Size, sex and quantity of Scylla serrata and Portunus pelagicus on
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Size, sex and quantity of Scylla serrata and Portunus pelagicus on
Size, sex and quantity of Scylla serrata and Portunus pelagicus on Inhaca Island, Mozambique 2007-2008 . Johan Florentzson Degree project for Master of Science (One Year) in Biology 60 hec Department of Marine Ecology University of Gothenburg Contribution number 530 0 Johan Florentzson Master Thesis in Marin Ecology University of Gothenburg Abstract Inhaca Island, located in Southern Mozambique, is a relatively unpolluted site and is often used as reference point in scientific investigations, as well as extensive research of the island itself because of its undisturbed status. This paper focuses on questions concerning two edible and commercially important crabs of the family Portunidae, with the main incentive being an estimation of their current status (population) and health (physiology). The investigations were performed in a creek surrounded by mangroves and mudflats, called Saco da Inhaca, where crabs were fished using baited cages. Results revealed larger amount of Scylla serrata being caught during the warm season compared to the cold. When looking at distribution over different substrates S. serrata showed no preferences, but Portunus pelagicus seemed to prefer substrates that did not dry out completely during low tides. The sexual distribution showed a slight dominance of males. Analysis of proteins in the blood indicated better health of crabs caught on Inhaca compared to Maputo, and plausible reasons for is discussed. P. pelagicus showed a faster degradation of haemolymph protein when starved, compared to S. serrata. A small literature study concerning aquaculture of these crabs is included. In conclusion a number of differences were detected both between sites, seasons and species but more extensive research is suggested should any commercial harvest of crabs begin and/or if aquaculture is possible on Inhaca. Introduction The purpose of this study was to investigate the amount and condition of two large portunid crabs, the Mud crab Scylla serrata (Forskål, 1755) and the Blue crab Portunus pelagicus (L. 1766) from two sites in southern Mozambique: Inhaca Island and Bairro dos Pescadores. These two species are commercially important in many parts of the Indowest pacific (e.g. Potter et al., 1983, Williams and Hill, 1982) and are subjected to increased exploitation in Mozambique (Macia et al. unpubl.). Scylla serrata is a common species all over the Indo-west pacific (Williams & Hill, 1982; Demopoulos, 2007), and the genus itself contains four species as determined by Keenan et al. (1998). The average internal carapace width (ICW) of S. serrata is previously reported to be 138 mm, with a maximum of 192 mm (Keenan et al., 1998). S. serrata is commonly associated with estuarine mangrove forests, but is also the most dominating of the Scylla species when salinities are above 34 i.e. more oceanic environments (Keenan et al. 1998). It is considered a nocturnal animal, and remains buried during the day, emerging at dusk, feeds during the night (provided the tide is in) and burrows again come dawn (Hill, 1976). According to investigations of gut-content (Hill 1976) over 50 percent contained mollusks and contents of fish was rarely found. This indicates that fish is not a natural diet for S. serrata but given the common usage of fish as bait for crabs one can suggest this to be a preferable food source to the crabs and therefore it attracts crabs in a satisfactory way. Use of fish could be considered preferable compared to their natural diet since this prevents surrounding food-sources to compete with what the cages have to offer (Fielder (1965). Portunus pelagicus (Portunidae) carries an important role in the fishing industry at many locations (Smith (1982) as cited by Potter et al. (1983)). The average carapace width (CW) for P. pelagicus was found to be 128 mm with a maximum size of 164 mm (Krishna and Balakrishnan, 1971). The species is often considered a benthic carnivore and eats mainly sessile mollusks and other invertebrates (Potter and Lestang, 2000; Hill 1976), but has also been known to prey on gobies (Potter and Lestang, 2000). An investigation performed in Australia showed that commercial fishing of P. pelagicus is seasonal and the largest catches are landed between January and April (Potter et al., 1983), which represents the summer season months. This was concluded to depend on the decreasing salinities which drove the crabs to migrate. P. pelagicus prefers salinities between 30 and 40 ppt (Potter et al. 1983) with the lower critical survival limit of juveniles being 5 ppt. Higher salinities resulted in faster growth rates with an optimum growth between 30 and 40 ppt; however when salinity above 40 ppt was tested decreases in growth was detected (Romano and Zeng, 2006). 