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Evaluation of hepatopancreas HP , pincher claw muscle PCM , crusher claw muscle CCM , and abdominal muscle AM lipid content in 86 recently caught, adult, male and female American lobsters, Homarus americanus , in the Northumberland Strait, PE, Canada, using the Folch method identified lipid as a significant metabolic reserve.

The HP was the primary lipid reservoir in male and non-berried females. Among five moult categories late inter-moult mc-1 , early pre-moult mc-2 , mid pre-moult mc-3 , late pre-moult mc-4 and post-moult mc-5 , devised for the study, the prominent trend observed was a post-moult depletion of lipid reserves in the HP, PCM, and AM.

An increased lipid index LIn in both CCM and PCM at late pre-moult suggested a differential distribution to provide needed energy in the immediate post-moult foraging period. Use of all three parameters in multiple linear regression model improved fit from 0. Hemolymph evaluation presents a non-lethal alternative to assess hepatopancreas lipid content in American lobsters when sex and moult stage are taken into account. These data emphasize the importance of timing and subject selection when using blood chemistry to aid in the monitoring of lobster populations, e.

Crustaceans have a cyclical life history punctuated with periods of marked physiological change associated with moulting and reproduction. A period of inanition prior to the moult is followed by a period of high-energy requirement Passano, ; McWhinnie et al.

The sequential accumulation and depletion of metabolic reserves during the moult cycle has already been identified in many decapod crustaceans including: Penaeus monodon Fabricius, Suneetha et al.

Adequate accumulation of metabolic reserves is integral to a successful moult. Condition in crustaceans has been defined as the extent to which nutrient reserves have been accumulated to allow for normal physiological function and growth Moore et al.

Lipids represent a significant source of energy in many decapod crustaceans Schirf et al. The hydrophobic, non-hydrated, nature of lipids allows for increased storage efficiency, i. The energetic efficiency of lipids makes them especially important in species that require large reserves to cope with extended periods of inanition Urich, Pronounced changes in hepatopancreas protein, lipid and carbohydrate content, have been documented in various crustaceans during the moult Chang, In vertebrates, changes in nutrition are generally coupled with alterations in physical appearance, i.

Non-lethal assessment methods allow for repetitive analyses of individuals. The biochemical composition of hemolymph can be highly influenced by environmental conditions including salinity, temperature, food availability, season, as well as life stage and physiologic responses such as stress Noga, ; Dove et al. Moult-induced changes in glucose concentrations were observed in the isopod Ligia exotica Roux, Parvathy, , the shrimp Litopenaeus vannamei Boone, Galindo et al.

Fluctuations in lipid triglyceride, cholesterol were noted in the Southern rock lobster Jasus edwardsii Hutton, Musgrove and Babidge, and the crab Carcinus maenas Linnaeus, Spindler-Barth, during the moult.

Hemolymph protein increased preceding the moult followed by a rapid decrease post moult Travis, b ; Stewart and Li, ; Bursey and Lane, ; Spindler-Barth, ; Mercaldo-Allen, Hemolymph metabolites may similarly correspond to nutritional status of the lobster. In this study, the natural fluctuations of lipid reserves of four tissues in H.

Correlations between these parameters and energy reserves could offer additional means of assessing lobster populations in the Northumberland Strait. Five moult categories were devised based on pleopod staging Aiken, and shell hardness Table 1: The goal was to collect 10 male and 10 female lobsters for each moult category for a total of lobsters from Lobster Fishing Area 26A in the Northumberland Strait, PE, Canada, from May through September of Standard wire commercial lobster traps were baited with mackerel and submerged overnight before hauling.

Poor trapping in necessitated continuing the sample collection in Summary counts of lobsters collected by moult category, year, and cohort. A cohort represents a group of lobsters beginning and progressing through a moult event together over a period of time.

Lobsters were tagged with a labelled cable-tie attached to a banded crusher or pincher claw. Pleopods were staged by assessing the development of the underlying new cuticle under direct light microscopy within 48 hours of collection using criteria established by Aiken, Physical data including carapace length, sex, and shell hardness were also recorded. Shell hardness was subjectively assessed by experienced field researchers based on compression of the carapace when digital pressure was applied in three areas of the thoracic carapace anterior dorsal, posterior dorsal, mid-ventral Scott-Tibbits, Shells were categorized as hard no compression , medium slight compression and soft easily compressed and fragile.

The relationship among pleopod stage, shell hardness and moult category is presented in Table 2. Summary of moult categories as defined by pleopod staging and shell hardness. Traditional Drach moult stages are depicted along with corresponding pleopod stage, shell hardness, and the devised moult category in which each would fall.

Table was adapted from Aiken, ; Aiken, Hemolymph was collected aseptically approx. The remainder was used to fill a 1. The supernatant plasma was placed into a new microfuge tube and stored in a refrigerated cooler. Caps were removed and the tops covered with gauze held in place with a rubber band. Aliquots of lyophilised HP mg , in triplicate, were mixed with 3. An additional aliquot of methanol was added 0. The weights of the lipid samples were measured every day until no changes in weight were observed approx.

