Physiological responses to salinity changes of the isopod Idotea chelipes from the Baltic brackish waters

Survival, haemolymph osmolality and the specific metabolic rate of the isopod Idotea chelipes from the brackish waters of the southern Baltic Sea (salinity 7 psu) were investigated after acclimation to different salinities in the range of 3-30 psu keeping other parameters constant (T=15 degrees C, full air saturation). Haemolymph osmolality of I. chelipes increased from 379+/-33 mmol kg(-1) at 3 psu to 935+/-12 mmol kg(-1) at 30 psu. In the salinity range of 3-11 psu I. chelipes exhibited a hyperosmotic pattern of haemolymph regulation, whereas from 14 to 25 it behaved like an osmoconformer. Isoosmosis occurred at 28.5 psu (920 mmol kg(-1)). The specific metabolic rate of I. chelipes was significantly (P<0.05) related to salinity, with maximum (2.4+/-0.8 J g(-1) wm h(-1)) and minimum values (0.6+/-0.3 J g(-1) wm h(-1)) at 7 and 20 psu, respectively. The changes in the specific metabolic rate and in the difference between the osmolality of haemolymph and external medium were significantly correlated for I. chelipes (P<0.05). Rates of change per salinity unit were, however, unequal varying according to salinity transition. These differences indicate that osmotic adjustment may be a more or less costly in terms of energy according to salinity.     

Metabolic response of juvenile gray snapper (Lutjanus griseus) to temperature and salinity: Physiological cost of different environments

Juvenile gray snapper (Lutjanus griseus) occupy a wide range of estuarine and nearshore habitats that differ in physico-chemical properties. To quantify the energetic cost of inhabiting these different habitats, routine metabolism of individual gray snapper was measured in the laboratory at 20 combinations of temperature (18, 23, 28, and 33 °C) and salinity (5, 15, 25, 35, and 45 psu). An open, flow-through respirometer was used, enabling trials to be run for long periods (∼16 h), while maintaining water quality (dissolved O₂>70% saturation), and providing fish sufficient time to habituate to the chambers undisturbed. Video recordings of fish in the respirometer chambers were analyzed to quantify the spontaneous activity rate of individuals. Analysis of covariance, using fish weight and mean activity rate as covariates, indicated significant temperature and salinity effects on oxygen consumption. Oxygen consumption was significantly higher at high salinities, and the salinity effect was temperature dependent. A polynomial equation describing oxygen consumption as a function of temperature and salinity indicated the increase due to salinity from 5 to 45 psu at high temperatures (30-33 °C) was equivalent to a 3 °C increase in temperature. At intermediate temperatures (24-26 °C), the increase due to salinity from 5 to 45 psu was less dramatic, equivalent to a 2 °C increase in temperature. At the lowest temperatures (18 °C), salinity did not have a significant effect on oxygen consumption. The increased metabolic costs in high salinities (∼7% at the high temperature) represent a significant energy cost for juveniles, that would need to be balanced by lower predation risk or greater food availability to result in similar juvenile production compared to lower salinity environments.     

The effect of temporal variability in salinity on the invasive red alga Gracilaria vermiculophylla

Non-native, invasive species are often characterized by being tolerant to environmental stressors, leaving them more fit relative to native species. The red alga Gracilaria vermiculophylla originates from the NW Pacific but has recently spread along the coastlines of Western Europe, where it has become abundant in many shallow, soft-bottom estuaries. Salinity is important for the local and regional distribution of algae. The distribution of G. vermiculophylla in Europe suggests that it thrives well in hyposaline environments and that it may be more fit than some native algae under such conditions. Little, however, is known about the ecophysiology of G. vermiculophylla and it is therefore difficult to predict its spread and future distribution. Laboratory experiments with G. vermiculophylla showed that steady-state salinity above 15?psu was optimal for growth and that the growth rate was reduced at salinities below 15?psu. Variable salinity reduced the growth rate and larger oscillations were more stressful than small ones. Exposure to very low salinity (0–5?psu) was stressful for the alga and algae exposed to these low levels for 2–4 days were unable to recover fully. Gracilaria vermiculophylla did not seem to perform better in hyposaline conditions than many native, estuarine species. The present distribution of G. vermiculophylla in Scandinavia can be explained well by its response to salinity, but this may not explain its present success relative to many naturally occurring algal species.     

