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New Zealand's green-lipped mussel (Perna canaliculus) is an ecologically and economically important species. Marine heatwaves are increasing in frequency around NZ's coastline, and these events are correlated with increased stress and mortality of some aquaculture species. This study aimed to identify general biomarkers of heat stress in P. canaliculus and to assess whether responses differed between genetically distinct selectively bred mussels. We exposed three families of selectively bred mussels (families A, B and C) to three seawater temperature regimes in the laboratory: 1) a "control" treatment (ambient 12°C), 2) a 26°C heat challenge with a subsequent recovery period, and 3) a sustained 26°C heat challenge with no recovery. We investigated whether the survival, immune response (hemocyte concentration and viability, oxidative stress and total antioxidant capacity), hemocyte gene expression and gill microbiome differed between the families during the temperature challenges. In the sustained heat-stress treatment, family A had the highest survival rate (42% compared with 25% and 5% for families C and B, respectively). Gene expression levels significantly shifted during thermal stress and differed between families, with family A more dissimilar than families B and C. Family C had substantially more genes impacted by temperature treatment and timepoint than the other families, while family B had very little genes/pathways that responded to thermal stress. Genes related to heat shock proteins and immune responses (e.g., AIF1, CTSC, TOLL8, CASP9, FNTA, AHCY, CRYAB, PPIF) were upregulated in all families during heat stress. Microbiome species-richness differed between families before and during heat-stress, with family A having a distinctly different microbiome flora than the other families. Microbial diversity changed similarly in all families exposed to prolonged heat-stress, with species of Vibrio and Campylobacter increasing in these mussels. Our study highlights the use of non-lethal sampling of hemocytes as a diagnostic tool to explore the immune response and gene expression of selectively bred mussels, to predict their response to ocean warming. This approach can identify potential thermotolerant candidates for further selective breeding, which may increase the resilience of the mussel aquaculture industry in a warming ocean. [ABSTRACT FROM AUTHOR]

The nutritional condition of seed mussels (spat) at the time of seeding is thought to play a major role in influencing spat losses on mussel farms, with nutritionally compromised spat thought to be more likely to be lost from aquaculture substrata than those in good condition. New Zealand's Greenshell™ mussel (Perna canaliculus) aquaculture industry relies almost exclusively (∼85%) on wild caught spat, with the vast majority, typically > 90%, lost shortly after being seeded onto coastal mussel farms. This study sought to quantify the extent of the variability in the nutritional condition (i.e., calorific, protein, lipid and carbohydrate content) of samples of spat used by the Greenshell™ industry between 2014 and 2021 (i.e., wild caught and hatchery produced) as a first step towards determining whether variability in nutritional condition may be a significant contributor to the marked variation observed in spat performance after it is seeded onto coastal farms. All parameters measured were highly variable among wild spat samples, with all but lipid content varying, including for samples of wild spat harvested at different times within the same month. Furthermore, there were no clear patterns in the nutritional condition of spat across months or years, suggesting environmental factors that limit feeding during the harvest and transport of spat may be contributing to marked differences in the quality of spat being used by the industry to seed farms. These results provide a baseline for assessing the nutritional condition of Greenshell™ spat and point towards a need to develop interventions to improve the quality of wild spat supply for this industry to help and prevent the ongoing massive losses of spat from farms. These measures could include, testing nutritional condition of spat at harvest or prior to seeding, development of nursery culture methods and development of artificial spat feeds that have the potential to replace live phytoplankton during nursery culture. [ABSTRACT FROM AUTHOR]

The Greenshell™ mussel, Perna canaliculus, is the most lucrative aquaculture species in New Zealand; however, losses of commercial juvenile mussels (spat) are common and can drastically reduce the yield of adults. Modification of seeding density is a potential method to manage loss of spat (~ 1–20-mm shell length), by reducing biofouling and intraspecific competition. Understanding the stress responses of spat in production systems during the early stages of aquaculture is a high priority for mussel farmers who aim to manage and optimise mussel production. Here, the effects of seeding density (low: 257, mid: 515, and high: 1030 mussels 10 cm−1 of culture rope), space (depth, site), and time (December 2019 and February 2020) on oxidative stress responses (total antioxidant capacity [TAC] and lipid peroxidation [LP] biomarkers) in spat and their viability (freshwater immersion test) were assessed in the Marlborough Sounds, New Zealand. The effect of seeding density on TAC was variable among depths, sites, and times, whereas seeding density had no impact on LP. In late summer (February 2020), oxidative stress appeared to be higher compared to early summer (December 2019). This indicates that mussels were exposed to more stressful conditions later in summer, although this did not reduce their short-term viability. This research found that seeding density had no major impact on oxidative stress biomarkers and suggests that stress responses and viability of Greenshell™ mussel spat during the summer months are not affected by the seeding density levels measured in this study. [ABSTRACT FROM AUTHOR]

