Critical areas for sea turtles in Northeast Brazil: a participatory approach for a data-poor context.

Fishing is one of the main threats to sea turtles due to the risk of entanglement in lost nets, vessel collision and mortality due to incidental catches. In Brazil, most of the studies regarding fishing interactions with sea turtles are focused on pelagic longline fisheries in the South and Southeast regions. However, their main reproductive areas in Southwest Atlantic RMU occur in Northeast Brazil, which overlaps small-scale coastal gillnet fisheries. Here, we aimed to use ethnobiology and participatory approaches as simple and cost-effective methods to identify areas for sea turtle conservation where impacts from small-scale fisheries are most likely. Expert captains were trained to recording sea turtle sightings during navigations from the landing port to the fishing grounds, informing their folk nomenclatures. By interpolation of environmental data (chlorophyll and bathymetry) and ecological data, we predicted habitats of 3,459.96 km² for Caretta caretta, Chelonia mydas, and Eretmochelys imbricata and fishing zones of 1,087 km² for management in 20 m and 50 m depths. Our results contributes to ongoing discussions of bycatch mitigation for sea turtle species and identification of habitats. We highlights the importance of considering particularities of overlapped areas in marine spatial planning and co-management arrangements.

Traces of oil in sea turtle feces.

In 2019, an oil spill hit the Brazilian Northeast coast causing impact to several ecosystems, including sea turtles' breeding and feeding areas. This study aimed to investigate whether sea turtles were impacted by this oil disaster, correlating the oil found inside feces with a sandy-oiled sample collected on the beach some days after the accident. The fecal samples were collected in the upper mid-littoral reef areas during three consecutive days in February 2020. The results suggested that sea turtles consumed algae contaminated by petroleum. Hydrocarbons composition of oil inside feces was similar to the sandy-oiled sample, suggesting they were the same. Lighter aliphatic and polycyclic aromatic compounds were missing, indicating both sandy-oiled and oil inside the feces had experienced significant evaporation prior to collection. Although the long-term damage is still unknown, the data are novel and relevant to support future research and alert authorities about the risks to sea turtles.

Endoscopy and rectal enema for fecal collection in wild sea turtles (Chelonia mydas, Eretmochelys imbricata) in a field setting.

Forty-seven free-ranging sea turtles (46- Chelonia mydas, 1- Eretmochelys imbricata) were examined via novel use of an endoscopy combined with a rectal enema to obtain large fecal sample volumes. The cloaca was insufflated using an endoscope, after which the bladder and rectum separated, allowing access to the colon. Environmental conditions and location influenced the performance of the procedure initially, but after several attempts the procedure was successfully initiated. In all cases, fecal samples were obtained, and the animals were released to their respective locations. Fecal sample collection using this approach enhances the ability to obtain diagnostic information and perform other scientific analyses of sea turtles.

Goldfish or aquatic turtle? Impact of two animal assisted interventions on children's pain, anxiety, and fear during IV catheterization: A randomized controlled trial.

PURPOSE: This study aimed to determine the effect of goldfish and aquatic turtle nursing interventions on children's pain, anxiety, and fear during IV catheter insertion. DESIGN AND METHODS: The randomized controlled trial included 98 children (control group = 32, goldfish group = 33, aquatic turtle group = 33) aged 5-6 years, attending outpatient clinic for blood sampling and catheterisation procedures. Pain was measured using the Wong Baker Facial Pain Scale, anxiety was measured using the Child Anxiety Scale-State, and child fear was measured using the Child Fear Scale, before, during, and after catheterization. RESULTS: When analyzing the pain, anxiety, and fear findings, no statistical difference was found between the scores of the mother, child, and researcher before the procedure. During the catheterization procedure, the pain, anxiety, and fear scores of the children in the fish and turtle groups increased statistically less than those in the control group. When analyzing the post-procedure findings, the pain, anxiety, and fear scores of the control group were similar to those before the procedure. In the fish and turtle groups, anxiety and fear scores were statistically the lowest (p < 0.05). CONCLUSION: It can be concluded that animal-assisted practices reduce anxiety and fear during and after the catheterization procedures while diverting attention from pain. PRACTICE IMPLICATIONS: Goldfish or aquatic turtle intervention improve children's attitudes toward pain, anxiety, and fear during IV catheter insertion. Pediatric nurses could consider incorporating, such as the animal assisted intervention, into their clinical practice to reduce the pain, anxiety, and fear experienced by children during medical/nursing procedures.