Osmotic and ionic haemolymph concentrations in the Baltic Sea amphipod Gammarus oceanicus in relation to water salinity

The aim of this work was to determine the osmotic and ionic (Na(+), K(+), Ca(2+), Mg(2+) and Cl(-)) haemolymph concentrations in Gammarus oceanicus at different salinity levels. Being a species of marine origin it inhabits brackish waters of the Baltic Sea. G. oceanicus specimens were collected in January 2003 from the Gulf of Gdansk (salinity 7 psu). The animals were gradually acclimated to eight different salinity levels (5, 7, 14, 20, 25, 30, 35 and 41 psu) at a temperature of 5 degrees C and 100% oxygen saturation. The haemolymph osmolalities correlated positively with external salinity, from 545.4+/-17.3 mOsm in 5 psu to 1185.9+/-34.6 mOsm in 41 psu. G. oceanicus hyperregulated within the 5-31.5 psu range; above 31.5 psu it hyporegulated its body fluids in comparison to the external medium. At 31.5 psu (1017 mOsm) the haemolymph concentration of G. oceanicus was isoosmotic with the habitat. The haemolymph concentrations of all the studied ions, except K(+), correlated positively with their concentrations at the various salinity.     

Effects of salinity and temperature on the expression of enzymatic biomarkers in Eurytemora affinis (Calanoida, Copepoda)

In order to establish effective enzymatic biomarkers that could provide in situ early warning of contaminant exposure in estuarine ecosystems, the potential effects of the principal abiotic factors (temperature and salinity) were investigated on common biomarkers, the acetylcholinesterase (AChE) and the glutathione S-transferase (GST) in Eurytemora affinis. Short term salinity stress effects simulated during an experimental tide indicated that enzymatic activities of this species are characterized by maximum expression related to an optimal salinity range (between 5 and 15 psu). Moreover, longer time exposure to various salinity tanks confirmed the effects of this factor on both AChE and GST activities. Therefore, optimal AChE activity was measured at 10 psu, while optimal GST activity was measured at 5 psu. Furthermore, significant effects of temperature were also recorded, particularly for AChE expression (slight effects were measured on GST expression) with an optimal condition at 11 degrees C. These experiments indicated a more pronounced effect of salinity over temperature especially on the AChE expression and confirmed the need to standardize sampling procedures in relation with environmental parameters for biomonitoring studies based on enzymatic analyses.     

Resistance patterns to salinity and temperature in an estuarine mysid (Mysidopsis bahia) in relation to its life cycle

Survival of the estuarine mysid Mysidopsis bahia was monitored in the laboratory from juvenile through maturation to an adult under various combinations of salinity (3–31 %₀ S) and temperature (19–31°C). Salinity had a greater influence on mysid survival than did temperature. Salinity tolerance was strongly modified by temperature with maximum tolerance at intermediate temperatures. Salinity-temperature interactions on mysid survival varied with age of the mysid. Canonical analysis of weekly resistance patterns produced absolute maximum survival combinations for salinity and temperature which changed over time. The salinity-temperature tolerance ranges described for this estuarine mysid correspond with previously described physiological capabilities and distribution patterns.     

Estuarine and scalar patterns of invasion in the soft-bottom benthic communities of the San Francisco Estuary

The spatial patterns of nonindigenous species in seven subtidal soft-bottom communities in the San Francisco Estuary were quantified. Sixty nonindigenous species were found out of the 533 taxa enumerated (11%). Patterns of invasion across the communities were evaluated using a suite of invasion metrics based on the abundance or species richness of nonindigenous species. Patterns of invasion along the estuarine gradient varied with the invasion metric used, and the ecological interpretation of the metrics is discussed. Overall, the estuarine transition community located in the estuarine turbidity maximum zone (mean 5 practical salinity unit (psu)), main estuarine community (mean 16 psu), and marine muddy community (mean 28 psu) were more invaded than two fresh-brackish communities (mean < 1 psu) and a marine sandy community (mean 27 psu). Nonindigenous species were numerically dominant over much of the Estuary, making up more than 90% of the individuals in two communities. The percentage of the total species composed of nonindigenous species increased at smaller spatial scales: 11% at the estuary (gamma) scale, 21% at the community (alpha) scale, and 42% at the grab (point) scale. Wider spatial distributions of nonindigenous species and a relatively greater percentage of rare native species may have resulted in this pattern. Because of this scale dependency, comparisons among sites need to be made at the same spatial scale. Native species were positively correlated with nonindigenous species in several of the communities, presumably due to similar responses to small-scale differences in habitat quality. The rate of invasion into the soft-bottom communities of the San Francisco Estuary appears to have increased over the last one to two decades and many of the new introductions have become numerically dominant.     