The physiology of the New Zealand sea urchin Evechinus chloroticus was evaluated through feeding, respiration, growth and gonad growth in adult animals acclimated for 90 days at 18 °C (annual mean temperature) and 24 °C (ambient summer temperature (21 °C) + 3 °C). Measured parameters with representative rates of assimilation efficiency were used to calculate scope for growth (SfG) for each treatment. All physiological parameters were negatively affected at 24 °C, showing a decrease in feeding rate which coincided with negative growth and gonad development at the end of the acclimation period, and a decrease in respiration rate suggesting metabolic depression. Histology of gonad samples after the acclimation period also showed no gametic material in animals acclimated at 24 °C. All animals acclimated at 24 °C had negative growth, differing from the calculated SfG which indicated that the animals had sufficient energy for production. The results suggest that calculated SfG in echinoderms should be used together with actual measurements of growth in individuals as, by itself, SfG may underestimate the actual effect of ocean warming when animals are exposed to stressful conditions. Overall, considering the total loss of reproductive output observed in E. chloroticus at higher temperatures, an increase in seawater temperature could dramatically influence the persistence of northern populations of this species, leading to flow-on effects in the subtidal ecosystem. [ABSTRACT FROM AUTHOR]

Temperature and salinity are important environmental factors affecting the normal functioning of marine animals, particularly animals such as sea urchins living in shallow waters and tide pools. Here, we evaluated the effect of different combinations of temperature and salinity on early embryos of the endemic New Zealand sea urchin Evechinus chloroticus. Animals were collected at Matheson's Bay (36º18′17′′S; 174º47′51′′E) in north-eastern New Zealand in February 2013. Embryos were exposed to five salinities (29, 31, 34, 35 and 37 ppt) and two temperatures (18 and 21 °C) during the first 24 h of development. Low salinity (29 ppt) affected all parameters (fertilization, development rate, gastrulation and normal development), with ca. 50 % of embryos surviving at 29 ppt, whereas seawater temperature only affected development rate and gastrulation. An increase in temperature from 18 to 21 °C minimized the negative effect of low salinity (≤31 ppt) on development rate and gastrulation of E. chloroticus. Overall, the results of this study suggest that early embryos of E. chloroticus have developmental plasticity to withstand reductions in salinity up to 29 ppt; however, it is still unknown whether the surviving embryos will be able to complete larval development at low salinities, particularly whether the embryos and larvae are carried into extreme environments such as estuaries where salinity is even lower. Multistressor studies are very important for climate change research as multiple environmental factors will act together in the wild, having major consequences for development and recruitment of marine invertebrates. [ABSTRACT FROM AUTHOR]

Seawater temperature is an important environmental factor for the early life stages of marine invertebrates. In this study we evaluated and described the effects of temperature during early development of E. chloroticus, identifying the optimum temperature range and upper thermal limit for successful development. The temperature range evaluated was between 15–24°C which included the normal seawater temperatures during the spawning season in northern New Zealand, as well as the highest temperature projected by the IPCC for this region due to global warming (1–3°C by the year 2100). Gametes from several females and males were used in the experiment. Fertilization was carried out at different temperatures and development was monitored at different time points after fertilization in each temperature. The development rate of E. chloroticus increased with an increase in seawater temperature. However, at temperatures higher than 21.5°C the amount of abnormal development reached ∼30%. The optimum temperature for early development was between 15–21°C, whereas the upper thermal limit was ∼24°C. Therefore, early development of E. chloroticus is negatively affected by an increase in seawater temperature of ∼3–4°C above current seawater temperature levels in northern New Zealand. The thermal sensitivity of early life stages of E. chloroticus could affect survival rates during early development of this species in a global warming scenario, which could impair recruitment in populations which are exposed to higher temperatures, leading to possible distributional shifts of this species. [ABSTRACT FROM AUTHOR]