Lipid extraction from muscle tissues was similar with two modifications. Due to the low lipid content of muscle and tissue availability, mg aliquots of tissue were extracted in duplicate and all solution volumes were doubled. Total lipid concentrations were standardized to take into account the differences in lobster size and water content as follows:.

The TP concentrations were determined colourimetrically biuret assay through the generation a copper-protein complex in an alkaline solution TP2 test kit, Roche Diagnostics. The small sample sizes warranted nonparametric analysis of plasma and lipid data. A total of 86 lobsters were ultimately sampled Table 1. Late pre-moult female lobsters mc-4 proved difficult to obtain in either year. The first cohort represented lobsters entering the initial stages of moult mc-1 in early June and moulting that summer.

The second cohort entered the initial stages in September. Multiple moult cohorts have been observed previously and attributed to limb regeneration Moriyasu et al. Lipid indices median and range for four tissues in male and female American lobsters, by moult category, year, and moult cohort.

Moult categories are defined as follows: All four tissues were available for analysis from all lobsters except for PCM from one mc-1 male in the primary cohort and CCM from one mc-3 female in on account of insufficient or missing tissue. Analysis of the primary moult cohort revealed no significant differences in the median LIn values of the HP from mc-1 through to mc-3 but, dropped significantly to Median values and ranges are reported in Table 3.

Although the LIn values were lower, similar trends were observed in the AM, where LIn values were also steady until a significant decrease in the post-moult. The CCM showed no significant changes in the lipid concentration. The median percent lipid by wet weight was included in Table 4 for comparison to previously published data. The general trend in percent lipid reserves was a gradual but not significant increase from mc-1 to mc-4 and a significant decrease in mc-5 for all tissues except the PCM and the HP where a decrease was observed in mc-4 Table 4.

Trends in water content were inverse to those observed in percent lipid data Table 4. Carapace length of lobsters ranged from 75 mm to 80 mm with only the mc-5 lobsters size being significantly greater than mc-3 and mc Tissue composition of combined male and female lobster collected from LFA 26A in and samplings at different moult stages. Percentages represent the median mg of lipid in mg of wet tissue. The effect of year on lipid concentrations in the crusher claw muscle CCM.

Lipid index values LIn for each lobster from the five moult categories: The secondary moult cohort of and the dataset contained data from lobsters in late pre-moult mc When added to the primary cohort data, the entire perimoult period was represented. Lipid concentrations in the HP were still noted to drop significantly in mc-5 with no statistically significant changes observed for mc-1 through mc The same trend was observed in the AM.

A similar, but not statistically significant pattern was noted for the PCM. Female lobsters demonstrated the best correlations between the HP lipid concentrations and the TP and Brix values only, with TP better correlated than Brix.

The glucose and uric acid concentrations showed poorer correlations in both males and females and varied between the two moult cohorts Table 5.

Use of all three parameters in multiple linear regression model improved from 0. When examining the data by moult category and sex separately, late inter-moult males showed the best correlations between HP LIn and hemolymph TG, TP and CHOL with decreasing correlation as the lobsters approached the moult. Females showed no significant correlations with any of these hemolymph metabolites for any of the moult categories Table 6 , Fig.

Lipids were identified as a substantial metabolic reserve in the American lobster. The HP constituted the largest reservoir with fluctuations occurring in all four tissues examined over the course of the moult. Hemolymph TG, TP, and CHOL concentrations were very well correlated to HP lipid content in late inter-moult mc-1 male lobster, while hemolymph TP concentration and Brix index provided a similar, although less robust, correlation in female lobsters as a group; however, correlations were even lower when moult category was considered.

Lobsters in this region generally moult mid-July Comeau and Savoie, Due to regional weather and ice conditions samples could only be collected between May and October, covering the periods before, during and immediately after the moult. To avoid any potential discrepancies that may have been caused by year or cohort variations in physiology, analysis of tissue lipid concentrations was first conducted on the primary moult cohort.

The significant decrease in the LIn of the HP and AM of post-moult lobsters suggests that lipid is being used during or shortly after ecdysis, most likely as an energy reserve, with the HP being the most significant source.

This corresponds well with what is typically reported in the literature where the HP is the primary organ involved in lipid storage with very low muscle lipid concentrations Chapelle, ; Dall, Hemolymph metabolites versus hepatopancreas lipid index for male lobsters, and Moult stage of lobster indicated by symbol type.

Addition of the secondary cohort and data allowed inclusion of the late pre-moult mc-4 lobsters when examining trends in tissue lipid stores. Significant changes were expected as the period of inanition begins at this time Dall, Although a gradual increase in median LIn in the HP and AM from mc-1 through mc-3, followed by a slight decrease during mc-4 was observed, statistically significant differences were still not detected.

A significant median decrease in the post-moult mc-5 , as noted for the primary cohort, remained. Natural variations of wild populations combined with small sample size of the current study may have confounded statistical detection of actual changes in metabolic reserves during mc-1 to mc The potential for year effects should also be noted.

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