Hematologic parameters and the effect of hemoparasites of wild anurans in Northern Sinaloa, Mexico.

BACKGROUND: Few hematologic profiles for free-ranging amphibians are available. Hematologic evaluation is a useful tool for determining the health of amphibian populations and providing further knowledge for conservation actions. OBJECTIVES: Hematologic variables and the presence and effect of hemoparasites in anuran species were evaluated in Northern Sinaloa, Mexico. METHODS: Blood samples were collected from wild anurans of eight species to perform blood cell counts, leukocyte differential counts, and serum protein concentrations using manual methods and refractometry. In addition, morphologic identification and quantification of the hemoparasites were performed on blood smears. RESULTS: Differences were observed by sex, age, and season for the hematologic values of Incilius alvarius (n = 23), Incilius mazatlanensis (n = 46), Rhinella horribilis (n = 64), Leptodactylus melanonotus (n = 46), Lithobates forreri (n = 135), Lithobates catesbeianus (n = 20), Smilisca fodiens (n = 42), and Scaphiopus couchii (n = 7). Intra- and extra-erythrocytic hemoparasites were found in 56.2% of amphibian hosts; the hemoparasite infection of R. horribilis and L. melanonotus was higher in the dry season, showing increases in erythroplastids and monocytes. For L. forreri, males were more infected than females, and increases in leukocytes were associated with infections of different types of hemoparasites species. CONCLUSIONS: Hematologic values, hemoparasite prevalence, and the response to hemoparasite infection vary among amphibian species, sex, and age, as well as on season and hemoparasite type. This highlights the importance of hematologic evaluations in wild amphibian populations to determine the subclinical effects of hemoparasite infections.

Trace elements concentration in blood of nesting Kemp's Ridley turtles (Lepidochelys kempii) at Rancho Nuevo sanctuary, Tamaulipas, Mexico.

The concentrations of trace elements including As, Zn, Cu, Se, Pb, Hg and Cd, were determined in the blood of nesting Kemp's ridley turtles (Lepidochelys kempii) at Rancho Nuevo sanctuary, Tamaulipas, Mexico during 2018-2020. The sequential concentrations analyzed were Zn> Se> Cu> As> Pb; while Cd and Hg concentrations were below the limits of detection (0.01 µg g-1). No significant differences were observed between the concentrations of trace elements (p> 0.05) by year, except Se levels, possibly resulting from recorded seasonal differences in turtle size. No relationships among turtle size vs elements concentration were observed. In conclusion, essential and toxic trace elements concentrations in the blood of nesting Kemp's ridley turtles may be a reflex of the ecosystem in which the turtles develop, that is, with low bioavailability of elements observed in the trophic webs in the Gulf of Mexico.

ISOLATION, CHARACTERIZATION, AND ANTIMICROBIAL SUSCEPTIBILITY OF BACTERIA ISOLATED FROM SEA LION (ZALOPHUS CALIFORNIANUS) PUPS IN NORTHWESTERN MEXICO.

Bacterial infections have been documented in marine mammals for decades, and some are considered emerging pathogens with zoonotic potential. The aerobic oral (n=16) and rectal (n=17) bacterial microbiota and their antimicrobial resistance were characterized for 17 apparently healthy California sea lion pups (Zalophus californianus) captured with a hoop net in Farallon Island, Sinaloa, Mexico, in 2016. Bacteriologic cultures, Analytical Profile Index, and PCR were used to identify bacterial species. The Escherichia coli phylogenetic groups were identified by PCR, Salmonella serotypes were identified, and resistance to antibiotics was evaluated. Overall, 39 bacterial species were isolated, including E. coli and Salmonella spp. (35.9% each) and Pseudomonas aeruginosa (28.2%). For E. coli, UNKNOWN phylogroup was the most prevalent (57.7%), followed by the A phylogroup (37.1%). Most Salmonella serotypes were identified as Newport (92.8%); serotype Saintpaul was also identified (7.2%). Sea lions with bacterial co-colonization included 24.2%, from which two bacterial species were isolated, and 3% with three species. Overall, 59% of bacteria were resistant to at least one antibiotic tested, and 25.6% were extensively drug resistant. Bacteria were highly resistant to ampicillin and cefotaxime. This study demonstrates the importance of characterizing the microbiome of sea lions, and the potential effect of pathogens with antimicrobial resistance on wildlife conservation and public health.