The New Zealand geoduck clam is a unique seafood delicacy, with animals selling for up to $US 220–330/kg. Stress accumulated during transport of juveniles to grow-out sites represent a bottleneck in the aquaculture process. In this study, the physiological responses of juvenile geoducks following emersion (3- and 8-h), and recovery (1- and 5-days) were investigated. An integrated approach of flow cytometry, osmolality and metabolomics, along with behavioural assessments was used. Both cellular and chemical haemolymph parameters and metabolite profiles were recorded for P. zelandica juveniles and are reported herein for the first time. An increase in haemolymph osmolality was experienced with an increase in emersion period, with significant differences seen between the 3- and 8-h emersion groups after 5 days of recovery. Viability measures of haemocytes varied insignificantly between experimental groups, creating baseline ranges. The proportion of haemocytes undergoing respiratory burst activity did not appear to be affected by emersion and re-immersion. Haemocyte mitochondrial membrane potential was highest following 1-day of recovery, likely linked to metabolic readjustment, and increased glycolysis, taking place following emersion. Metabolomics analyses suggest that protein, lipid and carbohydrate metabolite classes assist with energy production in geoducks. Activation of anaerobic metabolic pathways, with a high dependence on succinate, were prominent in the 8-h exposure group, with metabolic recovery still taking place following 5-days of immersion, mainly due to proteins restoring energy reserves. Elucidating the physiological responses of juvenile geoduck subjected to transport stress can aid cultivation methods already underway to develop a novel, high value aquaculture industry. [Display omitted] • Prolonged emersion (8 h) requires extended recovery to restore physiological parameters (>5d). • The presence of geoduck siphons post emersion is easy to apply and useful to guide recovery. • Emersion apparently supported by anaerobic energy production pathways. • Increased haemolymph osmolality resulting in increased amino acid production. • The haemocyte status was not significantly affected by emersion or recovery. [ABSTRACT FROM AUTHOR]

Seawater temperature is projected to increase globally due to climate change, affecting physiological responses, fitness and survival of marine organisms. Thermal tolerance studies are critical to determine the ability of animals to adapt to future environmental conditions. In this study, we aimed to determine if the thermal limits of the New Zealand Evechinus chloroticus would shift with animal's thermal history. We tested the effect of six thermal regimes on the righting ability, temperature of loss of righting (T LOR), median lethal temperature (LT 50), lethal temperature (LT) and the gene expression of the heat shock protein 70 (hsp70) of the New Zealand sea urchin E. chloroticus when exposed to a thermal shock of 1 °C day−1 (duration of 7–16 days depending on the treatment). Treatments consisted of laboratory acclimation for one and four weeks to 18 °C and 24 °C (mean winter (15 °C) and summer temperature (21 °C) + 3 °C of warming, respectively), compared to non-acclimated sea urchins collected during winter (14.6 °C) and summer seasons (20.4 °C). Thermal history did not have a significant effect on the righting ability of E. chloroticus (T LOR ranged between 28 and 29 °C for all treatments) and LT 50 (ranged between 29 and 30 °C for all treatments). However, LT of E. chloroticus collected during winter season was significantly lower than animals acclimated for one week at 18 ° C. Maximum expression of hsp70 mRNA (T max) was observed at around 27–28 °C regardless of treatment; however, relative hsp70 mRNA levels were significantly higher in animals acclimated for four weeks at 24 °C. Despite proving to be a thermotolerant species with LTs around 30 °C, E. chloroticus was unable to increase thermal tolerance and T max when acclimated to high temperatures, suggesting that E. chloroticus may have a limited adaptive capacity to modify its phenotype; however, evolutionary adaptations may allow E. chloroticus to adapt to future ocean temperatures. Unlabelled Image • E. chloroticus is a thermotolerant temperate species with thermal limits >28 °C. • Acclimation temperature or season did not affect thermal limits of E. chloroticus. • Maximum hsp70 gene expression was observed at ~27–28 °C regardless of treatment. [ABSTRACT FROM AUTHOR]

