2008, The Journal of Arachnology 36:464-467 SHORT COMMUNICATION Relationship between litter characteristics and female size in Tityus stigmurus (Scorpiones, Buthidae) Ana P.N. Aguiar, Pedro L. Santana-Neto, José R.B. Souza and Cleide M.R. de Albuquerque! Departamento de Zoologia / Universidade Federal de Pernambuco, Avenida Morais Rego sin, Recife ~ PE, CEP 50570-420, Brazil Abstract. Ttsus stigmrus (Thorell 1876) is one of the most medically important scorpion species in Brazil, but many basic aspects o its life history are unknown. Here the pattern of female reproductive investment was examined, along with {development of the P and 2" instars and the relationship betwcen 2nd instar mass and molting to the 3" insta. Relative {o other buthid scorpions. 7: seigrurus has a smaller litter (average 10 young) and a shorter [instar period (averase 4 days) and 2" instar period (average 68 days), Neither liter size nor offspring mass showed a relationship to female siz. A Significant positive correlation wis observed between total fitter mass and Titer size. The minimum mss required for socessful molting tothe 3° ins to tho production of more but not heavier ofispring, Keywords Dilerences in female reproductive investment are common in closely elated spovies of animals. A female may use her finite reproductive resourees to maximize her fitness by producing either large litters of small young or small liters of large young. Given that ‘the individual fitness of each offspring is generally coreated with larger maternal investment, the female's option of producing large Timers with sina young ereates a potential confit inthe fitness o the female and her young. Thus, the optimal distribution of pareatal resouress tends to be based on the cost-benefit relation for both paren and offspring (Smith & Fretwell 1974; Swear 1992: Fox & ‘Caesak 2000), In scorpions, studies on the female reproductive investment are still scarce, yet such studies have shown that different taxa show different tendencies. In species such as Centruroides exiicanda (Wood 1863), aejonsspinigerus (Wood 1868), Diplocenaruspelonclensis Francke 1975, nnd Pseuiouraciomss apacteamus (Gerisch & Solegad 1972) there is no relationship between female size and offspring production (Brown 2004). However, a higher reproductive investment in larger Titers was recorded inthe largest females in Centruroiesvittatus (Say 1821), Vaejovis spinigerus (Wood 1863), aad Ticrus columbian (Thorell 1876) (Formanowice & Sehalfer 1993; Lourengo eta. 1996; Brown 2004). Although Tinus species are capable of producing multiple ters from a single insemination (Kovoor etal. 1987; Polis & Sissom 1990; Lourengo ef al. 1996), litte is Known about female reproductive investment, including species involved in medically importaut eavenomations in Brazil, Population dynamics are also influenoad by the duration of postembryonie development, Within Buthidae, this vases from six ‘o 48 months depending on the species (Polis & Sissom 199). Both female reproductive investment ‘and postembryonic duration are unknown for Tityus stigmas (Thorll 1876), a species tha is widely listibuted in urban aceas of northeastern Brazil and responsible for many reported envenomations each year (Fickstedt 1983, 1984; Lis ‘daSilva etal. 2000; Barbosa et al. 2003). Is occurrence in urban settings refers its ability 10 Hive under roofs, among accumulated debris in the exterior areas of residences (Eickstedt 1983, 1984 Lourengo etal. 1996), and in cesspits This study analyzed the litter size, offspring mass, and total liter amass of T stfgomus and evaluated the relation of female size wo these ‘variables. Times of development for and 2" instars, as well a the ‘Corresponding author. E-malileide.ufpedmail.com Liter size, female sizs, reproductive investment, offspring mass, as 34. mg. Overall, female reproductive resources nT. stigmmms appear to be applied incemolt period relationship between mass and molting (0 the 3 developmental instar, were also assessed METHODS Animal sampling and maintenance. Tijus sigur adulis were colleied from residential ateas of Recife (°04'03'S, 3455'00W), Pernambuco State, The climate is predominantly hot and wet with mean temperature 25° 2° C and annual pluviometie precipitation ‘of 2.094 mim, Following capture, animals were individually housed in pst teraria (8.5 em diameter % 7.8 em height) where water and shelter were made available. Weekly, Peripfanera americana (Linnaeus 758) nymphs were offered as food. The animals were kept at 28° °C mean temperature and 12:12 h lightdark photoperiod. Data collection and analysis.