Microplastic ingestion by sea turtles around Tokyo Bay: Level of water pollution influences ingestion amounts.

We investigated the contents inside the esophagus and stomach of turtles inside and outside of the Tokyo Bay area, which face high and low risks of microplastic (MP) exposure, respectively. 65 synthetic particles were recovered from 8 out of 22 turtles, using ATR-FTIR followed by density separation with calcium chloride solution. Statistical analysis indicated that turtles in high-risk areas ingested significantly more MPs than those in low-risk areas. As the inflow of MPs from major rivers influences pollution levels in the ocean, the results of this study highlight the importance of major rivers for MP ingestion by turtles. Additionally, we discussed the current methodology's shortcomings and addressed scope for subsequent research, along with suggestions on future conservation.

Antibiotic Resistance of Bacteria Isolated from Clinical Samples and Organs of Rescued Loggerhead Sea Turtles (Caretta caretta) in Southern Italy.

Antimicrobial resistance affects all environments, endangering the health of numerous species, including wildlife. Increasing anthropic pressure promotes the acquisition and dissemination of antibiotic resistance by wild animals. Sea turtles, being particularly exposed, are considered sentinels and carriers of potential zoonotic pathogens and resistant strains. Therefore, this study examined the antibiotic resistance profiles of bacteria isolated from loggerhead sea turtles hospitalised in a rescue centre of Southern Italy over a 9-year period. Resistance to ceftazidime, doxycycline, enrofloxacin, flumequine, gentamicin, oxytetracycline and sulfamethoxazole-trimethoprim was evaluated for 138 strains isolated from the clinical samples or organs of 60 animals. Gram-negative families were the most isolated: Vibrionaceae were predominant, followed by Shewanellaceae, Pseudomonadaceae, Enterobacteriaceae and Morganellaceae. These last three families exhibited the highest proportion of resistance and multidrug-resistant strains. Among the three Gram-positive families isolated, Enterococcaceae were the most represented and resistant. The opportunistic behaviour of all the isolated species is particularly concerning for diseased sea turtles, especially considering their resistance to commonly utilised antibiotics. Actually, the multiple antibiotic resistance was higher when the sea turtles were previously treated. Taken together, these findings highlight the need to improve antimicrobial stewardship and monitor antibiotic resistance in wildlife, to preserve the health of endangered species, along with public and environmental health.

Are we even close? Five years marine litter ingestion monitoring in loggerhead turtles along Italian coast reveals how far we are from the Good Environmental Status.

The loggerhead sea turtle Caretta caretta has been chosen as bioindicator to monitor the amount of litter ingested by marine animals within the European Marine Strategy Framework Directive and the Barcelona Regional Sea Convention. European Member States and Contracting Parties are committed to achieve the Good Environmental Status (GES), which is reached when the quantity of ingested litter does not adversely affect the health of the species concerned. Although the monitoring strategy has been outlined for more than a decade, to date no threshold values have been adopted to verify GES achievement. After five years of extensive monitoring along the Italian coasts, this study evaluates the suitability of five different GES scenarios and proposes a new threshold value (i.e., "there should be less than 33% of sea turtles having more than 0.05 g of ingested plastic in the GI") for its implementation in the European seas and the Mediterranean basin.

Loggerhead Sea Turtles as Hosts of Diverse Bacterial and Fungal Communities.

Research on microbial communities associated with wild animals provides a valuable reservoir of knowledge that could be used for enhancing their rehabilitation and conservation. The loggerhead sea turtle (Caretta caretta) is a globally distributed species with its Mediterranean population categorized as least concern according to the IUCN Red List of Threatened Species as a result of robust conservation efforts. In our study, we aimed to further understand their biology in relation to their associated microorganisms. We investigated epi- and endozoic bacterial and endozoic fungal communities of cloaca, oral mucosa, carapace biofilm. Samples obtained from 18 juvenile, subadult, and adult turtles as well as 8 respective enclosures, over a 3-year period, were analysed by amplicon sequencing of 16S rRNA gene and ITS2 region of nuclear ribosomal gene. Our results reveal a trend of decreasing diversity of distal gut bacterial communities with the age of turtles. Notably, Tenacibaculum species show higher relative abundance in juveniles than in adults. Differential abundances of taxa identified as Tenacibaculum, Moraxellaceae, Cardiobacteriaceae, and Campylobacter were observed in both cloacal and oral samples in addition to having distinct microbial compositions with Halioglobus taxa present only in oral samples. Fungal communities in loggerheads' cloaca were diverse and varied significantly among individuals, differing from those of tank water. Our findings expand the known microbial diversity repertoire of loggerhead turtles, highlighting interesting taxa specific to individual body sites. This study provides a comprehensive view of the loggerhead sea turtle bacterial microbiota and marks the first report of distal gut fungal communities that contributes to establishing a baseline understanding of loggerhead sea turtle holobiont.