Bioaccumulation of mercury in Lake Michigan painted turtles (Chrysemys picta).

Mercury (Hg) contamination of aquatic environments can lead to bioaccumulation in organisms, but most previous work has focused on fish and not on semi-aquatic reptiles such as turtles that traverse both terrestrial and aquatic habitats. Here, we analyzed total Hg (THg) concentrations in 30 painted turtles (Chrysemys picta) collected from Lake Michigan (USA) coastal wetlands in 2013 to determine if (1) turtles bioaccumulated THg from the environment, (2) concentrations differed between turtle liver and muscle tissue, and (3) tissue concentrations were related to environmental concentrations (e.g., sediment THg). All individual turtles had detectable THg concentrations in both liver and muscle tissue. On average, THg concentrations were over three times higher in liver tissue compared to muscle tissue. We found a positive linear relationship between muscle THg concentrations and turtle body mass, a proxy for age, suggesting bioaccumulation in this species. Neither liver nor muscle THg concentrations followed the sediment contaminant gradient in the wetlands. Despite this, location was a strong predictor of tissue concentration in a linear model suggesting that other site-specific characteristics may be important. Overall, our results demonstrate that painted turtles accumulate mercury in liver and muscle tissues at different rates, which may be constrained by local conditions. Further research is needed to better understand the relationship between environmental mercury concentrations and body burdens in animals like turtles that traverse habitats. In addition, long-lived turtles could be incorporated into pollution monitoring programs to provide a more holistic picture of food web contamination and ecosystem health.

Effects of dietary chito-oligosaccharide and ß-glucan on the water quality and gut microbiota, intestinal morphology, immune response, and meat quality of Chinese soft-shell turtle (Pelodiscus sinensis).

Chito-oligosaccharides (COS) and ß-glucan are gradually being applied in aquaculture as antioxidants and immunomodulators. However, this study examined the effects of dietary supplementation of COS and ß-glucan on the water quality, gut microbiota, intestinal morphology, non-specific immunity, and meat quality of Chinese soft-shell turtle. To investigate the possible mechanisms, 3-year-old turtles were fed basal diet (CK group) and 0.1%, 0.5%, and 1% COS or ß-glucan supplemented diet for 4 weeks. Colon, liver, blood and muscle tissues, colon contents, water and sediment of paddy field samples were collected and analyzed after feeding 2 and 4 weeks. The results indicated that COS and ß-glucan altered microbial community composition and diversity in Chinese soft-shell turtles. The relative abundance of Cellulosilyticum, Helicobacter and Solibacillus were increased after feeding COS, while Romboutsia, Akkermansia and Paraclostridium were increased after feeding ß-glucan, whereas Cetobacterium, Vibrio and Edwardsiella were enriched in the control group. Furthermore, colon morphology analysis revealed that COS and ß-glucan improved the length and number of intestinal villi, and the effect of 0.5% ß-glucan was more obvious. Both ß-glucan and COS significantly improved liver and serum lysozyme activity and antibacterial capacity. COS significantly increased the total antioxidant capacity in the liver. Further, 0.1% ß-glucan significantly increased the activity of hepatic alkaline phosphatase, which closely related to the bacteria involved in lipid metabolism. Moreover, dietary supplementation with 1% COS and 1% ß-glucan significantly enhanced the content of total amino acids, especially umami amino acids, in muscle tissue, with ß-glucan exerting a stronger effect than COS. Additionally, these two prebiotics promoted the quality of culture water in paddy fields and reshaped the bacterial community composition of aquaculture environment. All these phenotypic changes were closely associated with the gut microbes regulated by these two prebiotics. In summary, the findings suggest that dietary supplementation with COS and ß-glucan in Pelodiscus sinensis could modulate the gut microbiota, improve intestinal morphology, enhance non-specific immunity and antioxidant capacity of liver and serum, increase meat quality, and improve the culture water environment. This study provides new insights and a comprehensive understanding of the positive effects of COS and ß-glucan on Pelodiscus sinensis.

