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The green turtle (Chelonia mydas) present in tropical seas, uses as a feeding area the coastal region of Peruíbe, has the skull as a relatively large and solid structure, and a strong jaw formed by the junction of small bones as it has very abrasive feeding. By applying scanning electron microscopy techniques, it was possible to identify the presence of a bone structure located in the hyoid in the ventral region of the skull along with the mandible of juvenile individuals of green turtles, and as there is no related research, it was necessary to perform a CT scan, decalcification and histology of the quelonian hyoid, to discover the morphological composition of this new structure, described only in the species Chelonia mydas. Thus, the morphology of the structures and its confirmation as a real bone, with characteristic of spongy bone, described as certobranchial II, was confirmed, thus helping researchers to seek other ways to understand the feeding processes of these animals that are going through a series of serious environmental problems and therefore perhaps having to change their eating habits to overcome the high level of pollution that we are finding in the oceans. RESUMO A tartaruga verde (Chelonia mydas) presente nos mares tropicais, utiliza como zona de alimentação a região costeira do Peruíbe, tem o crânio como uma estrutura relativamente grande e sólida, e uma mandíbula forte formada pela junção de pequenos ossos, uma vez que tem uma alimentação muito abrasiva. Aplicando técnicas de microscopia electrónica de varrimento, foi possível identificar a presença de uma estrutura óssea localizada no hióide na região ventral do crânio, juntamente com a mandíbula de indivíduos juvenis de tartarugas verdes, e como não existe investigação relacionada, foi necessário realizar um TAC, descalcificação e histologia do hióide quelónico, para descobrir a composição morfológica desta nova estrutura, descrita apenas na espécie Chelonia mydas. Assim, a morfologia das estruturas e a sua confirmação como um osso real, com característica de osso esponjoso, descrito como certobranchial II, foi confirmada, ajudando assim os investigadores a procurar outras formas de compreender os processos de alimentação destes animais que estão a passar por uma série de graves problemas ambientais e, por conseguinte, talvez tenham de alterar os seus hábitos alimentares para superar o elevado nível de poluição que estamos a encontrar nos oceanos.

Los embriones de tortugas marinas son susceptibles a perturbaciones ambientales, las cuales se traducen en anomalías en su fenotipo. Uno de los principales parámetros para la medición de estas perturbaciones es la asimetría en los carapachos (caparazones), que puede ser obtenida a través del Índice de Inestabilidad de Desarrollo (DIx, por sus siglas en inglés). La temperatura juega un rol importante, influenciando no solo en la futura supervivencia de los neonatos en temperaturas extremas (altas o bajas), sino en atributos específicos, tales como la determinación del sexo, o incluso puede ser la causa del incremento en el número de apéndices dérmicos (escudos) sobre la parte dorsal del carapacho. Este estudio evaluó la relación entre la temperatura de incubación conla asimetría de carapacho de los neonatos de Lepidochelys olivacea incubados en condiciones relocalizadas en el tortugario del Área de Usos Múltiples Hawaii en Santa Rosa, Guatemala. Para lograrlo, se replicaron las condiciones de anidación seleccionadas por las hembras anidadoras en nidos relocalizados, utilizando dispositivos termosensitivos para la toma de la temperatura durante el período de incubación. A través del cálculo del DIx en 210 neonatos, y mediante las pruebas de correlación de Kendall y Jockheere-Terpstra, se obtuvo que en valores de temperaturas extremas tanto altas (Kendall tau = .23, p < .001; JT = 2,891, p < .001), como bajas (Kendall tau u= -.21, p < .001; JT = 5,005, p < .001), así como en altos rangos de variación durante el período de desarrollo embrionario (Kendall tau = .23, p < .001; JT = 6,619, p < .05), los neonatos presentan una mayor asimetría.

We herein describe Paraharmotrema karinganiense n. gen., n. sp. Dutton & Bullard (Liolopidae Dollfus, 1934) from specimens infecting the intestine of the serrated hinged terrapin (Pelusios sinuatus), east African black mud turtle (Pelusios subniger) (both Nwanedzi River, Mozambique), and South African helmeted terrapin (Pelomedusa galeata) (North-western Zululand, KwaZulu-Natal Province, South Africa). The new genus can be easily differentiated from the other accepted liolopid genera (Liolope Cohn, 1902; Helicotrema Odhner, 1912; Harmotrema Nicoll, 1914; Dracovermis Brooks & Overstreet, 1978) by the combination of having a linguliform body approximately 6–9 × longer than wide, tegumental spines/scales, a minute ventral sucker located in the anterior 1/7−1/8 of the body, deeply lobed testes that are transverse and abut the caeca (spanning the intercaecal space), a uterus that is lateral to the anterior testis (not ventral to the anterior testis), a lobed ovary that is dextral and nearest the posterior testis, and a vitellarium that does not extend anteriad to the level of the ventral sucker and that does not fill the intercaecal space. Nucleotide sequences of large subunit ribosomal DNA (28S) and internal transcribed space region (ITS2) from all analyzed specimens of the new species were identical, respectively; the 28S sequences differed from that of Liolope copulans Cohn, 1902 and from that of Harmotrema laticaudae Yamaguti, 1933 by 103 (8%) and 105 (8%) nucleotides, respectively. The 28S phylogenetic analysis recovered the new genus sister to a clade comprising L. copulans and H. laticaudae. A key to liolopid genera is provided herein. The present study comprises the first nucleotide-based phylogenetic placement of Harmotrema and first record of a liolopid from South Africa or Mozambique. It is the first proposal of a new liolopid genus in 43 yrs, and it documents a second liolopid genus from P. subniger while tripling the number of liolopid turtle hosts reported from the continent of Africa., Graphical abstract Image 1, Highlights • Description of Paraharmotrema karinganiense n. gen., n. sp. Dutton & Bullard (Liolopidae Dollfus, 1934) from African turtles. • First nucleotide-based phylogenetic placement of Harmotrema and first record of a liolopid from South Africa or Mozambique. • Documents a second liolopid genus from P. subniger. • Triples the number of liolopid turtle hosts reported from the continent of Africa. • Comments on Brooks and Overstreet 1978.

The Eastern Pacific green turtle (EPGT) has been differentiating from the green turtle and is currently being considered a separate population. In this study, values for hematology and biochemistry of 85 green turtles captured from 2010 to 2014 were analyzed. These animals correspond to a resident foraging population at Golfo Dulce, Costa Rica. All of them underwent a physical examination and were deemed apparently healthy. Hematological analyses were done using manual methods, while seric metabolites were determined with automated and semi-automated spectrophotometry. According to the curved carapace length (CCL) measurements, the subadult individuals were the most frequent (n = 74). The predominant cells were heterophils, followed by lymphocytes. Basophils were observed in 75% of the slides and in a proportion as high as 43%. Up to 80 samples were analyzed for the following seric parameters: total protein (2.6–6.2 g/dL), albumin (1–2.7 g/dL), globulin (1.2–4.1 g/dL), glucose (54–145 mg/dL), cholesterol (58–336 mg/dL), ureic nitrogen (0–12 g/dL), creatinine (0.1–0.7 mg/dl), phosphorus (3.4–17 mg/dl), calcium (4.9–10.2 mg/dl), magnesium (5.2–12.8 mg/dl), uric acid (0.1–2.8 mg/dL), triglycerides (14–743 mg/dl), alanine aminotransferase (0–13 U/L), aspartate aminotransferase (82–289 U/L) and seric alkaline phosphatase (41–1263 U/L). These data can aid health status and conservation programs for the EPGT in this and nearby, similar locations.