Osteolytic Lesions in a Sub-Adult Loggerhead Sea Turtle (Caretta caretta): A Case Report.

Osteolytic lesions in loggerhead sea turtles (Caretta caretta) during rehabilitation are attributed to multiple causes, including gas embolism, hypothermia, and osteomyelitis due to bacterial or fungal infection. This study reports the appearance of osteolytic lesions in a sub-adult loggerhead sea turtle with involvement of the right fore and hind flippers, visible swelling of the elbow and knee joints, and accompanied by lameness after 45 days of rehabilitation. Radiographs and computed tomography revealed multiple lytic bone lesions. This was the fourth rehabilitation admission of the turtle after being accidentally captured by trawler ships (bycatch) in 2019, 2020, 2022, and 2023. Potential causes were dysbaric osteonecrosis due to a past decompression sickness event and hypothermia with osteomyelitis from bacterial infection. Blood cultures and antibiotic susceptibility testing led to the isolation of Ewingella americana responsive to enrofloxacin. This study investigates extensive fore and hind flipper involvement in a sub-adult loggerhead turtle, aiming to determine causes and risk factors. The pathogenesis and significance of these lesions is discussed.

Deep vs shallow: GPS tags reveal a dichotomy in movement patterns of loggerhead turtles foraging in a coastal bay.

BACKGROUND: Individual variation in movement strategies of foraging loggerhead turtles have been documented on the scale of tens to hundreds of kilometers within single ocean basins. Use of different strategies among individuals may reflect variations in resources, predation pressure or competition. It is less common for individual turtles to use different foraging strategies on the scale of kilometers within a single coastal bay. We used GPS tags capable of back-filling fine-scale locations to document movement patterns of loggerhead turtles in a coastal bay in Northwest Florida, U.S.A. METHODS: Iridium-linked GPS tags were deployed on loggerhead turtles at a neritic foraging site in Northwest Florida. After filtering telemetry data, point locations were transformed to movement lines and then merged with the original point file to define travel paths and assess travel speed. Home ranges were determined using kernel density function. Diurnal behavioral shifts were examined by examining turtle movements compared to solar time. RESULTS: Of the 11 turtles tagged, three tracked turtles remained in deep (~ 6 m) water for almost the entire tracking period, while all other turtles undertook movements from deep water locations, located along edges and channels, to shallow (~ 1-2 m) shoals at regular intervals and primarily at night. Three individuals made short-term movements into the Gulf of Mexico when water temperatures dropped, and movement speeds in the Gulf were greater than those in the bay. Turtles exhibited a novel behavior we termed drifting. CONCLUSIONS: This study highlighted the value provided to fine-scale movement studies for species such as sea turtles that surface infrequently by the ability of these GPS tags to store and re-upload data. Future use of these tags at other loggerhead foraging sites, and concurrent with diving and foraging data, would provide a powerful tool to better understand fine-scale movement patterns of sea turtles.

Heart rate reduction during voluntary deep diving in free-ranging loggerhead sea turtles.

Air-breathing vertebrates exhibit cardiovascular responses to diving including heart rate reduction (diving bradycardia). Field studies on aquatic mammals and birds have shown that the intensity of bradycardia can vary depending on diving behaviour, such as the depth of dives and dive duration. However, in aquatic reptiles, the variation in heart rate during deep dives under natural conditions has not been fully investigated. In this study, we released five loggerhead sea turtles (Caretta caretta) outfitted with recorders into the sea and recorded their electrocardiogram, depth, water temperature and longitudinal acceleration. After 3 days, the recorders automatically detached from the turtles. The heart rate signals were detected from the electrodes placed on the surface of the plastron. The mean (±s.d.) heart rate of 12.8±4.1 beats min-1 during dives was significantly lower than that of 20.9±4.1 beats min-1 during surface periods. Heart rate during dives varied with dive depth, although it remained lower than that at the surface. When the turtle dived deeper than 140 m, despite the relatively high flipper stroke rate (approximately 19 strokes min-1), the heart rate dropped rapidly to approximately 2 beats min-1 temporarily. The minimum instantaneous heart rate during dives was lower at deeper dive depths. Our results indicate that loggerhead sea turtles show variations in the intensity of diving bradycardia depending on their diving behaviour, similar to that shown by marine mammals and birds.

Marine turtles as bio-indicators of plastic pollution in the eastern Mediterranean.

