Hematologic and serum biochemical characteristics of rare minnow (Gobiocypris rarus) in two different culture environments

doi:10.21203/rs.3.rs-2250555/v1

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Hematologic and serum biochemical characteristics of rare minnow (Gobiocypris rarus) in two different culture environments

https://doi.org/10.21203/rs.3.rs-2250555/v1

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Hematological and serum biochemical parameters are essential for the evaluation of both structural and function status of fish species due to the involvement of culture environment and sex. To obtain a basic knowledge about the variation in hematological and serum biochemical parameters of conventional (CV) and specific pathogen-free (SPF) rare minnow (Gobiocypris rarus) with respect to sex, blood samples collected from 720 healthy and adult rare minnow (♀:♂=1:1) are analyzed by using standard methods and techniques. The results indicated that hemoglobin (Hb), hematocrit (HCT) and mean corpuscular volume (MCV) values in female CV rare minnow were 82.4 ± 9.5 g/L, 41.6 ± 8.4%, 408.3 ± 81.6 fl, which were significantly lower than those in SPF rare minnow (p < 0.05). Alkaline phosphatase (ALP) differed significantly between these two cultured rare minnows (p < 0.05), and no significant differences were observed in other serum biochemical parameters (p > 0.05). In addition, MCV values of female rare minnow are significantly higher than male individuals (p > 0.05). These data demonstrated that hematological and serum biochemical parameters of rare minnow were affected by culture environment and sex, and that a clean culture environment could contribute to the health physiological status of rare minnow.

hematological parameters

serum biochemistry

conventional rare minnow

specific pathogen-free rare minnow

Hematological and serum biochemical parameters have been widely used as vital indices to determining the healthy status of fish (Cao and Wang 2010; Houston 1997; Tang et al. 2015). Numerous studies have demonstrated that culture environment and sex can influence the hematological and serum biochemical profile in fish species (Santos et al. 2009; Shigdar et al. 2009; Zhang et al. 2011). For example, white blood cells (RBC), hemoglobin (Hb) and mean corpuscular volume (MCV) revealed significant difference between male and female walking catfish, Clarias batrachus (Linnaeus, 1758) (Gayatri Acharya 2014). Similarly, statistically significant differences in the various parameters, such as red blood cells (RBC), hematocrit (HCT), neutrophils, alkaline phosphatase (ALP) and high density lipoprotein (HDL), were observed between Tian-E-Zhou Oxbow Yangtze Finless Porpoises (YFPs) (Neophocaena asiaeorientalis ssp. asiaeorientalis) and Poyang Lake YFPs (Nabi et al. 2017). Studies on the comparison of hematology and serum biochemistry have focused mainly on comparison between wild and wild fish or wild and hatchery fish, and little is known about how different culture environments affect hematological and serum biochemical parameters in fish species.

Rare minnow, Gobiocypris rarus Ye et Fu, 1983, a freshwater cyprinid (Ding 1994; Wang and Cao 1997), is commonly cultured in laboratory using as a conventional experimental model in various studies, such as toxicology, water-monitoring, and so on (Fang et al. 2010; Wang and Cao 2017; Wang et al. 2013). To accelerate its development as an experimental animal and help ensure that the results of studies using them are reliable, specific pathogen-free (SPF) stocks of rare minnow have been established according to a set of standards. These standards for conventional (CV) and SPF rare minnow consist of five parts, including microbiological standards and monitoring, parasitological standards, genetic quality control, nutrition and food, and water environment and facilities standards. Preliminary reports have confirmed that these standards are rigorous, and declared them to be the local standard in Hubei Province for establishing and cultivating SPF rare minnows (Su et al. 2020). Research on the species’ hematology and serum biochemistry is limited to the CV individuals reported by Shao et al. (2006), and these parameters of SPF rare minnow have not yet been reported.

In the present study, 12 hematological indices and 10 serum biochemical indices of CV and SPF rare minnows in relation to sex, performed by the routine method of blood physiology and automated biochemical analyzer, are reported. The objectives of the study are to: 1) describe the knowledge of hematological and serum biochemical parameters of rare minnows in different culture environments; 2) evaluate similarities and dissimilarities in the hematology and serum biochemistry between these two inbred rare minnows to determine whether the clean environment affects the its healthy, and 3) consider how the sex affect these parameters.