Effects of dietary sodium butyrate on growth performance, immune function, and intestinal microflora of Chinese soft-shelled turtle (Pelodiscus sinensis).

The Chinese soft-shelled turtle (Pelodiscus sinensis) has become increasingly susceptible to frequent diseases with the intensification of farming, which severely impacts the development of the aquaculture industry. Sodium butyrate (SB) is widely used as a feed additive due to its promotion of growth, enhancement of immune function, and antioxidative properties. This study aimed to investigate the effects of dietary SB on the growth performance, immune function, and intestinal microflora of Chinese soft-shelled turtles. A total of 300 Chinese soft-shelled turtles (mean weight: 11.36 ± 0.21g) were randomly divided into four groups with three parallel sets in each group. Each group was fed a diet supplemented with 0%, 0.005%, 0.01%, or 0.02% SB for 60 days. The results demonstrated an upward trend in weight gain rate (WGR) and specific growth rate (SGR) with increasing SB supplementation, and the experimental group fed with 0.02% SB showed a significant increase in WGR and SGR compared to other groups (P< 0.05). These levels of SB also decreased the levels of feed conversion ratio (FCR) and the total cholesterol (TC) content of Chinese soft-shelled turtles, and the 0.02% SB was significantly lower than that of other groups (P< 0.05). The activity of complement protein in vivo increased with increases in SB content, and the activities of complement C3 and C4 reached the highest level with 0.02% SB. The species abundance of the experimental group D fed with 0.02% SB was significantly higher than that of other groups (P< 0.05). Furthermore, the relative abundance of Clostridium sensu stricto 1 was significantly increased with 0.02% SB (P< 0.05). In conclusion, adding 0.02% SB to the diet improves the growth performance, feed digestion ability, and intestinal microbiota of Chinese soft-shelled turtles.

Understanding contaminant exposure risks in nesting Loggerhead sea turtle populations.

Queensland loggerhead turtle nest numbers at Mon Repos (MR) indicate population recovery that doesn't occur at Wreck Island (WI). Previous research illustrated that MR and WI turtles forage in different locations, potentially indicating risks differences. Blood, scute, and egg were collected from turtles nesting at MR and WI, with known foraging sites (from concurrent studies). Trace element and organic contaminants were assessed via acid digestion and in vitro cytotoxicity bioassays, respectively. WI turtles had significantly higher scute uranium and blood molybdenum compared to MR turtles, and arsenic was higher in WI turtles foraging north and MR turtles foraging south. Egg and blood titanium, manganese, cadmium, barium, lead, and molybdenum, and scute and egg selenium and mercury significantly correlated. Blood (75 %) extracts produced significant toxicity in vitro in turtle fibroblast cells. In conclusion, reducing chemical exposure at higher risk foraging sites would likely benefit sea turtles and their offspring.

Molecular characterization and potential function of Rxrγ in gonadal differentiation of Chinese soft-shelled turtle (Pelodiscus sinensis).