The loggerhead turtle (Caretta caretta) has been suggested as a bio-indicator species for plastic pollution. However, detailed investigations in the eastern Mediterranean are limited. Here, we present data from loggerhead turtles (2012-2022; n = 131) of which 42.7 % (n = 57) had ingested macroplastic (pieces ≥ 5 mm). Frequency of occurrence (%) was not found to have changed over time, with body size (CCL cm), between stranded or bycaught turtles, or with levels of digesta present. The characteristics of ingested plastic (n = 492) were largely sheetlike (62 %), clear (41 %) or white (25 %) and the most common polymers identified were Polypropylene (37 %) and Polyethylene (35 %). Strong selectivity was displayed towards certain types, colours and shapes. Data are also presented for posthatchling turtles (n = 4), an understudied life stage. Much larger sample sizes will be needed for this species to be an effective bio-indicator, with the consideration of monitoring green turtles (Chelonia mydas) for the eastern Mediterranean recommended allowing a more holistic picture to be gathered.

Iatrogenic gas embolism in a juvenile loggerhead sea turtle (Caretta caretta).

A mild pneumocoelom was diagnosed by computed tomography in a stranded juvenile loggerhead sea turtle (Caretta caretta). After gas extraction by ultrasound-guided puncture, the animal did not improve and was subjected to hyperbaric oxygen therapy (HBOT). After HBOT, the turtle developed marked subcutaneous emphysema and was found dead the following morning. Gross lesions included a distended right atrium with numerous gas bubbles within the epicardium, gas bubbles in the hepatic, gastric and mesenteric veins, a small gas-filled bulla in the left lung and diffuse haemorrhages in the encephalon. Histological lesions included gas-like emboli in the lumen of the right atrium with myocardial necrosis, gas-like emboli in the lumina of intestinal, pulmonary and renal blood vessels and severe meningeal haemorrhages. From a forensic pathology perspective, the subcutaneous emphysema of immediate onset after HBOT and the greater severity of the histological lesions in blood vessels, heart, lung and brain differentiate this case from other cases of gas embolism in turtles due to incidental capture. Two factors contributed to this outcome: the existence of a probably unresolved pneumocoelom and the application of HBOT without an initial diagnosis that accurately indicated its use. Therefore, as in human medicine, the use of HBOT in sea turtles with lung lesions and pneumocoelom is discouraged. This is the first description of an iatrogenic gas embolism in a sea turtle.

Citrobacter freundii-associated lesions in stranded loggerhead sea turtles (Caretta caretta).

Citrobacter freundii, an opportunistic bacterial pathogen belonging to the family Enterobacteriaceae, has been sporadically reported in sea turtles. Here, the authors describe 3 unusual lesions associated with C. freundii infection in 3 loggerhead sea turtles stranded on the coast of Gran Canaria Island, Spain. It is possible that these 3 distinct lesions played a major role in the death of these turtles. The first turtle had caseous cholecystitis, a lesion not previously described in sea turtles. The second turtle had large intestinal diverticulitis, a rare condition in loggerheads. The third turtle had bilateral caseous salt gland adenitis. Histologically, numerous gram-negative bacilli were observed at the deepest edge of inflammation in all cases. Pure cultures of C. freundii were obtained from these 3 lesions. Molecular detection of C. freundii DNA in formalin-fixed paraffin-embedded samples from the lesions of the 3 turtles confirmed the microbiological isolation. These cases, in addition to expanding the limited body of knowledge on bacterial infections in sea turtles, highlight the potential pathogenic role of C. freundii in loggerhead turtles.

Assessing microplastics contamination in unviable loggerhead sea turtle eggs.

Sea turtles, in comparison with marine mammals, sea birds, and fishes, are the most affected by microplastics in terms of number of individuals impacted and concentration within each organism. The ubiquitous nature and persistence of microplastics in the environment further compromises sea turtles as many species are currently vulnerable, endangered, or critically endangered. The objective of this study was to quantify microplastic contamination in unviable loggerhead sea turtle eggs (Caretta caretta). Eggs were collected from seven locations along the northwest coast of Florida. A total of 70 nests and 350 eggs were examined. Microplastics (n = 510) were found in undeveloped loggerhead sea turtle eggs across all seven sites, suggesting that maternal transference and/or exchange between the internal and external environment were possible. The frequency found was 7.29 ± 1.83 microplastic pieces per nest and 1.46 ± 0.01 per egg. Microplastics were categorized based on color, shape, size, and type of polymer. The predominant color of microplastics were blue/green (n = 236), shape was fibers (n = 369), and length was 10-300 µm (n = 191). Identified fragments, films, beads and one foam (n = 187) had the most common area of 1-10 µm2 (n = 45). Micro-Fourier Transform Infrared (µ-FTIR) spectroscopy analysis demonstrated that polyethylene (11 %) and polystyrene (7 %) were the main polymer types. For the first time microplastics were found in unviable, undeveloped loggerhead sea turtle eggs collected in northwest Florida. This work provides insight into the distribution patterns of microplastic pollutants in loggerhead sea turtle eggs and may extend to other species worldwide.