Experimental animals and fish rearing

Approximately 2000 fertilized eggs were collected from a closed colony (Ihb:IHB) of SPF rare minnow broodstock held in the National Aquatic Biological Resource Center (NABRC) of the Institute of Hydrobiology, Chinese Academy of Sciences. Half of the eggs were cultured in an open environment (OE), and the others were housed in an isolate (clean) environment (IE) in which the air was filtered through low-, medium- and high-efficiency filters. Rare minnows cultured in the open environment were free of acute or highly infectious diseases such as virulent Aeromonas, Mycobacterium marinum, Vibro cholera and Flavobacterium colummare. These were designated CV rare minnows. In addition, SPF rare minnows reared in the isolation (clean) environment were also free of Pseudomonas fluorescens, Streptococcus agalactiae and Edwardsiella ictaluri. To keep the isolation environment clean, the facility manager sterilized the tools that were used daily. In both environments, the water temperature, pH, and dissolved oxygen were monitored daily (Hach, HQ30d, Loveland, CO., USA), and ammonia nitrogen, nitrite nitrogen (NO₃-N), nitrate nitrogen (NO₂-N), and total hardness were determined monthly. The parameters of rearing water in both culture environments, along with their standard and measure values, are shown in Table 1. Other than that, they were both fed to satiety with sterilized pellet feed twice daily.

Table 1

Water parameters in the open and isolation environments
Parameters	Standard values		Measured values
Parameters	Open environment (OE)	Isolation (clean) environment (IE)	Open environment (OE)	Isolation (clean) environment (IE)
Temperature (°C)	20–30	22–30	23.2 ± 0.52	25.8 ± 0.85
Dissolved oxygen (mg/L)	> 5	> 6	7.72 ± 0.986	9.04 ± 0.519
Conductivity (µS/cm)	150–1000	150–650	383.00 ± 22.517	291.97 ± 85.110
pH	7.0-8.5	7.0-8.5	8.25 ± 0.395	7.79 ± 0.069
Total hardness (mg/L CaCO₃)	≤ 450	≤ 450	183.44 ± 2 1.119	55.59 ± 6.443
NO₃-N (mg/L)	≤ 0.1	≤ 0.1	0.041 ± 0.0049	0.046 ± 0.0026
NO₂-N (mg/L)	≤ 30	≤ 30	0.0015 ± 0.0019	0.0347 ± 0.0480
Ammonium-N (mg/L)	≤ 0.5	≤ 0.5	0.33 ± 0.017	0.41 ± 0.128

Blood collection

After 12 months of feeding, 720 health and mature rare minnow (360 CV and 360 SPF individuals) were used in this study. For whole blood acquisition, 180 female and 180 male rare minnows in each environment were euthanized with 200 mg/L tricaine methanesulfonate (MS-222, Sigma-Aldrich, St Louis, MO). After euthanized, rare minnows were weight, measured, and blood samples were immediately collected from the diagonal incision just between the anal fin and caudal fin by a pipettor (20 µL). Blood yields from individual fish ranged from 5 to 20 µL. A small amount of the whole blood was immediately smeared on a slide for differential leukocyte account and the rest was collected for another hematologic analysis. For serum acquisition, whole blood from groups of 10 rare minnows was pooled in a 0.5 mL microcentrifuge tube, left to clot at room temperature for 1 h and spun for 5 min at 2500 rpm; then serum was pipetted off the top (Murtha et al. 2003). The extracted serum was kept at − 20°C until further analysis. The blood sampling procedure was also reviewed and approved by the Research Ethic Committee of Institute of Hydrobiology, Chinese Academy of Science (Permit Number 20180516).