Retinoid X receptor (RXR) is a member of the ligand-dependent nuclear receptor family. Previous studies revealed that RXRs are involved in reproduction in vertebrates. However, information on the function of RXRs in turtles is scarce. In this study, the Rxrγ cDNA sequence of Pelodiscus sinensis was cloned and analyzed, and a polyclonal antibody was constructed. RXRγ protein showed a positive signal in both mature and differentiated gonads of the turtle. Subsequently, the function of the Rxrγ gene in gonadal differentiation was confirmed using short interfering RNA (RNAi). The full-length cDNA sequence of the Rxrγ gene in P. sinensis was 2152 bp, encoding 407 amino acids and containing typical nuclear receptor family domains, including the DNA-binding domain (DBD), ligand-binding domain (LBD), and activation function 1 (AF1). Moreover, gonadal Ps-Rxrγ showed sexual dimorphism expression patterns in differentiated gonads. Real-time quantitative PCR results revealed that the Rxrγ gene was highly expressed in the turtle ovary. RNAi treatment increased the number of Sertoli cells in ZZ embryonic gonads. Furthermore, RNA interference upregulated Dmrt1 and Sox9 in ZZ and ZW embryonic gonads. However, Foxl2, Cyp19a1, Stra8, and Cyp26b1 were downregulated in embryonic gonads. The results indicated that Rxrγ participated in gonadal differentiation and development in P. sinensis.

The mediation of the AHR/IL-22/STAT3/IL-6 axis by soft-shelled turtle (Pelodiscus sinensis) peptide and Chinese pond turtle (Chinemys reevesii) peptide contributed to their amelioration effects on intestinal mucosal immunity in immunosuppressed mice.

This study investigated the immunomodulatory effects of soft-shelled turtle (Pelodiscus sinensis) peptide (TP) and Chinese pond turtle (Chinemys reevesii) peptide (TMP) on the intestinal mucosal immune system (IMIS). The results demonstrated that TP and TMP improved holistic immunity by restoring the vital immune organ atrophy and proliferation capacity of spleen immune cells. Moreover, TP and TMP significantly increased the serum content of IgA and cytokines that are responsible for immune cell activation and antigen clearance. TP and TMP promoted intestinal B cell activation, class switching recombination, and antibody secreting processes in a T cell-independent manner to increase the SIgA content. Furthermore, TP and TMP enhanced the intestinal barrier by increasing the protein expression of tight junctions (TJs) and adhesion junctions (AJs) and ameliorating the intestinal morphology. Mechanistically, TP and TMP activated the AHR/IL-22/STAT3/IL-6 axis to enhance the IgA response and improve the intestinal barrier, indicating their potential in intestinal health modulation.

B-esterase measurements and other blood related biomarkers in loggerhead sea turtles (Caretta caretta) as indicators of health status.

The loggerhead sea turtle (Caretta caretta) has been selected as sentinel species by the Marine Strategy Framework Directive (MSFD) descriptor 10 in relation to marine litter. In this, and other protected species, there is a need to develop conservative pollution biomarkers equally informative of chemical exposures to those traditionally carried out in metabolic organs, such as the liver. With this aim, plasma from turtles undergoing rehabilitation at the Fundació Oceanogràfic rescue centre (Arca del Mar) were selected and tested for B-esterase measurements. Hydrolysis rates of acetylcholinesterase (AChE), butyrylcholinesterase (BChE) and carboxylesterases (CEs) using four commercial substrates were undertaken on 191 plasma samples. Results indicated that acetylthiocholine was the most adequate substrate of cholinesterases and butyrate esters for CE measures. The correlation of these parameters with well-established blood biochemistry measurements was analysed. B-esterase measures in wild specimens were discussed in relation to age group, pathology on admission to the rescue centre and season; moreover, contrasts with long-term resident turtles were also made. Although this study provides baseline data on B-esterase measures in a large sample size for this species, more complementary information is still needed in terms of population genetics, chemical exposures, and in relation to other biochemical parameters before they can be confidently applied in wild specimens within the regulatory MSFD.

Screening of anti-liver fibrosis peptides from turtle shell protein using two-enzyme hydrolysis by molecular docking.