In the family Asterinidae, development through a planktonic lecithotrophic brachiolaria larva is common and has evolved independently several times. Here, we describe the lecithotrophic development of the asterinid Stegnaster inflatus, a species endemic to New Zealand. Early development through the blastula and gastrula stages is short, with hatching at the brachiolaria stage occurring within 48 hr. After hatching, larvae are negatively buoyant, and without aeration remain near the bottom of the culture containers. The settled benthic juvenile stage was reached in ~2 weeks. The brachiolaria of S. inflatus shares common characteristics with the planktonic brachiolaria of other asterinids in that the brachiolar attachment apparatus comprises three brachia and a central adhesive disc, although the latter is thin and appears to be reduced. Mortensen (1925, Videns kabelige Meddelelser fra Dansk naturhistorisk Forening i København, 79(15), 261‐420) had hypothesized that individuals of S. inflatus might brood within the "cave" formed in the interambulacral space between the arms. We found no evidence for brooding, but hypothesize that S. inflatus may have demersal development, on or near the bottom, which has implications for larval dispersal and population structure. [ABSTRACT FROM AUTHOR]

Marine heatwaves (MHW) are projected for the foreseeable future, affecting aquaculture species, such as the New Zealand green-lipped mussel (Perna canaliculus). Thermal stress alters mussel physiology highlighting the adaptive capacity that allows survival in the face of heatwaves. Within this study, adult mussels were subjected to three different seawater temperature regimes: 1) low (sustained 18 °C), 2) medium MHW (18–24 °C, using a +1 °C per week ramp) and 3) high MHW (18–24 °C, using a +2 °C per week ramp). Sampling was performed over 11 weeks to establish the effects of temperature on P. canaliculus survival, condition, specific immune response parameters, and the haemolymph metabolome. A transient 25.5–26.5 °C exposure resulted in 61 % mortality, with surviving animals showing a metabolic adjustment within aerobic energy production, enabling the activation of molecular defence mechanisms. Utilisation of immune functions were seen within the cytology results where temperature stress affected the percentage of superoxide-positive haemocytes and haemocyte counts. From the metabolomics results an increase in antioxidant metabolites were seen in the high MHW survivors, possibly to counteract molecular damage. In the high MHW exposure group, mussels utilised anaerobic metabolism in conjunction with aerobic metabolism to produce energy, to uphold biological functions and survival. The effect of exposure time was mainly seen on very long-, and long chain fatty acids, with increases observed at weeks seven and eight. These changes were likely due to the membrane storage functions of fatty acids, with decreases at week eleven attributed to energy metabolism functions. This study supports the use of integrated analytical tools to investigate the response of marine organisms to heatwaves. Indeed, specific metabolic pathways and cellular markers are now highlighted for future investigations aimed at targeted measures. This research contributes to a larger program aimed to identify resilient mussel traits and support aquaculture management. [Display omitted] • Thermal history and current temperature interact to influence survival at 24 °C. • Aerobic and anaerobic energy supplies are vital for defence mechanisms and survival. • Oxidative stress is managed by antioxidant metabolites and haemocyte functions. • Mussel condition index decreased with an increase in temperature. • Marine heatwave duration and rate of onset determine net physiological impact. [ABSTRACT FROM AUTHOR]

Increasing seawater temperatures coupled with more intense and frequent heatwaves pose an increasing threat to marine species. In this study, the New Zealand green-lipped mussel, Perna canaliculus, was used to investigate the effect of genetics and ontogeny on thermal resilience. The culturally and economically significant mussel P. canaliculus (Gmelin, 1971) has been selectively-bred in New Zealand for two decades, making it a unique biological resource to investigate genetic interactions in a temperate bivalve species. Six selectively-bred full sibling families and four different ages, from early juveniles (6, 8, 10 weeks post-fertilisation) to sub-adults (52 weeks post-fertilisation), were used for experimentation. At each age, each family was exposed to a three-hour heat challenge, followed by recovery, and survival assessments. The shell lengths of live and dead juvenile mussels were also measured. Gill tissue samples from sub-adults were collected after the thermal challenge to quantify the 70 kDa heat shock protein gene (hsp70). Results showed that genetics, ontogeny and size influence thermal resilience in P. canaliculus, with LT 50 values ranging between 31.3 and 34.4 °C for all studied families and ages. Juveniles showed greater thermotolerance compared to sub-adults, while the largest individuals within each family/age class tended to be more heat sensitive than their siblings. Sub-adults differentially upregulated hsp70 in a pattern that correlated with net family survival following heat challenge, reinforcing the perceived role of inducible HSP70 protein in molluscs. This study provides insights into the complex interactions of age and genotype in determining heat tolerance of a key mussel species. As marine temperatures increase, equally complex selection pressure responses may therefore occur. Future research should focus on transcriptomic and genomic approaches for key species such as P. canaliculus to further understand and predict the effect of genetic variation and ontogeny on their survival in the context of climate change., (© 2024. The Author(s).)