Hematologic analysis

Blood smears were stained with Wright-Giemsa (Wuhan, China) stain for differential leukocyte count (Gao et al. 2007). Hematological values except for erythrocyte osmotic fragility (EOF) were performed manually on heparin anticoagulated blood specimens. Red blood cell count (RBC) and white blood cell count (WBC) were determined by Beckman-Coulter Z2 cell counter (USA) in red cell dilute and white cell dilute (Haling, Shanghai, China), respectively. The hematocrit (HCT) was obtained by filling a micropipette tip with blood and this was spun at 3000×g for 10min and then read with a vernier caliper (GUANGLU, Guilin, China). For EOF, one drop blood (with no anticoagulant) was added to a number of 1mL NaCl solutions containing concentrations from 0.05–0.5% at 0.05% intervals. After 1h, the concentration of starting partial hemolysis was taken as the value of EOF (Tang et al. 2015). The hemoglobin (Hb) was determined by sharli colorimetry (Shao et al. 2006). The HCT, EOF, Hb, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) was calculated as followed (Gao et al. 2007a):

MCV (fL) = HCT (%) × 1000/RBC (10⁶uL⁻¹),

MCH (pg) = Hb (g/L)/RBC (10⁶uL⁻¹),

MCHC (g/L) = Hb (g/L) ×100/HCT (%)

Serum biochemical analysis

Liver function parameters; Alkaline Phosphatase (ALP), Alaline amino Transferase (ALT), lipid profile; Total Cholesterol (TCHO), Triglyceride (TG), High Density Lipoprotein cholesterol (HDLC), enzymes; (Lactate Dehydrogenase (LDH), and other biochemical parameters such as Total Protein (TP), Albumin (ALB), Glucose (GLU), Blood Urea Nitrogen (BUN) were measured using the automated biochemical analyzer (CHEMIX-800, Sysmex, Japan). The analyzer was calibrated before each assay.

Statistical analysis

An unpaired Student’s t-test was used to compare CV rare minnows to SPF individuals using SPSS 19.0. A one-way ANOVA followed by the post hoc test was used to investigate changes in the mean level of each parameter across all groups in each environment. Normality was assessed using the Shapiro-Wilk test. All data were presented as mean ± SD. Significance was accepted at p < 0.05.

Hematological parameters

Reference intervals for hematological analytes in these two different inbred rare minnows are summarized in Table 2 and Fig. 1. Blood parameters like RBCs, WBCs, Hb and HCT were statistically significantly higher in rare minnow living in IE compared to those living in OE (Duncan multiple-comparison tests, p < 0.05) (Table 2).

Table 2

Hematological parameters of rare minnow in two different environments
Parameters	Female		Male
Parameters	CV (n = 20)	SPF(n = 20)	CV(n = 20)	SPF(n = 20)
Total length (mm)	40.11 ± 3.73	33.78 ± 2.804	36.48 ± 2.811	32.71 ± 2.613
Body weight (g)	1.410 ± 0.382	0.917 ± 0.216	0.947 ± 0.202	0.743 ± 0.143
RBC (10⁴/mm³)	105.9 ± 28.7^a	138.0 ± 31.8^b	121.6 ± 33.2^ab	144.6 ± 32.4^b
WBC (10⁴/mm³)	31.8 ± 12.7^a	45.5 ± 18.0^b	26.2 ± 9.8^a	40.7 ± 20.3^ab
Hb (g/L)	82.4 ± 9.5^a	95.7 ± 14.9^b	81.0 ± 13.8^a	98.8 ± 16.8^b
HCT (%)	41.6 ± 8.4^a	48.3 ± 9.5^c	43.3 ± 5.9^a	48.6 ± 8.7^d
EOF (%)	0.3 ± 0.0^a	0.3 ± 0.1^ab	0.3 ± 0.1^a	0.2 ± 0.1^b
MCV (fl)	408.3 ± 82.6^a	350.1 ± 69.0^b	355.8 ± 48.3^b	336.4 ± 60.4^c
MCH (pg)	77.9 ± 9.0^a	69.4 ± 10.8^b	66.6 ± 11.4^b	68.3 ± 11.6^b
MCHC (g/L)	198.1 ± 22.9^a	198.1 ± 30.8^a	187.3 ± 32.0^a	203.2 ± 34.6^a

Different superscript letters in the same column indicate significant difference (P < 0.05).