Turtle shell as a food residue of Pelodiscus sinensis (a type of edible aquatic animal) is widely used in Traditional Chinese Medicine for hepatic fibrosis therapy. Previous studies have demonstrated that the peptides (<6 kDa) derived from turtle shells are considered effective components. The protein of turtle shells has important potential as a source of bioactive peptides which may play a role as ingredients in functional foods. In the present study, the protein of turtle shell was hydrolyzed using a two-enzyme combination. It was found that the hydrolysates obtained by a combination of pepsin and trypsin showed the highest anti-liver fibrosis activity relative to other combinations in a cell viability assay. The hydrolysates were separated and purified by ultra-filtration (<6 kDa), gel filtration chromatography (GFC) and high-performance liquid chromatography (HPLC). Subsequently, the sequences of purified peptides were analyzed by liquid chromatography-mass spectrometry (LC-MS/MS). Molecular docking was used to analyze the interaction of these peptides with the transforming growth factor-ß1 (TGF-ß1) receptor. Two (GPPGVPGPGPL, TSLPVPAPV) of these novel peptides displayed lower binding energies to the TGF-ß1 receptor (-8.18 kcal mol-1, -8 kcal mol-1). Finally, the above two peptides were synthesized chemically and their in vitro anti-liver fibrosis activity was verified by MTT assay. Among them, GPPGVPGPGPL showed a better in vitro anti-liver fibrosis activity (IC50: 80.13 µM). We established a method to obtain anti-liver fibrosis peptides from turtle shells by using bioactivity-guided isolation with molecular docking. Turtle shell protein is an excellent source of anti-liver fibrosis peptides which can offer therapeutic and commercial benefits as an ingredient in functional foods.

Distocia em tigre d'água-de-orelha-vermelha, Trachemys scripta elegans, wied (1839): relato de caso / Dystocia in red-eared slider, Trachemys scripta elegans, wied (1839): case report / Distocia en tortuga de orejas rojas, Trachemys scripta elegans, wied (1839): reporte de caso

Atualmente muitos répteis se tornaram animais de companhia e são mantidos como pet's exóticos. A espécie Trachemys scripta elegans, Wied (1839) é um animal exótico da América do Norte, sua identificação é realizada pelas marcas avermelhadas encontradas lateralmente a sua cabeça. Na rotina clínica as principais enfermidades que acometem os quelônios são as de origem reprodutiva, como a estase folicular e distocia. O objetivo deste trabalho foi relatar um caso recorrente de distocia em um tigre d'água fêmea, para isso, a anamnese, o histórico da paciente, e seus sinais clínicos, em conjunto com os exames complementares de imagem foram essenciais para se obter diagnóstico definitivo. O tratamento foi realizado com a indução medicamentosa utilizando borogluconato de cálcio, seguida da aplicação de ocitocina, esta trouxe resultados positivos para a eliminação dos ovos. Porém devido ao histórico do paciente, optou-se pela intervenção cirúrgica de ovariossalpingectomia, sendo está a maneira permanente de resolução da patologia. O protocolo terapêutico escolhido proporcionou um resultado satisfatório e bem estar ao animal.(AU)

Currently, many reptiles have become companion animals and are kept as exotic pets. The species Trachemys scripta elegans, Wied (1839) is an exotic animal from North America, and its identification is based on the reddish markings found laterally on its head. In routine clinical practice, the main diseases that affect chelonians are those of reproductive origin, such as follicular stasis and dystocia. The aim of this study was to report a recurrent case of dystocia in a female red-eared slider turtle. For this purpose, the patient's anamnesis, history, and clinical signs, along with complementary imaging exams, were essential to obtain a definitive diagnosis. The treatment involved medical induction using calcium borogluconate, followed by the administration of oxytocin, which yielded positive results in egg elimination. However, due to the patient's history, surgical intervention in the form of ovariosalpingectomy was chosen as the permanent solution to the pathology. The chosen therapeutic protocol provided a satisfactory outcome and improved the animal's well-being.(AU)