1 Johan Florentzson Master Thesis in Marin Ecology University of Gothenburg Inhaca Island, my primary study site, lies in 0 0 southern Mozambique (25 56´40’’-26 04´57’’ S, 0 0 32 54´36’’-32 57´33’’ E) approximately 30 km offshore from the capital Maputo. It is a small Island with a population estimate of roughly 5000 (Olavo pers. com.). The island is 14 km long and 7 km wide (map, appendix I) and with a marine research station located on the west side. The second site outside of Maputo consist of large mangrove forests along the shore i.e. they have direct contact with the ocean. S. serrata and P. pelagicus are both harvested in these two study areas and most of the collected animals are used as food locally (de Boer and Longamane, 1996). Since S. serrata is a highly attractive food-source all over the Indo-west Pacific, the populations are feared to be put under dangerous exploitation pressure in places were regulations of the fisheries does not exist (Robertson and Kruger, 1994, Vay, 2001). S. serrata has been shown to move and feed less as temperature drops below 20 °C, resulting in decreased catches during the cold season (Hill 1980), and similar variations have been shown in Australia (Hill 1982). However, there might be other factors than temperature influencing the abundance of S. serrata, one of which is suggested to be the migration of berried females offshore in times of spawning (Hill 1975 & 1994). The offshore spawning was suggested to increase dispersal of larvae (Hill 1994). Temperature was shown to be a critical factor for growth and survival of juvenile S. serrata (Ruscoe et al., 2004). However in their study they refer to previous experiments showing salinities to have effect on juvenile survival, so the area is still debated of sorts. Genetic stability and mixing of the gene-pool have previously been investigated at several sites along the South African coast, also one site in southern Mozambique and one on Madagascar. The results showed that populations from estuaries close to each other share a large proportion of their genes. This is likely to be because of the offshore release of eggs by gravid females (Davis et al., 2003), which strengthens the argument concerning offshore spawning since mixing of larvae would provide different estuaries with genetic material from other nearby estuaries. Hyland et al. (1984) argued that S. serrata with access to mud flats and mangroves don’t migrate since they have everything they need in their habitat. It has also been suggested that S .serrata moves onto mudflats with the high tides in order to forage (Hill et al. 1982). It is, however, not known whether they have any substrate preferences. Concerning the habitats themselves the harvesting of large predators have shown to lead to an increase in the number of opportunistic species, which in turn might cause changes in the compositions of the habitats i.e. heavy harvesting of S. serrata and P. pelagicus might change the habitat in which these are found (de Boer and Prins, 2002). Aquaculture is commonly used to enhance the market value of crabs sold in different ways. Different aquaculture methods for Mud crabs are being used around the world, and a study from South Africa shows good possibilities to keep and breed S. serrata in captivity (Davis, 2004). Currently no aqua culturing of S. serrata (only existing Scylla species in Mozambique) exists in southern Mozambique. Aqua culturing, however, is still dependant on wild caught crabs which are used mainly in three different ways and only one of these were caught crabs are used for egg rendering decreases the stress on the natural population. The different ways of using crabs in aquaculture are fattening, soft shell timing and as breeding stock. Fattening is when undersized crabs are caught and kept in cages or pens until they reach the market demands for size and weight. Secondly, for the soft shelled market crabs are caught and kept until they molt at which point they are preserved and sold before their shells hardens (Shelley, 2008). Last is as breeding stock, egg producers providing starting batches for cultivation of juvenile crabs. One goal of aqua culturing is to lessen the stress induced by harvesting of the existing population and since humans increase in number the harvesting is bound to increase which is why aqua culturing is thought to help secure the continued existence of natural crab population. The primary objective of this study was to investigate whether there are any seasonal variations in catches and whether the crabs show 2 Johan Florentzson Master Thesis in Marin Ecology University of Gothenburg any preference towards certain habitats/substrates. Physiological studies were also included into the investigations and crabs were collected from a second site outside Maputo called Bairro dos Pescadores. In order to return to the purpose of the work performed here additional physiological studies were performed during winter season. Blood was sampled and analyzed for total content of protein, in order to investigate the general health of individuals. During well fed conditions crabs build up protein and store as reserves that can be used later if needed (Subhashini and Ravindranath, 1982). Specimens for this analysis were sampled at Inhaca Island and Bairro do Pescadores outside of Maputo, to see if health was site related. The haemolymph system is complex and regulated by many factors, for e.g. the molting cycle. when the haemolymph is diluted, as water is accumulated in the crabs e.g. after molting (Chen & Chia, 1997). All crabs investigated where hard-shelled and actively foraging, and thus assumed to be in intermoult stage. A small experiment, following the basic principles from the study by Subhashini and Ravindranath (1982), was set up with the attempt to enlighten possible differences in blood protein degradations rates between S. serrata and P. pelagicus. The aim of this experiment was to clarify whether the two species investigated differed in adaptation to starvation and capture (i.e. immediate and more long-term response to human handling and lack of food). Reason for this experiment was based mainly on morphological differences between S. serrata and P. pelagicus, and since the latter is more shaped to move in the open water to collect food and has the ability to move to better areas should this become scarce. More active species are likely to have higher metabolic rates and therefore require more energy. It was therefore believed that they would be less adaptable to starvation. Hypotheses 1. There are seasonal effects on crab abundance due to different climate conditions such as rainfall and temperature fluctuations following different seasons. 