DNA barcode analysis of the endangered green turtle (Chelonia mydas) in Mexico1.

Technological and analytical advances to study evolutionary biology, ecology, and conservation of green turtles (Chelonia mydas) are realized through molecular approaches including DNA barcoding. We characterized the usefulness of COI DNA barcodes in green turtles in Mexico to better understand genetic divergence and other genetic parameters of this species. We analyzed 63 sequences, including 25 from green turtle field specimens collected from the Gulf of Mexico and from the Mexican Pacific and 38 already present in the Barcode of Life Data Systems (BOLD). A total of 13 haplotypes were identified with four novel haplotypes from the Pacific Ocean and three novel haplotypes from the Atlantic Ocean. Intraspecific distance values among COI gene sequences by two different models were 0.01, demonstrating that there is not a subdivision for green turtle species. Otherwise, the interspecific distance interval ranged from 0.07 to 0.13, supporting a clear subdivision among all sea turtle species. Haplotype and total nucleotide diversity values of the COI gene reflect a medium genetic diversity average. Green turtles of the Mexican Pacific showed common haplotypes to some Australian and Chinese turtles, but different from the haplotypes of the Mexican Atlantic. COI analysis revealed new haplotypes and confirmed that DNA barcodes were useful for evaluation of the population diversity of green turtles in Mexico.

Essential and trace metals in a post-nesting olive ridley turtles (Lepidochelys olivacea) in Ceuta beach, Sinaloa, Mexico.

Trace metals have been found in sea turtle blood and tissues and may represent a threat to these endangered species. Essential trace metal (Cu, Zn Cd, Pb, As, and Hg) concentrations were determined in blood of adult female, post-nesting olive ridley turtles Lepidochelys olivacea (n = 35) on Ceuta beach, Sinaloa, Mexico. Essential metals (Zn and Cu) analyzed were found in higher concentrations than toxic metals (Cd and Pb), while As and Hg concentrations were below the limits of detection (0.01 µg g-1). Low Pb concentrations (0.09 µg g-1) were previously observed in sea turtles in the Gulf of California. There were no significant correlations found between curved carapace length (61.00-71.00 ± 2.29) vs metal concentrations (p > 0.05). Cd levels were relatively high when compared to other species and populations of sea turtles worldwide and Cd may represent the greatest risk for sea turtles in the Mexican Pacific. Such concentrations of Cd may pose a further risk to sea turtles through bioaccumulation from the nesting female to offspring which may affect embryo development.

Molecular Characterization of Chelonid Alphaherpesvirus 5 in a Black Turtle (Chelonia mydas) Fibropapilloma from Baja California Sur, Mexico.

During routine monitoring in Ojo de Liebre Lagoon, Mexico, a juvenile black turtle (Chelonia mydas) was captured, physically examined, measured, weighed, sampled, and tagged. The turtle showed no clinical signs suggestive of disease. Eleven months later, this turtle was recaptured in the same area, during which one lesion suggestive of fibropapilloma on the neck was identified and sampled for histopathology and molecular analysis. Histopathology revealed hyperkeratosis, epidermal hyperplasia, acanthosis, papillary differentiation and ballooning degeneration of epidermal cells, increased fibroblasts in the dermis, and angiogenesis, among other things. Hematological values were similar to those reported for clinically healthy black turtles and did not show notable changes between the first capture and the recapture; likewise, clinicopathological evaluation did not show structural or functional damage in the turtle's systems. The chelonid alphaherpesvirus 5 (ChHV5) UL30 gene was amplified and sequenced for phylogeny; Bayesian reconstruction showed a high alignment with the genus Scutavirus of the Eastern Pacific group. This is one of the first reports of ChHV5 in a cutaneous fibropapilloma of a black turtle in the Baja California peninsula.

Presence of chelonid herpesvirus 5 (ChHV5) in sea turtles in northern Sinaloa, Mexico.