Actualmente muchos reptiles se han convertido en animales de compañía y se mantienen como mascotas exóticas. La especie Trachemys scripta elegans, Wied (1839) es un animal exótico de América del Norte, su identificación se realiza por las marcas rojizas que se encuentran lateralmente a su cabeza. En la rutina clínica, las principales enfermedades que afectan a los quelonios son las de origen reproductivo, como la estasis folicular y la distocia. El objetivo de este trabajo fue reportar un caso recurrente de distocia en una hembra de tigre de agua, para ello la anamnesis, la historia de la paciente y sus signos clínicos, junto con los exámenes imagenológicos complementarios fueron fundamentales para obtener un diagnóstico definitivo. El tratamiento se realizó con inducción farmacológica con borogluconato de calcio, seguido de la aplicación de oxitocina, que arrojó resultados positivos con la eliminación de huevos. Sin embargo, debido a los antecedentes de la paciente, se optó por la intervención quirúrgica de ovarialpingectomía, que es la forma definitiva de resolución de la patología. El protocolo terapéutico elegido proporcionó un resultado satisfactorio y bienestar al animal.(AU)

Soft-Shelled Turtle Peptides Extend Lifespan and Healthspan in Drosophila.

In traditional Chinese medicine, soft-shelled turtle protein and peptides serve as a nutraceutical for prolonging the lifespan. However, their effects on anti-aging have not been clarified scientifically in vivo. This study aimed to determine whether soft-shelled turtle peptides (STP) could promote the lifespan and healthspan in Drosophila melanogaster and the underlying molecular mechanisms. Herein, STP supplementation prolonged the mean lifespan by 20.23% and 9.04% in males and females, respectively, delaying the aging accompanied by climbing ability decline, enhanced gut barrier integrity, and improved anti-oxidation, starvation, and heat stress abilities, while it did not change the daily food intake. Mechanistically, STP enhanced autophagy and decreased oxidative stress by downregulating the target of rapamycin (TOR) signaling pathway. In addition, 95.18% of peptides from the identified sequences in STP could exert potential inhibitory effects on TOR through hydrogen bonds, van der Walls, hydrophobic interactions, and electrostatic interactions. The current study could provide a theoretical basis for the full exploitation of soft-shelled turtle aging prevention.

Survival outcomes of rehabilitated riverine turtles following a freshwater diluted bitumen oil spill.

Rehabilitation is often used to mitigate adverse effects of oil spills on wildlife. With an increase in production of alternatives to conventional crude oil such as diluted bitumen (dilbit), emergency spill responders and wildlife rehabilitators need information regarding the health and survival of free-ranging vertebrates exposed to dilbit under natural conditions. In 2010, one of the largest freshwater oil spills in the United States occurred in the Kalamazoo River in Michigan, when over 3.2 million liters of spilled dilbit impacted 56 km of riverine habitat. During 2010 and 2011 cleanup efforts, thousands of northern map turtles (Graptemys geographica) were captured from oiled stretches of the river, cleaned, rehabilitated, and released. We conducted extensive mark-recapture surveys in 2010, 2011, and 2018-2021, and used this dataset to evaluate the monthly survival probability of turtles 1-14 months post-spill and 8-11 years post-spill based on whether turtles were temporarily rehabilitated and released, overwintered in captivity and then released, or were released without rehabilitation. We found that rehabilitated or overwintered turtles had a higher probability of survival 1-14 months post-spill than non-rehabilitated turtles; however, 8-11 years post-spill the among-group differences in monthly survival probability had become negligible. Additionally, following the oil spill in 2010, nearly 6% of northern map turtles were recovered dead, died during rehabilitation, or suffered injuries that precluded release back into the wild. Our results demonstrate that exposure to dilbit in free ranging turtles causes direct mortality, while effort spent on the capture and rehabilitation of oiled freshwater turtles is important as it increases monthly survival 1-14 months post-spill.

Bioaccumulation of total mercury, copper, cadmium, silver, and selenium in green turtles (Chelonia mydas) stranded along the Potiguar Basin, northeastern Brazil.