For CV rare minnow, the reference intervals were determined as: RBCs,70.9-205.8 and 66.0-193.9 10⁴/mm³; WBCs, 13.0–59.0 and 9.9–49.9 10⁴/mm³; Hb, 65.3-101.5 and 58.0-110.2 g/L; HCT, 22.7–59.5 and 31.1–53.0%; EOF, 0.25–0.35 and 0.20–0.35%; MCV, 223.0-583.9 and 255.6-436.1 fl; MCH, 61.6–95.8 and 47.7–90.6 pg; MCHC, 156.8-243.9 and 134.1-254.7 g/L in females and males, respectively (Fig. 1). And only MCV and MCH were statistically significantly higher for CV rare minnow (t-test, p < 0.05) (Fig. 1). For SPF rare minnow, except for EOF and MCV, other parameters did not show significant difference with respect to sex (p > 0.05) (Fig. 1).

Leukocyte differential counts

There were five types of WBCs, lymphocytes, neutrophils, monocytes, eosinophils and basophils in rare minnow (Fig. 2). Lymphocytes were round to ovoid in shape, with a large nucleus and thin rim of cytoplasm (Fig. 2B). Monocytes were the largest type of WBC in rare minnow, with abundant blue cytoplasm. The nucleus of monocytes was mainly irregular (Fig. 2E). Neutrophils were of several sizes, but represented the largest nucleus. Usually, the nucleus was observed as round or ovoid (Figs. 2A and 2F). Basophils was round or oval cells with a kidney shaped nucleus, with light blue granules (Fig. 2C). In addition, basophils were observed only rarely in the blood of rare minnow. Eosinophils was of round shape. The granules in the granulocytes stained a light red in pale gray cytoplasm (Fig. 2D).

Counts from the smears of female and male rare minnow (n = 20) in two different environments indicated predominantly lymphocytes and neutrophils, with smaller numbers of monocytes, eosinophils and basophils (Table 3). Neutrophils counts in CV rare minnow were higher than SPF rare minnow, but the opposite was true for lymphocytes counts. And only female rare minnow showed statistically significant differences (p < 0.05). CV rare minnow had higher level of monocytes, eosinophils and basophils compared to SPF rare minnow, while there was no significant difference between these two inbred rare minnows (p > 0.05).

Table 3

White blood cell differential counts of rare minnow in two different environments
Parameters	Female		Male
Parameters	CV (min-max, n = 20)	SPF (min-max, n = 20)	CV (min-max, n = 20)	SPF (min-max, n = 20)
Neutrophils(%)	48.7 ± 17.6^a (18-80.8)	37.5 ± 16.3^b (10–70)	38.6 ± 10.5^b (20–59)	35.1 ± 11.2^b (14–51)
Lymphocytes(%)	45.4 ± 19.5^a (13.46-80)	59.9 ± 15.7^b (30–88)	59.2 ± 11.0^b (36–79)	63.9 ± 11.2^b (47–86)
Basophils(%)	4.2 ± 2.4^a (1-9.2)	2.2 ± 2.9^ab (0–10)	1.4 ± 1.3^b (0–5)	0.7 ± 1.5^b (0–6)
Eosinophils(%)	1.1 ± 1.5^a (0–4)	0.3 ± 0.7^a (0–2)	0.4 ± 0.5^a (0–1)	0.2 ± 0.5^a (0–2)
Monocytes(%)	0.6 ± 1.1^a (0-3.06)	0.1 ± 0.5^a (0–2)	0.5 ± 1.2^a (0–5)	0.1 ± 0.3^a (0–1)

Different superscript letters in the same column indicate significant difference (p < 0.05).

For CV rare minnow, except for lymphocytes, other types of WBCs all showed higher levels in females than those in males. And basophils showed significant differences between females and males. For SPF rare minnow, the differences in relation to sex were similar to those of CV fish, except that there was no significant difference in the count of lymphocytes (t-test; p = 0.380) (Fig. 3).

Serum biochemical parameters

The results of serum biochemical parameters for female and male rare minnow in two inbred environments were shown in Table 4 and Fig. 4. Generally, enzymes, like ALP, were statistically significantly higher in SPF rare minnow versus those in CV individuals, but lower for ALT. And the other biochemical parameters (TCHO, HDCL, TG, LDH, TP, ALB, GLU and BUN) showed no significant differences between these two inbred rare minnows (p > 0.05) (Table 4).