The aim of this study was to assess whether possible metal contamination in the sediment of the nests of giant Amazon river turtle, Podocnemis expansa, could contaminate eggs and hatchlings, triggering genotoxic damage. Therefore, sediments of P. expansa nests from two sites in the Brazilian Amazon were evaluated, with the first being collected at Araguaia River and the second at Crixás-Açu River. Newly hatched offspring, eggs, and sediments were collected from the beaches of these two rivers and the quantification of metals (Al, Cd, Co, Cr, Cu, Fe, Mn, Pb, and Zn) was carried out by atomic absorption spectroscopy. All targeted metals were found in both sediment and P. expansa biological samples collected on the beaches presenting higher concentrations in the sediment of Crixás-Açu River. Metals found in the eggshells before nesting and in the egg contents were maternally transferred. Moreover, augmented concentration of metals led by metal transfer from the nests sediments were detected in the eggshells after nesting (ENH) and in the newly hatched offspring (H). Probably this metal relocation to the newly hatchlings augmented the frequency of micronuclei in their blood, presenting 15.25‰ in hatchling found in Crixás-Açu River beaches and 10‰ in newly hatched animals from Araguaia River beaches. These results indicate the occurrence of maternal transfer of metals (essential or not) to the eggs in testudines as well as a transference from the sediments to the nesting eggs, triggering genotoxic effects on the hatchlings.

A new glossiphoniid leech species, Placobdelloides tridens sp. n., is discovered on the Malayan Giant Turtle (Orlitia borneensis) at the Nakhon Ratchasima Zoo in Thailand. The morphological study of this new species revealed that it is distinguished from P. siamensis, a turtle leech species that can be found commonly in Thailand. Placobdelloides tridens presented the following diagnostic morphological characteristics: a pear-shaped and triannulate body, well-developed rod-like papillae on the dorsal surface, smooth posterior and anterior suckers with nominal pits inside, a single pair of dark contiguous eyes, light yellow-brown to greenish dorsal color, absence of median line, male and female gonopore separated by a single annulus and a unique trident shape at the tip of the crop ceca. The phylogenetic relationships of P. tridens sp. n., was clarified, and shown to be a sister clade to the P. siamensis and P. sirikanchanae clade. Furthermore, this is a new host record for P. siamensis, which was found on O. borneensis, Batagur affinis and B. borneoensis in the Khao Kheow Open Zoo, Chonburi, Thailand.

SOX9 (SRY-related HMG box gene 9) is an essential regulator of male sex determination and testis differentiation in many vertebrate species. However, the functional role of Sox9 in testis differentiation has not yet been identified in any reptilian species. Herein, Sox9 knockdown in the red-eared slider turtle (Trachemys scripta) embryos at a male-producing temperature led to complete male-to-female sex reversal, characterized by the formation of an ovary-like structure, disappearance of male marker AMH, and ectopic expression of ovarian regulator FOXL2, as well as a female distribution pattern of germ cells. Conversely, in-ovo overexpression of Sox9 at a female-producing temperature resulted in partial masculinization of putative female embryos, with the co-existence of AMH and FOXL2. Our study provides the first direct evidence that Sox9 is indispensable for testicular differentiation in a reptilian species, further confirming the conserved role of Sox9 in vertebrate sexual development.

Many egg-laying reptiles possess temperature-dependent sex determination (TSD) in which outcome of gonadogenesis is determined by incubation temperature during a temperature-sensitive period of development. Prior studies on Malayemys macrocephala showed that incubation temperatures influence gonadal development and suggested that M. macrocephala exhibits TSD. However, information on the temperature-sensitivity period in this species was unknown until the current study. Turtle eggs were collected from rice fields in central Thailand from December 2016 to February 2017. In the laboratory, eggs were incubated at male-biased temperature (26 °C) and shifted to female-biased temperature (32 °C), or vice versa. Single shift experiments were performed systematically during embryonic stages 13-21. After hatching, sex of individual turtles was determined by histological analysis. We found that the sex determination of M. macrocephala is affected by temperature up to stage 16 of embryonic development.

The black marsh turtle (Geoemydidae: Siebenrockiella crassicollis) is a freshwater turtle that occurs in equatorial tropical climates in South East Asia. The semen of S. crassicollis was investigated by electroejaculation. The spermatozoa of S. crassicollis are filiform in shape with curved heads. The entire length, midpiece to tail length, tail width and tail length of the spermatozoa were 71.33 ± 1.55 μm, 49.92 ± 1.13 μm, 0.43 ± 0.02 μm and 48.53 ± 0.25 μm, respectively. The head length, head width across the middle and head width across the base were 14.00 ± 0.38 μm, 0.79 ± 0.03 μm and 0.91 ±0.0.03 μm, respectively. The acrosomal region of the S. crassicollis spermatozoa was narrower than the head, with an acrosomal length and width at the annulus of 2.90 ± 0.13 μm and 0.43 ± 0.01 μm, respectively. The midpiece of the S. crassicollis spermatozoa was narrower than the head and contained 30-40 mitochondrial balls, each with a ball diameter of 0.16 ± 0.002 μm. The midpiece length, midpiece width and tail length were 4.92 ± 0.16, 0.78 ± 0.03 and 48.53 ± 0.25 μm, respectively. This study presents the characteristic appearance of a freshwater turtle spermatozoa in Southeast Asia, as observed under an electron microscope. The spermatozoa of Siebenrockiella crassicollis are morphologically different from those of other freshwater turtles from other regions described in previous studies.

Kinosternon scorpioides, popularly known as scorpion mud turtle (jurará in Brazil), is a fresh water species. There is little information about its reproduction and the present study aims to morphologically characterize the reproductive organs of male K.scorpioides bred in captivity in two seasons of the year. The reproductive tracts of adult animals under went macroscopic ultrastructural analysis of the lumen, as well as scanning electron and transmission microscopy. Macroscopically, the male genital organs consist of a pair of testicles, epididymis, the vas deferens and a penis. Testicles, epididymis and deferents ducts were characterized by reproductive activity during the rainy season and reproductive inactivity in the dry period. The morphometry regarding the tubular and luminal diameter and epithelial height of the testicles, epididymis and deferents ducts showed changes along the studied periods. The rainy season presented higher averages than the dry period. The penis did not show any changes during both periods. It was concluded that K. scorpioides exhibits reproductive seasonality., The male genital organs consist of a pair of testicles, epididymis, vas deferens and a penis. The seminiferous tubules had irregular arrangements in the interstitium. Reproductive seasonality in the K. scorpioides in captivity

RESUMEN Kinosternon chimalhuaca (Casquito de Jalisco) es una tortuga kinosternida endémica de México. La talla máxima de longitud de caparazón (CL) es de 160 mm en machos y de 130 mm en hembras. Tiene un pequeño rango de distribución en la costa del Pacífico de Jalisco y Colima (México), desde el río Tuito en Jalisco hasta el río Cihuatlán en el noroeste de Colima. Este nuevo registro en Puerto Vallarta, Jalisco, extiende su distribución al menos 32 km al norte de la localidad más septentrional conocida. Se registra un nuevo tamaño máximo (CL) para la especie de 172,00 mm (un macho). ABSTRACT Kinosternon chimalhuaca (Jalisco Mud Turtle) is a kinosternid turtle endemic to Mexico. The previously recorded maximun carapace length (CL) is 160 mm in males and 130 mm in females. It has a small distribution range along the Pacific coast of Jalisco and Colima, Mexico, from the rio Tuito in Jalisco to the rio Cihuatlán in northwestern Colima. This new record in Puerto Vallarta, Jalisco, extends its distribution at least 32 km north from the northernmost known locality. A new maximun size (CL) for the species of 172.00 mm (one male) is recorded.