Sea turtles face several threats and pollution has become a major concern for their conservation worldwide. We analyzed samples of the liver, muscles, and kidneys of 38 Chelonia mydas stranded along the Potiguar Basin, northeastern Brazil, between 2015 and 2018 to determine the total Hg concentration (THg), as well as the concentrations of Cu, Cd, Ag, and Se. The relation between turtle size and element concentrations revealed a negative correlation for THg and Se (liver, muscles, and kidneys), Cu and Cd (liver and kidneys) and a positive correlation for Ag in the three organs analyzed. Concentrations of THg, Cu, Ag, and Se were high in the liver, highlighting the Cu concentration (median = 25.1150 µg g-1 w.w.), while the kidneys had the highest Cd levels (median = 12.2200 µg g-1 w.w.). There was significant difference between element concentrations and the three organs analyzed, except for Ag and Se concentrations in the muscle and kidney samples. Our study showed that green turtles found in Potiguar Basin, northeastern Brasil, have bioaccumulated inorganic elements which indicate the need of further investigations on the environmental quality of the region.

Evolutionary Dynamics of Host Organs for Microbial Symbiosis in Tortoise Leaf Beetles (Coleoptera: Chrysomelidae: Cassidinae).

Diverse insects host specific microbial symbionts that play important roles for their growth, survival, and reproduction. They often develop specialized symbiotic organs for harboring the microbial partners. While such intimate associations tend to be stably maintained over evolutionary time, the microbial symbionts may have been lost or replaced occasionally. How symbiont acquisitions, replacements, and losses are linked to the development of the host's symbiotic organs is an important but poorly understood aspect of microbial symbioses. Cassidine leaf beetles are associated with a specific gammaproteobacterial lineage, Stammera, whose reduced genome is streamlined for producing pectin-degrading enzymes to assist the host's digestion of food plants. We investigated the symbiotic system of 24 Japanese cassidine species and found that (i) most species harbored Stammera within paired symbiotic organs located at the foregut-midgut junction, (ii) the host phylogeny was largely congruent with the symbiont phylogeny, indicating stable host-symbiont association over evolutionary time, (iii) meanwhile, the symbiont was not detected in three distinct host lineages, uncovering recurrent losses of the ancient microbial mutualist, (iv) the symbiotic organs were vestigial but present in the symbiont-free lineages, indicating evolutionary persistence of the symbiotic organs even in the absence of the symbiont, and (v) the number of the symbiotic organs was polymorphic among the cassidine species, either two or four, unveiling a dynamic evolution of the host organs for symbiosis. These findings are discussed as to what molecular mechanisms and evolutionary trajectories underpin the recurrent symbiont losses and the morphogenesis of the symbiotic organs in the herbivorous insect group. IMPORTANCE Insects represent the biodiversity of the terrestrial ecosystem, and their prosperity is attributable to their association with symbiotic microorganisms. By sequestering microbial functionality into their bodies, organs, tissues, or cells, diverse insects have successfully exploited otherwise inaccessible ecological niches and resources, including herbivory enabled by utilization of indigestible plant cell wall components. In leaf beetles of the subfamily Cassininae, an ancient symbiont lineage, Stammera, whose genome is extremely reduced and specialized for encoding pectin-degrading enzymes, is hosted in gut-associated symbiotic organs and contributes to the host's food plant digestion. Here, we demonstrate that multiple symbiont losses and recurrent structural switching of the symbiotic organs have occurred in the evolutionary course of cassidine leaf beetles, which sheds light on the evolutionary and developmental dynamics of the insect's symbiotic organs and provides a model system to investigate how microbial symbionts affect the host's development and morphogenesis and vice versa.

Health and nutrition of loggerhead sea turtles (Caretta caretta) in the southeastern United States.

