNA genes. The SHA was used to provide semi-quantitative data showing the intermittent presence of the species during a 13-month period in Monterey Bay. Samples that showed a variety of responses in the SHA (negative as well as the highest) were then subjected to the PCR assay in an attempt to confirm species identification using an independent DNA probe method that employs cell homogenates; samples included those from Monterey Bay and one from a red tide event in San Francisco Bay. SHA and PCR assays agreed on the presence or absence of H. akashiwo. Gene products from two field samples positive for H. akashiwo by PCR were cloned and sequenced and found to be identical to those of that species in GenBank. When the same samples were viewed by light microscopy, however, H. akashiwo cells were only seen in the sample with the highest abundance of that species, as evidenced by SHA. It was extremely difficult to recognize naturally occurring H. akashiwo using light microscopy in field samples that had been preserved with Lugol's iodine, including samples that gave positive results by cell homogenate methods. Results of this study indicate that H. akashiwo is present along the open California coast and could easily be missed in routine phytoplankton surveys. Despite its presence, H. akashiwo does not appear to routinely bloom with sufficient densities to cause harmful outbreaks of the frequency and severity documented in some other coastal environments. Molecular identification techniques may be the preferred approach over light microscopy when there is a need to rapidly screen many samples for fragile, harmful species and those that are otherwise problematic to identify based on their gross morphology alone.     

赤潮过程浮游植物与营养物质时间变化率研究

利用2000年大亚湾澳头海域赤潮定点连续调查资料及其多年现场调查资料,采用灰色回归模型,综合分析赤潮发生过程水体中浮游植物细胞密度与营养物质(NO3-、NH4+、PO4^3-、SiO3^2-、Fe)的时间变化率关系,分析了叶绿素a含量与浮游植物细胞密度相互关系．结果表明,预测值与实测基值本一致,复相关系数范围在0．51～0．83．当水体叶绿素浓度为5．8μg·dm-3,预示可能发生赤潮,通过采样分析水体叶绿素a含量或利用水色卫星遥感资料反演水体叶绿素浓度,计算浮游植物细胞密度,为赤潮的预测预报提供简便有效的方法．此外．本水域初级生产力由磷控制．     

大鹏湾维生素B1、B12与赤潮

海湾及近海水域 ,因某些浮游生物爆发性繁殖引起的赤潮异常现象时有发生 ,已构成水产资源的严重破坏。为弄清赤潮发生的机理 ,已有许多作者进行过研究 ,现已知水温、盐度、溶解氧、ｐＨ、Ｎ、Ｐ ,以及Ｆｅ、Ｍｎ、Ｓｅ等微量元素都是发生赤潮的重要环境因子[1 ]。近年来通过生态实验研究 ,发现维生素Ｂ1 、Ｂ1 2 (以下简称Ｂ1 、Ｂ1 2 )亦有促进赤潮生物生长繁殖的作用[2 ]。本文报道 1 991年对大鹏湾海域水样中Ｂ1 、Ｂ1 2 含量的测定结果 ,并对赤潮发生与其相关性进行探讨。1　采样布点和分析方法1 .1　采样布点大鹏湾盐田海域的Ｓ0 、Ｓ1…     

Method for enumeration of 5-cyano-2,3-ditoyl tetrazolium chloride (CTC)-active cells and cell-specific CTC activity of benthic bacteria in riverine, estuarine and coastal sediments

Bacteria are the most abundant and active organisms in marine sediments and are critical for nutrient cycling and as a food source to many benthic and pelagic organisms. Bacteria are found both as free-living cells and as particle-associated cells, which can make investigations of these communities difficult. We found that common procedures for extracting bacteria from sediments leave the bacteria clay particle-associated and the clay particles clump, which reduce the reproducibility of direct counts. We optimized a sonication/surfactant method that produces a homogeneous suspension of bacterial cells against a uniform background of clay particles, which results in reproducible samples for epifluorescence microscopy. We developed a method to estimate CTC-positive cells and cell-specific CTC content in intact cores of surficial sediment communities from riverine, estuarine and coastal sites. Benthic bacterial abundances averaged 4.9x10(8) cells/g dry wt sediments in Apalachicola River, Florida sediments, 4.9-13.8x10(9) cells/g dry wt sediments in a variety of Apalachicola Bay sediments and 3.6x10(8) cells/g dry weight in shallow, anoxic Gulf of Mexico sediments. Percent CTC-positive cells ranged from low values of 9-10% CTC-positive cells in Apalachicola River and Apalachicola Bay sediments to high values of 25% CTC-positive cells in anoxic Gulf of Mexico sediments. After correction for abiotic CTC reduction and chlorophyll interference, estimates of cell-specific CTC reduction ranged from 0.15 to 0.55 fmol CTC(red)/active cell in the Apalachicola Bay sediments to 1.6 to 3.8 fmol CTC(red)/active cell in anoxic Gulf of Mexico sediments.     