Neusticemys neuquina(Fernández and de la Fuente, 1988) is a turtle from the Upper Jurassic of the Neuquén Basin, Patagonia, Argentina. Here we describe in detail a new skull, lower jaw, and a vertebra, utilizing both traditional anatomical description and computed tomography (CT). New diagnostic cranial characters ofNeusticemys neuquinaare: a round depression on the ventral surface of the basisphenoid, a relatively larger oval foramen nervi trigemini, and reduced and steepened triturating surfaces on both the maxilla and dentary. The new morphological information presented in this study was included in a phylogenetic analysis, the primary result of which was recovery ofNeusticemys neuquinawithin Thalassochelydia. Characters recognized as synapomorphies of this clade include: (1) anterolateral recess of the anterior surface of the quadrate positioned lateral to the processus trochlearis oticum, (2) presence of a fossa on the supraoccipital-opisthotic-exoccipital contact area, (3) foramina anterius caroticus cerebralis located close together but independently perforating the basisphenoid, and (4) the presence of the splenial in the mandible. Two contrasting dispersal scenarios could explain how this species of Thalassochelydia can be found outside of Europe. The presence ofNeusticemys neuquinain the Neuquén Basin could be the consequence of an early dispersion event, for which we lack intermediate forms, or it could be the result of a later event once the clade was already established in Europe.

First described from the Mascarenes (Rodrigues Island, Indian Ocean), Olifantiella gorandiana Riaux-Gobin was later observed from several other oceanic basins. We here examine the morphology of this taxon from a scraping of a juvenile green turtle Chelonia mydas Linnaeus, named ‘G5-Manahere’, from Moorea Island (Society Archipelago, South Pacific). In this sample, a great range of morphologies, not previously described for the species, were encountered, demonstrating a high degree of polymorphism with regard to valve shape and also stria pattern. These morphological differences appear to be associated with size, as smaller cells become more rounded. The fine structure (e.g., stria density and pattern) may also be associated with changes in shape. These specimens may all belong to the same and unique taxon: Olifantiella gorandiana. This high degree of polymorphism is described, and put into the context of the constraining epizoic conditions. This study permits furthers the description of the fine structure of O. gorandiana, using focused ion beam (FIB) and scanning electron microscopy (SEM) techniques.

Caracterización haplotípica de la tortuga golfina (Lepidochelys olivacea) en el noroeste de México: el límite septentrional de su distribución La tortuga golfina (L. olivacea) tiene una distribución pantropical. En el Pacífico oriental, los límites oficiales de su zona de reproducción son la península de Baja California y el sur de Sinaloa, en México. La playa de Ceuta en Elota, Sinaloa, México ha servido de sitio de protección para L. olivacea durante más de tres decenios. En este estudio, se caracterizó genéticamente la población de L. olivacea de la playa de Ceuta. Concretamente, se amplificó un fragmento de 712 pb de la región de control del ADNmt de 32 tortugas golfinas. Se identificaron ocho haplotipos (siete tras reducir a ~468 pb) y se incluyeron dos haplotipos nuevos (Lo–T7 y Lo–T8) y cinco haplotipos que se habían identificado anteriormente en otras playas de anidación. El haplotipo Lo–T2 era dominante (~60 %) en las muestras: h = 0,6048 (± 0,0974) y π = 0,002212 (± 0,001504). Si bien este estudio se realizó en el límite septentrional de la zona de anidación de la tortuga golfina en el Pacífico oriental, el grupo estudiado presenta una diversidad genética moderada y pertenece a una población que, en la escala evolutiva, ha pasado recientemente por una expansión demográfica. Debido a que la tortuga golfina del Pacífico oriental se considera resiliente a la variación ambiental, es necesario estudiar las zonas de anidación en el noroeste de México., The olive ridley sea turtle (L. olivacea) has a pantropical distribution. In the Eastern Pacific, the official limits of its reproduction area are south of the Baja California peninsula and south of Sinaloa, Mexico. Ceuta beach in Elota, Sinaloa, Mexico, has served as a protection site for L. olivacea for over three decades. In this study, the L. olivacea population from Ceuta beach was genetically characterized. Specifically, a 712–bp fragment from the control region of mtDNA was amplified from 32 olive ridley turtles. Eight haplotypes (seven after cutting to ~468 bp) were identified, and these included two novel haplotypes (Lo–T7 and Lo–T8) and five haplotypes that were previously identified in other nesting beaches. The Lo–T2 haplotype was dominant (~60 %) in the samples: h = 0.6048 (± 0.0974) and π = 0.002212 (± 0.001504). Although this study was conducted in the northernmost limit of the olive ridley turtle nesting distribution in the eastern Pacific, the sampled group presents moderate genetic diversity and belongs to a population that, on an evolutionary scale, only recently underwent demographic expansion. Because the olive ridley turtle in the eastern Pacific is considered resilient to environmental variation, nesting area studies in northwest Mexico are necessary.

The western painted turtle (Chrysemys picta bellii) can survive extended periods of anoxia via a series of mechanisms that serve to reduce its energetic needs. Central to these mechanisms is the response of mitochondria, which depolarize in response to anoxia in turtle pyramidal neurons due to an influx of K+. It is currently unknown how mitochondrial matrix pH is affected by this response and we hypothesized that matrix pH acidifies during anoxia due to increased K+/H+ exchanger activity. Inhibition of K+/H+ exchange via quinine led to a collapse of mitochondrial membrane potential (Ψm) during oxygenated conditions in turtle cortical neurons, as indicated by rhodamine-123 fluorescence, and this occurred twice as quickly during anoxia which indicates an elevation in K+ conductance. Mitochondrial matrix pH acidified during anoxia, as indicated by SNARF-1 fluorescence imaged via confocal microscopy, and further acidification occurred during anoxia when the F1Fo-ATPase was inhibited with oligomycin-A, indicating that ΔpH collapse is prevented during anoxic conditions. Collectively, these results indicate that the mitochondrial proton electrochemical gradient is actively preserved during anoxia to prevent a collapse of Ψm and ΔpH.

From 2000 to 2002, the reproductive ecology of the endangered Hainan four eye-spotted turtle (Sacalia insulensis) was studied on Hainan Island, China. A total of 147 adult females were captured, and their reproductive status was evaluated by palpation, X-ray imaging, and ultrasound. Twenty-two gravid females were under observation when they laid their eggs. We observed gravid females and nesting behavior along stream banks between December and April. Nest dimensions ranged from 5 to 6 cm in diameter and from 2 to 3 cm in depth. No females in our study produced more than 1 clutch per season. Clutch size average was 1.9 eggs (range = 1–3 eggs, SD = 0.56, n = 22) with an average egg mass of 16.05 g (range = 11.8–21.0 g, SD = 2.417, n = 21), an average egg width of 2.36 cm (range = 2.1–2.7 cm, SD = 0.135, n = 21), and an average length of 4.65 cm (range = 3.9–5.3 cm, SD = 0.400, n = 21). Hatching success rate was 53%, and hatchlings emerged after an average of 120.9 d (range = 98–148 d, SD = 14.94, n = 21) in late June to early July. The mean nesting site ground temperature was 24.43°C (range = 15.0°C–30.2°C, SD = 2.712, n = 7).