— The relationship between maternal carapace length (CT) and liter ize (LT) was analyzed ina sample of | 25 pregnant females that gave birth to young. In 4 broods, it was possible o analyze 1" instar duration and 10 determine toval liter mass (TLM) and mean offspring mass (MOM). TLM sas taken as a measure of reproductive investment and was calculated 2s the sum of individual masses of juveniles aeconting to the methodology of Brown (2004), Juveniles were weighed to the nearest 0.1 mg with an analytical balance (Meter AE 260 DeliaRange) as soon as they moved down from the mother’s dorsum, After oflspring dispersal female CL was measured with digital calipers to the nearest 0. mm under a dissecting microseope (Leiea MZ), Liter size was estimated by counting juveniles on the female's dorsum within 24 ho birth and again immediately aftr dispersal. Al ‘dead and living juveniles were counted except for these exnnibalize Due to a femak’s habit of cannibalizing young at birth and the superimposed distribution of young on her dorsum, an avcurate titer size was difficult to obtain. Eight gravid females were disected and the number of young recorded for Titer size comparison. Females chosen Tor dissestion showed signs of imminent parturition such as Tack of movement and an enlarged mesosoma with offspring visible through the integument. In general, females presenting these traits save birth within two weeks ANOVA was used (0 test the differences between brood sizes. The correlation coefficient (Pearson's 1) was used to evaluate the relationships between female CL and liter size, mean offspring mass ‘of TLM, It was also used to examine the relationship between TLM. and liter size or MOM. The significance of these corelations was assessed with a estAGUIAR ET AL.—FEMALE-YOUNG INTERACTIONS IN 7: STIGMURUS as alg - FA Bo[2 Se « 2. . g : 2 i aoe 2° Bol + Bal él see Sal. : 2 : : Female Size (CL mm) Female Size (CL mm) | %on] Balae * 02s: 3 ode . = is gx ot se] 2 | 2: « awl ® ac “| Ba rat : 5 | oa = . ga}. oe : 2, : Fomale Size (CL mm) TLM (mg) . — 8, : als * ar] = -. Pa a se = 3 . 2° 4. ° 5 al = foe & . . . TUM (m9) Litter Size Figures 1-6. Comelation results between carapace Fength (CL) aud brood characterises of Tiyus sgmarus. 1. Liter size (LT); 2. Mean ‘offspring mass (MOM) 3. Total liter mass (TLM): 4. Correlation between total liter mass and mean offspring mass. 5,6. Correlation between total litter mass (TLM) against litter size and litter size, respectively, with mean offspring mass in Tiyus sigmurus uration of fist and second instars an mass variation in the late. ‘The duration of the I” instr stage was determined for 25 Kitts by counting the days between bieth and the Fest moll. A second sample fof 19 nesly dispersed individuals was used to determine the period hetween the Ist and 2nd molts. Hetween mots, the animals were fe weekly with nymphs of P. americana with mean mass of 7.8 mg. Masses of 2nd instars were determined 24h ater cach Feeding in ‘order to estimate variation in mass und to determine any relationship ‘between mass and molting. The difference bauween the intial mass (MD, recorded up to 24 alter dispersal, and the final mass (MD, recorded immediately prior to the 2nd molt, was sed as an estimate For the mass variation inthe 2nd instar, The lowest MY was used as un cstimate ofthe minimum mass needed to mole succesful RESULTS Tutyusstignwns showed gxcat variation in female size, iter size, and mean offspring mass. Female size (CL) averaged 6.00 + 0.44 mm, varying from 482 to 6.59 mm, The liter size (LS. » = 25 females) ‘veraged 10.40 + 2.90 and was unrelated 10 female size (Fig. 1) Similar data were obtained in the group used to analyze total liter ‘mass (TLM) and mean offspring mass (MOM) (= 1 females) (CL = 608 = O48 mm and LS = 10.14 » 2.11, respectively), MOM in this group averuged 27.67 = 4.8 mg.and was not related to female size (Fig. 2), The mean brood size obined from females dissected before partutition (n = 8; 1187 + 408, range 8-21) was not significantly silerent from those that gave birth to hve young (Fg ian = 0.6%: P 55) ‘The reproductive investment (TLM) showed a postive, but not Sgnifcant, correlation with female size (Fig. 3) and mean ollspring ‘mass (Fig. 4), However, significant postive correlation was found between TEM and liter size Fig. 5}. A negative but not significant correlation was found between litter size and mean offspring mass Fig. 6. Female mass hefore parturition (until after the last feeding) ranged, fiom 9754 10. 