2. There is a difference in blood protein content in crabs on Inhaca Island compared to those from Maputo. 3. Scylla serrata and Portunus pelagicus shows habitat preference because the species prefers a certain substrate to reside on and use as feeding grounds. 4. There is a difference in blood protein degradation rate between S. serrata and P. pelagicus because the species have different physiological adaptations to their environment. Method Field collection Scylla serrata and Portunus pelagicus were caught on Inhaca Island during two periods, November December 2007 and April – June 2008, correlating to the warm and cold season (average water temperatures: 27,4°C & 21,9°C) and hereby referred to as summer and winter seasons respectively, using cages locally produced in Maputo. The cages were built as rectangular parallelepipeds with two entrances, one at each short side. Nine cages were used during the summer season and twelve during winter season. The cages varied somewhat in construction between seasons due to different manufacturers (details: appendix II). Cages were placed randomly on three different substrates (mangrove, mud and river) in the Saco bay area on Inhaca Island 0 0 (26 01’56’’S, 32 54’53’’, details: appendix I) and were baited with small fish bought locally on the island. Efforts were made to keep the size of bait similar and when possible also stick to the same species of fish (Terapon jarbua were commonly used). The cages were left to fish for one high tide (or on occasion two due to inaccessibility caused by weather). The catches were collected, cages rebaited and randomly placed on their substrates. Crabs from Bairro dos Pescadores near Maputo were bought due to inability to fish in this area, partly from a local fisherman and partly at the fish 3 Johan Florentzson Master Thesis in Marin Ecology University of Gothenburg market. No P. pelagicus were investigated from this area since they were sold dead and would not have given any reliable blood samples. Crabs were measured in two ways over the dorsal carapace using slide calipers with mm accuracy, CW (Carapace Width) & ICW (Internal Carapace Width) as described by Keenan et al. (1998) (se appendix III). The measurement ICW was determined to be most trustworthy for S. serrata since a number of specimens had lost one or two of their ninth anterolateral spines which made the CW measure less reliable. P. pelagicus have centimeter long spines (compared to millimeters for S. serrata) and none of the animals in this study had broken spines. Since CW is commonly used in other studies as well as in the fishing industries it was determined to use the CW measurements for the P. pelagicus. Sex was determined by observing abdomen width, which differs between females and males (se appendix III). Crab haemolymph was collected in the field by inserting a 1 ml syringe into the joint of the most posterior leg. The blood was transferred from the syringe to eppendorph vials containing an 1 anticoagulant buffering solution . Syringes were also flushed with the buffer prior to sampling in order to prevent coagulation in needle or syringe. Initially 0,5 ml of blood was mixed with equal amount of anticoagulant, however this proved insufficient, since coagula formed in many samples, and the proportions were changed to 0,3 ml blood with 0,6 ml anticoagulant. The samples were immediately put on ice to keep them cold during transport, which took approximately 45 minutes. Upon arrival at the laboratory samples were frozen at -18 °C. Some of the crabs caught were tagged with cable ties around one of the legs to get an estimate on recapture rates. These crabs were only sampled for blood at the first capture. Water temperature in the river was measured during sampling using Oxi 330/SET probe equipment. 1 Anticoagulant: 0,45M NaCL, 0,1M Glucose, 30mM Tri Na Citrate *2 H2O, 26mM Citric acid, 2ml 0,5M EDTA/100ml solution. Adaptation/starvation experiment A total number of 23 crabs were brought back to the laboratory on Inhaca (9 S. serrata & 14 P. pelagicus) and placed in an aquarium with running seawater. Salinity and temperature varied since the water was pumped from the ocean into a large elevated tank from which altitude pressure made the water flow. The crabs were starved for eight days. Blood was sampled from each crab at the following intervals; field, arrival at the station followed by interval testing on hours 2, 6, 12, 24, 48, 96 (4 days) and 184 (8 days). The blood samples were frozen immediately after extraction. Analysis of blood The blood samples collected on Inhaca Island were analyzed at the Eduardo Mondlane University in Maputo. Haemolymph analysis were performed using a revised version of the Coomassie Blue method (Spector 1978, modified by Hernroth) In case of coagula formation in the blood sample the liquid to be investigated was taken with care to avoid this. Samples were diluted with distilled water to fit within the standard curve. The Coomassie reagent was mixed with the diluted blood samples in the proportion 50µl sample to 950µl reagent, after five minutes the samples were placed in a spectrophotometer (JENWAY 6105UV/Vis Spectrophotometer)) and read at λ595nm. To get the absorbance of the reagent a blank of 50µl distilled water and 950µl reagent was also measured. Results from the spectrophotometric analysis were compared to that of a standard curve constructed from a dilution of BSA (Bovine Serum Albumin- base solution 500mg BSA/50ml sterile water) which was corrected to a pH level of 4,6. Coagulation in samples was treated with a sonic homogenizer with the attempt to dissolve clots, however this proved rather insufficient. Statistics Statistical comparisons were performed using single factor ANOVA for comparing differences in haemolymph and also degradation of these. Multifactorial ANOVA (MANOVA) was used to compare catches over tides, seasons and different substrates. Histograms were used to compare classes of animals and to draw result figures. 