Development and mortality of the first naupliar stages of Eurytemora affinis (Copepoda, Calanoida) under different conditions of salinity and temperature

As a prevalent species complex in temperate estuaries and salt marshes of the Northern Hemisphere, populations of Eurytemora affinis that inhabit these environments must be adapted to salinity fluctuations. Some populations have invaded freshwater environments. In this work, we focus on the combined effects of temperature and salinity fluctuations on mortality rates and development time of the first naupliar stages under starvation. Two temperatures (10 and 15 °C) and eight salinities, ranging from 0 to 35 psu are investigated. We show (i) that among all experimental conditions the optimal temperature and salinity for naupliar survival and development are 15 psu and 15 °C, and (ii) that only the most extreme salinities (i.e. 0 and 35 psu) have a negative effect on naupliar survival. Nauplii develop faster and reach a higher developmental stage at 15 than at 10 °C, independent of salinity. The relevance of this metabolic adaptive pattern is discussed in the general framework of in situ behavior, tidal forcing and biogeographic variability, as well as the potential sources of the observed individual variability.     

Changes of photosynthetic performances in mature thalli of the red alga Gelidium amansii (Gelidiaceae) exposed to different salinities

Photosynthetic parameters, including net O₂ evolution, pigment content and fast chlorophyll a (Chl a) fluorescence kinetics, were studied in mature thalli of Gelidium amansii, a marine agar-producing red seaweed, exposed to different salinities (0–35?psu) for 10 days. The results showed that the net O₂ evolution at 25–32?psu was unchanged, but significantly decreased at either lower or higher salinities. Hypo-saline (15?psu and below) and hyper-saline (35?psu) conditions induced significant losses of Chl a, carotenoids and phycobiliproteins, which correlated with the decrease in the absorption flux per cross-section of fronds (ABS/CS_o). Polyphasic fluorescence transients revealed that salinities at 10 and 35?psu both caused multiple effects on photosynthetic electron transport. Algae exposed to low salinity at 10?psu showed extensive damage to the donor side of the oxygen-evolving complex (OEC), reaction centre and acceptor side of PSII. Data on net O₂ evolution showed that 35?psu salinity was more destructive than 10?psu, with the absence of marked osmotic injury to the OEC. The results of this study indicate the possibility of G. amansii cultivation in estuarine waters with 20–32?psu salinity.     

Temperature and salinity effects on post-marsupial growth of Neomysis integer (Crustacea: Mysidacea)

There has been an increasing interest in using the brackish water mysid Neomysis integer as a toxicological test species for Western European estuarine systems. In this respect, more data on growth, moulting and development in this species is needed. The influence of prevailing environmental variables (e.g. temperature, salinity) and age on these processes as well as their optimal range have to be known in order to develop optimal laboratory cultures and to differentiate between chemically induced variability and natural variability in toxicity testing. Individual post-marsupial growth (size, intermoult period, growth factor) was studied from first day neonates until adulthood at eight environmentally relevant temperature-salinity conditions. Three salinities (5, 15 and 30 psu) were tested at 15 and 20 °C, and two more extreme temperatures (8 and 25 °C) were tested at a salinity of 5 psu.Survival and growth of N. integer were detected within the whole range tested, but sexual maturation was only possible in the narrower range of 15-25 °C and 5-15 psu. The size at maturity of N. integer increased with decreasing temperature and increasing salinity. Salinity seems to have a stronger effect than temperature on the duration of maturation. The sigmoid von Bertalanffy growth model was fitted to the individual and pooled data, except for the 8 °C experiment where growth was linear. Estimates from pooled data were comparable with individually based estimates, but generally underestimated the asymptotic length. Temperature was negatively correlated with the asymptotic length and positively correlated with the growth constant K. Higher temperatures caused smaller intermoult periods but had no effect on the growth increment, while salinity effects were less straightforward and dependent on the water temperature. A tool is provided to estimate the age, moult number, intermoult period, growth factor and growth rate from the body standard length of N. integer. Experimentally derived von Bertalanffy parameter estimates resulted in a higher growth performance index compared with field-based estimates for the Schelde estuary and Galgenweel populations of N. integer.     

