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Background: Chinese soft-shell turtle (Pelodiscus sinensis) is an important commercial species for their high nutritional value and unique taste, but it has been a vulnerable species due to habitat loss. In this study, homologous juvenile turtles were allocated to lake, pond and paddy field to investigate the habitat effects on turtles., Results: The growth, morphology and gut microbial communities were monitored during the 4 months cultural period. It showed higher growth rate of turtles in paddy field and pond. The appearance, visceral coefficients, gut morphology and microbial communities in turtles were distinct among different habitats. The microbial community richness on Chao1 was obviously lower in initial turtle guts from greenhouses, whereas it was relative higher in turtle guts sampled from paddy fields than ponds and lake. Significant differences on dominant microbes were found among initial and subsequent samples from different habitats. Firmicutes was the most abundant phylum in the guts of turtles sampled from the greenhouse initially, while Proteobacteria was the most abundant phylum after cultivation in different habitats, followed by Bacteroidetes. The microbial composition were distinct in different habitats at 60d, and the appearance of dominant phyla and genera was more driven by sampling time than habitats at 120d. Both the sampling time and habitats affected the appearance of dominant phyla and genera during the cultivation. The functional predictions indicated that both habitat type and sampling time had significant effects on metabolic pathways, especially amino acid and carbohydrate metabolism., Conclusions: The turtles could adapt to natural lakes, artificial ponds and paddy fields. The gut microbial abundance was different among the habitats and sampling time. The species of microbes were significantly more diverse in paddy field specimens than in those from ponds and lakes. Rice-turtle coculture is a potential ecological and economic farming mode that plays important roles in wild turtle protection and food security.

Peptidoglycan recognition proteins (PGRPs), which are discovered in invertebrates and vertebrates, play an important role in antibacterial immunity. However, the function of PGRPs is largely uninvestigated in reptiles. In the present study, a short-type PGRP gene, designed as C-turtle-PGRP-S, was identified in the Chinese soft-shelled turtle, Pelodiscus sinensis. The C-turtle-PGRP-S contains a highly conserved PGRP domain and has close relationship with PGRP-S orthologues in other species according to sequence and phylogenetic analyses. C-turtle-PGRP-S gene was constitutively expressed in all detected tissues and was induced by Edwardsiella tarda. Additionally, recombinant C-turtle-PGRP-S showed PGN binding activity and antibacterial function against E. tarda. Therefore, it is suggested that the function of PGRP-S is likely to be conserved in reptile vertebrates, as observed in other vertebrates, shedding light on the evolutionary conservation of PGRPs., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Muscle is an important structural tissue in aquatic animals and it is susceptible to bacterial and fungal infection, which could affect flesh quality and health. In this study, Chinese soft-shelled turtles were artificially infected with two pathogens, Proteus vulgaris and Elizabethkingia meningoseptica and the effects on muscle nutritional characteristics, oxidative stress and autophagy were assayed. Upon infection, the muscle nutritional composition and muscle fiber structure were notably influenced. Meanwhile, the mRNA expression of Nrf2 was down-regulated and Keap1 up-regulated, thus resulting in a decrease in antioxidant capacity and oxidative stress. However, with N-acetylcysteine treatment, the level of oxidative stress was decreased, accompanied by significant increases in antioxidant enzyme activities and the mRNA levels of SOD, CAT, GSTCD, and GSTO1. Interestingly, there was a significant increase in autophagy in the muscle tissue after the pathogen infection, but this increase could be reduced by N-acetylcysteine treatment. Our findings suggest that muscle nutritional characteristics were dramatically changed after pathogen infection, and oxidative stress and autophagy were induced by pathogen infection. However, N-acetylcysteine treatment could compromise the process perhaps by decreasing the ROS level and regulating Nrf2-antioxidant signaling pathways.

Providencia rettgeri infection has occurred occasionally in aquaculture, but is rare in turtles. Here, a pathogenic P. rettgeri strain G0519 was isolated from a diseased slider turtle (Trachemys scripta) in China, and qPCR assay was established for the RTX toxin (rtxD) gene. Histopathological examination showed that many inflammatory cells were infiltrated into heart, liver and intestine, as well as the necrosis of liver, kidney and spleen. The genome consisted of one circular chromosome (4.493 Mb) and one plasmid (18.8 kb), and predicted to contain 4170 and 19 protein-coding genes, respectively. Multiple pathogenic and virulence factors (e.g., fimbria, adhesion, invasion, toxin, hemolysin, chemotaxis, secretion system), multidrug-resistant genes (e.g., ampC, per-1, oxa-1, sul1, tetR) and a novel genomic resistance island PRI519 were identified. Comparative genome analysis revealed the closest relationship was with P. rettgeri, and with P. heimbachae closer than with other Providencia spp. To our knowledge, this was first report on genomic characterization of multidrug-resistant pathogenic P. rettgeri in cultured turtles.

Salmonella Pomona, a highly pathogenic serotype, can cause severe human salmonellosis, especially in children. Turtles and other reptiles are reservoirs for S. Pomona, and these cold-blooded animals remain a source of human Salmonella infections. Since the 1980s, this serotype has become a significant public health concern because of the increasing number of cases of S. Pomona infection in humans. To date, outbreaks of Salmonella Pomona infection in humans have mainly occurred in the United States, with some in other countries (e.g. Belgium, Germany, Canada), and most of the infections in humans were associated with turtles and other reptiles. In China, S. Pomona was first isolated from the feces of an infant in Shanghai in 2000, and two further cases of S. Pomona infection in humans were later found in Guangzhou. No one knew the source of S. Pomona in China. In this study, for the first time we isolated S. Pomona from free-living exotic red-eared sliders in the wild in China. Salmonella serotype (S. Pomona) was isolated from 16 turtle samples. The total carrying rate of S. Pomona in the collected red-eared sliders was 39% (n = 41) overall: 40% (n = 25) in juveniles and 38% (n = 16) in adult turtles. This study suggests that the widespread exotic red-eared sliders may impact on public health and ecosystems of China by transmitting S. Pomona. Additional steps should be considered by the governments and public health agencies to prevent the risk of turtle-associated Salmonella infections in humans in China., (© 2013.)