The presence of fibropapilloma and its associated chelonid herpesvirus 5 (ChHV5) was assessed in 82 wild sea turtles. Olive ridley turtles Lepidochelys olivacea (n = 58) were caught in the pelagic Area of Marine Influence (AMI) (off the coast of Guasave, Sinaloa), and black turtles Chelonia mydas agassizii (n = 24) were captured in the Navachiste Lagoon System. The apparent physical condition was evaluated as 'good' or 'poor' by physical examination. The population structure and general health status was determined by condition index, hematocrit and total plasma protein. Detection of ChHV5 from skin samples was done by PCR. The overall physical condition of black turtles was good and all the individuals were tumor-free. Likewise, the physical condition of most olive ridley turtles was good, except for 10 individuals with poor condition. Of these, 4 had fibropapilloma-like tumors. PCR analyses showed that 3 tumors were ChHV5-positive. The DNA sequence showed 96% identity with ChHV5. All other skin samples from black or olive ridley turtles were ChHV5-negative. This is the first report of fibropapillomatosis-ChHV5 in foraging grounds off northern Sinaloa. The virus was present in a small proportion of L. olivacea individuals, a free-ranging species. It is suggested that infected turtles acquired the virus at a different location somewhere during their development before arriving in the AMI zone. This finding makes the case for setting up a health monitoring program for turtle populations in the area, enforcing sanitary measures to reduce the spread of the pathogen.

Isolation, characterization, and antibiotic resistance of Vibrio spp. in sea turtles from Northwestern Mexico.

The aerobic oral and cloacal bacterial microbiota and their antimicrobial resistance were characterized for 64 apparently healthy sea turtles captured at their foraging grounds in Ojo de Liebre Lagoon (OLL), Baja California Sur (BCS), Mexico (Pacific Ocean) and the lagoon system of Navachiste (LSN) and Marine Area of Influence (MAI), Guasave, Sinaloa (Gulf of California). A total of 34 black turtles (Chelonia mydas agassizii) were sampled in OLL and eight black turtles and 22 olive ridley turtles (Lepidochelys olivacea) were sampled in LSN and MAI, respectively from January to December 2012. We isolated 13 different species of Gram-negative bacteria. The most frequently isolated bacteria were Vibrio alginolyticus in 39/64 (60%), V. parahaemolyticus in 17/64 (26%), and V. cholerae in 6/64 (9%). However, V. cholerae was isolated only from turtles captured from the Gulf of California (MAI). Among V. parahaemolyticus strains, six O serogroups and eight serovars were identified from which 5/17 (29.4%) belonged to the pathogenic strains (tdh (+) gene) and 2/17 (11.7%) had the pandemic clone (tdh (+) and toxRS/new (+)). Among V. cholerae strains, all were identified as non-O1/non-O139, and in 4/6 (66%) the accessory cholera enterotoxin gene (ace) was identified but without virulence gene zot, ctxA, and ctxB. Of the isolated V. parahaemolyticus, V. cholerae, and V. alginolyticus strains, 94.1, 33.4, and 100% demonstrated resistance to at least one commonly prescribed antibiotic (primarily to ampicillin), respectively. In conclusion, the presence of several potential (toxigenic) human pathogens in sea turtles may represent transmission of environmental microbes and a high-risk of food-borne disease. Therefore, based on the fact that it is illegal and unhealthy, we discourage the consumption of sea turtle meat or eggs in northwestern Mexico.

Perceptions of fishers to sea turtle bycatch, illegal capture and consumption in the San Ignacio-Navachiste-Macapule lagoon complex, Gulf of California, Mexico.

In this study, 10% of all registered fishermen in the coastal towns of Navachiste in Sinaloa, in northwestern Mexico, answered a survey designed to collect data on their perceptions of the following topics: the impact of turtle meat consumption; human health; bycatch; illegal turtle fishing; the illegal sea turtle market; the local economy; pollution; environmental education; the success of protective legislation; and sea turtle-based ecotourism. Perceptions were analyzed using the fuzzy logic method through classification into 5 fuzzy membership sets: VL, very low; L, low; M, moderate; H, high; VH, very high. The 9 topics generated decision areas upon applying fuzzy inference that revealed the membership level of the answers in each fuzzy set. The economic potential of sea turtle-based ecotourism and the economic profitability of the illegal turtle meat market were perceived as VL. Conservation legislation was perceived as H, although inefficiently applied due to corruption. Ecotourism and impacts on sea turtles were perceived as VL, because they were deemed unprofitable activities at the individual and community levels. Environmental education was perceived as L, because it centers on nesting, hatching and releasing turtles and is directed at elementary and middle-school students. While fishers perceive a serious negative impact of fishing activities on sea turtles in the San Ignacio-Navachiste-Macapule area, they do not see themselves individually as part of the problem. Achieving sea turtle conservation in this region requires: suitable ecotourism infrastructure, government investments in promotion, and studies to estimate the minimum number of tourists needed to assure profitability.