At the time of hatchling emergence from a nest laid on Juno Beach, Florida, USA, by a normally pigmented green turtle (Chelonia mydas), 23 albino hatchlings and 75 normally pigmented hatchlings were observed. This condition is rarely seen in sea turtles, and little is known about blood analytes and genetics of albino wildlife to date. Therefore, the objective of our study was to assess and compare morphometric measurements (mass, minimum straight carapace length, body condition index), carapacial scute anomalies, a suite of hematologic and plasma biochemical analytes, and two glucose analysis methodologies (glucometer and dry chemistry analysis) in albino (n=20) versus normally pigmented (n=24) hatchlings from this nest. Genetic analyses were completed to identify paternal contributions of hatchlings and to test Mendelian inheritance assumptions. Although morphometric measurements, scute anomalies, and leukocyte morphology were similar between albino and normally pigmented hatchlings, several differences were observed in blood analyte data: immature erythrocytes, packed cell volume, heterophil:lymphocyte ratio, and glucose concentrations (by both methodologies) were significantly higher, whereas absolute immature heterophils, absolute lymphocytes, number of erythrocyte micronuclei, sodium, and chloride were significantly lower in albino hatchlings compared with normally pigmented hatchlings. Considerations for these differences include a stress response from sampling (e.g., timing of procedures or possibly from photosensitivity or reduced visual acuity in albinos) and different osmoregulation, which may reflect physiologic variations or stress. There was a small positive bias (0.10 mmol/L) with glucose by glucometer, similar to reports in other sea turtle species and confirming its suitability for use in hatchlings. All albino hatchlings analyzed (n=10) were from the same father, but the normally pigmented hatchlings (n=24) were from two other fathers. These findings provide insight into the physiology and genetics of albinism in sea turtles.

A recent study by Norell et al. (2020) described new egg specimens for two dinosaur species, identified as the first soft-shelled dinosaur eggs. The authors used phylogenetic comparative methods to reconstruct eggshell type in a sample of reptiles, and identified the eggs of dinosaurs and archosaurs as ancestrally soft-shelled, with three independent acquisitions of a hard eggshell among dinosaurs. This result contradicts previous hypotheses of hard-shelled eggs as ancestral to archosaurs and dinosaurs. Here we estimate the ancestral condition for dinosaur and archosaur eggs by reanalyzing the original data from Norell et al. and that from a recent study on reptile eggshells (Legendre et al., 2020) with the addition of these new dinosaur specimens. We show that the recovery of dinosaur eggs as ancestrally soft-shelled is conditioned by the discretization of a continuous character (eggshell thickness), the exclusion of turtle outgroups from the original sample, and a lack of branch length information. When using a larger sample, calibrated trees, and a definition of hard-shelled eggs referencing their unique prismatic structure, we recover dinosaur and archosaur eggs as either hard-shelled or uncertain (i.e. equal probability for hard- and soft-shelled). This remaining ambiguity is due to uncertainty in the assessment of eggshell type in two dinosaur species, i.e. ∼1% of the total sample. We conclude that more reptile egg specimens and a strict comparative framework are necessary to decipher the evolution of dinosaur eggs in a phylogenetic context.

Placobdelloides siamensis is a glossiphoniid leech with a short life cycle. In a laboratory setting, ten mature P. siamensis left their host (a turtle) after feeding for approximately three days and initiated copulation. The adults spent 3–4 days gestating before depositing eggs (272.8±62.9 eggs/clutch; range: 186–359 eggs/clutch). The eggs then changed from a creamy white to a creamy brownish color before hatching. Hatching occurred after incubation on the parent's ventral surface for 5–7 days. The transparent brood, with a single pair of red eyes, spent a couple of weeks under the venters of their parents. After this period, they left their parents and grew to maturity in 10–15 days; leeches were considered mature when their color was similar to that of their parents and they performed their first copulation. In addition, the mature leeches survived for 163 days on one feeding.

This study investigated 12 trace elements in paired green turtle (Chelonia mydas) eggshell composites and coral sand samples to examine within-habitat heavy metal pollution from 40 nesting sites in the Xisha Islands. The concentrations of the elements (μg·g−1) found in the eggshells ranged as follows: Sr (41.3) > Zn (20.3) > Cu (12.8) > Fe (4.92) > Al (4.37) > Se (2.44) > Mn (0.91) > Cr (0.81) > Ba (0.44) > Pb (0.14) > As (0.08) > Cd (0.02). Significant correlations were observed between the levels of Cd and Se and the levels of Zn, Cu, and Pb in eggshells. The concentrations of Mn, Zn, Se, As, Cd, and Pb in C. mydas eggshells were significantly correlated with those in coral sand sediments. Cu concentrations in the eggshells exceeded the toxic reference value for bird eggs and Se concentrations were between the worst- and best-case scenario hazard quotients.

The objective of this study was to verify the accumulation of trace metals in eggs and hatchlings of Chelonia mydas, evaluating if metal accumulation is originated from maternal transfer and/or from the incubation environment. Other assessments were also performed, as metal distribution in different tissues (blood, kidney, liver, muscle, and turtle shells) of newly hatched turtles, and genotoxic analysis, to verify possible damages caused by the presence of metals.The assessments were carried out by quantifying Cd, Ni, Pb, Mn and Fe in egg sample collected during laying time (eggshells (ELT) and egg content (EC)), eggshells from newly hatched turtles (ENH), hatchlings tissues (H - blood, kidney, liver, muscle, and shell)) (n = 18 for each biological sample - 3 of each nest) and nest sediments (n = 6, one of each nest). Comparative analysis were made between ELT and ENH, as well as between egg content (EC) and the sum of tissue samples from hatchlings, using Mann-Whitney hypothesis test (p0,05). The amount of metals in different hatchling was quantified and followed by the Dunn post-test. A principal component analysis (PCA) was also employed.Metals studied were found in all investigated samples. The concentration of a great amount of investigated metals was significantly higher (P=0.001) in eggshells from ENH than in ELT. An increase in Cd (2.16-fold), Pb (3.47-fold), Fe (6.83-fold) and Mn (195.57-fold) concentration was noticed in ENH. We also observed an increase in Fe (1.59-fold), Mn (1.74-fold) and Ni (1.59-fold) concentration in hatchling, when compared with EC, due to transfer from nest sediments. In relation to the hatchling's tissues, blood was shown to accumulate higher concentrations of Ni and Pb, while shells accumulated more Cd and Fe, and Mn is more associated with liver and kidney. Fe was the highest accumulated metal in both tissues, and muscles presented discrete concentrations of Ni, Mn, and Pb. A mean concentration of 1.25‰ MN was obtained in C. mydas hatchlings, indicating that the accumulation of metals in hatchlings didn't cause toxicology effects.Hatchlings accumulate metals through the maternal and sediment transfer, although the levels of metal accumulation were not enough to cause genotoxic damage.

Low pH-shifting was firstly applied in the black turtle bean (Phaseolus vulgaris L.) protein isolate treatment by acidic (pH 1.0–3.0) buffer incubation for 8 h, then was adjusted to pH 7.2 and kept 3 h for protein stabilizing. Mild loss of secondary structure was confirmed in the protein isolate after low pH-shifting treatment by CD and FT-IR analyses. Intrinsic fluorescence, UV spectra, surface hydrophobicity, SH content and SDS-PAGE analyses indicated the protein conformation was unfolded with the exposure of much more buried hydrophobic residues, which would result in the enhancement of emulsifying properties, foaming properties and fat holding capacity, and lead to the reduction of solubility and water holding capacity. Furthermore, lower immunoreactivity was observed by the ELISA, and improved digestibility was found in in vitro digestion assay. Our results suggested the low pH-shifting treatments would broaden the application of bean protein isolate with better hydrophobic processing functions and safety.