温度和盐度突变对菲律宾蛤仔斑马蛤耗氧率和排氨率的影响

为研究温度和盐度对蛤仔新品种斑马蛤耗氧排氨的影响,以野生蛤仔为对照,实验设置15、20、25、30和35℃五个温度梯度和20、25、30、35和40五个盐度梯度,结果表明:温度和盐度对斑马蛤的耗氧率和排氨率影响显著（P < 0.05）。在温度15-35℃内,随着温度的增加,耗氧率和排氨率整体上呈增加的趋势。在20-40盐度内,耗氧率随着盐度的升高先减少后增加,排氨率随着盐度的升高先增加后减少,在盐度为30时达到最高值。在水温为15℃,盐度20-40内,斑马蛤的O:N为9.534-62.008;在盐度为35,水温在15-35℃内,斑马蛤的O:N是20.700-74.138。与野生蛤仔比较,斑马蛤的耐高温能力要强于野生蛤仔,从Q₁₀的变化反映出斑马蛤对温度的敏感性相对较弱,适应温度变化的能力比较强;斑马蛤的耐低盐和耐高盐能力强于野生蛤仔。研究结果为进一步完善蛤仔斑马蛤的人工养殖技术提供参考。     

Eco-physiological responses to salinity changes across the freshwater-marine continuum on two euryhaline bivalves: Corbicula fluminea and Scrobicularia plana

The influence of global climate change will potentially alter the salinity of aquatic ecosystems. This represents a tremendous challenge for societies worldwide. Different sources of salinization (natural or anthropogenic) amplify the introduction of salt in rivers and streams, causing an increase of salt flowing down to estuarine and coastal areas. In this study, Corbicula fluminea and Scrobicularia plana have been selected because of their large tolerance for salinity variation (euryhaline organisms). They will allow the study of effect on the whole spectrum of salinity from fresh to marine waters respectively. The aim was to study the impact of experimental salinity stress at physiological, biochemical and behavioral levels by exposing both species to a salinity close to their limit range of tolerance, 15 practical salinity unit (psu), and at their field salinity (1.5 psu and 30 psu for C. fluminea and S. plana respectively) in the presence or absence of food during 2 and 7 days of exposure. Negative impacts of hyper saline condition for C. fluminea (15 psu) and hypo saline condition for S. plana (15 psu) have been measured at biochemical, physiological and behavioral levels. At sub-individual and individual levels, structural and energetic parameters and behavioral impairments seemed to be suitable biomarkers to assess salinity stress on C. fluminea and S.plana. After exposure to the limit of salinity tolerance (15psu) for both organisms, fitness modifications could appear, and may participate in endangering populations.     

Metabolic responses to salinity changes in the subantarctic notothenioid teleost <Emphasis Type="Italic">Eleginops maclovinus</Emphasis>

Eleginops maclovinus is an endemic, subantarctic Notothenioidei species. This study examined the influence of different environmental salinities (5, 15, and 45 psu; and 32 psu as a control) on energy metabolism in E. maclovinus over a period of 14 days. Metabolite contents and enzymatic activities related to carbohydrate, amino acid, and lipid metabolisms were evaluated in metabolic (liver) and osmoregulatory (gill and kidney) tissues. At extreme salinities (5 and 45 psu), the liver showed a high consumption of energy reserves, mainly as amino acids and carbohydrates. Carbohydrate metabolism in the gills did not change under different salinities, but increased lactate levels were found, suggesting that this tissue may use lactate as an energy substrate. Amino acid metabolism in the gills decreased at 5 psu but increased at 45 psu, and lipid metabolism increased at 5 and 15 psu during the first days of the trial, indicating a possible use of lipids as energy. Kidney carbohydrate catabolism and amino acid metabolism increased after 14 days at 45 psu, while lipid metabolism did not vary in relation to salinity changes. Together, these results suggest that the liver is most affected by salinity changes, probably due to its role as a supplier of energetic substrates. The gills and kidney, osmoregulatory tissues, maintained their energy metabolism levels with minor modifications. In conclusion, E. maclovinus exhibits metabolic adjustments to adapt to different salinities, showing the best responses in isosmotic environmental salinities.     

