Martinez, E. The Distribution and Shell Utilization of the Female Hermit Crab Abstract Two intertidal hermit crabs, Pagurus granosimanus and Pagurus samuelis, were studied at the Hopkins Marine Station. The two species demonstrate definite zonation along the intertidal, but intraspecies differences are not known. Past studies have shown that shell characteristics, such as damage, fouling, and adequacy have varying influences on the reproductive success of female. Ovigerous females, non-ovigerous females, and males were examined at three different locations. The study of the distribution and the shell utilization of each of the hermit crabs revealed that males were more predominant than females. Non-ovigerous females were more common than males. Shell fouling had no effect on the shell utilization by ovigerous, non-ovigerous and male hermit crabs. Ovigerous females were in shells with less damage. Shell adequacy assignments showed that females were more likely to be ovigerous if they occupied smaller shells than the preferred size calculated from a shell adequacy index. Clutch size was minimally correlated to other measurements. Martinez, E. The Distribution and Shell Utilization of the Female Hermit Crab Introduction Ovigerous female hermit crabs must rely on their shell for survival and for the survival of their egg clutch. Factors such as shell availability and the habitat in which a female hermit crab lives may affect the success of reproduction and egg maintenance. Furthermore, reproduction and energy expenditure may influence the proportions of ovigerous females, non-ovigerous females, and males located at different lcoations and at different heights along the intertidal zone. Shell size affects a hermit crab’s reproductive effort, i.e., how many eggs are brooded, but the effect varies from species to species. For one species, C. vittatus, a large shell provides more room for brooding eggs (Fotheringham 1976), but for others, such as the Calcinus obscurus, increased shell size results in greater energy expenditure in growth rather than reproductive effort, resulting in fewer eggs (Bertness 1980). Shell damage can be a variable in the susceptibility to predation for hermit crabs. One of the local predators, Cancer antennarius, can more easily eat hermit crabs occupying a damage shell than an intact shell (personal observation). However it is unclear whether shell damage influences how shells are distributed among the non-ovigerous, ovigerous and males. A study of Pagurus longicarpus indicated that shell fouling and shell damage may have an affect on the clutch size depending on the habitat and the size of the hermit crab(Wilber 1989). The distributions of Pagurus granosimanus and Pagurus samuelis have been identified for the Hopkins Marine Station (Belknap, Robert and John Markham 1965). P. samuelis occupy the higher portions of the intertidal, and P. granosimanus occupy the intertidal zone below, with some overlap. However, Martinez, E. The Distribution and Shell Utilization of the Female Hermit Crab distribution studies have not reviewed the breakdown of non-ovigerous, ovigerous, and male hermit crabs. P. granosimanus and P. samuelis are susceptible to dessication and thermal stress (Taylor 1981), but how those stresses affect the proportion of non-ovigerous, ovigerous, and males along the intertidal is uncertain. The purpose of this study was to determine how a habitat affects the proportions of females, ovigerous females, and males. Öther elements such as relative tidal height were examined to indicate if the females, ovigerous females, and males were more likely to remain in particular parts of the intertidal to alleviate thermal and dessication stress. Also the type of damage on a shell was examined for differences between the shell utilization of the non-ovigerous, ovigerous, and males. Methods and Materials Two species of hermit crabs, Pagurus granosimanus and Pagurus samuelis were collected from the intertidal zone of the Hopkins Marine Station, Pacific Grove, California. Collection took place during May 11-13, 1994, during low tide. Three locations were chosen for study: West Beach, an exposed area that receives direct wave action; Hewett Transect, a semi-protected area; and Agassiz Beach, a protected area which receives less wave action of the three (Fig 1). At each location a transect running perpendicular to the shore from the high intertidal to the lowest exposed position (approximately -0.5 m MLLW) was sampled. Each transect was divided into five collection sites 2 meters apart horizontally. For each site, approximately 0.25 m2, the first fifty (50) hermit crabs were collected or until thirty (30) minutes had elapsed. If fifty crabs were not Martinez, E. The