Chelonians (Turtle and Tortoise) are kept in captivity as pet animals. Testudo. graeca called Spur thighed tortoise for its large conical tubercle present on each Thigh.T. graeca considered endangered across its entire range. They were illegally imported from Libya to Egypt. The aim of this work is to determine normal Hematological parameters and gastrointestinal parasite in Spur-thighed tortoise. Blood samples were taken from 37 tortoises, 13 female and 24 Male. Fecal samples were collected from 33 tortoises. Feces were examined by qualitative and quantitative methods. RBCs Count, WBCs Count, PCV, HB, MCV, MCH, MCHC value and DLC% were measured. Strongylid and Oxyurid eggs founded. Mean RBCs count in tortoise was 0.68 106 /µL, Hemoglobin (Hb) concentration was 8.26 g/dl, Packed cell Volume (PCV) was 25.12%. Lymphocytes % were 30.3 %. Monocytes (%) were 25.03. Heterophils (%) were 40.44%. Eosinophils (%) were 5.87%. Basophils (%) were 0.05%.We report hematological parameters in normal and parasitic infection. In addition, we investigate parasites inSpur-thighed tortoise.

Based on current knowledge of the ecology and distribution of Snapping Turtle (Chelydra serpentina), both in eastern Canada and elsewhere, we conclude this species is native to Cape Breton Island. Seventy-two reports of Snapping Turtle from Cape Breton (1999–2017) indicate a range centred in the area south of Bras d’Or Lake. Date of oviposition ranged from 19 June to 10 July (median = 26 June) among 26 nests observed during 2012–2014. Clutch size for these nests was 23–65 eggs (mean = 46) and among 25 protected nests average rate of hatchling emergence was 21.5%. Time from oviposition to emergence of hatchlings (n = 256) was 75–120 days (mean = 87.2; SD = 9.0) among 20 nests. First emergence ranged from 9 September to 20 October (75–114 nest days; mean = 90) and last emergence ranged from 13 September to 28 October (86–120 nest days; mean = 100). Duration of emergence ranged from one day (i.e., synchronous emergence; five nests) to 37 days (mean = 11 days). The number of days on which hatchlings emerged at a nest ranged from one to nine days (mean = 4 days). Maximum carapace length was 25.0–31.8 mm (mean = 29.0 mm) and maximum carapace width was 23.5–30.0 mm (mean = 27.0 mm) for 256 hatchlings that emerged from 20 protected nests. Mass of hatchlings was 4.9–9.9 g (mean = 7.8 g).

New shell material of a trionychid turtle from the Upper Cretaceous (upper Campanian) Fossil Forest Member of the Fruitland Formation of northwestern New Mexico represents a new species,Gilmoremys gettyspherensis. The material consists of right costals I–III, V, VI, and VIII, left costals V, VII, and VIII, the left half of the entoplastron, the right hypo- and xiphiplastron, and the left hyo-, hypo-, and xiphiplastron. The specimen shows great similarities to the Late Cretaceous (Maastrichtian) trionychidGilmoremys lancensis(Gilmore, 1916) by having a relatively thin shell, carapacial sculpturing consisting of fine pits combined with extended sinusoidal ridges or grooves, free costal rib ends, presence of a preneural, a distally constricted costal I and distally expanded costal II, two lateral hyoplastral processes, low hyoplastral shoulders, and full midline contact of the elongate xiphiplastra, but differs by being smaller, having raised sinusoidal ridges on the carapace instead of grooves, less distally expanded costals II, and less elongate xiphiplastra. Phylogenetic analysis placesGilmoremys gettyspherensisn. sp. as sister toGilmoremys lancensisnear the base of the clade Plastomenidae. Like the majority of previously described plastomenid materials, the type specimen ofGilmoremys gettyspherensisn. sp. was collected from a mudstone horizon, suggesting a preference for ponded environments.UUID:http://zoobank.org/e7165061-d86b-46b7-a1f8-c31f5a8ed628

The incidence of multiple paternities in two populations of Podocnemis expansa of Brazilian Amazon both rose in captivity and in natural habitat, was verified by using five microsatellite loci. Three hundred twenty-seven offspring from two different sampling sites were genotyped. The analysis revealed a 100% rate of multiple paternities in both populations. The Mendelian distribution of the alleles found in each nest was used to determine the number of contributing males. It was estimated that at least ten males contributed to each brood in captivity and nine contributed to each brood in the wild. These values are the highest ever recorded for the number of contributing P. expansa males. The findings have considerable implications regarding the conservation of this species, given that multiple paternities is important to the maintenance of genetic variability and has important consequences in increasing the effective size of a population in comparison to single paternities. Normal 0 21 false false false PT-BR X-NONE X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Tabela normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin-top:0cm; mso-para-margin-right:0cm; mso-para-margin-bottom:10.0pt; mso-para-margin-left:0cm; line-height:115%; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:"Times New Roman"; mso-bidi-theme-font:minor-bidi;}

Made available in DSpace on 2018-12-11T17:18:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-05-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) Based on morphological, morphometric, and molecular data, we describe a new hemoparasite of the genus Haemogregarina Danilewsky 1885, isolated from the Brazilian aquatic turtle Podocnemis unifilis (Testudines: Podocnemididae). The new species, Haemogregarina podocnemis sp. nov. (Apicomplexa: Haemogregarinidae), is characterized by small trophozoites with a single cytoplasmic vacuole on one side; pre-meronts with nuclear chromatin dispersed in the cytoplasm, with or without cytoplasmic vacuoles; meronts that are usually broad and slightly curved (kidney-shaped), with an average of eight small rectangular nuclei; immature gamonts (bean-shaped) with two morphological types: one with nuclear chromatin dispersed in the cytoplasm and the other with nuclei in the middle of the cell; mature gamonts of two morphological types: one with a length equal to or greater than that of the erythrocyte and the width of the nuclei similar to that of the hemoparasite and the other smaller than the erythrocyte with the width of the nuclei less than that of the hemoparasite. This is the first hemogregarine species described that infects the Brazilian turtle Po. unifilis. These findings highlight the need for further studies of Haemogregarina spp. to better determine the biodiversity of this understudied parasite group. Laboratório de Parasitologia Instituto de Ciências Biomédicas Universidade Federal de Uberlândia, Campus Umuarama Instituto de Biociências Campus de Botucatu Departamento de Parasitologia UNESP-Univ Estadual Paulista, Distrito de Rubião Junior Laboratório de Ensino e Pesquisa em Animais Silvestres Faculdade de Medicina Veterinária Universidade Federal de Uberlândia Faculdade de Matemática-FAMAT Centro de Ciências e Tecnologia Universidade Federal de Uberlândia—UFU Instituto de Biociências Campus de Botucatu Departamento de Parasitologia UNESP-Univ Estadual Paulista, Distrito de Rubião Junior

The effects of dopamine D2-class receptor blockade by sulpiride on the electroretinographic (ERG) b-wave (ON response) and d-wave (OFF response) were investigated in light-adapted turtle (Trachemys scripta elegans) and frog (Rana ridibunda) eyecups. For turtle ERG, sulpiride (240 μM) produced an amplitude increase of the b- and d-waves, while the 40 μM and 120 μM of sulpiride were ineffective. Alternatively, for frog ERG, a well-developed and dose-dependent b- and d-wave amplitude decrease was obtained with 40 μM and 240 μM sulpiride. In both species, 240 μM sulpiride significantly affected the maximal voltage range of the ERG responses without altering their relative sensitivity (determined by the semi-saturation point). The absolute sensitivity of the ON and OFF responses (evaluated by threshold estimation) was not significantly altered for turtle ERG, but it was decreased for frog ERG. The time characteristics of the ERG responses were unchanged in both species. Our results show important differences between dopamine D2-class receptor-mediated pathways in turtle and frog retina (revealed by ERG).