Effects of nitrogen and phosphate enrichment on the activity of nitrate reductase of <Emphasis Type="Italic">Ulva prolifera</Emphasis> in coastal zone

As one of the main species causing “green tides”, Ulva prolifera always inhabits in estuarine areas with changes in salinity and nutrients. Reduced salinity may affect directly or indirectly the processes of uptake and assimilation of nitrate, in which the nitrate reductase (NR) activity play the crucial roles. In this experiment, we investigated the different effects of enriched nitrogen and phosphate on NR activity of Ulva prolifera at salinity 30, 15, and 5 psu. The results showed that when salinity being lowered NR activity decreased under no enrichment (CT) or PO₄ ^3? enrichment condition. NO₃ ^? or combination with PO₄ ^3? could significantly enhance NR activity at three salinities, among which the highest value occurred at 15 psu. Enrichment of NH₄ ⁺ significantly decreased NR activity at 30 and 15 psu, but not at 5 psu. The results suggested NR of Ulva prolifera could be triggered by NO₃ ^?, especially at middle salinity, and keep low when exposed under hyposaline or NH₄ ⁺ enrichment for long term to rapidly respond to pulse of NO₃ ^? in estuarine areas.     

Effects of Salinity and Nutrient Addition on Mangrove Excoecaria agallocha

Effects of salinity on seed germination and growth of young (1 month old) and old (2-year old) seedlings of Excoecaria agallocha were investigated. Combined effects of salinity and nutrient level were also examined on old seedlings. Seed germination was best at 0 and 5 psu salinity. 15 psu salinity significantly delayed root initiation and decreased final establishment rate. All seeds failed to establish at 25 psu salinity. Young seedlings performed best at 0 and 5 psu, but growth was stunned at 15 psu, and all seedlings died within 90 days at 25 psu. Old seedlings grew best at salinities below 5 psu and they survived the whole cultivation at 25 psu. This indicated that E. agallocha increased salt tolerance over time. Gas exchange was significantly compromised by salinities above 15 psu but evidently promoted by high nutrient. Proline accumulated considerably at high nutrient, and its contents increased from 0 to 15 psu but decreased at 25 psu salinity. Lipid peroxidation was aggravated by increasing salinity beyond 15 psu but markedly alleviated by nutrient addition. These responses indicated that E. agallocha was intolerant to high salinity but it can be greatly enhanced by nutrient addition.     

The effect of salinity on experimental infections of a Hematodinium sp. in blue crabs, Callinectes sapidus

The parasitic dinoflagellate Hematodinium sp. parasitizes blue crabs along the Atlantic seaboard of the United States. Infections in blue crabs have only been reported from waters where salinity is >11 practical salinity units (psu). Blue crabs maintain a hyperosmotic internal concentration at low salinities (0-5 psu), roughly comparable to 24 psu, and should be capable of maintaining an infection in low-salinity waters even if Hematodinium spp. cells are intolerant of low salinities. We tested this notion by observing the effect of low salinity on the progression of disease in crabs experimentally infected with the parasite. Blue crabs were acclimated to 5 psu or 30 psu salinity treatments. They were inoculated with Hematodinium sp. and necropsied 3, 7, 10, and 15 days post-inoculation. The low-salinity treatment did not have an effect on the proliferation of Hematodinium sp. infections in blue crabs; moreover, a greater proportion of infections in crabs in the low-salinity treatment developed dinospore stages than did those in the high-salinity treatment, indicating that salinity may affect the development of the parasite. However, dinospores from in vitro cultures rapidly became inactive when held in salinities <15 psu. Our experiments indicate that Hematodinium spp. can develop in blue crabs at low salinities, but that the parasite is incapable of transmission in this environment, which explains the lack of natural infections in crabs at low salinities.     

A response-surface analysis of the relative importance of the temperature,salinity and body weight on the respiratory metabolism of the white shrimp Litopenaeus vannamei (Boone, 1931)

We used a response-surface analysis to determine the importance of different factors affecting the resting routine metabolic rate (QO₂) of the white shrimp Litopenaeus vannamei. The oxygen consumption rates were estimated using a multi-factorial design with 28 combinations of different salinities (15, 20, 25, 30, 35, 40 and 45 psu) and temperature (20, 25, 30 and 35 °C) values. The response-surface analysis produced a quadratic function showing that temperature more profoundly affects the oxygen consumption rate. Response-surface curves were generated to predict the optimal conditions resulting in oxygen consumption to better understand the successful growth of this species.     