Distribution and Shell Utilization of the Female Hermit Crab collected within the 0.25 m2 area, collection was expanded to the surrounding area, but did not exceed 1.0 m2. Specimen collection was done while the site was partially submerged to facilitate locating motile hermit crabs and to reduce the chances of missing hidden individuals. After collection, crabs were placed in holding tanks with running sea water. Crabs were fed on a diet of the brown seaweed Pelvetia. Hermit crabs were removed from their shells by piercing the whorl of the shell with the edge of a small, flat screwdriver, and gently proding the abdomen with a thin piece of magnet wire. Two other methods, heating the shell with a soldering iron, and placing the crab in mixture of freshwater and seawater were tried but were not consistently succesful. Out of 400 crabs, piercing the shells and proding resulted in 5 deaths. Piercing the shell resulted in loss of a small portion, usually no bigger than 1.5 mm2. Since these fragments were only a small proportion of the total weight, shell weight measurements were considered reliable. After crabs were removed from their shells, they were placed in small, plastic cylindrical containers with running sea water. Some individuals burrowed under the shell rubble substrate of the containers. The following data were recorded for each crab: species, sex, shield length, wet weight and number of eggs. Sex was determined by locating the gonopores on the third pereiopods for females and on the fifth pereiopods for males. Shield length, from the tip of the rostrum to the edge of the hard carapace, was measured to the nearest 0.1 mm with Venier calipers under a 7X magnification dissection microscope. Specimens were blotted dry before wet weight measurements. Eggs were removed from females by clipping the three pleopods to which eggs are attached, and the eggs were counted under 7X magnification. Seven clutches were removed and the eggs individually counted Martinez, E. The Distribution and Shell Utilization of the Female Hermit Crab and egg weight was measureed. A linear regression determined an egg number:weight ratio that was later used to determine the number of eggs for some clutches for which the eggs were not counted. The following data were recorded for the shells occupied by the hermit crabs: species, diameter, weight, fouling, and degree of damage. Shell diameter was measured with Vernier calipers to the nearest 0.1 mm. Shells were dried before weighing in a microwave for two minutes on the high setting. Shell weight included weight of any fouling organisms. The gastropods Spirorbis spp., the polychaetes Cripidula adunca and Cripidula planca, and the non- geniculate coralline alga Lithothamnion spp. on the underside of the shell and on the whorl were noted. Damage of the lip, body, and apex was also noted. A shell adequacy index (SAI) which measures deviation from a calculated preferred shell size, (Kim 1994) was assigned to each hermit crab and its shell. The SAI was derived by placing 50 P. samuelis and 57 P. granosimanus individuals into tanks with an overabundance of shells (1:8 crab to shell ratio). The crabs were given 5 days to acclimate to their new environment and select shells. At the end of the five days, it was assumed that the crab was in the shell size it preferred, and the crab’s wet-weight and the diameter of its shell were measured. A linear regression was used to analyze the relationship between crab weight and shell diameter to derive the SAI. Wet weight of the crab was compared to its shield length and a linear regression between the two measurements was determined. This regression was used to determine either the wet-weight or shield length for a few individuals («10) whose wet-weight or shield length measurement was not recorded. Results Martinez, E. The Distribution and Shell Utilization of the Female Hermit Crab A total of 136 Pagurus granosimanus and 180 Pagurus samuelis were collected. Pagurus hirsuiticulus were collected as well but a limited sample size (n = 36) was not sufficient for a similar study of this species. The smallest ovigerous P. granosimanus was 4.3 mm in shield length, and the largest had a shield length of 6.8 mm. The females ranged from 2.3 mm to 5.9 mm, while the males were 1.8 mm to 7.2 mm in shield length. The smallest ovigerous P. samuelis was 4.0 mm in shield length and the largest was 5.8 mm. Females ranged from 2.5 mm to 5.6 mm and the males ranged from 2.2 mm to 6.5 mm. The different ranges and overlapping of sizes for non-ovigerous, ovigerous and males indicates that the ovigerous females, non-ovigerous females and males are not necesssarily competing for the same shells (Fig 2). Distribution At West Beach, the Hewett