Complete Colombian Caribbean loggerhead turtle mitochondrial genome: tRNA structure analysis and revisited marine turtle phylogeny / Genoma mitocondrial completo de la tortuga caguama del Caribe colombiano: análisis de la estructura del tRNA y revisión de la filogenia de las tortugas marinas / Genoma mitocondrial de tartaruga-cabeçuda do Caribe colômbiano completo: análise de estrutura de tRNA e filogenia revisada de tartarugas marinhas

Abstract The loggerhead marine turtle, Caretta caretta, is a widely distributed and endangered species that is facing critical population decline, especially in Colombian Caribbean rookeries. Mitochondrial DNA sequence data are of great importance for the description, monitoring, and phylogenetic analyses of migratory turtle populations. In this study, the first full mitochondrial genome of a loggerhead turtle nesting in the Colombian Caribbean was sequenced and analyzed. This mitochondrial genome consists of 16 362 bp with a nucleotide composition of T: 25.7 %, C: 27 %, A: 35 % and G: 12 %. Sequence annotation of the assembled molecule revealed an organization and number of coding and functional units as reported for other vertebrate mitogenomes. This Colombian loggerhead turtle (Cc-AO-C) showed a novel D-Loop haplotype consisting of thirteen new variable sites, sharing 99.2 % sequence identity with the previously reported Caribbean loggerhead CC-A1 D-Loop haplotype. All 13 protein-coding genes in the Cc-AO-C mitogenome were compared and aligned with those from four other loggerhead turtles from different locations (Florida, Greece, Peru, and Hawaii). Eleven of these genes presented moderate genetic diversity levels, and genes COII and ND5 showed the highest diversity, with average numbers of pair-wise differences of 16.6 and 25, respectively. In addition, the first approach related to t-RNAs 2D and 3D structure analysis in this mitogenome was conducted, leading to observed unique features in two tRNAs (tRNATrp and tRNALeu). The marine turtle phylogeny was revisited with the newly generated data. The entire mitogenome provided phylogenetically informative data, as well as individual genes ND5, ND4, and 16S. In conclusion, this study highlights the importance of complete mitogenome data in revealing gene flow processes in natural loggerhead turtle populations, as well as in understanding the evolutionary history of marine turtles.

Resumen La tortuga marina caguama, Caretta caretta, es una especie ampliamente distribuida pero que enfrenta una crítica reducción de su población en las colonias del Caribe colombiano. Los datos de las secuencias de DNA mitocondrial son de gran importancia para la descripción, monitoreo y análisis de la filogenia de las tortugas migratorias. En este estudio se secuenció y analizó por primera vez el genoma mitocondrial completo de la tortuga caguama que anida en el Caribe colombiano. Este genoma tiene un tamaño de 16.362 pb con una composición de nucleótidos de T: 25.7 %, C: 27 %, A: 35 % y G: 12 %. La anotación de la secuencia de la molécula reveló una organización y número de unidades codificantes y funcionales como los reportados para mitogenomas de otros vertebrados. Esta tortuga caguama colombiana (Cc-AO-C) mostró un nuevo haplotipo D-Loop que contiene trece nuevos sitios variables, que comparten el 99.2 % de identidad de secuencia con el haplotipo CC-A1 D-Loop previamente reportado para la tortuga caguama del Caribe. Los trece genes que codifican proteínas en el mitogenoma Cc-AO-C se compararon y alinearon con los de otras cuatro tortugas caguama de distintas localidades (Florida, Grecia, Perú y Hawái). Once de estos genes presentaron niveles moderados de diversidad genética, y los genes COII y ND5 mostraron las diversidades nucleotídicas más altas, con un número promedio de diferencias entre pares de secuencias de 6.6 y 25, respectivamente. Adicionalmente, se llevó a cabo la primera aproximación relacionada con el análisis de la estructura 2D y 3D de t-RNAs en este mitogenoma, lo cual condujo a la observación de características únicas en dos tRNAs (tRNATrp y tRNALeu). La filogenia de las tortugas marinas fue revisada a la luz de la nueva información mitogenómica. El mitogenoma, así como los genes individuales ND5, ND4 y 16S, proporcionan datos filogenéticamente informativos. En conclusión, este estudio resalta la importancia de los datos del mitogenoma para revelar procesos de flujo génico en las poblaciones naturales de tortuga caguama, así como para entender la historia evolutiva de las tortugas marinas.

