Effects of water temperature on growth performance, digestive enzymes activities, and serum indices of juvenile Coreius guichenoti.

This study was conducted to determine the optimal temperature for Juvenile Coreius guichenoti, which is critically endangered in China. Test fish with an initial body weight range of 17.60-18.78 g were reared in 5 independent recirculating aquaculture systems (RAS) with three tanks (50 fish/tank) for 8 weeks. The water temperature of the RAS was set at 14 °C, 18 °C, 22 °C, 26 °C, and 30 °C, respectively. At conclusion of the feeding trial, the effects of water temperature on C. guichenoti were analyzed based on survival, growth performance, digestive enzymes activities, and antioxidant capacity. The results showed that the fish survived in all temperature groups. The fish in the groups (26 °C and 30 °C) had higher values of body length, body weight, specific growth rate, weight gain rate, and daily gain than those in the group (22 °C). The lowest values of growth performance were observed in the groups (14 °C and 18 °C) among the 5 groups. The lowest values of feed conversion ratio (FCR) were observed in the group (26 °C). Regression analysis identified the minimum FCR at 26.55 °C. The feeding rates (FR) were increased gradually with the increase of temperature. Regression analysis identified the maximum FR at 28.33 °C. The activities of protease and lipase in the gut were higher than those in the liver, while amylase activity was lower in the gut than that in the liver. There were no significant differences in activities of protease and lipase among different parts of the intestinal tract at the temperatures range from 18 to 26 °C. When the temperature was out the optimal range, C. guichenoti would be under chronic stress, resulting in decreased lysozyme activity and cortisol concentration. Inconclusion, the optimal temperature for juvenile C. guichenoti is 26.55-28.33 °C based on the maximal growth performance and feed utilization.

Temperature Acclimation Alters the Thermal Tolerance and Intestinal Heat Stress Response in a Tibetan Fish Oxygymnocypris stewarti.

Numerous studies have shown that thermal tolerance and intestinal heat resistance are strongly associated with temperature acclimation. However, few reports have successfully conducted similar research on fishes from the Qinghai-Tibetan Plateau, an area that is facing the threat of climate warming. Therefore, the present study determined the growth, thermal tolerance, and intestinal heat stress (exposure to 30°C) responses in juveniles of a Tibetan fish, Oxygymnocypris stewarti, acclimated to three temperature levels (10°C, 15°C, and 20°C, named as T10, T15, and T20, respectively) for 30 days. The fastest growth was recorded in the T15 group. At 1°C/30 min heating rate, the critical thermal maximum (CTMax ) ranged from 31.3°C to 32.3°C, and the lethal thermal maximum (LTMax ) ranged from 31.8°C to 32.6°C among the three acclimation temperatures. According to the results of thermal tolerance tests, the heat stress temperature was set to 30°C. When the water temperature reached 30°C, the expression of the intestinal heat shock protein 70 (HSP70) gene as well as the intestinal microbiome and histology of experimental fish were monitored at 0, 2, 6, and 12 h. The expression of HSP70 reached the highest level at 2 h in all three temperature treatments. The histological analysis showed damage to intestinal cells, including diffuse infiltration of lymphocytes, villi epithelial cell swelling, decrease of intestinal villi length, and cytoplasmic light staining at 2 h in all three temperature treatments. In terms of the intestinal microbiome, phyla Proteobacteria and Firmicutes dominated the treatments at each monitored time in the T10 and T15 groups and at 0 h in T20 group, while phyla Fusobacteria, Proteobacteria, and Cyanobacteria were dominant in treatments at 2, 6, and 12 h in the T20 group. The overall results indicated that acclimation temperature could affect the growth, thermal tolerance, and intestinal heat stress response of O. stewarti juveniles. As the first report on intestinal heat stress response associated with temperature acclimation in a Tibetan fish, this study will help to understand the potential effects of climate change on highland fishes.

Assessment of eutrophication and nitrogen and phosphorus carrying capacity before and after removing pen culture (2013-2018) in Lake Changhu, China.

In this study, the eutrophication levels and nitrogen and phosphorus carrying capacities of Lake Changhu in Jingzhou, Hubei Province, China, were measured using the trophic level index (TLI) and Dillon model for the first time. The measurements were taken before (2013 and 2015) and after (2017 and 2018) the removal of pen aquaculture from the lake. The lake was divided into three districts: Lake Haizihu, Mahongtai Channel, and Lake Dahu. The results showed total nitrogen (TN), chemical oxygen demand (COD), chlorophyll a (Chl-a), and total suspended solid values were significantly higher in 2017 than in the other years. The Lake Haizihu district was predicted to be more seriously polluted than the other districts. In the sediment, the organic matter, STN (TN in sediment) and STP (TP in sediment) contents increased from 2013 to 2018. The mean TLI values ranged from 62.99 to 78.93 in the studied years, and the eutrophication level was highest in 2017. According to the Dillon model, when the target water quality was level III (GB 3838-2002, Ministry of Environmental Protection of China, 2002), the remaining TN and TP loading capacities were -1470.72 t/a and -182.74 t/a, respectively, in 2015, and 320.03 t/a and -111.14 t/a, respectively, in 2018. Our results provide valuable and integrated information about the water conditions of Lake Changhu, thus laying a foundation for the theoretical study of water eutrophication process in lakes and paving the way for informed decision-making for managing water environments to ensure the safety of ecology.

