Record of Carcharocles megalodon in the Eastern Guadalquivir Basin ( Upper Miocene , South Spain ) Registro de Carcharocles megalodon en el sector oriental de la Cuenca del Guadalquivir ( Mioceno superior , Sur de España )

Tortonian diatomites of the San Felix Quarry (Porcuna), in the Eastern Guadalquivir Basin, have given isolated marine vertebrate remains that include a large shark tooth (123.96 mm from apex to the baseline of the root). The large size of the crown height (92.2 mm), the triangular shape, the broad serrated crown, the convex lingual face and flat labial face, and the robust, thick angled root determine that this specimen corresponds to Carcharocles megalodon. The symmetry with low slant shows it to be an upper anterior tooth. The total length estimated from the tooth crown height is calculated by means of different methods, and comparison is made with Carcharodon carcharias. The final inferred total length of around 11 m classifies this specimen in the upper size range of the known C. megalodon specimens. The palaeogeography of the Guadalquivir Basin close to the North Betic Strait, which connected the Atlantic Ocean to the Mediterranean Sea, favoured the interaction of the cold nutrient-rich Atlantic waters with warmer Mediterranean waters. The presence of diatomites indicates potential upwelling currents in this context, as well as high productivity favouring the presence of large vertebrates such as mysticetid whales, pinnipeds and small sharks (Isurus). These large vertebrates recorded in the Eastern Guadalquivir Basin were potential prey of C. megalodon.


Introduction
The Carcharocles megalodon is the largest macropredatory shark to have ever lived.However, the fossil record of this predator is restricted to isolated teeth and vertebral centra due the cartilaginous skeleton of most of the sharks.The most complete specimen is an associated column of around 150 vertebral centra from the Miocene of the Antwerp Basin, Belgium (Leriche, 1926).The record of C. megalodon teeth ranges from 17 to 2 Ma (middle Miocene to Pleistocene) (Gottfried et al., 1996;Purdy, 1996;Pimiento et al., 2010).
Based on tooth size, C. megalodon reached a maximum inferred total length around 16 m (Randall, 1973;Gottfried et al., 1996), more than twice the reported maximum length of the white shark Carcharodon carcharias with 6.4 m (Bigelow & Schroeder, 1948;Randall, 1973;Compagno, 1984).The largest tooth of C. megalodon measures 168 mm (specimen PF 1168, Field Museum of Natural History of Chicago).
Data on C. megalodon from the Iberian Peninsula are relatively scarce, sometimes reported only as news in the daily press, without scientific revision.Descriptions of remains of C. megalodon from the Iberian Peninsula are those of Areitio (1877) and Meseguer Pardo (1924) from Messinian diatomites of the Lorca Basin (Southeastern Spain); Balbino (1995) from uppermost Miocene of the Alvalade Basin (Portugal); and García et al. (2009) from the Arenas Fm (Huelva, Spain) in the western Guadalquivir Basin.
In this work a complete tooth of C. megalodon from the Eastern Guadalquivir Basin (South Spain) is reported and examined in detail, including comparisons with other shark teeth of C. megalodon and C. carcharias described in the literature.