Differential response of varying salinity and temperature on zoospore induction, regeneration and daily growth rate in Ulva fasciata (Chlorophyta, Ulvales)

Seaweed cultivation is imperative to augment increasing industrial demand. Ulva fasciata Delile is a potential seaweed for cultivation with applications in food industries. There is a renewed interest in large-scale aquaculture of this species in India due to its envisaged demand in snack food products. In the present study, we have successfully demonstrated the possibility of inducing zoospores in vegetative tissue, effective regeneration and improved growth in this seaweed by manipulating salinity (from 15 to 30 psu) and temperature (from 15 to 35°C). The optimum salinity and temperature requirement for zoospores induction were found to be 15 psu and 25°C, respectively. The quadriflagellate zoospores showed negative phototaxis and the settlement and germination pattern similar to several other green seaweeds. The optimum regeneration (78.53?±?10.05%) was recorded at 25°C and 30 psu salinity. The maximum daily growth rate (16.1?±?0.28%) was at 25°C and 30 psu salinity which corresponded to the field conditions. This method could be further refined at nursery culture to achieve artificial seeding essential for the success of commercial cultivation of this seaweed.     

Embryonic encapsulation and maternal incubation: Requirements for survival of the early stages of the estuarine gastropod Crepipatella dilatata

During their reproductive period, females of Crepipatella dilatata deposit their embryos in capsules that they then brood in the pallial cavity until juveniles emerge several weeks later, after passing through a transient veliger “larval” stage. Artificially excapsulated veligers of this species experimentally exposed to a wide range of salinities (5, 10, 15, 20, 25, and 30 psu) for six hours showed reduced activity at salinities of 15 and 20 psu, whereas encapsulated veligers exposed to those same salinities showed no reduction of activity. Artificially excapsulated veligers showed high mortality at salinities of 5 and 10 psu; encapsulated embryonic stages also showed high mortalities at 5 psu and serious sublethal effects at 10 psu in tests excluding maternal protection, showing that encapsulation alone does not provide complete protection from low salinity stress. Natural tidal cycles in the Quempillén River estuary also reduced embryonic survival at salinities of ≤ 10 psu when the capsules were exposed without maternal protection. In contrast, encapsulated embryos protected by their mothers survived well regardless of the salinity to which they were exposed, under both natural and laboratory-simulated estuarine tidal cycles. C. dilatata are able to develop in the estuary only because of maternal protection, since salinity levels in this environment sometimes decline to as low as 7 psu. Successful embryonic development in this estuary reflects the capacity of C. dilatata adults to detect dangerously low salinity levels and then seal themselves off from the environment for up to 50 hrs (O. Chaparro pers. obs.) when the salinity drops below 22.5 psu, allowing salinity to remain above this level within the pallial cavity despite continued salinity declines in the surrounding seawater.     

Factors influencing the sporulation and cyst formation of Aphanomyces invadans, etiological agent of ulcerative mycosis in Atlantic menhaden, Brevoortia tyrannus

Oomycete infections caused by Aphanomyces invadans occur in freshwater and estuarine fishes around the world. Along the east coast of the USA, skin ulcers caused by A. invadans are prevalent in Atlantic menhaden, Brevoortia tyrannus. From laboratory observations low salinities appear crucial to transmission of the pathogen. To better understand aspects of transmission, we characterized sporulation and cyst formation of secondary zoospores of two isolates of A. invadans at different salinities and temperatures. Sporulation occurred only at low salinities. At room temperature (ca. 20-22 C), using "pond water" augmented with artificial sea salts, the endemic strain WIC and the Thailand strain PA7 of A. invadans produced free-swimming secondary zoospores at salinities of 0, 1 and 2 psu (practical salinity unit = per thousand), but not at 4 psu or higher. Secondary zoospores of another species, ATCC-62427 (Aphanomyces sp.), were observed at 1, 2, 4 and 8 psu but not at 0 and 12 psu. Secondary zoospores of all three isolates, especially WIC, were abundant and motile 1-2 d postsporulation. Sporulation was temperature dependent and occurred over a relatively narrow range. No sporulation occurred at 4, 30 or 35 C for either WIC or PA7. For both strains zoospore production within 1-3 d after the initiation of sporulation was more prolific at 25 C than at 20 and 15 C. At 15 C production of zoospores was sustained over 11 d for WIC and 5 d for PA7. At room temperature single WIC secondary zoospores remained motile 12-18 h. Salinities exceeding 4 psu or vigorous shaking caused immediate cyst formation of WIC secondary zoospores. Exposure to menhaden tissue, but not tissues of other fishes to secondary zoospores (WIC), caused rapid (2 h) cyst formation. Cysts were capable of excysting when transferred to 1 psu water within 2-3 h of cyst formation. Cysts that had remained encysted in 6.5 psu for 24 h did not excyst when transferred to 1 psu water. Salinity and temperature requirements for sporulation indicate that juvenile menhaden must acquire infections during rain or in low salinity oligohaline waters.