4 No. Scylla serrata Results Catch and size 18 Summer 2007 Spring tide 16 Summer 2007 Neap tide 14 Winter 2008 Spring tide 12 Winter 2008 Neap tide 10 8 6 4 2 0 5 7 9 11 13 15 Size-class ICW (cm) Figure 1. Histogram based data showing distibution of Scylla serrata between different size-classes (5-7cm, 7-9cm, 9-11cm, 11-13cm, 13-15cm internal carapace width (ICW)) on Inhaca Island , Mozambique, for different seasons and tides. Fig. 1 compares total number of Scylla serrata caught in different size-classes caught during spring/neap tide and summer/winter season. More crabs were caught during spring tides than neap tides of the summer season (p=0,003). Differences in size showed borderline significance between spring and neap-tides during the summer season, and between spring tides for the two seasons (p=0,057 & p=0,058 respectively). Average size (and Standard Deviation (SD)) of S. serrata was 99,4 (±20) mm and the largest individual measured was 148 mm (ICW). Average catch, not comparing -1 tides or substrates, was 1,5 crabs day for the -1 summer season and 0,93 crabs day for the winter season, this difference was not significant (p=0,064). No. Portunus pelagicus 12 10 8 6 4 2 0 5 7 9 11 13 15 Size-class CW (cm) Figure 2. Histogram data showing distribution of Portunus pelagicus between different size classes (5-7cm, 7-9cm, 9-11cm, 11-13cm, 13-15cm caparapace width (CW)) on Inhaca Island , Mozambique for different seasons and tides. Note the reverse order of series compared to those of S. serrata, this due to eaiser visibillity . As can be seen in fig. 2 there are no large differences between total catches of Portunus pelagicus during the four different periods. A slight peak in catch can be seen for spring tide of the summer season, when larger crabs appear to be more frequent and the opposite is noted for spring tide during winter season. Average size (±SD) of P. pelagicus was 113 ±16 mm, with a maximum size of 144 mm (CW). Average catch, without -1 comparing substrates or tides, was 0,73 crabs day -1 for the summer season and 1,76 crabs day for the winter season (p=0,045). 5 Johan Florentzson Master Thesis in Marin Ecology University of Gothenburg Substrates 1 Catch day-1 Scylla serrata Mangrove 0,8 Mud River 0,6 0,4 0,2 Catch day-1 Portunus pealgicus 1 Mangrove 0,8 Mud River 0,6 0,4 0,2 0 0 Summer Summer Winter Figure 3 a). Comparisons of catches (no. crabs/day) between different substrates for Scylla serrata on Inhaca Island , Mozambique, during summer (2007) and winter (2008). Error bars represented by standarad deviation. In fig. 3 the average catch per day of S. serrata (a.) and P. pelagicus (b.) caught on different substrates (Mangrove, Mud & River) is shown, comparing the two investigated seasons. Analysis of S. serrata reveals no significant differences between substrates or season, high variances hid possible differences. P. pelagicus differs between all substrates comparing seasons, mud &river showed p=0,000 and mangroves p= 0,031. Winter Figure 3 b). Comparisons of catches (no. crabs) between different substrates for Portunus pelagicus on Inhaca Island, Mozambique, during summer (2007) and winter (2008). Error bars represented by standard deviation. Ten previously caught S. serrata were re-caught during this investigation of which seven had moved onto another substrate compared to where they were first caught, for P. pelagicus four were caught again of which two had moved onto other substrates. No statistics were performed on these data. Sex 100% % of sex 18 5 4 5 6 7 6 7 female male 50% 25 12 8 9 17 12 4 8 0% S:ST S:NT W:ST Scylla serrata W:NT S:ST S:NT W:ST Portunus pelagicus W:NT Figure 4. Distribution between sexes for catches of Scylla serrata and Portunus pelagicus on Inhaca Island, Mozambique. S:ST- Summer Spring Tide, S:NT- Summer Neap Tide, W:ST- Winter Spring Tide, W:NT- Winter Neap Tide. As shown in fig. 4 a majority of males were caught on all occasions apart from one for P. pelagicus during the summer season neap tides. There were no significant differences in sex distribution for the two species. Similar results were shown in studies where only slightly higher number of male P. pelagicus were caught (Potter and Lestang 2000) and equal amount male and female S. serrata (Hill et al. 1982). 