Clinical and Bacterial Characteristics of Klebsiella pneumoniae Affecting 30-Day Mortality in Patients With Bloodstream Infection.

Background: There is a paucity of studies using clinical characteristics and whole-genome sequencing together to fully identify the risk factors of patients with Klebsiella pneumoniae (KP) bloodstream infection (BSI). Methods: We retrospectively analyzed the clinical and microbiological characteristics of patients with KP BSI. Isolates were processed using Illumina NGS, and relevant bioinformatics analysis was conducted (multi-locus sequence typing, serotype, phylogenetic reconstruction, detection of antibiotic resistance, and virulence genes). A logistic regression model was used to evaluate the risk factors of hosts and causative KP isolates associated with 30-day mortality in patients infected with KP BSI. Results: Of the 79 eligible patients, the 30-day mortality rate of patients with KP BSI was 30.4%. Multivariate analysis showed that host-associated factors (increased APACHE II score and septic shock) were strongly associated with increased 30-day mortality. For the pathogenic factors, carriage of iutA (OR, 1.46; 95% CI, 1.11-1.81, p = 0.002) or Kvar_1549 (OR, 1.31; 95% CI, 1.02-1.69, p = 0.043) was an independent risk factor, especially when accompanied by a multidrug-resistant phenotype. In addition, ST11-K64 hypervirulent carbapenem-resistant KP co-harbored acquired blaKPC-2 together with iutA (76.5%, 13/17) and Kvar_1549 (100%, 17/17) genes. Comparative genomic analysis showed that they were clustered together based on a phylogenetic tree, and more virulence genes were observed in the group of ST11-K64 strains compared with ST11-non-K64. The patients infected with ST11-K64 strains were associated with relatively high mortality (47.2%, 7/17). Conclusion: The carriage of iutA and Kvar_1549 was seen to be an independent mortality risk factor in patients with KP BSI. The identification of hypervirulent and carbapenem-resistant KP strains associated with high mortality should prompt surveillance.

Optimal Empiric Polymyxin B Treatment of Patients Infected with Gram-Negative Organisms Detected Using a Blood Antimicrobial Surveillance Network in China.

BACKGROUND: Few pharmacodynamics studies to date have evaluated the efficacy and safety of polymyxin B (PMB) in treating patients with bloodstream infections (BSIs) in China. METHODS: Patients with BSIs were identified using an antimicrobial surveillance network, and their pathogens were isolated. Patients were treated with a loading dose of PMB followed by either a weight-based or weight-independent maintenance dose. Monte Carlo simulation was utilized to calculate the probability of target attainment (PTA) and cumulative fraction of response (CFR) against Gram-negative organisms in patients with normal or decreased renal function. RESULTS: A total of 10,066 Gram-negative organisms, including 5500 Escherichia coli (Eco), 2519 Klebsiella pneumoniae (Kpn), 501 Acinetobacter baumannii (Aba), were isolated from patients with BSIs. Although these strains were highly resistant to carbapenem, they remained susceptible to PMB. Among patients with renal impairment (mean CrCL, 42 mL/min), a PMB 2.5 mg/kg loading dose followed by a maintenance dose of 60 mg q12h reached ≥90% PTA against isolates with an MIC of 2 mg/L, with a low risk of toxicity. Among patients with normal renal function (mean CrCL, 123 mL/min), all simulated regimens showed PTAs of 25-80%. A weight-based loading dose followed by either a weight-based or weight-independent maintenance dose showed a promising CFR, especially in patients with renal impairment, with CFRs ≥90% against carbapenem-resistant Eco, Kpn, and Aba. Simulated regimens showed a disappointing CFR (<80%) against carbapenem-resistant Pae in patients with normal renal function. Based on the optimal balance of efficacy and toxicity, a fixed maintenance dose of 60 mg q12h among patients with renal impairment yielded a CFR similar to regimens based on total body weight and was associated with a probability of toxicity (12.5%) significantly lower than that of simulations based on total body weight. Among patients with normal renal function, a weight-based maintenance dose of 1.25 mg/kg q12h achieved a higher CFR than a fixed maintenance dose, without significantly increasing toxicity. CONCLUSION: A 2.5 mg/kg loading dose of PMB is optimal, regardless of renal function. A fixed maintenance dose of 60 mg q12h is recommended for empirical treatment of patients with renal impairment infected with Eco, Kpn, and Aba, whereas a weight-based maintenance dose of 1.25 mg/kg is recommended for patients with normal renal function.

Expression Profiles of dmrt1 in Schizothorax kozlovi, and Their Relation to CpG Methylation of Its Promoter and Temperature.