Transect, and Agassiz Beach, 50, 31, and 55 P. granosimanus were collected, respectively. Of the Pagarus granosimanus collected, males (avg 64%) were more abundant than the ovigerous and non- ovigerous. Non-ovigerous females were more common than ovigerous females. There was a slight increase in males and a decrease in ovigerous females as the studied area shifted from the exposed area, West Beach, to the protected area, Agassiz beach (Fig 3). However, there was no statistical difference among the proportions of non-ovigerous, ovigerous, and male Pagurus granosimanus at the three transects (p = 0.82). A larger sample size may be needed to accurately describe a possible trend between the habitats. At West Beach, the Hewett Transect, and Agassiz Beach, 22, 56, and 102 Pagurus samuelis were collected, respectively. At the three transects, males Martinez, E. The Distribution and Shell Utilization of the Female Hermit Crab were more abundant than ovigerous and non-ovigerous crabs (Fig 4). At West Beach and the Hewett Area, there were more non-ovigerous females than ovigerous females. But at Agassiz Beach, there was a lower percentage of males and a higher percentage of ovigerous females than non-ovigerous females. There was no statistical difference among the proportion of non-ovigerous, ovigerous and males at the three transects however (p = 0.16). Trends across the sites (up/down along the intertidal) in a transect were inconclusive due to some sites having a sparse population of hermit crabs. Therefore, population distribution analysis focused on trends between the transects and not the sites within the transects. Shell Damage Overall shell damage observations showed that ovigerous P. granosimanus had shells that were more intact than the shells occupied by the non-ovigerous and males (Fig 5). At the three transects, ovigerous P. granosimanus were in shells that had no whorl or apex damage (Fig 6). Among the non-ovigerous, ovigerous and males, the shells in the Hewett Area had the most damage, whereas Agassiz Beach had least shell damage. Despite this, damage differences among the non-ovigerous females, ovigerous females and males were not found to be significant(p = 0.33). Shells occupied by P. samuelis in the Hewett Transect were more damaged than the shells found in the other two transects. The transect with the lowest amount of shell damage was West Beach. Overall, 86% of ovigerous P. samuelis were in shells that were intact (Fig 7). Non-ovigerous females were present in shells that were more damaged than the ovigerous. Males had most of the shells with lip, whorl and apex damage. The difference between the non-ovigerous Martinez, E. The Distribution and Shell Utilization of the Female Hermit Crab and ovigerous’ shell damage was significant (p = 0.01). The difference between the ovigerous’ and males’ shell damage was significant too (p = 0.04) Fouling The predominant fouling organisms were Spirobis spp. and Crepidula planca , which were commonly found together. There was no significant difference in the proportion of fouling for shells occupied by P. granosimanus non-ovigerous, ovigerous, and males (p = 0.93) (Fig 8). The same was true for shells inhabited by P. samuelis (p = 0.60) (Fig There was a difference in fouling of shells (p = 0.04) between the two species of hermit crabs (Fig 10), which may be related to the difference in intertidal heights in which the crabs live and where fouling organisms live along the intertidal. Adequacy Non-ovigerous P. granosimanus were in shells larger than their preferred size at all three sites (Fig 11). The ovigerous were in shells smaller than the preferred size. At West Beach, ovigerous females were in shells 25% smaller than preferred. For P. samuelis, all were in shells larger than they preferred, except for males at West Beach (Fig 12). At Agassiz Beach and West Beach, ovigerous females were in shells smaller than the shells inhabited by the non-ovigerous. At the Hewett Transect, the situation reversed, and the ovigerous female had a larger shell than the preferred size shell of the non-ovigerous. Shell adequacy did have an effect on the reproductive potential of a female (Fig 13 & 14). To examine this relationship, females below the first Martinez, E. The Distribution and Shell Utilization of the Female Hermit Crab observed shield length of an ovigerous female were omitted. The remaining non-ovigerous and ovigerous females were separated into adequacy bins, and the percentage of ovigerous females of the total females for each bin was calculated. For both Pagurus granosimanus and Pagurus samuelis, as shell size increased from a small fit to a larger fit, the percentage of ovigerous females out of the total number of females