Resumo A tartaruga marinha Caretta caretta (Cc) é uma espécie amplamente distribuída e ameaçada de extinção que enfrenta um declínio crítico da população, especialmente nas colônias do Caribe colombiano. Marcadores moleculares, como sequências de DNA mitocondrial (mtDNA), são de grande importância para a descrição, monitoramento e análise filogenética de populações migratórias de tartarugas. Este estudo mostra a obtenção e análise do genoma mitocondrial de uma tartaruga-cabeçal Cc aninhada na costa Caribe da Colômbia. O genoma mitocondrial é constituído por 16.362 pb, com uma região não codificante (D-Loop), 13 genes codificadores de proteínas (13 PCG), 22 genes tRNA e 2 rRNA (16S e 12S) e uma frequência nucleotídica de T: 25.7 % , C: 27 %, A: 35 % e G: 12,2 %, todos organizados de forma semelhante à maioria dos mitogenomos de vertebrados. Esta tartaruga Cc colombiana apresentou um novo haplótipo D-Loop com treze sítios polimórficos quando comparado ao haplótipo CC-A1.1 (96 %). Além disso, onze genes codificadores de proteínas entre as tartarugas marinhas de diferentes origens apresentaram uma diversidade genética semelhante, exceto os genes COII e ND5 que apresentaram o maior número médio de diferenças entre pares de seqüências (16.600 e 25.000, respectivamente). Aqui relatase a primeira abordagem relacionada à análise de estruturas 2D e 3D para Cc e descrevese as diferenças em dois tRNAs (tRNATrp, tRNALeu). As inferências bayesianas e os métodos de máxima verossimilhança explicam melhor a filogenia das tartarugas marinhas quando utilizamse mitogenomes completos, assim como os genes ND5, ND4 e 16S. Os genes marcadores ATP8, ND4L e ND1 apresentaram relação filogenética pouco suportada. Como conclusão, este estudo apresenta o uso de mitogenomes completos como uma alternativa para melhorar a análise filogenética em tartarugas marinhas e é a primeira análise genética de mitogenomes completos de nidificação na Colômbia.

Análisis de haplotipos de la tortuga cabezona Caretta caretta (Testudines: Cheloniidae) en dos playas del Caribe colombiano / Analysis of haplotypes of the loggerhead turtle, Caretta caretta (Testudines: Cheloniidae), on two beaches of the Colombian Caribbean / Análise de haplotipos da tartaruga cabeçona, Caretta caretta (Testudines: Cheloniidae) em duas praias do Caribe colombiano

Resumen Introducción. La tortuga Caretta caretta habita los mares tropicales y subtropicales. Es una especie en vía de extinción que anida las playas en Colombia y hace extensas migraciones. Los haplotipos mitocondriales de esta tortuga se han utilizado para estudios de genética poblacional, filogeografía y estado de las especies con el objetivo de desarrollar planes de conservación de la especie. Objetivo. Identificar haplotipos mitocondriales en tortugas cabezonas anidantes del Caribe colombiano. Materiales y Métodos. Se recolectaron muestras de sangre periférica de esta especie en dos sitios del Caribe colombiano: Don Diego (playa de anidación) y la Isla San Martin de Pajarales (localidad de alimentación). El ADN total fue extraído a partir de las células sanguíneas, y utilizado para amplificar por PCR la región control mitocondrial (398 pb). Estos productos fueron purificados y secuenciados. Se realizó un alineamiento básico buscando regiones de similitud local entre las secuencias obtenidas y las descritas previamente para la especie. Se hicieron análisis filogenéticos utilizando los criterios de máxima parsimonia (MP) y máxima verosimilitud (ML). Resultados. Se identificaron tres haplotipos, CC-A1 y CC-A2 comúnmente encontrados en poblaciones reproductivas de México, el Mediterráneo y el sudeste de Estados Unidos, y un nuevo haplotipo CC-SM1 en la playa Don Diego (Magdalena). Los árboles filogenéticos muestran relación de una porción de los individuos anidantes y de forrajeo de las agregaciones del Caribe colombiano con las súper-agregaciones del Atlántico y el Mediterráneo, sugiriendo que estas podrían ser algunas de las fuentes importantes de individuos presentes en Colombia. Conclusiones. Es necesario estudiar una muestra más grande para poder confirmar hipótesis planteadas. Se identificó un nuevo haplotipo denominado CC-SM1. Este es el primer estudio sobre haplotipos mitocondriales de C. caretta realizado en Colombia.