Objective: Vibrio cholera was extremely rare in Sichuan province(no cases in 2008). Any outbreak could indicate contamination through the food supply system. In July 2009, a hospital reported a cluster of 7 diarrhea patients;all attended the same banquet. One patient was confirmed to have Vibrio cholera (O139). We conducted this investigation to identify the source of this possible cholera outbreak., Methods: We defined a suspect case as any banquet attendee with diarrhea (≥ 3 times/day). A confirmed case was a suspect case with a positive Vibrio cholera culture. We took stool samples or rectal swabs from all attendees for cholera culture and interviewed 272 banquet attendees about foods they ate at the banquet and kitchen workers about food preparation., Results: 7.1% (24/337) of attendees developed cases within an average of 65 hours after eating. Three meals were served. All patients had the lunch whereas no patients only ate breakfast and/or dinner. Of 180 attendees who ate turtle meat 12% were case-patients, compared to 3.3% of 92 attendees who did not (RR = 3.6, 95%CI: 1.1 - 12). Of the 150 attendees who ate peanuts 13% were cases compared to 4.1% of 122 attendees who did not eat peanuts (RR = 3.1, 95%CI: 1.2 - 8.0). During preparation, the same utensil was used for fresh turtle meat and peanuts without washing in-between the process. Turtle meat and peanuts were stored for > 16 hours at room temperature after cooking before consumption. All 33 turtles originated from commercial production in another province., Conclusion: This outbreak was likely caused by poor food handling of commercially produced turtles. We proposed that to improve microbiologic monitoring of aquatic food animals, and raise the awareness of good handling practices at mass gathering in rural China.

Objective: To study the correlation between Vibrio cholerae strains isolated from natural enviroment and fishery products and the source of infection during V. cholerae outbreaks., Methods: Cholera toxin gene was detected by polymerase chain reaction (PCR) amplification. Pulsed-field gel electrophoresis (PFGE) was used to subtype the isolates. Results of PFGE were analyzed and clustered by BioNumerics software (Version 4.0)., Results: During the outbreaks, a total number of thirty O139 V. cholerae related serogroups were collected from patients, carriers, sewage and fishery products were identified and proved to be toxigenic. They could be clustered into four PFGE patterns when digested by Not I. These two V. cholerae outbreaks were caused by the same source of infection because of the following reasons: (1) PFGE patterns of the predominant strains isolated from two outbreaks were identical; (2) they were identical to the PFGE patterns of the strains isolated from the green turtle and rana catesbiana which were bought from the same wholesale store., Conclusion: Green turtle and rana catesbiana that were contaminated by toxigenic O139 V. cholerae strains seemed to be the source of infection causing the O139 V. cholerae outbreaks in Jiangxi province. Rapid laboratory surveillance and epidemiologic investigation were important in identifying the source of infection during the outbreaks of V. cholera.

Objective: To understand the pollution rates of vibrio cholera (V. cholera) in different seafood, aquatic products and their circulatory processes, so as to help making measures for cholera control and prevention., Methods: Different seafood, aquatic products and breed water specimen collected from 12 provinces of China were tested from July to September in 2005., Result: A total of 12 104 samples of seafood and aquatic products were tested and the average pollution rate of vibrio cholera was 0.52%. The positive isolate rate of turtle sample (1.72%) was the highest among all samples. The second higher isolated rate was 1.14% in water specimen of turtle breed pool. The positive rate of bullfrog was 0.50%. The percentage of toxin strains was 47.54% and 79.31% of them were isolated from turtle and water samples of turtle breed pool. The important sector of the pollution of vibrio cholera was in turtle breed pool (2.38%)., Conclusion: The average pollution rate of vibrio cholera in seafood and aquatic products in 12 provinces of China was low. It should be very necessary to supervise the sanitation in turtle breed for controlling and preventing the vibrio cholera.

Objective: To study the characteristics of molecular typing and phylogenic relationship among the Vibrio cholerae serogroup O139 strains isolated from environment and sea food samples during cholera outbreaks, in Sichuan province in 2004 and to trace the source of infections so as to support the ascertainment of epidemic control strategy., Methods: Cholera toxin gene was detected by polymerase chain reaction amplification. Pulsed-field gel electrophoresis (PFGE) was used for subtyping of isolates and clustering of patterns was analysed with the software BioNumerics., Results: In all the 72 strains under analysis, 68 appeared to be toxigenic while 4 from river water derived isolates were toxin gene negative. Sixty-seven strains were clustered into 16 PFGE patterns when digested with Not I. The patterns of toxigeinc O139 strains isolated from turtles in the markets were identical with the patterns of strains appeared in the outbreaks respectively. The PFGE patterns of isolates from different outbreaks were inconsistent., Conclusion: The sources of infection causing these outbreaks were complicated. Contaminated turtles might also be one of the major sources of outbreaks when being served at the dinner parties in Sichuan in 2004.