1635.0 mg, showing a TLM-to-femalesmass ratio higher than 0.20 for most females (n = 9/12, ranging from (1.13 to 042),‘THE JOURNAL OF ARACHNOLOGY Table 1.—Data on reproductive investment and frst instar traits of 12 broods of Tityusstignusus. CL. = maternal carapace length (mm Litter size instar Period (days) ‘Mean aflspring mass (me) “Total liter mase (ma) ear ir 4 MS 19 220 584 nL 5 238432 2614 oat 6 4 343230 2089 651 2 4 2362 Lo 2837 482 0 3 DSR +26 2s18 631 a 5 329841 3023 631 2 5 2630 Bis 64 in 3 Mos 3I 03 560 5 4 239421 1909) 58 5 4 B30 m6 69 2 3 250405 o07 580 3 7 227 16 2044 602 0 3 99423 Doss 581 1 3 24 308 68.6 Birth, frst instar and pre-dispersl phase. The duration ofthe fist instar was investigated in a sample of [42 individuals from diferent Tiers. This period was on average 407 + 1.14 days (range 3-7) (Table 1) and the molt occurred simultaneously in offspring of the same litter. After the first molt the immature individvals remained on the mothers dorsum for 5-6 more days, Dispersil occurred 5-10 days following birth. Individual young occasionally climbed down from the female's dorsum, particularly inthe nocturnal period, then returned the mother. While off the female, they searched for water ‘and food in th environment After the transition period from dorsum to the environment, they remained apart fom the mother. though aggteyated in the shelters placed in the terara ‘Second instar phase. The intrmoht period between the 2 and 3 instar (n = 19) was approximately 16 times longer than the duration of the instar. The simultanety observed inthe frst molt did not occur inthe second, and the duration ofthis phase of development was on ‘#vcrage 67.81 = 18.80 days (ranging = 42-107 days). The morality for the 2” instar was 47.37%, and among these 33.33% died during the intermolt period (one due to cannibalism) and 66.67% ded ducing the ‘molting proces (Table 2), No relationship was observed between mass ‘of the offspring before a molt and survival during the molt, The ‘minimal pre-molt mass of a surviving individual was 34.0 mg and the maximum was $9.0 mg. Hovtever, many immatures with, masses ranging between 494 mg and 56.0 mg died duting o afer the molt DISCUSSION ‘Tipps stignurus can vary greatly in female size, litter size, mean ‘ollspring sie, and total liter mass, as described for other scorpion species both within and among populations (Brown 2001, 2003). These teats are important components of female reproductive investment, ‘and impact the fimess of both the Female and her offspring. Based on data summarized by Polis & Sissom (1990) and Lourengo 2007), 7. stigauras had lower brood sizes (mean of 10 young) ‘computed to most buthid species (average 32) and other species of Tuble 2.—-Development length (ays) and mortality rate in Tips sugmurus 2° insta. ‘Second instar period, Mortality Litter size ‘mean = SD (range) @ 7 B22 = 22.70 (82-107) 8571 2 645 = 20551 (50-79) 0 1 6 0 8 68.12 © 17.00 (50-95) 25.00 1 2 10 9 6781 18.80 4131 Tigyus (mean variation = 15-25), Our results show that females that allocated more resources to reproduction (measured as TLM) had ‘more but not heavier offspring, and that TLM was not significantly related to female size (CL). This may’ explain the ack of correlation between CL and liter size. There are few studies on the reproductive investment in Tipus species, although Lourengo etal. (1996) found Positive correlation tween liter size and female size in both parthenogenetic and sexual populations of T: colonbiamas. 1 is Important to note that our study used females that Tad been maintained in the laboratory for atleast a year nd that data such 3s prey availablity and specimen age were not controled. Thus. vvaration in prey availability in the environment may have led to dllerential resource allocation by females resulting in large variation in TLM. A shift in offspring size from nevative to postive values in V- ‘vorbis! Stahnke 190 was attributed (oan inereasein prey availabilty fn the environment, which allowed females 10 invest more in reproduction (Brown 2001), In addition, old females tend to reduce the numberof follicles (Lourengo 1979) which may lead to small ter sizes. Therefore, lick of correlation between most of our vatiables may have been influenced by factors such as accessibility of food in the environment and the age of Females, In contrast, in