Abstract Introduction. The Caretta caretta turtle inhabits tropical and subtropical seas. It is an endangered species that nests on Colombian beaches and makes long migrations. The mitochondrial haplotypes of this turtle have been used for population genetics, phylogeography and species status studies with the aim of developing species conservation plans. Objective. Identify mitochondrial haplotypes in nesting loggerhead turtles from the Colombian Caribbean. Materials and Methods. Peripheral blood samples from this species were collected in two sites of the Colombian Caribbean: Don Diego (nesting beach) and San Martin de Pajarales island (feeding ground). The total DNA was extracted from blood cells and used to amplify the mitochondrial control region by PCR (398 bp). These products were purified and sequenced. A basic alignment was performed looking for local similarity regions between the sequences obtained and those previously described for the species. Phylogenetic analyses were conducted by using the maximum parsimony (MP) and maximum likelihood (ML) criteria. Results. Three haplotypes were identified: CC-A1 and CC-A2, which are commonly found in breeding populations in Mexico, the Mediterranean and southeast U.S.; and a new haplotype, CC-SM1, on the Don Diego beach (Magdalena). The phylogenic trees show a relationship between a portion of the nesting and feeding individuals from the Colombian Caribbean aggregations and the Atlantic and Mediterranean super-aggregations, suggesting that these could be some of the important sources for individuals inhabiting Colombia. Conclusions. It is necessary to study a larger sample to be able to confirm the proposed hypotheses. A new haplotype called CC-SM1was identified. This is the first study on C. caretta mitochondrial haplotypes conducted in Colombia.

Resumo Introdução. A tartaruga Caretta caretta habita os mares tropicais e subtropicais. É uma espécie em via de extinção que enraíza as praias na Colômbia e faz extensas migrações. Os haplotipos mitocondriais desta tartaruga se há utilizado para estudos de genética populacional, fílogeografía e estado das espécies com o objetivo de desenvolver planos de conservação da espécie. Objetivo. Identificar haplotipos mitocondriais em tartarugas cabeçonas enraizada no Caribe colombiano. Materiais e Métodos. Se coletaram amostras de sangue periférica desta espécie em dois lugares do Caribe colombiano: Don Diego (praia de enraizamento) e a Ilha San Martin de Pajarales (localidade de alimentação). O DNA total foi extraído a partir das células sanguíneas, e utilizado para amplificar por PCR a região controle mitocondrial (398 pb). Estes produtos foram purificados e sequenciados. Se realizou um alinhamento básico buscando regiões de semelhança local entre as sequências obtidas e as descritas previamente para a espécie. Se fez análise filogenéticos utilizando os critérios de máxima parcimônia (MP) e máxima verossimilitude (ML). Resultados. Se identificaram três haplotipos, CC-A1 e CC-A2 comumente encontrados nas populações reprodutivas do México, o Mediterráneo e o sudeste de Estados Unidos, e um novo haplotipo CC-SM1 na praia Don Diego (Magdalena). As árvores filogenéticos mostram relação de uma porção dos indivíduos enraizados e de forragem das agregações do Caribe colombiano com as super-agregações do Atlântico e o Mediterrâneo, sugerindo que estas poderiam ser algumas das fontes importantes de indivíduos presentes na Colômbia. Conclusões. É necessário estudar uma amostra maior para poder confirmar hipótese proposta. Se identificou um novo haplotipo denominado CC-SM1. Este é o primeiro estudo sobre haplotipos mitocondriais de C. caretta realizado na Colômbia.

AMDAR y PCR-extra-rápida para la identificación de la tottuga cabezona Caretta caretta (Testudines: Cheloniidae) utilizando el gen mitocondrial citocromo c oxidasa I (COI) / AMDAR and PCR-Extra-fast for Molecular identification of the loggerhead sea turtle Caretta caretta (Testudines: Cheloniidae) using the mitochondrial gene cytochrome c oxidase I (COI) / AMDAR e PCR extra-rápida para a identificação da Tartaruga-Comum Caretta caretta (Testudines: Cheloniidae), utilizando o gene mitocondrial citocromo c oxidase I (COI)

Para identificación molecular de tortuga cabezona Caretta caretta, se utilizó Amplificación Mitocondrial DNA y Análisis de Restricción y PCR Extra-rápida. Se obtuvo muestras de sangre de juveniles de C. caretta de playa Don Diego (Magdalena; n=4) e Islas Del Rosario (Bolívar; n=2) en el Caribe colombiano. Se aisló DNA, se estandarizó la amplificación del gen mitocondrial citocromo c oxidasa I (COI) por PCR extra-rápida, obteniendo fragmentos de 650 pares de bases, disminuyendo en un tercio el tiempo de reacción. El producto amplificado de COI se analizó con enzimas HindIII, HyCH4III y MseI generando perfil electroforético que al compararlo in silico con secuencias para otras especies de tortugas marinas; permitió identificar patrón específico para la tortuga cabezona. Las secuencias nucleótidicas se analizaron con BLAST y similaridad entre 97-99 % con C. caretta en cinco secuencias y 92 % en otra. La información de tortugas muestreadas fue integrada en base datos BOLD y se generó el código de barras. La metodología descrita para identificación de C. caretta es procedimiento rápido y bajo costo que minimiza el tiempo de PCR mejorando su especificidad.