Geological setting
The Guadalquivir Basin is the foreland basin of the Betic Cordillera and it opens directly to the Atlantic to the WSW (Gulf of Cádiz).Autochthonous units to the north and allochthonous units to the south may be differentiated in this basin (Fig. 1A).The allochthonous units are composed by olistostrome deposits and olistoliths, located at the frontal part of the Subbetic nappes, which are mixed with Miocene clays and marls.
The shark tooth was collected in diatomaceous marl blocks obtained from the old quarry of San Félix, located 2 km south of Porcuna (coord.37° 51′ 21.4″ N; 4° 11′ 21.6″ W) (Fig. 1B).The diatomaceous marls are white to yellowish, sometimes laminated, and they alternate locally with greyish marls without visible stratification.These deposits belong to the highest part of the Valenzuela Formation (Aquitanian-Torttonian in age, Tjalsma, 1971), whose stratotype was established in the San Félix Quarry.The marls contain abundant diatoms and subordinately sponge spicules, silicoflagellates and radiolarians.The sediments of the type section (San Felix Quarry area) have a planktic foraminiferal assemblage with Globorotalia menardii, Globigerina globorotaloidea, Globigerina nepenthes and Globigerinoides obliquus, among others (Tjalsma, 1971).This foraminiferal assemblage belong to the zone N16 of planktic foraminifera (Tortonian) equivalent with most of the middle part of the stratotype of the Tortonian.The main diatom assemblages consist mostly of planktic (Thalassionema, Thalassiotrix, Thalassiosira) and scarce benthic (Delphineis) forms typical of cold oceanic upwelling waters (Molina et al., 1987;López-García & Bustillo, 1994).The outcrops belong to the third and last episode of diatomitic sedimentation defined in this area by Bustillo & López-García (1997) of the Thalassiosira yabei Zone (Middle Tortonian).
In this area of the North Betic Strait, which connected the Atlantic Ocean to the palaeo-Mediterranean sea, high concentrations of diatomaceous deposits were produced because it was more oceanographically dynamic than the Atlantic.The Guadalquivir Basin was an Atlantic basin connecting with the palaeo-Mediterranean through small basins (e.g.Granada Basin and Guadix Basin) to the south by straits.
Flood-tidal currents as well as temperature and salinity gradients favoured movement of sea-water (e.g.Reolid et al., 2012;García-García et al., 2014;Martín et al., 2014).The interaction of the cold nutrient-rich Atlantic waters with the gulfs, bays, small basins and bottom topography of the North Betic Strait caused upwelling, and high productivity of biogenic silica (Molina et al., 1987).The narrowing of the strait from the Langhian to the Tortonian could have caused the progressive upwelling deduced from the study of the diatomites.This narrowing could also explain the evolution of the upwelling from open sea conditions to near shore conditions (Bustillo & López-García, 1997).
The youngest part of the Valenzuela Fm, in which the area of the San Felix Quarry is included, has to be considered parautochthonous of the syn-olistostromic sequence of the middle-late Tortonian times (Riaza & Martínez del Olmo, 1996), while the older parts of the Valenzuela Fm belong no doubt to the allochthonous basin filling that have travelled longer distances.

Methods
The tooth analysed, coming from diatomites of the Eastern Guadalquivir Basin, is now displayed in the Museo Obulco (Porcuna, Jaén province).This tooth is described following traditional measurements used by previous authors working on C. megalodon and C. carcharias (e.g.Applegate & Espinosa-Arrubarrena, 1996;Hubbell, 1996;Mollet et al., 1996).
The main measurements for describing the tooth and estimating the total length of the shark are the crown height and the crown width (Fig. 2).According to Gottfried et al. (1996) and Hubbell (1996), the crown height or tooth height is the vertical distance between a straight line touching the lower extensions of the enamel adjacent to the root at the base of the crown and a parallel line touching the tip of the enamel.For these authors, the crown width is the widest part of the enamel at the base of the crown (Fig. 2).
The Recent white shark Carcharodon carcharias is the closest living relative of C. megalodon, and therefore several authors have used skeletal material of C. carcharias as the primary basis for comparison with C. megalodon and to infer lifestyle (Uyeno et al., 1989;Applegate, 1991;Gottfried et al., 1992Gottfried et al., , 1996;;Applegate & Espinosa-Arrubarrena, 1996;Adnet et al., 2010;Pimiento et al., 2010).In order to better approximate the total length of the specimen of C. megalodon from Porcuna, we considered data on crown height and crown width of upper anterior teeth from 102 specimens of C. carcharias (85 specimens) and C. megalodon (17 specimens) available in the literature (Table 1).