several species suet as Centnovides exiicaula, Vacjovisspinigerus, Diploceneras pelonetnss, Pseudowroctonus ape cheanus (Brown 2004) and Centrioides vrtatus (Formanowice & Shafer 1993), larger females invest more in reproduction. In these specs, positive correlation between female size and total iter mass has been deseribed. Moreover, iter size was often positively related with CL and TEM, showing that the largest females invest more in reproduction through production of more offspring. In all of these species, as well asin 7: suigmunes, there was no correlation between Female size and mean offspring mass, indicating that large females do not produce larger offspring. Offspring mass in 7. stigmas was also not correlated with litter size, which may suggest a Tack of trade-off between mass and number of individuals. Similar resulls were ‘obiained by Salomon et al. (2005) in the spider Stezodypus linear Lawcille 1817, Offspring mass in this spider showed no correlation ‘with clutch size, indicating that each variable may be determined independently. In addition. offspring mass at hatching. (and cconsequently egg size) appeared to be relatively constant and independent of female size ind body mas, In all of the observed liter, juveniles remained unfed om the ‘mother's dorsum during the entire instar phase and underwent the 1M ecdysis simultaneously, a phenomenon characteristic of many seorpion species (Sisom & Francke 198%; Polis & Sissom 1990; Brown 1997, 200M; Loutengo 2000; Farley 2005; Lourengo & Goodman 2006). Mean firstinstar duration in 7. stigmaurus (4 days) was shorter than other Buthidae, including Ceniruoides gracilisAGUIAR ET AL. FEMALE-YOUNG INTERACTIONS IN T. STIGMURUS a6 (Latrville 180 at 8 days (Francke & Jones 1982), Conruoides exticanda (Wood 1863) (= C.seudpurates) at 7 days (Brown. 2004), and Grogphus hirtus Kracpelin 1901 at 14. days (Lourengo & Goodman. 2006). The fiestinstar period in 7 sitenaras is even shorter than that of eongenerie species (Polis & Sisson 1990) Similarly, the 2nd instar of 7. stigoes lasted 67 days an average, ‘which wats shorter than in other species of Buthidae. Toscano-Gades (2004) confirmed 321 days for 7 trivia and Lourengo & Goodman (2006) described 112 day period for Ghia. ATonger developmental petiod for T.stigmurus was described by Mathiesen (1971), who found an average of $0 and 148 days in 660 broods of five young each ‘produced bya sinle female. Scorpions from both studies are likely to have come from different populations singe they were obtained from cites separated by 209 km. According to Brown & Formanowicz (1995), individuals from different populations may face an adaptation {o mietovariation inthe environment, and this fact may reflect genetic «liferences that might explain the differences relative to our Findings. ‘Mortality inthe period botwoen the 2 and 3° stages was high, as ‘only $2.63% of the intial simple survived. Most of these deaths, 166,67%, occurred during cedysis, and the body mass atthe end ofthe 2 sage was not associated with the survival of juveniles. In their experiments with Vacjovisbilinasus Pocock 1898, Sssom & Francke (1983) noted that only 43% ofthe inital sample survived the 2" molt and suggested dehydration as a posible cause. Tinus stigrarus ‘young also appeared to be desicated after dying during molting, with ‘the imtevument being dried and fal, In toial the data gathered in this study suggest that femle 7: stigmaras make a significant investment in reproduction (aver 20% of the body mass), Te i important to note that the allocation is ‘underestimated given that the nconate seorpions decreas in mass uring the first instar period and ther frst molt Polis & Sissom 1950). With larger investments in reproduction, there is a trend toward ‘increasing the number but not mass of ofsprine, Moreover thistrai is not influenced by the mothers size, so that factors such a5 age of the female and food availabilty in the environment ate likely 10 have Fundamental importance in the reproductive investment made by this scorpion species. The comparatively short developmental period of this species may promote rapid populational growth when environmental ‘conditions are favorable. Considering that this analysis is restricted to ‘observations ofthe [and 2" instars, studs of other developmental stages should be conducted to test this hypothess, ACKNOWLEDGMENTS ‘We are especially grateful tothe two anonymous reviewers andthe editor for their comments and very helpful suggestions to the manuscript. Voucher specimens are deposited at Entomological Collection at Laboratory of Terrestrial Invertebrats, Universidade Federal de Pernambuco, Brazil LITERATURE CITED Barbosa, MGR, ME, Bavia, CE, Pinto da Silva & FR, Barbosa. 