Viral mortality in the final stage of Heterosigma akashiwo (Raphidophyceae) red tide

During a bloom of Heterosigma akashiwo (Raphidophyceae) in northernHiroshima Bay, Japan, in June 1993, the proportion of H.akashiwocells containing virus-like particles (VLPs) was monitored bytransmission electron microscopy. Until 3 days before the terminationof the red tide, no VLP-containing cells were detected, andthe proportion of VLP-containing cells was <1% on the last2 days of the red tide. However, the sample collected on thelast day, continuously incubated for 26 h at 22°C, revealeda high frequency (11.5%) of VLP-containing cells. These findingssuggest that viral mortality occurs in quite a short time andmay play an important role in regulating the disintegrationof H.akashiwo red tide.     

Daily variation of marine cladoceran densities in a tropical bay – Brazil

This contribution presents the daily variation of marine cladoceran densities in Guanabara Bay. The samples were obtained by vertical tows with a 200 m mesh-size conical net at different times over a 4-day period in September, 1995. Penilia avirostris was the most common species, followed by Evadne tergestina and E. spinifera. These organisms presented higher densities at nightfall. Physical factors like tide and light are responsible for the highest densities. High tide and nightfall time are requested conditions for the P. avirostris and E. tergestina abundance. Temperature and salinity appear to have no relation with cladoceran densities.     

Vertical microscale patchiness in nano- and microplankton distributions in a stratified estuary

Microscale (decimetre) vertical heterogeneity in the distributionof nano- and microplankton was studied on August 23, 1999 attwo different sites (separated by ~40 m) in Aarhus Bay, Denmark.At the time of sampling, the water column was stratified withrespect to both temperature and salinity and a subsurface fluorescencemaximum (corresponding to ~10 µg chlorophyll a l^-1) occurredimmediately below the primary pycnocline (9 m). Samples forspecies identification were taken at the surface, near the bottomof the water column and at 20 15-cm intervals in and aroundthe depth of maximum fluorescence. The plankton communitiesrecorded in the three different regions of the water columndiffered dramatically from one another. In addition, significantdifferences were found in the distribution patterns of speciesand functional groups in the region of sampling around the fluorescencepeak. The same patterns in vertical species distributions wereobserved at the two stations. In the region surrounding thesubsurface fluorescence peak, diatoms were, generally, regularlydistributed, although the total diatom biomass decreased slightlywith depth. Dinoflagellate species were mainly non-regularlydistributed and could be divided into two groups: (i) autotrophicor potentially mixotrophic species (Dinophysis norvegica, Dinophysisacuminata, Dinophysis cf. dens, Prorocentrum micans, Gymnodiniumchlorophorum and Ceratium macroceros) that mainly decreasedin numbers with depth or were aggregated in their distribution;and (ii) heterotrophic or potentially mixotrophic species (Ceratiumlineatum, Ceratium longipes, Ceratium furca, Ceratium tripos,Ceratium fusus, Protoperidinium curtipes, Protoperidinium steinii,Diplopsalis spp. and Katodinium glaucum). In the latter group,the species mainly increased in numbers with depth or were randomlydistributed. Most ciliates were uniformly distributed verticallyin the water column. However, small cells of the photosynthesizingciliate Mesodinium rubrum were most abundant at the depth ofmaximal fluorescence while large M. rubrum cells were equallyabundant at all the depths sampled, suggesting the two sizegroups of this organism may differ ecologically. Overall, thestudy demonstrates that traditional plankton sampling methodsmay lead to misinterpretations of speciesco-existence and interactions.     

TRENDS IN ABUNDANCE OF ALASKA HARBOR SEALS, 1983–2001

We estimated trends in abundance of harbor seals ( Phoca vitulina richardsii ) using over dispersed, multinomial models and counts obtained during aerial surveys conducted during 1983–2001 in the Ketchikan, Sitka, Kodiak, and Bristol Bay areas of Alaska. Harbor seal numbers increased significantly at 7.4%/yr during 1983–1998 and 5.6%/yr during 1994–1998 in the Ketchikan area, and 6.6%/yr during 1993–2001 in the Kodiak area. Counts were stable (trends not significant) during 1984–2001 (0.7%/yr) and 1995–2001 (-0.4%/yr) in Sitka, and during 1998–2001 (-1.3%/yr) in Bristol Bay. The influence of covariates ( e.g. , survey date, tide height) on trend estimates was significant and varied among areas and across years, demonstrating the need to include covariates in statistical analyses to accurately estimate trend. Our increasing trend estimate for Kodiak represents the first documented increase in harbor seal numbers over a relatively expansive area in the Gulf of Alaska. However, the trend for the Gulf of Alaska stock is equivocal due to the continued decline in Prince William Sound. Similarly, the trend for the Southeast Alaska stock is equivocal based on our increasing (Ketchikan) and stable (Sitka) trend estimates, and a recent decline reported for Glacier Bay. The Bering Sea stock appears stable after a period of possible decline.