Results
This tooth presents the diagnostic characters of C. megalodon such as large size, triangular shape, fine serration on the cutting edges, a convex lingual  face, a slightly convex to flat labial face, and a large v-shaped neck (Fig. 3).Two root lobes are welldifferentiated and rounded on the basal root edge.The crown height is 92.2 mm and the crown width is 81.3 mm.The total height of the tooth measured perpendicular from the apex of the tooth to the baseline of the root is 123.96mm.The tooth is symmetrical and nearly perpendicular, only slightly slanted (2.1°).For this reason, the mesial crown edge length is 102.0 mm while the distal crown edge length is 94.8 mm.The size of the teeth varies within the jaw, anterior teeth being large and symmetrical, whereas the large posterior teeth are asymmetrical and feature slanted crowns (Applegate & Espinosa-Arrubarrena, 1996;Hubbell, 1996;Pimiento et al., 2010).Taking into account the symmetry and the size, this tooth probably corresponds to the upper anterior (A2 or A3 s.Hubbell, 1996; or I or III s.Applegate & Espinosa-Arrubarrena, 1996) or the second upper lateral position.The cutting edge presents 135 serrations in the mesial crown edge and 126 serrations in the distal crown edge (Fig. 3D).There are approximately 13 serrations per cm, and they are smaller and denser close to the root and the apex.The points of basal serrations are oriented 90° with respect to the cutting edge.
We follow the first hypothesis, where C. megalodon is separated from genus Carcharodon on the basis of tooth features such as large size, absence of lateral denticles, and fine serrations-as opposed to a small size, large serrations and the presence of lateral denticles in C. carcharias.However, Carcharocles and Carcharodon are included in the Order Lamniformes, and in the absence of living members of Otodontidae, Carcharodon carcharias is the most analogous species available.

Size estimation
Among the different measurements on C. carcharias, the crown height of the upper anterior tooth A2 presents a higher correlation with the total length of the specimen (r=0.92)than the crown width (r=0.53) (Fig. 4).In addition, the correlation is extremely good between the crown width and crown height in the case of C. megalodon (r=0.94),better than in C. carcharias (r=0.83) (Fig. 5).For these reasons, commonly, for the estimation of total length of C. megalodon from fossil teeth in comparison with the most analogous specie C. carcharias, it is used the crown height only.
The largest tooth specimen of C. megalodon is the first upper anterior tooth of the Field Museum of Natural History of Chicago (specimen PF 1168), measuring 168 mm in total height and 125 mm in total width, and the 2nd upper lateral tooth of the British Museum of Natural History (specimen P10725), with 154 mm total height and 134 mm total width.Applegate & Espinosa-Arrubarrena (1996) estimated 12 m length for the individual having the first upper anterior tooth of the Field Museum (Chicago).This estimation was based on comparison with the same tooth of C. carcharias, of known length.C. megalodon dentition is apparently similar in morphology and the number of tooth rows to that of C. carcharias (Uyeno et al., 1989).However, the jaws of C. megalodon must be somewhat more robust, larger and thicker, having more massive muscles than those of the C. carcharias (Gottfried et al., 1996).Correspondingly to great jaws and teeth, the head would have most likely been massive as well, with a large branchial region for respiration.In turn, a large stomach and intestine would be necessary for processing large prey, meaning a morphology similar to C. carcharias but more robust.
Previous authors assume that the ratio of upper A2 tooth size to total length of the C. megalodon is similar to that in white sharks, and that tooth size increased along with the total length of the shark in both species.The upper A2 tooth in Recent white sharks ranges in size from 56-63 mm height and 40-50 mm width for adult females, with a total length of 490-600 cm (Gottfried et al., 1996).A simple method for obtaining a relative total length ratio of the C. megalodon specimen from the Guadalquivir Basin with respect to the white shark consists of determining the tooth height ratio of the C. carcharias and C. megalodon from the Guadalquivir Basin.In the case of the C. carcharias, Gottfried et al. (1996) propose an average height for the upper A2 tooth of 59.2 mm, corresponding to an average total length of 550 cm.550 cm ×92.2 mm/59.2mm =856.6 cm According to this method, the studied tooth belonged to a specimen around 8.5 m in length.
Using the upper A2 height measurements from 85 specimens of C. carcharias (Table 1; Fig. 4), a linear least-squares regression was calculated following Gottfried et al. (1996).where a and b, as calculated, were −0.001 and 0.119, respectively (r 2 =0.85;Fig. 4).Using 92.2 mm as the crown height for the C. megalodon specimen of Porcuna, we obtain −0.001+0.119(92.2 mm) =10.97 m.Shimada (2002) proposes a linear least-square regression based on relationship between crown height and total length in C. carcharias, but taking into account all the positions of the tooth set.This relation is calculated as: Total Length (cm) =a+b (Crown Height (mm)) with a being a constant and b being the slope of regression line.Shimada (2002) propose a and b values for predicting the total length of any tooth from C. carcharias.
Bearing in mind that the tooth from Porcuna (92.2 mm) is probably an upper anterior tooth A2, a=−2.160 and b=12.103(Shimada, 2002): That is, the tooth of Porcuna belonged to an adult, and is in the large size range of specimens of C. megalodon (Fig. 6).
According to Shimada´s (2002) method, the largest tooth of C. megalodon, the upper anterior A2 of the Field Museum of Natural History of Chicago (specimen PF 1168), which reaches a crown height of 168 mm, would have belonged to a shark 20.31 m long (Fig. 6); and the second upper lateral tooth of the British Museum of Natural History (specimen P10725), reaching a crown height of 154 mm, belonged to a shark 20.73 m long.