2003." Aspectos epidemio\dgicos dos acidentes eseorpidnicos em Salvador, Bahia, Brasil. Citncin Animal Brasicia 42:15S-162, Brown, C.A. 1997, Growth rates in the scorpion Pscudouractonus reddel’ (Scorpionidéa, Vacjovidae). Journal of Arachnology 25288294, Brown, CA. 2001. Allometry of offspring size and number in scorpions. Pp. 307-315, Scompions 2001. In Memoriam Gary A. Polis. (V. Fet & P.A, Selden, eds). British Arselnological Society, Burnham Besches, Buckinhamshire, UK. Brown, CA, 2003, Offspring size-number trao-off in seompions: un empirical test of the van Noordwik and de Jong model. Evolution 57184-2190, Brown, C.A. 2004. Life histories of four species of scorpion in three families (Buthidac, Diplocentridae, Vaejovidae) from Arizona and [New México, Journal of Arachnology 32:193-207, Brown, C.A. & DR. Formanowicz, 1995, Variation in reproduetive investment among and within populations of the scorpion Centruroies vinaius. Oscologia 03:140-147 Fickstedt, V.R.D. 1983.84. Escorpionismo por Tityus stigmurus no ‘Nondoate do Brasl(Scorpiones; Buthidac). Memorias do Instituto Batantans 47486 133- 137 Farley, R.D, 2005, Developmental changes in the embrye, pronymph, and fist moult ofthe scorpion Comruroides vittatus (Scorpiones: Buthidao). Journal of Morphology 2681177 Formanowiez, DR, & LR, Shafer 1993, Reproductive i inthe scorpion Cenirarldes vinaws, Oecologia 94:368-37 Fox, CW, & ME. Czesak, 2000, Evolutionary ecology of progeny size in arthropods. Annual Review of Entomology 45°41" 369, Francke, OF, &S.K, ones. 1982. The ile history of Cenruroies grails (Scorpionesies Buthidae) Journal of Arachnolopy 10.223-239 Kovoor,., W.R. Lourengo & A, Muioz-Cuevas, 1987. Conservation ‘des sperinatozoides dans les voies aénitales des femelle et bolo de la reproduction des Scorpions (Chélcérates), Comptes Rendus de PAgidemie des Scignce, Paris Ser 1 304:259.264, Lire-DaSiha, RIM. A.M. de Amorim & TK. Brazil, 2000, Envenenamento por Tijus stigmas (Scorpiones, Buthidae) no cstado da Bahia. Revista da Sociedade Brasileira de Medicina “Tropical 33:239-245, Lourengo, WR, 1979. La biologie sexuslle et Ie développement ppostemiryonnaite da Scorpion Buthidae: Tityessrivinanus fascox lous, Pesoa 1935. Revista Nordestna de Biologia 29-56 Lourenjo, W.R. 2002. Reproduction in seorpions, with special reference to parthenogenesis. Pp. 71-85, fx European Arichnology 2000. (S. Toft & N. Scharff, eds). Aarhus University Press ‘Aarhus, Denmark, Lourengo, W.R. 2007. Litter size in mioro-buthoid_seorpions (Chelicerata, Scorpiones). Boletin Sociedad Entomolégica Arago- esa 0473 477 Lourengo, W.R.,J-L. Cloudsley-Thompson, O. Cuellar, VLD. Von ickstedt, B, Burra Viera & M.B. Knox. 1996, The evolution of scorpionism in Bruil in recent years. Journal of Venomous ‘Animnals and Toxins 2121-134 Lourengo, WR., 0. Cuéllar & FRM. La Cruz, 1996, Variation of reproductive effort between parthenogenetic and sexual popute tions ofthe scorpion Tis cohmbianus.Fournal of Biogeography 235681686. Lourengo, W.R, & SM. Goodman. 2006. Notes on the postembry- ‘nie development and ecology of Groyphus hirsus(Kraepelin, 1901) from the Pare National d Ankarafantsika, northwest Madagascar. Zoologischer Anzeiger 244:181-185 Matthiensen, FA. 1971. Observations on four species of Brazilian ‘scorpions in captivity. Revista Brasileita de Pesquisas Médicas ¢ Biolosicas 4301 302. Polis, G.A. & WD. Sissom. 1990. Life history. Pp. 161-228. In The Biology of Scorpions. (G.A. Polis, ed). Stanford University Pres, Stanford, California Salomon, MJ. Schneider & Y. Lubin. 2005. Matemal investment in spider sith suicidal maternal cate Stegedypnsfneatus Araneae, Eresidac). Oikos 108-614-622 Sissom, W.D. & OF. Francke, 1983, Post birth development of Vaejovis bilineatus Pocock (Ssorpiones: Vaciovidae). Journal of Arachnology 11:69-75, Smith, C.C. & S.D. Fretwell, 1974, The optimal balance bewwoen size and number of offspring. American Naturalist 108:499-506. Stearns, SC. 1992, The Evolution of Life Histories. Oxford University Press, Oxford, UK. 264 pp. ‘Teseano-Gadea, CA. 208, Confirmation of parthenogenesis in ‘Tuyus sriitatus Kracpein, 1898 Scorpiones, Buthidac). Journal of Arachnology 32:866- 80). Manuscript received 15 December 2007, revised 22 July 2008,