The present study investigated the population of the xanthid brachyuran crab, Macromedaeus distinguendus (De Haan, 1835), at a toxic hydrogen sulfide, shallow hydrothermal vent area at Turtle Island, off northeastern Taiwan. This is the first record of this species from this extreme environment, and M. distinguendus is only the second brachyuran discovered in this area. The wet weight of male and female M. distinguendus varied from 0.11 to 1.22 g (mean ± standard deviation: 0.56 ± 0.39 g) and 0.06 to 0.62 g (0.37 ± 0.15 g), respectively. The wet weight: carapace width ratio was significantly correlated in both sexes. The local distribution patterns of M. distinguendus were different from those of the obligate hydrothermal crab Xenograpsus testudinatus N. K. Ng, J.-F. Huang & Ho, 2000 (Xenograpsidae) in this area. An analysis of resource use and habitat partitioning revealed that the two crab species were able to co-exist within the study area.

We studied the sexual dimorphism in body size and shape of Beal's eyed turtle, Sacalia bealei, by measuring 15 morphological characteristics of 68 adult individuals (40 females, 28 males) collected from Fujian Province, China. Females were slightly larger than males in carapace length with a sexual dimorphism index of 0.09. This translated into greater absolute values of females for all the traits we measured except for tail length, preanal tail length, and analia to supracaudal junction length. Comparisons between the sexes of size-corrected morphological traits indicated that females exhibited a longer plastron and higher carapace relative to carapace length, thereby providing a larger internal volume. Females also exhibited relatively wider heads. In males, the plastron was smaller and more indented than in females and the openings between the plastron and the carapace were also more developed. Overall, females were larger in size and displayed a more voluminous shell than did males while male...

Trachemys scripta elegans, as a freshwater invasive species, can survive and lay eggs in brackish water, which may lead to the expansion of its potential invasion range due to freshwater salinization. Our previous studies have shown that high salinity leads to the accumulation of serum lipid content, which may induce endoplasmic reticulum stress (ERS) in the turtle. To better understand whether ERS is triggered by salinity, and in turn whether the turtles promote the protection mechanism, we exposed the turtles to the freshwater (CK), 5‰ salinity water (S5) and 15‰ salinity water (S15), and sampled at 6 h, 24 h and 30 d. 13 differentially expressed genes (DEGs) related to ERS pathways were found in the comparison of CK vs. S15 by transcriptomics analysis. Then, the mRNA and protein expression of ERS and its related activation pathways were further investigated. ERS marker glucose regulated protein 78 kD (GRP78) increased significantly (p 0.05) in both the transcript and protein levels after exposure to 15‰ salinity water, which clearly indicated that salinity could induce ERS in T. s. elegans. Meanwhile, the three unfolded protein response (UPR) including transducers protein kinase RNA (PKR)-like endoplasmic reticulum kinase (PERK), inositol-requiring enzyme 1α (IRE1α) and activating transcription factor-6 (ATF6) were promoted by salinity, suggesting that the turtle might promote physiological process to eliminate damaged cells and cope with unfolded proteins accumulation induced by ERS. Our results provide new insight into the mechanism of salinity adaptation in T. s. elegans and salt-tolerant biological invasion.

A new species of pelomedusoid turtle: Elkanemys pritchardi gen. et sp. nov. from Upper Cretaceous (Cenomanian) is described. The specimen MMCH-PV 73 outcrops at Lago Ezequiel Ramos Mexia, near El Chocon town, Rio Negro Province, Argentina. The holotype is only represented by a partial carapace and plastron, which has been included in a cladistic analysis, resulting Elkanemys pritchardi as a member of Cearachelyini tribe of the clade Bothremydidae. This new taxon corresponds to the first record of a Bothremydidae in a Cretaceous horizon of Neuquen Basin. The results here presented highlight the diversity of Pelomedusoides in the Cretaceous of Patagonia allowing the inclusion of this area in further paleobiogeographic history of Bothremydidae clade.

The present study evaluates the nutraceutical composition of muscle and calipash of Yangtze and Japanese strains of Chinese soft-shelled turtle, Pelodiscus sinensis. In general, both strains possessed high protein content (16.62–22.18 g 100 g−1 wet weight) and negligible fat content (0.22−0.95 g 100 g−1 wet weight). The ratios of total essential amino acids to total amino acids and to total nonessential amino acids in muscle of two strains were about 40 % and 84 %, while the ratios in calipash were about 20 % and 29 %, respectively. Both the values of amino acid scores and in vitro digestible indispensable amino acid scores in muscle of two strains were in the range of good protein. The unsaturated fatty acids/saturated fatty acids ratios and n-3/n-6 ratios of two strains were 1.04−2.08 and 1:3.78−1:1.11, respectively. We also found that the atherogenicity index, thrombogenicity index and hypocholesterolemic/hypercholesterolenic fatty acid values in muscle varied between 0.35−0.44, 0.41−0.46 and 2.16–2.80, respectively, while in calipash were 0.62−0.72, 1.02–1.23 and 1.26−1.56. Taken cumulatively, these results indicate that consumption of muscle of Yangtze and Japanese strains of P. sinensis is favorable to human health, while moderation needs to be exercised for the calipash consumption.

Hatchling turtles are rare in the fossil record. Here we report two incomplete juvenile specimens of the genus Araripemys from the Aptian (ca. 115 Ma) Crato Formation (Araripe Basin, Brazil). Although the description of this material does not completely elucidate the ontogeny of this taxon, the analysis of these specimens yield relevant information about diagnostic features of the genus, showing their presence in hatchling such as: skull with nearly oval shape in dorsal view; closely spaced orbits; cervical vertebrae with long vertebral body indicating the presence of a long neck; the extension and the angle of curvature of the axillary (obtuse angle); and unguals arrow-shaped. The small size of the specimens (40–50 mm) and their poor degree of ossification including unfused costal bones indicate that both represent hatchling individuals. The paleoenvironment of the Crato Formation was similar to mangroves, which is corroborated by the presence of juvenile turtles and fishes, anurans and insects. Araripemys barretoi was also recorded in the Romualdo Formation, which represents a lagoon. The fact that this turtle is found in these quite distinct paleoenvironments suggests that this species could be tolerant to distinct salinities levels.