6 Johan Florentzson Master Thesis in Marin Ecology University of Gothenburg Blood mg protein per ml haemolymph 160 In fig. 5 the amount of blood proteins were compared between areas and species. These data were based on samples with no (or small amounts of) coagulum. No statistical difference was detected between the three groups (p=1) Scylla serrata 140 120 Portunus pelagicus 100 80 60 n=10 40 20 n=12 n=27 0 Maputo Site Figure 5. Differences in Blood protein concentration (mean mg/ml ±SD) of Scylla serrata between Maputo and Inhaca Island, and between Scylla serrata and Portunus pelagicus on Inhaca Island, Mozambique. Blood protein concetration (mg/ml) Inhaca Island 200 180 160 140 120 100 80 60 40 20 0 y = 0.28x + 116 a) 0 160 b) R² = 0,0003 2 4 c) 6 0 6 0 d) 50 100 150 200 50 100 150 200 y = -5.12x + 110 140 R² = 0.23 120 100 80 60 40 20 0 0 2 4 Hours after capture Figure 6. Degradation of blood protein for Scylla serrata (a & b) and Portunus pelagicus (c & d) held in captivity without food over a maximum period of eight days at Inhaca research station. Blood samples were taken at predetermined intervals (X-axis) to compare the haemolymph concentration. Figures a) & c) describes what occurred during the initial six hours of the experiment and figures b) & d) shows the results of the whole experiment. Fig. 6 show the degradation of haemolymph proteins for S. serrata (a & b) and P. pelagicus (c & d). S. serrata show a slower degradation rate of the proteins, as can be seen by comparing trend line slope. However this could only be seen as a trend for the whole 180 hour captivity, but for the first six hours the difference in degradation is statistically significant (p=0,005). 7 Johan Florentzson Master Thesis in Marin Ecology University of Gothenburg Coagulum % samples with different coaugla ammount 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 0% None Some Half Much Very much Fig. 7 describes the amount of coagula formed from proteins in the samples. The samples from Inhaca, that had been stored in a freezer for 1-5 weeks, contained a lot more coagula compared to samples taken in Maputo, which were only frozen and immediately defrosted to create a similar treatment effects. Unkown Inhaca P. Pelagicus Inhaca S. Serrata Maputo S. Serrata Site and species Figure 7. amount of coagulum in Scylla serrata and Portunus pelagicus, Inhaca island (29april-13june 2008) and for Scylla serrata from Bairro dos Pescadores, Maputo (26/6 2008) Temperature Measurements of temperature showed a difference (p=0) between seasons where the daytime water temperature in the summer season averaged 27,4 ± 1,4 °C (n=28) and during winter season an average of 21,9 ± 1,2 °C (n=15) was measured. 8 Johan Florentzson Master Thesis in Marin Ecology University of Gothenburg Discussion Local fishermen on Inhaca refer to spring tides as “La mare vida”, an expression that translates to “the living ocean”, and describes how maritime harvest and fishing give higher yields during these periods. This statement describes a part of the current investigation as to whether Scylla serrata or Portunus pelagicus were predominant during any specific tide. Scylla serrata – Mud crab S. serrata was more abundant during the spring tides compared to neap tide during the summer season. Animals that were caught during spring tides were also slightly larger than animals caught at neap tides. In addition trends suggest larger crabs being caught during the spring tides of the summer season compared to spring tides in the winter season. This appears to concord with an earlier study from South Africa where lower catches were found during their “winter season” (Hill 1975). Given the proximity between Mozambique and South Africa one could expect seasons to be called the same, but the article does not discern which months is considered winter or summer seasons. However temperature were lower in South Africa during both seasons, with winter season temperatures ranging from 13 to 0 0 15 C and summer season between 20 and 25 C (Hill 1975). The largest crabs, which were females, were as previously mentioned caught during the summer season, which according to Hill (1994) should be the time for large females to migrate offshore in order to spawn. However since no gravid crabs were caught in this study they would have little use of migrating offshore, since the main reason for this would be better larval dispersal. The size of the Scylla serrata in this study (mean 99.4 mm) was smaller compared to the mean of 138.4 mm found by Keenan et al (1998) when samples from the whole Indo-West Pacific were taken. No significant differences could be detected between catches of S. serrata on different substrates, however trends suggest that more crabs were caught during the summer season. Concerning the substrates no conclusions can be made but other authors have suggested that S. serrata move onto mudflats and mangroves in order to feed (Hill et al., 1982). There are studies showing S. serrata to be relatively sedentary, with movement limited to 1 square kilometer, provided they have sufficient living conditions (Demopoulos et al., 2007) and the currently investigated area was a little over one square kilometer. Also as mentioned in the introduction crabs with access to mud flats and mangroves, both of which are present in the Saco, show no great movement (Hyland et al. 1984). Population size estimates and movement patterns are calculated on amount of recaptured animals which was the original plan of this investigation. However since there was a low amount of recapture no conclusions could be drawn from these data. When regarding the continuous catch found in this study, it appears as though the population of S. serrata in Saco bay is stable with current harvest pressure. However, no predictions can be made for what increased exploitation pressure might lead to. Haemolymph Investigations comparing blood protein concentration of S. serrata populations on Inhaca Island with those of Bairro dos Pescadores provides no reliable results since variations were too high. However one could argue that Inhaca Island should be a healthier place for the crabs to live due to its less polluted state. When looking at water movements, Maputo Bay receives inland water from three large rivers (Espírito Santo, Maputo River and N´komati), emptying large amounts unprocessed water into the bay. Therefore Maputo Bay is thought to be more polluted compared to Inhaca which receives large parts of its surrounding waters directly from the