To elucidate the role of dmrt1 in sex differentiation of a teleost fish Schizothorax kozlovi, the full-length sequences of its cDNA and promoter were cloned by rapid amplification of cDNA ends (RACE) and genome walking. The relative mRNA expression levels were determined by quantitative real-time PCR (RT-PCR). The 1095-bp dmrt1 cDNA was predicted to encode a protein of 264 amino acids. It was expressed only in the gonads, and the expression was 17-times higher in the testis than in the ovary. The 1215-bp promoter sequence of dmrt1 was cloned and analyzed to detect sex-related differences in its methylation levels. A significant negative relationship between the dmrt1 expression and CpG methylation of its promoter were found in the testes and ovaries of S. kozlovi. Significant differences in dmrt1 expression levels were also found between the larval and juvenile stages. No significant differences in expression were found during the entire larval stage, and in the individuals among three different temperature groups (10°C, 14°C, and 18°C). Considering that the sex of sampled larval fish cannot be distinguished, correlations between dmrt1 expression and effects of temperature on sex differentiation in S. kozlovi need further study.

Expression profiles of amh and foxl2 in Schizothorax kozlovi, and their response to temperature during the early developmental stage.

To elucidate the role of amh and foxl2 in sex differentiation of the teleost fish Schizothorax kozlovi, the full-length cDNAs were cloned from the mature testis and ovary by rapid amplification of cDNA ends (RACE), and their relative mRNA expression levels were determined by quantitative real-time polymerase chain reaction among tissues and temperature groups. The complete amh and foxl2 cDNAs of S. kozlovi were 2060 bp and 1750 bp, which encoded 568 and 306 amino acids, respectively. The amh were expressed only in gonads, while foxl2 was expressed in the gills, brain and gonads, both exhibiting relatively high tissue specificity. The amh exhibited sex-specific expression pattern in the gonads. No sex differences in the foxl2 expression were observed in the brain and gonads, but significant sex differences were found in the gills. No significant differences were found in the foxl2 expression, from the larval to the juvenile stage, and also between different temperature groups. However, significant differences were found in the expression levels of amh from the larval (12-63 days posthatching (dph)) to the juvenile stage (190 dph), and also among the 18°C and 10°C groups at 31 dph. This result suggested that amh plays an important role in male sex differentiation of S. kozlovi during the early developmental stage, but no similar effect was observed in foxl2.

The influence of weight and gender on intestinal bacterial community of wild largemouth bronze gudgeon (Coreius guichenoti, 1874).

BACKGROUND: Largemouth bronze gudgeon (Coreius guichenoti) is of economic importance in China, distributed in upstream regions of the Yangtze River in China. But it has recently dramatically declined and is close to elimination. However, there is little knowing about the character of its intestinal microbiota. This study was conducted to elucidate the intestinal microbiota of wild largemouth bronze gudgeon with different body weight and gender. RESULTS: Thirty wild largemouth bronze gudgeon were measured for body length and body weight, and identified for male and female according to gonadal development, and thereafter the intestinal microbiota's were assessed by MiSeq sequencing of 16S rRNA genes. The results revealed that phyla Proteobacteria and Tenericutes were dominant in wild largemouth bronze gudgeon intestine independent of the body weight. Shannon's and Inverse Simpson's diversity indexes were significant (P < 0.05) different between male and female fish. The phylum profile in the intestine of male fish revealed that phylum Proteobacteria was dominant, in contrast to female fish where five phyla Tenericutes, Proteobacteria, Firmicutes, Fusobacteria and Spirochaetes were dominant. The genus profile revealed that genera Shewanella and Unclassified bacteria were dominant in male fish, while genus Mycoplasma was dominant in female fish. CONCLUSIONS: Our results revealed that the intestinal microbial community of wild largemouth bronze gudgeon was dominated by the phyla Proteobacteria and Tenericutes regardless of the different body weight, but the communities are significant different between male and female fish. These results provide a theoretical basis to understand the biological mechanisms relevant to the protection of the endangered fish species.

Effect of rearing temperature on growth and thermal tolerance of Schizothorax (Racoma) kozlovi larvae and juveniles.

Effect of rearing temperature on growth and thermal tolerance of Schizothorax (Racoma) kozlovi Nikolsky larvae and juveniles was investigated. The fish (start at 12d post hatch) were reared for nearly 6 months at five constant temperatures of 10, 14, 18, 22 and 26°C. Then juvenile fish being acclimated at three temperatures of 14, 18 and 22°C were chosen to determine their critical thermal maximum (CTMax) and lethal thermal maximum (LTMax) by using the dynamic method. Growth rate of S. kozlovi larvae and juveniles was significantly influenced by temperature and fish size, exhibiting an increase with increased rearing temperature, but a decline with increased fish size. A significant ontogenetic variation in the optimal temperatures for maximum growth were estimated to be 24.7°C and 20.6°C for larvae and juveniles of S. kozlovi, respectively. The results also demonstrated that acclimation temperature had marked effects on their CTMax and LTMax, which ranged from 32.86°C to 34.54°C and from 33.79°C to 34.80°C, respectively. It is suggested that rearing temperature must never rise above 32°C for its successful aquaculture. Significant temperature effects on the growth rate and thermal tolerance both exhibit a plasticity pattern. Determination of critical heat tolerance and optima temperature for maximum growth of S. kozlovi is of ecological significance in the conservation and aquaculture of this species.