Loggerhead sea turtles (Caretta caretta) are opportunistic carnivores that feed primarily on benthic invertebrates and fish. Sea turtle rehabilitation requires provision of a species-specific, balanced diet that supplies nutrition similar to that of a wild diet; this can be challenging because free-ranging loggerheads' diets vary depending on their life stage and geographic location, with predominant prey species dictated by local availability. The goal of this study was to better understand the nutritional needs of subadult and adult loggerheads in rehabilitation. This was accomplished by conducting a retrospective survey of stomach contents identified during gross necropsy of 153 deceased loggerheads that stranded in coastal Georgia, USA. A total of 288 different forage items were identified; the most frequently observed prey items belong to the subphylum Crustacea (N = 131), followed by bony fish (Osteichthyes; N = 45), gastropod mollusks (N = 40), bivalve mollusks (N = 23), and Atlantic horseshoe crabs (Limulus polyphemus; N = 15). The proportions of certain prey items differed significantly with turtle size; adult turtles ate proportionately more gastropods (p = 0.001), and subadults ate proportionately more fish (p = 0.01). Stomach contents information was used to determine common local prey items (blue crab, cannonball jellyfish, horseshoe crab, whelk), which were evaluated for nutritional content. Additionally, we compared hematology and plasma biochemistry profiles (including proteins, trace minerals, and vitamins) between four cohorts of loggerhead turtles, including free-ranging subadults and adults, nesting females, and loggerheads undergoing rehabilitation. This information was applied to inform a regionally specific, formulated diet for tube feeding, and a supplement containing vitamins and minerals for captive loggerheads, to more closely approximate the nutritional content of their natural diet. Assessing the regional and temporal variability in loggerhead diets is an important component in their effective conservation because resultant data can be used to help understand the impacts of environmental perturbations on benthic food webs.

Effect of dietary soybean saponin Bb on the growth performance, intestinal nutrient absorption, morphology, microbiota, and immune response in juvenile Chinese soft-shelled turtle (Pelodiscus sinensis).

Soybean meal is widely applied in the aquafeeds due to the limitation of fish meal resources. Numerous studies have manifested that dietary soybean saponin, an anti-nutrient factor in soybean meal, may slow growth and induce intestinal inflammation in aquatic animals, but the possible causes are unclear. The juvenile Pelodiscus sinensis (mean initial body weight: 6.92 ± 0.03 g) were fed basal diet (CON group) and 2.46% soybean saponin Bb-supplemented diet (SAP group) for 35 days to further explore the effects of dietary soybean saponin Bb on the growth performance, apparent digestibility coefficients, intestinal morphology, the gut microbiota, intestinal transporters/channels, and immune-related gene expression. The results indicated that dietary soybean saponin Bb significantly decreased final body weight, specific growth rate, protein deposition ratio, and apparent digestibility coefficients (dry matter, crude protein, and crude lipid) of nutrients in Pelodiscus sinensis, which may be closely correlated with markedly atrophic villus height and increased lamina propria width in the small intestine. In addition, plasma contents of cholesterol, calcium, phosphorus, potassium, lysozyme, and C3 were significantly decreased in the SAP group compared with the control group. Soybean saponin Bb significantly downregulated the mRNA levels of glucose transporter 2, fatty acid binding protein 1 and fatty acid binding protein 2, amino acid transporter 2, b0,+-type amino acid transporter 1, and sodium-dependent phosphate transport protein 2b in the small intestine. At the same time, the expressions of key transcription factors (STAT1, TBX21, FOS), chemokines (CCL3), cytokines (TNF-α, IL-8), and aquaporins (AQP3, AQP6) in the inflammatory response were increased by soybean saponin Bb in the large intestine of a turtle. Additionally, dietary supplementation of SAP significantly reduced the generic abundance of beneficial bacteria (Lactobacillus, Bifidobacterium, and Bacillus) and harmful bacteria (Helicobacter and Bacteroides). In a nutshell, dietary supplementation of 2.46% soybean saponin not only hindered the growth performance by negatively affecting the macronutrients absorption in the small intestine but also induced an inflammatory response in the large intestine possibly by damaging the intestinal morphology, disturbing the intestinal microbiota and decreasing intestinal epithelial cell membrane permeability.