Skeletal remains of a new early Miocene (Ottnangian, MN 4 mammal zone) monitor lizard, Varanus mokrensis sp. nov., are described from two karst fissures in the Mokrá-Western Quarry (1/2001 Turtle Joint; 2/2003 Reptile Joint), Czech Republic, providing the first documented example of a European varanid for which osteological data permit a well-supported assignment to the genus Varanus. The new species is morphologically similar to the Recent Indo-Asiatic varanids of the Varanus bengalensis group. It differs from all other Varanus species on the basis of a single autapomorphy and a combination of 11 characters. As a distinguishing feature of V. mokrensis, the parietal and squamosal processes of the postorbitofrontal form a narrowly acute angle. The teeth show distinct, smooth cutting edges along the mesial and distal margins of the apical portion of their crowns. This feature is not observed in most extant Asiatic Varanus species and may represent a plesiomorphic condition. The results of parsimony phylogenetic analyses, with and without character reweighting, reveal poor resolution within Varanus. A Bayesian analysis shows V. mokrensis to be closely related to extant representatives of the Indo-Asiatic Varanus clade, with close affinities to the V. bengalensis species group. The topology of the Bayesian tree supports the hypothesis that Miocene monitors from Mokrá are representatives of a lineage that is ancestral to the well-defined clade of extant African varanids, including the early Miocene V. rusingensis. In addition, our results support a Eurasian origin for the varanid clade. The extant African Varanus species probably originated in the late Oligocene. The radiation of African varanids probably occurred during the late Oligocene to early Miocene time interval. The occurrence of Varanus in the early Miocene of Mokrá-Western Quarry corresponds to the warm phase of the Miocene Climatic Optimum. Remains of a diverse aquatic and heliophobe amphibian fauna at the 2/2003 Reptile Joint site indicate more humid conditions than those at the 1/2001 Turtle Joint site.

This paper describes a new species of Camallanus found in the freshwater turtle Trachemys dorbigni. Sixty hosts collected in Southern Brazil were examined. All hosts (100%) were parasitized by a new species of Camallanus, which was described as Camallanus emydidius n. sp. The new species differs from other Camallanus species of freshwater turtles mainly because of the morphology of the right spicule, the number of male precloacal and postcloacal papillae, and the presence of “mucrons” in the female posterior extremity., Graphical abstract Image 1, Highlights • A new species of Camallanidae (Nematoda) is described from freshwater turtle. • All hosts (n = 60) were parasitized for a new species of Camallanus. • The new nematode is the first species of Camallanidae described in freshwater turtle Emydidae from South America.

Knowledge of embryonic development is important for choosing strategies for nest conservation, and reducing negative effects on eggs and embryos of turtle species subject to human and natural interference, as is the case of Podocnemis expansa in the Amazon. Nine P. expansa nests located in Balbina, Amazonia, Brazil, were used to describe embryonic development. An embryo was collected each day, totaling 240 embryos. The specimens were fixed in Bouin’s and Karnovsky’s solutions for macroscopic analysis and scanning electron microscopy, respectively. The incubation period lasted between 58 and 64 days with a mean nest incubation temperature of 30.3 °C. The main characteristics used for description and comparison of embryonic development were: development of the eyes, the mandibular process, limbs, carapace and plastron. We found that the timing of appearance and disappearance of structures in P. expansa were more advanced than in other turtle species. Furthermore, P. expansa displays some unique characteristics during its normal development, such as craniofacial morphology, frontonasal groove and carapace and plastron characteristics.

Survival of prolonged anoxia requires a balance between cellular ATP demand and anaerobic ATP supply from glycolysis, especially in critical tissues such as the brain. To add insight into the ATP demand of the brain of the anoxia-tolerant red-eared slider turtle (Trachemys scripta) during prolonged periods of anoxic submergence, we quantified and compared the number of Na+-K+-ATPase units and their molecular activity in brain tissue from turtles acclimated to either 21°C or 5°C and exposed to either normoxia or anoxia (6h 21°C; 14days at 5°C). Na+-K+-ATPase activity and density per g tissue were similar at 21°C and 5°C in normoxic turtles. Likewise, anoxia exposure at 21°C did not induce any change in Na+-K+-ATPase activity or density. In contrast, prolonged anoxia at 5°C significantly reduced Na+-K+-ATPase activity by 55%, which was largely driven by a 50% reduction of the number of Na+-K+-ATPase units without a change in the activity of existing Na+-K+-ATPase pumps or α-subunit composition. These findings are consistent with the "channel arrest" hypothesis to reduce turtle brain Na+-K+-ATPase activity during prolonged, but not short-term anoxia, a change that likely helps them overwinter under low temperature, anoxic conditions.

Five 2-week feeding trials were conducted to investigate five stimulants on P. sinensis. Two isonitrogenous and isoenergetic diets were formulated, one contained 600 g kg–1 fishmeal (FM diet) and the other contained 420 g kg–1 fishmeal and 225 g kg–1 animal protein blend (APB diet), and each tested one stimulant with four levels. The feeding stimulant candidates were betaine, 2-carboxy-ethyl dimethyl sulphonium bromide (DMPT), inosine-5′-monophosphate (IMP), taurine and squid extract. In the experiment on each stimulant, turtles (10.85 ± 0.10 g) were equally divided into FM and APB groups and fed with an equal mixture of corresponding diets containing four levels of stimulants, respectively. Each diet contained a unique rare earth oxide as inert marker. Turtles were fed twice daily (8:00 and 17:00), and faeces were collected. Preference for each diet was estimated based on the relative concentration of each marker in the faeces. In the FM group, only IMP showed the enhanced attraction. But in APB groups, all the stimulants, except IMP, showed higher preference than basal diets for at least one inclusion level, and the optimum level was 10 g kg–1 for betaine, 0.1 g kg–1 for DMPT, 0.1 g kg–1 for IMP, 5 g kg–1 for taurine and 10 g kg–1 for squid extract, and the squid extract had the strongest stimulating effect among the stimulants.

A new species of Eimeria Schneider, 1875 (Apicomplexa: Eimeriidae) collected from an ornate box turtle, Terrapene ornata (Agassiz) from Arkansas, USA, is described. Oocysts of Eimeria doddi n. sp. are ovoidal to ellipsoidal with a smooth, light to darker brown, bi-layered wall, measure 21.1 × 14.0 µm, and have a length/width (L/W) ratio of 1.5; both micropyle and oocyst residuum are absent, but a polar granule is present. Sporocysts are ellipsoidal, 9.9 × 6.1 µm, L/W 1.6; the Stieda body is present, but the sub-Stieda and para-Stieda bodies are absent and the sporocyst residuum is composed of small granules in a cluster. Sporozoites have a spheroidal anterior refractile body, a subspheroidal posterior refractile body, and one centrally-located nucleus. This is the third description of an eimerian from the turtle genus Terrapene Merrem and the second from T. ornata. In addition, we report Eimeria ornata McAllister & Upton, 1989 from T. ornata from Texas.

Temperature dependent sex determination (TSD) is the process by which the environmental temperature experienced during embryogenesis influences the sex of an organism, as in the red-eared slider turtle Trachemys scripta elegans. In accord with current paradigms of vertebrate sex determination, temperature is believed to exert its effects on sexual development in T. scripta entirely within the middle third of development, when the gonad is forming. However, whether temperature regulates the transcriptome in T. scripta early embryos in a manner that could influence secondary sex characteristics or establish a pro-male or pro-female environment has not been investigated. In addition, apart from a handful of candidate genes, very little is known about potential similarities between the expression cascade during TSD and the genetic cascade that drives mammalian sex determination. Here, we conducted an unbiased transcriptome-wide analysis of the effects of male- and female-promoting temperatures on the turtle embryo prior to gonad formation, and on the gonad during the temperature sensitive period. We found sexually dimorphic expression reflecting differences in steroidogenic enzymes and brain development prior to gonad formation. Within the gonad, we mapped a cascade of differential expression similar to the genetic cascade established in mammals. Using a Hidden Markov Model based clustering approach, we identified groups of genes that show heterochronic shifts between M. musculus and T. scripta. We propose a model in which multiple factors influenced by temperature accumulate during early gonadogenesis, and converge on the antagonistic regulation of aromatase to canalize sex determination near the end of the temperature sensitive window of development.