Indian Ocean (de Boer et al. 2000). Also Inhaca Island is often used as a reference site in local scientific investigations (Macia pers. comm.) which might say something about its cleanliness. Pollutants have been suggested to affect multiple factors in crustaceans such as blood glucose and hormone regulation (Fingerman et al. 1998), direct effect on haemolymph by pollutants have been reported by Watson et al. 2004 who detected changes in the haemolymph of Carcinus maenas after these were exposed to PAH's (Poly Aromatic Hydrocarbons). However the results found for 9 Johan Florentzson Master Thesis in Marin Ecology University of Gothenburg haemolymph were not as great as from urinary analysis after the same treatment and therefore the latter was considered a more reliable investigative parameter. Since no specific pollution parameters were tested in this investigation and also since no results differentiating Maputo from Inhaca Island were detected little can be said about the pollution in Maputo bay other than the hypothetical problems mentioned above. Portunus pelagicus – Blue crab The abundance of P. pelagicus shows tendencies to be smaller during the summer season (NS) which might be explained by its preference for higher salinities of 30-40 ppt (Potter et al. 1983). Since the amount of freshwater runoff during the summer season is larger due to seasonally heavier rainfall P. pelagicus could be believed to relocate to habitats less estuarine. Potter and Lestang (2000) found that P. pelagicus was more likely to spawn in the bay area outside their investigated estuary as suggested by Anon (1983); this might be because of better larval dispersal and better survival of juveniles in higher salinities (Romano and Zeng 2006). Like for S. serrata berried females were scarce, with only one ovigerous P. pelagicus caught during both seasons, thus making patterns of migration for females impossible to determine. As for S. serrata P. pelagicus gave no re-catches reliable enough to conclude anything about these. P. pelagicus seems to prefer the river and to some extent mud substrates compared to the mangroves. Only 1,5% of total P. pelagicus catches (i.e. summer and winter seasons combined) were caught in the mangroves. S. serrata showed no such habitat preference, being evenly distributed over all three substrates. The reason behind this difference between the two species could be that P. pelagicus is less adapted to desiccation than S. serrata. P. pelagicus caught in cages above the water at low tide were less active and in a few cases even dead, while S. serrata seemed to bury despite the mesh and did not seem less alert. Also P. pelagicus caught in dry cages regained activity quickly when submerged. Another indication of the difference in desiccation tolerance was that no living P. pelagicus were to be found at the fish market in Maputo while the S. serrata were still lively. Therefore it is suggested that P. pelagicus prefers substrates that are lower relative to the zero-tide level compared to that of S. serrata. Concerning P. pelagicus caught in the two other substrates the largest amount was caught in the river, but the largest specimens was found on the mud. This might be explained by the higher exposure to predators on the mud flats, i.e. birds, and could be a reason for smaller animals to stay away from this area. Not only other species prey on small crabs, cannibalism is also frequently occurring (Marshall et al. 2005). Scylla serrata & Portunus pelagicus Looking at sex distribution for both S. serrata and P. pelagicus, overall more males were caught. This might be because of the assumed migration of female’s offshore (Hill, 1994; Potter et al. 1983), but since this migration is seasonally dependant it cannot explain the overall majority of males caught during all periods of this investigation. However since the difference is mostly minute there is no reason to interpret too much into these results. Haemolymph degradation The proteins in the haemolymph of starved Portunus pelagicus degraded faster compared to Scylla serrata. The biggest difference was found during the initial six hours after capture where P. pelagicus showed a significantly greater loss of haemolymph proteins compared to S. serrata. This is of interest because it indicates the different species immediate reaction to capture/captivity. This in turn shows that testing of S. serrata in the field is less important since they remain relatively stable during transport, while P. pelagicus needs immediate blood extraction in order to obtain reliable values. P. pelagicus is more active when compared to S. serrata which mostly stays still and endures, this might be the reason the previous shows greater drops in blood protein levels. Faster more active animals have a higher metabolism and therefore P. pelagicus is likely to metabolize the proteins faster than S. serrata. Some relations to this could be seen in the work by Paterson & Spanoghe (1997) who found that Rock lobsters (Panulirus cygnus) shows metabolic degradation of haemolymph 10 Johan Florentzson Master Thesis in Marin Ecology University of Gothenburg when exposed to stressful environments such as transportation. Low levels of oxygen and poor environmental conditions in the aquarium, as well as the lack of food, could lead to similar effects and depending on the species said degradation should differ. The oxygen levels of the water flowing through the aquarium was unknown and in wild S. serrata used burrows which are likely to be subjected to decreased oxygen levels and increased temperature during low tide. Therefore suggestions that S. serrata is more adapt to stress caused by oxygen depletion