We molecularly identified the loggerhead turtle Caretta caretta using high-speed PCR amplification and restriction analysis of mitochondrial DNA. We isolated the DNA from blood from juvenile C. caretta from Don Diego beach (Magdalena; n=4), in Islas Del Rosario (Bolívar; n=2) in the Colombian Caribbean. By using high-speed PCR amplification of mitochondrial cytochrome c oxidase I (COI), we reduced reaction by one third and obtained fragments of 650 base pairs. We analyzed the amplified IOC product using enzymes HindIII, HpyCH4III and MseI and generated an electrophoretic profile, which compared in silico to other sea turtle species sequences, revealed the loggerhead's specific pattern. We found similarity between 97-99% with C. caretta in five of the BLAST analyzed nucleotide sequences and 92% in another. We generated a bar code for the sampled turtle information and sequences and stored them in the BOLD database. The methodology described for the identification of C. caretta is a fast and inexpensive procedure that minimizes time and improves PCR specificity.

Identificámos molecularmente a tartaruga-comum Caretta caretta usando PCR amplificado de alta-velocidade e análise de ADN mitocondrial restrito. Isolámos o ADN do sangue de juvenis C. carreta das praias Don Diego (Magdalena ; n=4), das Ilhas del Rosario (Bolívas; n=2) no Caribe Colombiano. Ao utilizar a amplificação por PCR de alta velocidade da citocromo c oxidase I (COI ), reduzimos a reacção a um terço, e obtivemos fragmentos de 650 pares de bases. Analisámos o produto IOC amplificado com as enzimas de restrição HindIII , HyCH4III e MseI e gerámos um perfil eletroforético, que comparado em silico com outras seqüéncias de espécies de tartarugas marinhas, revelou padrão específico à tartaruga-comum. Encontramos semelhanças entre 97-99 %, com C. caretta em cinco das seqüéncias de nucleotídeos BLAST analisados e 92% com outro. Gerámos um código de barras para a informação e seqüéncias das tartaruga amostradas e foram armazenados na base de dados BOLD. A metodologia descrita para a identificação de C. caretta é um procedimento rápido e barato, que minimiza o tempo e melhora a especificidade da PCR.

Serum biochemistry profile determination for wild loggerhead sea turtles nesting in Campos dos Goytacazes, Rio de Janeiro, Brazil / Determinação do perfil bioquímico sérico de tartarugas marinhas de vida livre da espécie em nidação em Campos dos Goytacazes, Rio de Janeiro, Brasil

Sea turtles are threatened to the point of extinction. The major goal of rehabilitating sick individuals is to eventually reintroduce them back into their habitat. In this way, they contribute to species preservation, as well as maintaining equilibrium of the ecosystems. Biochemical analysis is a commonly used test to detect illness and evaluate the general health of the animals. However, the data in the literature on sea turtles are scarce and the majority of studies used small sample sizes, being the majority of animals in captivity. The aim of the present study is to establish baseline biochemical profile values for free-ranging, nesting, female loggerhead turtles (Caretta caretta). The baseline values can then be used for comparison in the overall evaluation, physiologic status and disease diagnostics of diverse populations of sea turtles. Twenty-eight females in their reproductive period were used from Farol de São Thomé (21°45'15"S - 41°19'28"W), city of Campos dos Goytacazes, north-fluminense region. The samples were collected without anticoagulant through venapuncture of the dorsal, cervical sinus. The average values determined were calcium, phosphorus, cholesterol and triglycerides, demonstrating a correlation with vitellogenesis and egg formation. The fact that females reduce feeding in the period preceding egg laying, influenced the average concentration of urea (35.25mg dL-1), sodium (147mEq L-1), potassium (28mEq L-1), uric acid (0.6mg dL-1) and lipids. Carapace length and width, and the weight of the turtles showed a positive correlation with liver enzymes ALT and AST, suggesting that animals with larger hepatic volume have greater enzymatic activity.