Table 4

Serum biochemical parameters of rare minnow in two different environments
Parameters	Female		Male
Parameters	CV (n = 10)	SPF (n = 12)	CV (n = 10)	SPF (n = 12)
ALP (U/L)	158.0 ± 15.5^ab	106.0 ± 19.7^c	173.5 ± 0.7^b	133.6 ± 8.1^a
ALT (U/L)	13.5 ± 0.2^a	18.8 ± 2.2^bc	13.2 ± 0.2^a	19.1 ± 3.5^ac
TCHO (mmol/L)	3.0 ± 0.2^a	3.7 ± 0.5^a	3.4 ± 0.2^a	4.0 ± 0.9^a
HDCL (mmol/L)	24.3 ± 1.5^a	14.8 ± 8.5^a	20.0 ± 4.2^a	15.2 ± 8.6^a
TG (mmol/L)	53.7 ± 2.1^a	36.4 ± 20.9^a	46.5 ± 7.8^a	34.6 ± 19.6^a
LDH (U/L)	1005.3 ± 29.1^a	1099.8 ± 473.4^a	803.5 ± 208.7^a	914.8 ± 577.2^a
TP (g/L)	1.6 ± 0.1^a	1.5 ± 0.3^a	1.4 ± 0.2^a	2.3 ± 0.3^b
ALB (g/L)	9.9 ± 1.0^a	13.4 ± 2.3^a	10.5 ± 0.3^a	10.3 ± 2.4^a
GLU (mmol/L)	25.4 ± 1.1^a	37.6 ± 7.1^b	27.3 ± 3.6^a	36.9 ± 2.5^b
BUN (mmol/L)	7458.0 ± 244.3^a	7366.0 ± 520.3^a	7693.0 ± 199.4^a	8129.7 ± 489.2^a

Different superscript letters in the same column indicate significant difference (p < 0.05).

For CV rare minnow, ALP, TCHO, ALB, GLU and BUN values in males were higher than those in females, but the opposite is true for the other serum biochemical parameters. And there were no significant differences between female and male rare minnow (t-test; ALP, p = 0.273; ALT, p = 0.494; TCHO, p = 0.222; HDLC, p = 0.701; TG, p = 0.594; LDH, p = 0.143; TP, p = 0.410; ALB, p = 0.372; GLU, p = 0.484; BUN, p = 0.225) (Fig. 4). For SPF rare minnow, certain enzymes, lipid profile, and some other parameters were determined as: ALP, 82–135 and 126–145 U/L; ALT, 503–1720 and 288–1670 U/L; TCHO, 3.0-4.3 and 3.2–5.2 mmol/L; HDLC, 16.7–21.7 and 13.6–22.6 mmol/L; TP, 39–52 and 38–46 g/L; BUN, 1.1–1.8 and 1.8–2.5 mmol/L in females and males, respectively. The mean values of these serum biochemical parameters in female rare minnow showed lower than those in male individuals, but only ALP and BUN showed statically significantly differences (t-test; p < 0.05) (Fig. 4).