decreased. Clutch Size The number of eggs to egg weight ratio was calculated from 7 clutches of hermit crabs (p = .947) (Fig 15). Note that these weights intermixed the eggs from both species, and did not take into account any variability in eggs between the species. For each species, clutch size was compared to crab wet weight, size, shell fouling, shell damage, and distribution at each three transects. None of these variables were correlated to the number of eggs that an ovigerous female carried. One possible correlation found was with the shell adequacy index for P. gransimanus. As the shell adequacy index decreased for the ovigerous P granosimanus, i.e., the hermit crab inhabited a shell smaller than the calculated preferred shell size, the egg number increased (Fig 16). Although the p-value is not considered significant (p « .07), more collection of ovigerous females may be able to reveal a significant trend. Discussion Martinez, E. The Distribution and Shell Utilization of the Female Hermit Crab The proportions of P. granosimanus non-ovigerous, ovigerous, and males do not appear to be strongly correlated to wave exposure. No trend was significant but a greater sample size may be needed for a further analysis. The lack of a significant difference may be that the overall habitat, i.e., the protected, exposed, and semi-protected areas, are indeed different, but the micro-habitats of where the hermit crabs live may not make one type of crab, say an ovigerous female, more or less susceptible to predation, dessication, and thermal stress. Also, courtship involves two hermit crabs being in close proximity for sometimes as long as two days (Coffin 1960), while the male goes through various movements, still holding onto the female’s shell. If an exposed area really provided significantly higher water turbulence without the convenience of a micro-habitat, it would be difficult to have courtship for a long time without the loss of some chelipeds. At each transect, the difference in relative tidal height did not correlate with the number of non-ovigerous, ovigerous, and males found. This suggests that ovigerous females did not seek different habitats than non-ovigerous females, at least in vertical height. However, more collections at different transects with higher numbers of hermit crabs would be required to resolve any subtle patterns, such as high temperatures, which cause eggs to fall from the pleopods prematurely. At most of the transects, ovigerous females had shells with less damage than the non-ovigerous females’ shells. For the avoidance of broken shells by the ovigerous female, there are at least two possibilities for this trend: (1) females are more likely to become reproductive when occupying an intact shell, or (2) ovigerous females prefer to move into intact shells. Courtship can take between one and two days, and since the male holds on to the female’s shell during all that time, the lip of the shell must still be Martinez, E. The Distribution and Shell Utilization of the Female Hermit Crab grippable. Even though there was a significant amount of lip damage found on the shells of ovigerous females, none of it seemed so severe as to hamper the efforts for courtship. Whorl and apex demage, however, can lead to unsuccessul reproduction. Since female hermit crabs must hold their eggs in their shell until they become attached to their pleopods (Coffin 1960), any holes in the shell are possible areas for eggs to be lost. In terms of shell use, this may imply that females, once ovigerous, must be in an intact shell to ensure attachment of the eggs to her pleopods. For these reasons, it appears unlikely that ovigerous females would ever want to change shells. Furthermore, observations in the lab made it clear that ovigerous females would have a harder time entering a shell with a clutch attached. The fouling data suggests that the fouling of shells does not appear to influence the choices of shells for non-ovigerous, ovigerous, and male hermit crabs. Therefore, the visual examination of shells as potential homes may be the same for ovigerous, non-ovigerous and male hermit crabs. Although not statiscally significant, shell adequacy appeared to affect the clutch sizes of P. granosimanus hermit crabs. These results agree with the findings of Bertness, in which he found that as the shell size adequacy decreased, the number of eggs increased. As shown by the shell adequacy index for the three crabs, the ovigerous females are living in shells that are at least 15% smaller than their preferred size. So it is possible that the Pagurus granosimanus is doing what Bertness described for another species: ovigerous females change the aim of their energy expenditure from finding a larger