As tartarugas marinhas são animais ameaçados de extinção. Sendo assim, o maior objetivo em reabilitar os indivíduos doentes é posteriormente reintroduzi-los em seu habitat. Dessa forma, contribui-se para a preservação das espécies e para a manutenção do equilíbrio dos ecossistemas. As análises bioquímicas são exames bastante utilizados na detecção de doenças e na avaliação do estado geral dos animais. Apesar disso, os dados na literatura para tartarugas marinhas ainda são escassos, e a maioria dos estudos utiliza um pequeno número de animais, sendo a maior parte deles de cativeiro. O presente estudo tem por objetivo realizar o perfil bioquímico de indivíduos do sexo feminino de tartarugas marinhas de vida livre da espécie Caretta caretta em processo de nidação, bem como estabelecer a correlação do perfil bioquímico destes animais com seu tamanho. Os valores médios obtidos poderão ser utilizados posteriormente como comparativo para avaliação do estado geral e para o diagnóstico de doenças de diversas populações de tartarugas marinhas. Foram utilizadas 28 fêmeas em período reprodutivo, em Farol de São Thomé (21°45'15"S - 41°19'28"W), no município de Campos dos Goytacazes, região norte-fluminense. As amostras foram coletadas sem anticoagulante, através de venopunção do seio venoso cervical dorsal. Os valores médios encontrados para as variáveis cálcio, fósforo, colesterol e triglicerídeos demonstram a correlação desses componentes com o processo de vitelogênese e a formação do ovo. O fato de as fêmeas reduzirem a alimentação, no período que antecede a postura dos ovos, influenciou a concentração média de uréia (35,25mg dL-1), sódio (147mEq L-1), potássio, ácido úrico e lipídios. As variáveis vinculadas ao tamanho das tartarugas apresentaram correlação positiva com enzimas ALT e AST, sugerindo que animais com maior volume hepático apresentam maior atividade enzimática.

Condiciones óptimas de cultivo de linfocitos y análisis parcial del cariotipo de la tortuga cabezona, Caretta caretta (Testudines: Cheloniidae) en Santa Marta, Caribe Colombiano

Lymphocyte culture and partial karyotype of the marine turtle Caretta caretta (Testudines: Cheloniidae) in Santa Marta, Colombian Caribbean. Over the past few years an important reduction in the number of nesting marine turtle Caretta caretta individuals has been registered in the Colombian Caribbean, raising the question of a possible extinction in the medium term. A conservation plan is needed. We studied the culture requirements for C. caretta lymphocytes and preliminary karyotype analysis for cytogenetic identification, immunological study and toxicology without the need to kill individuals. Peripheral blood samples were obtained from 47 individuals in Santa Marta, Colombia and tests were made until optimal conditions were established for lymphocyte culture. The karyotype had 56 chromosomes, 32 macrochromosomes and 24 micro-chromosomes. An ideogram showed that C. caretta has four groups of chromosomes. Sexual chromosomes were not observed. These results do not coincide with the karyotype described from the Pacific (Japan). The present study is the first to include a complete description of the chromosome morphology of turtles from the Atlantic Ocean. it is possible that one of the adaptive strategies of this species is genetic interchange with other species of the family, producing viable hybrids. individuals in this study might be viable hybrids of C. caretta and further molecular studies are needed. Rev. Biol. Trop. 56 (3): 1459-1469. Epub 2008 September 30.

En los últimos años se ha registrado una disminución importante en el número de individuos de la tortuga marina Caretta caretta anidantes en el Caribe colombiano, situación que pone en evidencia la posibilidad de su extinción a mediano plazo. Por esto, es necesario implementar planes para su manejo y conservación. En este estudio se determinaron los requerimientos del cultivo de linfocitos de Caretta caretta para la obtención de cariotipos que permitan la identificación citogenética, el estudio inmunológico y toxicológico de individuos sin necesidad de sacrificarlos. Se muestrearon 47 individuos de C. caretta de Santa Marta, Colombia obteniendo sangre periférica con la que se realizaron ensayos de las diferentes variables hasta obtener las condiciones óptimas para el cultivo convencional de linfocitos. El cariotipo obtenido presentó 56 cromosomas: 32 macrocromosomas y 24 microcromosomas. El ideograma mostró que C. caretta tiene cuatro grupos de cromosomas: el grupo A compuesto por doce (12) pares de cromosomas de mayor tamaño. El Grupo B compuesto por cuatro (4) pares de cromosomas medianos y pequeños y el Grupo C conformado por 12 pares de microcromosomas. No se observaron cromosomas sexuales. Estos resultados están en desacuerdo con el cariotipo descrito por Kamesaki (1989), debido posiblemente a que las muestras analizadas en ese estudio fueron colectadas en el Océano Pacifico (Japón). El presente estudio es el primero realizado con tortugas del Océano Atlántico que cuenta con la descripción completa de la morfología cromosómica. Es posible, que una de las estrategias adaptativas de esta especie sea el intercambio genético con otras especies de la familia, que produce individuos híbridos viables. En este aspecto se ha descrito la hibridación de tortugas Caretta caretta con Eretmochelys imbricata, Chelonia mydas y Lepidochelys kempii, esto sugiere la posibilidad que los individuos caracterizados en este estudio sean híbridos viables de C. caretta, por lo tanto, se hace necesario realizar estudios a nivel molecular.