is suggested since their health was better than that of P. pelagicus. However since P. pelagicus is a smaller crab they might be affected by the amount of blood sampled even though the volumes were small Coagulum Even though blood samples were mixed with anticoagulant buffering solution many samples formed coagula. The main reason for this is the storage time which was longer for samples taken on Inhaca du to the lack of proper equipment for immediate testing. This was strengthened by the observation that blood samples from Maputo formed very small amounts of coagula, despite similar treatment, with the only big difference between the sites being said storage time. Triggering factors of clotting are suggested to depend on the levels of fibrinogen in the blood, which sets of the coagulation process and is suggested to have similar chemical composition and exist in different amount in all decapods (Ghidalia et al. 1980). However, since the attempt to solve the coagulum by sonic homogenization failed, the conclusion concerning blood samples is that they should not be stored frozen for extended periods of time and if possible should be processed immediately. Surrounding factors The introduction mentions lower activity caused by lower temperature with the critical level being 20 °C (Hill 1980), however there is still a possibility that the water temperature gave a lowered activity during the winter season when mean measured 0 temperature was 21,9 C . Lowered activity may results in fewer catches which could be a reason for lower catches during the winter season. The investigation of recapture collided somewhat with the continuous blood sample investigation since the latter required crabs to be brought to the research station and kept there, while recapture needed these animals to be set free in order for possible recapture to occur. Information that cages placed within 100 meters of each other (Williams and Hill, 1982) was discovered after the investigation was done and thereby no actions to avoid this was taken. And lastly since levels of water filtration for the aquarium on the research station was unknown starvation might be questioned since food particles could possibly be transported with the water. Loo et al. (1993) investigated the clearing rate of suspended food particles by Nephrops norvegicus and Hommarus gammarus and found that some nutritional benefits can be detected when animals are allowed only to filter for food. Since these two also are large decapods one could suspect similar results should they be performed on S. serrata and P. pelagicus. Aquaculture Since no aqua culturing of S. serrata is presently applied in southern Mozambique the following text is based solely on literature research. As mentioned in the introduction there are three currently working ways of keeping S. serrata (only species existing in Mozambique). Of these the rearing of larvae should be the most economically viable since it would produce more crabs, while the other two methods only deliver the crabs already caught in the wild. Should interest of aqua culturing on Inhaca shown up the subject must be thoroughly investigated beforehand. Acknowledgments My thanks to everyone who helped me with this thesis, to mention a few: my supervisor Susanne Eriksson for great help with lab work and writing. Adriano Macia who supervised my work in Mozambique. Stina Larsson who helped me with pretty much everything. Olavo and Olga for practical help at the research station on Inhaca Island and Peter Tiselius for help with statistics. 11 Johan Florentzson Master Thesis in Marin Ecology University of Gothenburg Cited Literature 1. Brian D. Paterson and Patrick T. Spanoghe. Stress indicators in marine decapod crustaceans, with particular reference to the grading of western rock lobsters (Panulirus cygnus) during commercial handling. Marine and Freshwater Research 48(8) 829 834 (1997). 2. Chen J and Chia P, Oxyhemocyanin, protein, osmolality and electrolyte levels in the hemolymph of Scylla serrata in relation to size and molt cycle Journal of Experimental Marine Biology and Ecology, 217, 93-105 (1997). 3. Davis J.A., van Blerk L.L., Kirby R and Hecht T. Genetic variation in the mud crab Scylla serrata (Forskål 1775) (Crustacea: Portunidae) in South African estuaries. African Zoology 38 (2), 343-350 (2003). 4. Davis A. J., Churchill G. J., Hecht T and Sorgeloos P. Spawning characteristics of the South African Mud crab Scylla serrata (Forskål) in captivity. Journal of the world of aquaculture society. Vol. 35 No. 2 (2004). 5. de Boer W. F. and Longamane F. A. The exploitation of intertidal food resources in Inhaca bay, Mozambique, by shorebirds and Humans. Biological conservation 78, 295-303 (1996). 6. de Boer W. F., Rydberg L. and Saide V. Tides, tidal currents and their effects on the intertidal ecosystem of the southern bay, Inhaca Island, Mozambique. Hydrobiologia, 428 187-196 (2000). 7. de Boer W. F and Prins H. H. T. The community structure of a tropical intertidal mudflat under human exploitation. ICES Journal of Marine Science 59, 1237-1247 (2002). 8. Demopoulos A. W.J, Cormier N., Ewel K.C. and Fry B. Use of multiple chemical tracers to define habitat of Indo-Pacific Mangrove crab, Scylla serrata (Decapoda: Portunidae). Costal and Estuarine Research Federation (2007). 9. Fielder DR. The spiny lobster, Jasus lalandei (H. Milne-Edwards), in South 10. 11. 12. 13. 14. 15. 16. 17. 