Temperature-dependent sex determination, or TSD, is a widespread phenomenon in reptiles. The shape of the relationship between constant incubation temperature and sex ratio defines the TSD pattern. The TSD pattern is considered a life-history parameter important for conservation because the wider the range of temperatures producing both sexes, the more resilient the species is to climate change impacts. We review the different published equations and methodologies that have been used to model TSD patterns. We describe a new flexible model that allows for an asymmetrical pattern around the pivotal temperature, which is the constant temperature producing both sexes in equal proportions. We show that Metropolis-Hastings with Markov chain produced by a Monte Carlo process has many advantages compared to maximum likelihood and is preferred. Finally, we apply the models to results from incubation experiments using eggs from the marine turtle Lepidochelys olivacea originating in Northeast Indian, East Pacific, and West Atlantic Regional Management Units (RMUs) and find large differences in pivotal temperatures but not in transitional ranges of temperatures.

El microhábitat de Chelonia mydas en el arrecife Cabezo, está compuesto por siete tipos de fondo arrecifal. Los parámetros oceanográficos registrados, consisten en aguas someras menores a 2.4 m de profundidad, temperatura del agua de 24.8 a 30.3 °C, concentraciones de oxígeno disuelto de 3.6 a 7.7 mg/L y salinidad de 25.4 a 38.1 ppt. La temperatura es el parámetro con variación significativa en relación al fondo arrecifal. La mayor abundancia de tortugas se distinguió en temporada de lluvias.

The yellow pond turtle, Mauremys mutica, is an economically important aquaculture species and is widely cultured in China because of its high value as a food and in traditional Chinese medicine, as well as being a popular pet. However, M. mutica shows poor tolerance to heat stress and frequently experiences mass mortality caused by infectious diseases during late spring and early summer, when high temperatures occur frequently and various pathogens begin to propagate rapidly. In this study, we performed a comparative transcriptome analysis of M. mutica in response to continuous heat stress (3 h, 9 h, 24 h and 48 h) at 32 °C. Compared to the control group (28 °C), a total of 813, 1488, 1856 and 2001 differentially expressed genes (DEGs) were identified in the four heat-stressed groups respectively, indicating that heat stress has a significant influence on gene expression in M. mutica and that the degree of influence gradually increases with heat stress duration. Interestingly, the immunity-associated category for the complement and coagulation cascade pathway was uniquely enriched in the latter three heat stress groups (9 h, 24 h and 48 h) and all DEGs (e.g. C3, C4, C6, C7, C8, C9 and MASP1/2) in this pathway showed down-regulation as the heat stress duration increased, suggesting that immunity continuously decreases with increased heat stress duration. These observations indicate that potential dysfunction of the immune response under heat stress may contribute to the mass mortality of M. mutica during late spring and early summer. This study provides insights into the molecular mechanisms underlying the mass mortality syndrome in M. mutica and is helpful for proposing effective strategies to alleviate the damage caused by heat stress.

The new Pleurodira turtle Itapecuruemys amazonensis gen. et sp. nov. from the Itapecuru Formation (Parnaiba Basin, Brazil) is described. The new species is represented only by its holotype, which consists of an almost complete carapace and plastron, with an oval-shaped outline. The most peculiar characters of Itapecuruemys amazonensis are: neural plates six and seven are separated by costal six, and the seventh neural plate contacts the sixth, seventh and eighth costal plates and a suprapygal. The phylogenetic hypothesis proposed in this paper suggests that Itapecuruemys amazonensis, together with Cearachelys and Galianemys spp., form a monophyletic assemblage and also widen the paleoherpetological diversity of the Itapecuru Formation in the Parnaiba Basin (Brazil).

Metabolic heating has been poorly investigated in eastern Mediterranean coastline of Turkey, which host some of the most important Green Turtle (Chelonia mydas) nesting sites in the Mediterranean. We studied the effects of clutch size and embryo numbers on nest temperature and discuss the feminizing effect of metabolic heating. Two test sites were conducted in Sugozu Beaches (Turkey). Data loggers were placed in eight nests with different clutch sizes. Nest temperature was strongly correlated with embryo numbers and metabolic heating produced by embryos was calculated to be 0.019°C per late stage embryo and 0.020°C per hatchling. Metabolic heating was calculated to be 0.6°C in the middle third of the incubation period during which sex is determined. It was estimated that metabolic heating increased 10.4% of female hatchlings. The heat produced by embryos should be taken into consideration while estimating sex ratios indirectly by nest and sand temperatures. Additionally, the metabolic heating value should...

In this study, we use geometric morphometric method to establish morphological differences between natural populations of Nile tilapia from two extreme environmental conditions (high temperature and salinity) in Kenya, and compare them to two populations from regions experiencing less extreme conditions. To determine genetic influence on morphology, we correlated genetic data with morphological data. The study observed significant morphological differences between all studied populations, including three closely related hot spring populations (Bogoria, Chelaba and Turtle Springs), and two populations with similar genetic background inhabiting saline environments in Lake Turkana basin (Turkana and Crocodile Lake populations). In addition, allometric growth patterns of the seven populations differed significantly, demonstrating that developmental changes (plastic or adaptive) operated between these populations. Positive correlation between morphometric and genetic data confirmed the influence of genetic factors on morphology. All observed differences were attributed either to genetic and/or environmental factors, which seemed to play a major role in influencing morphology of wild Nile tilapia populations. We recommend further studies to be carried out under controlled conditions to confirm the role of temperature, pH and salinity in morphological diversification of Nile tilapia.

Embryonic development is strongly affected by incubation temperature in most animals, and the effects of incubation temperature continue after partition. However, relatively little is known about the effects of incubation temperature on the development of morphological features in reptiles. As a result, whether developmental rates of different traits change uniquely in response to incubation temperature remains unknown. In this study, we compared embryonic development of an Australian turtle incubated at different temperatures. We examined 43 internal and external morphological features of embryos at each week of development and compared presence and absence of each feature within and across weeks and temperature treatments, using a multivariate statistical approach. We identified morphological features that develop at different rates in turtle embryos incubated at different temperatures. Embryos incubated at 30°C hatched 3 weeks earlier, grew to hatchling size faster, and developed advanced morphological features earlier than those incubated at 26°C. Furthermore, embryonic features that appear late in development appeared progressively earlier in embryos incubated at 30°C. Embryos incubated at 26°C also retained some early embryonic features for longer periods of time during development, especially from weeks 2–4. Despite temperature effects, some traits appeared at the same week of development in embryos from both treatments, including a clearly distinguishable carapace and distinct gut villi at week 3. Variation in appearance and retention of features means that not all external features consistently appear in the same order, depending on incubation temperature. From a practical perspective, cool- and warm-incubated embryos may exhibit different sets of embryological features, which may not be captured by previously published embryological staging guides.

Blood levels of progesterone (P4), estradiol-17β (E2), calcium (Ca), triglyceride (TG), and total protein (TP) were monitored in a captive female loggerhead turtle throughout a nesting season that included three 13- or 14-d internesting intervals. We observed the turtle's ovaries to monitor the appearance of shelled eggs, using ultrasonography, and compared the results of blood analyses and ovary observations. The blood P4 level remained low (