Palaeoecology
Tooth structure and size indicate that C. megalodon began primarily as a fish feeder and evolved to feed on marine mammals (Applegate & Espinosa-Arrubarrena, 1996).The almost worldwide distribution of the C. megalodon in the Miocene to Pliocene seas in nearshore deposits at moderately high temperature latitudes indicates environmental requirements close to those favouring living C. carcharias.These environments are nutrient rich shelf areas where large marine mammals are relatively abundant.The record of bite marks of C. megalodon on fossil cetacean remains indicates that whales and dolphins were regular prey (Demèrè & Cerutti, 1982;Cigala-Fulgosi, 1990;Purdy, 1996;Ehret et al., 2009b).In the Guadalquivir Basin, and Estudios Geológicos, 71(2), julio-diciembre 2015, e032, ISSN-L: 0367-0449.doi: http://dx.doi.org/10.3989/egeol.41828.342concretely in the sector between Porcuna and Bailén, numerous remains from large marine vertebrates have been recorded, in cluding Isurus, indeterminate pinnipeds and whales (Reolid et al., 2014), which could be potential prey of C. megalodon.The palaeogeography of this basin, as an Atlantic branch connecting with the Mediterranean Sea, probably favoured the hunting techniques of C. megalodon.Obviously, this is very speculative due the scarce record of marine vertebrate fossils from Upper Miocene in this basin (Fig. 7).However, the record of C. megalodon is congruent in a context as the North Betic Strait where high productivity is reported (Molina et al., 1987;Bustillo & López-García, 1997) favorable for marine mammals, also recorded in the same basin (Reolid et al., 2014).

Conclusions
A large shark tooth is recorded in Tortonian diatomites of the Eastern Guadalquivir Basin (Porcuna, Jaén).The large size (92.2 mm crown height), triangular shape, broad serrated crown, lingual face convex, labial face flat, large neck, and robust, thick angled root determine that this specimen corresponds to Carcharocles megalodon.The symmetry with a low slant indicates this is an upper anterior tooth.The total length estimated from the tooth crown height is around 11 m.
In the North Betic Strait, which connected the Atlantic Ocean to the palaeo-Mediterranean sea, the interaction of the cold nutrient-rich Atlantic waters with the palaeogeography and the bottom topography caused upwelling, and a high productivity of biogenic silica (diatomites).High nutrients and productivity favoured the presence of large vertebrates such as mysticetid whales, pinnipeds and small sharks (Isurus) recorded in the Upper Miocene deposits of this area.These large vertebrates constitute potential prey of C. megalodon.The palaeogeography of this basin, as an Atlantic branch connecting with the Mediterranean Sea, was probably well suited to the hunting techniques of C. megalodon.(Sendra, 1997;Sendra & Bajo Campos, 2013;Toscano et al., 2013;Reolid et al., 2014) and the C. megalodon teeth studied.

Fig. 1 .
Fig. 1.-Geological setting.A. Location of the Guadalquivir Basin with the autochthonous and allochthonous units.B. Geological sketch of the Porcuna area with the location of the San Félix Quarry.

Fig. 2 .
Fig. 2.-Tooth measurements employed in the description of the tooth of Carcharocles megalodon.
Fig. 5.-Comparison of crown width vs. crown height in C. carcharias and C. megalodon.

Fig. 4 .
Fig. 4.-Linear relationship between the different crown measurements of the upper second anterior tooth (crown height and crown width) and the total length in Carcharodon carcharias.

Fig. 6 .
Fig. 6.-Linear relationship between the crown height of the upper second anterior tooth and the total length in Carcharodon carcharias and Carcharocles megalodon.Note total length in C. megalodon calculated according to regression equation of Shimada (2002).