18. Australia. III. Food, feeding, and locomotor activity. Australian Journal of Marine and Freshwater Research 16(3) 351- 368 (1981). Fingerman M., Jackson N. C. and Nagabhushanam R. Hormonallyregulated functions in crustaceans as biomarkers of environmental pollution. Comparative Biochemistry and Physiology part C 120, 343-350 (1998). Ghidalia W., Vendrely R., Montmory Y., Coirault Y. and Brouard M. O. Coagulation in Decapoda Crustacea. Comparative study of the clotting process in species from groups A, B and C. Journal of Comparative Physiology. B. 142: 473-478 (1981). Hill B.J. Abundance, Breeding and Growth of the Crab Scylla serrata in Two South African Estuaries. Marine Biology 32, 119-126 (1975). Hill B.J. Natural food, foregut clearance-rate and activity of the crab Scylla serrata. Marine biology 34, 109-116 (1976). Hill B.J. Effects of Temperature on Feeding and Activity in the Crab Scylla serrata. Marine Biology 59, 189-192 (1980). Hill B. J., Williams M. J. and Dutton P. Distribution of Juvenile, Sub adult and Adult Scylla serrata (Crustacea: Portunidae) on Tidal Flats in Australia. Marine Biology 69, 117-120 (1982). Hyland S. J., Hill B. J., and Lee C. P. Movement within and between different habitats by the portunid crab Scylla serrata. Marine Biology 80, 57-61 (1984). Keenan C. P., Davie P. J.F., Mann D.L. A revision of the genus Scylla de Haan, 1833 (Crustacea: Decapoda: Brachyura: Portunidae). The Raffles Bulletin of Zoology 46(1): 217-245 (1998). Krishna K. P. and Balakrishnan N. The annual reproductive cycles of Uca anulipes, Portunus pelagicus and Metapenaeus affinis (Decapoda: Crustacea) from the South-west coast 12 Johan Florentzson Master Thesis in Marin Ecology University of Gothenburg 19. 20. 21. 22. 23. 24. 25. 26. of India. Marine Biology 11, 152-166 (1971). Loo L-O., Baden S.P. and Ulmestrand M. Suspension feeding in adult Nephrops norvegicus (L.) and Hommarus gammarus (L.) (Decapoda). Netherland Journal of Sea Research 31, issue 3, 291-297 (1993). Marshall S., Warburton B., Paterson B and Mann D. Cannibalism in juvenile blue-swimmer crabs Portunus pelagicus (Linnaeus, 1766): effects of body size, moult stage and refuge availability. Applied Animal Behaviour Science 90, issue 1, 65-82 (2005). Shelley C. Capture based aquaculture of mud crabs (Scylla spp.).Fisheries Technical Paper no. 508, 255-269 (2008). Smith S.S and Sumpton W.D. Behavior of the commercial sand crab Portunus pelagicus (L.) at trap entrances. Asian fisheries science 3, 101-113 (1989). Spector T. Refinement of the Coomassie blue method of protein quantitation. A simple and linear spectrophotometric assay for ≤0.5 to 50 μg of protein. Analytical Biochemistry Volume 86, Issue 1, May, 142-146 (1978). Subhashini M.H. and Ravindranath M.H, Significance of periodic fluctuations in the haemolymph proteins and their catabolic products during starvation and repeated injury in Scylla serrata (Forskal). The journal of experimental zoology 222, 27-35 (1982). Potter C., Chrystal P. J., and Loneragan N. R. The biology of the blue manna crab Portunus pelagicus in an Australian estuary. Marine Biology 78, 75-85 (1983). Potter I.C. and Lestang de S. Biology of the blue swimmer crab Portunus pelagicus in Leschenault estuary and 27. 28. 29. 30. 31. 32. Koombana Bay, south-west Australia. Journal of the royal society of Western Australia 83, 443-458 (2000). Robertson W. D. and Kruger A. Size at maturity, mating and spawning in the Portunid crab Scylla serrata (Forskål) in Natal, South Africa. Estuarine, Coastal and Shelf Science. 39, 185200 (1994). Romano N. and Zeng C. The effects of salinity on the survival, growth and haemolymph osmolality of early juvenile blue swimmer crabs, Portunus pelagicus. Aquaculture 260, 151-162 (2006). Ruscoe I. M., Shelley C. C. and Williams G. R. The combined effects of temperature and salinity on growth and survival of juvenile mud crabs (Scylla serrata Forskål). Aquaculture 238, 239-247 (2004). Watson M. G., Andersen O-K., Galloway S. T. and Depledge H. M. Rapid assessment of polycyclic aromatic hydrocarbon (PAH) exposure in decapod crustaceans by flurometric analysis of urine and haemolymph. Aquatic toxicology 67, 127-142 (2004). Vay. L. Le Ecology and management of Mud crab Scylla spp. Asian Fisheries Science 14: 101-111 (2001). Williams M.J. and Hill B.J. Factors influencing pot catches and population estimates of the Portunid crab Scylla serrata. Marine Biology 71,187-192 (1982). Web pages 1. http://www.hkfish.net/eng/database/crabs/structur al.htm 2008-10-15 Sex determination Of Scylla serrata. 2. http://www.fishsa.com/crabs.php 2008-10-15 Sex determination of Portunus pelagicus. 13 Johan Florentzson Master Thesis in Marin Ecology University of Gothenburg Appendix I Map of Inhaca With details of Saco Bay Mangrove Mud River Sand 2 Scale 3 0 1 Fished area k m Map modified from Salmaõ Bandeira’s map of Segrass dispersal on Inhaca Island (With permission) 14 Johan Florentzson Master Thesis in Marin Ecology University of Gothenburg II 40 cm Detailed description of the cages 15cm 40 cm D=20cm 20cm 90 cm The nine cages used during fall were equipped with a hatch on top which cages used during spring lacked. This is not believed to affect the catches in any way. The twelve cages used during spring-season all had shorter entrances as shown by the right entrance (15cm) compared to that of the cages used during the fall which measured 20cm in tube-length (left side of picture). The mesh size used was 2cm2tion of cages 15 Johan Florentzson Master Thesis in Marin Ecology University of Gothenburg III Description CW, ICW and Sex Curtsey of Keenan et al. 1998 Scylla serrata male Picture from http://www.hkfish.net/eng/database/crabs/structural.htm Portunus pelagicus male Picture from http://www.fishsa.com/crabs.php Scylla serrata female Picture from http://www.hkfish.net/eng/database/crabs/structural.htm Portunus pelagicus female Picture from http://www.fishsa.com/crabs.php 16