shell, to just producing a larger number of eggs. This trend was not true for the P. samuelis crabs. Since most of the shells were slightly larger than predicted, it is difficult to determine how ovigerous Martinez, E. The Distribution and Shell Utilization of the Female Hermit Crab females may decide to expend their energy in growth effort or reproductive effort. However, since the adequacy data suggests that P. samuelis is not shell stressed, this trend may be irrelevant. Summary Examination of the different areas along the intertidal at the Hopkins Marine Station, Pacific Grove, CA, has shown that the difference in habitats, from an exposed area to a protected area did not have a strong correlation with the makeup of non-ovigerous, ovigerous and male hermit crabs of either species. Differences were noted along each of the sites of the transects but no definite trend was found to describe the distribution of female hermit crabs. Damage of shell measurements reflected that ovigerous females were in shells that were more intact than shells occupied by non-ovigerous females. This was true for both species. This finding may be of a preference of ovigerous females or a sign that reproduction may be facilitated when a shell is intact. The latter seems more probable but additional studies need to be conducted. Females were more likely to be ovigerous if the shell size was smaller than the preferred size suggested from a shell adequacy index. Reproductive effort was only affected by the adequacy of the shell, as no variables regarding habitat, shell fouling, or tidal height had any correlation. My findings agree with some other species of hermit crab, i.e., as shell size becomes smaller than the preferred size, ovigerous females put more effort into reproduction than growth, rather than expend energy on pursuing a shell that may be only minimally larger. Acknowledgements Martinez, E. The Distribution and Shell Utilization of the Female Hermit Crab 1 would like to thank James Watanabe for his invaluable assistance and patience. 1 would also like to thank Joe Wible for relentlessly tracking down journals. And thanks to Maggie for getting me off of my butt. Literature Cited Belknap, Robert and John Markham. 1965. The intertidal and subtidal distribution of four species of Pagurus. Unpublished student paper. Hopkins Marine Station. Bertness, Mark D. 1980. Pattern and plasticity in tropical hermit crab growth and reproduction. The American Naturalist 177:754-773. Coffin Harold. 1960. The ovulation, embryology, and developmental stages of the hermit crab Pagurus samuelis. Walla Walla College Publications 25. Fotheringham, Nick. 1976. Population consequences of shell utlilization by hermit crabs. Ecology 57: 570-578. Taylor, Phillip R. 1981. Hermit crab fitness: the effect of shell condition and behavioral adaptations on environmental resistance. J. exp. mar. Biol. Ecol. 52: 205-218. Wilber, T. Payson. 1989. Associations between gastropod shell characteristics and egg production in the hermit crab Pagurus longicarpus. Oecologia 81: 6- Figures Figure 1. Map of the Hopkins Marine Station Figure 2. Shield length of Pagurus granosimanus Shield length of Pagurus samuelis Figure 3. Pagurus granosimanus distribution at each transect Figure 4. Pagurus samuelis distribution at each transect Figure 5. Overall damage of shells inhabited by Pagurus granosimanus and Pagurus samuelis Figure 6. Shell damage of Pagurus granosimanus at each transect Figure 7. Shell Damage of Pagurus samuelis at each transect Figure 8. Fouling of shell inhabited by Pagurus granosimanus Figure 9. Fouling of shell inhabited by Pagurus samuelis Figure 10. Overall Shell Fouling Figure 11. Shell adequacy of Pagurus granosimanus Figure 12. Shell adequacy of Pagurus samuelis Figure 13. Affect of shell adequacy on percentage of ovigerous females. Pagurus granosimanus Figure 14. Affect of shell adequacy on percentage of ovigerous females. Pagurus samuelis Figure 15. Number of eggs versus clutch weight Figure 16. Affect of shell adequacy on th reproductive output of Pagurus granosimanus 5 . V 1 8 S G At Ju O 22 3. 27 - . 8. o S . S 8 - 8 8 Shield Length (mm) wAd Hermit Crabs (%) 18.5 60.5 25.8 12.9 61.3 ENE 8 a 2 Hermit Crabs (%) 22.7 18.2 59 25 61 18.2 30.3 51.5 E 21 1 % intact % intact Number P 25 d avwd Number ooON Number a awa Number 88 PT 5 Number Number N08 L r awd number 5 ESE 3 Powpa number — ENE SESESE 2 5 0 0 n va 0.745 Adequacy - 1.009 1.029 1.213 0.94 0.953 0.958 1.037 0.932 ESE 10 9 wa Adequacy 0.926 1.027 .023 1.068 1.077 .066 1.124 1.148 E 2 +1 w Ovigerous females % (out of all females in ovigerous size range) n Vud Ovigerous females % (out of all females in ovigerous size range) 8 8 id 2pwe 8 Number 31 H Egg Number 8 L