Hematological and serum biochemical characteristics of healthy rare minnow

In the present study, the values of RBC and Hb for female CV and SPF rare minnow were 105.9 ± 28.7 and 138.0 ± 31.8 (10⁴/mm³), 72.4 ± 9.5 and 95.7 ± 14.9 g/L. respectively. The results of this study showed that five types of WBCs were observed in rare minnow. Meanwhile, the levels of ALP, TCHO and ALB for male CV and SPF rare minnow were 173.5 ± 0.7 and 133.6 ± 8.1 U/L, 3.7 ± 0.5 and 4.0 ± 0.9 mmol/L, 10.5 ± 0.3 and 10.3 ± 2.4 g/L, respectively. To our knowledge, this is the first systematic report on CV and SPF rare minnow defining hematological and biochemical profiles. Therefore, this study may be important as references values for evaluation of inadequate management conditions. In general, the hematological and biochemical values of fish species can provide an effective tool in the assessment of their health status (Ahmed and Sheikh 2019; Nigam et al. 2020). On the other hand, these values of healthy fish species also can support valuable information for their physiological status. Numerous studies indicated that the hematological values in fish species showed highest in carnivorous fish, middle in herbivore fish and lowest in omnivorous. For example, Fazio et al. (2013) showed levels of glucose to be higher in Sparus aurata (carnivorous fish) and lower in Mugil cephalus (herbivore fish). Similarly, as a carnivorous fish (Wang and Cao 2017), the values of GLU of male CV and SPF rare minnow were 37.6 ± 7.1 and 36.9 ± 2.5 mmol/L, respectively. Regardless of culture environment, the GLU values of rare minnow is not lower than 10 mmol/L, which is higher than those in grass carp (Lin et al. 1985). In addition, Ahmed et al. (2019a) indicated that RBC greater than 10⁶ is considered high and indicative of high oxygen carrying capacity of blood. Thus, it is not difficult to speculate that rare minnow is a type of high activity fish. Although the hematological and serum biochemical variation was observed between different species, the same fish species may also result in variation in hematological and serum biochemical values due to different measurement methods (Prasad and Charles 2010). That may be the reason why the results of this study are different from those of Shao et al. (2006).

Effect of culture environment on the hematological and biochemical characteristics

Hematological and serum chemical parameters of fish species are easily influenced by culture environment (Fang et al. 2014; Shigdar et al. 2009; Zhang et al. 2010). For laboratory animal, changes in the culture environment are usually associated with variability in water chemistry parameters and degree of bacterial control. The present study showed that the RBC, Hb and MCH were significantly higher in SPF rare minnows than CV ones. It was reported that the blood parameters of fish increased with increasing temperature within certain limits, as was evident from a significant reduction in RBC and WBC of snow trout (Schizothorax labiatus) in winter (Jan and Ahmed 2021). Moreover, water flow velocity also had a significant effect on the physiological paremeters for Hemiculter leucisculus (Lu 2015). Thus, the different hematological parameters that we observed between CV and SPF rare minnows may have been caused by the culture environments, which differed in temperature, and water flow velocity. Similarly, Kuang et al. (2012) found that the clean culture conditions significantly affected physiological status. Therefore, it could be argued that the cleaner culture environment contributed to the health physiological status, although the reasons for this requires further study.

It is widely accepted that clean culture environment has an adverse role on the immunity of fish species. For example, Zang et al. (2013) demonstrated that pond-cultured Japanese flounder had higher serum immunity level than indoor tank-cultured individuals. However, the present results disagree with that research and show that the values of ALP and lymphocytes in CV rare minnow are statistically lower than those in SPF individuals. Satheeshkumar et al. (2011) demonstrated that salinity could stimulate the production of leukocytes, especially lymphocytes. In truth, the pH of aquaculture water of SPF rare minnow is occasionally low due to the lack of mother liquid or machine failure. Therefore, it is not difficult to speculate that the higher ALP and lymphocytes levels of SPF rare minnow is caused by salinity.

Effect of sex on the hematological and biochemical characteristics

It has been reported that sex has a great influence on the hematological and serum biochemical parameters of fish species (Jan and Ahmed 2020; Sharma et al. 2016). Generally, the values of RBC in female fish species were lower than those in male individuals, but the opposite was true for the values of WBC. This phenomenon applied to Hypophthalmichthys molitrix (Ahmed et al. 2019), Paralichthys olivaceus (Zang et al. 2013), as well as rare minnow. Sharma et al. (2016) demonstrated that sex may be related to differences in blood cell constituents which caused higher metabolic activity of male than female fish. Thus, the regular different RBC values and WBC values that we observed between female and male rare minnows may be caused by sex. In addition to this regular difference, sex had also irregular difference in the hematological and serum biochemical parameter of fish. For example, the higher values of MCV, neutrophils and TP and the lower levels of HCT, ALP in female rare minnows were observed in this study. The irregular difference between female and male species may be due to sexual maturity.

This study explored the hematological and biochemical parameters of CV and SPF rare minnow. On the one hand, it could provide a theoretical basis for relevant physiological parameters of CV and SPF rare minnow. On the other hand, hematological and serum biochemical parameters of rare minnow were affected by culture environment and sex, as evidenced by significantly higher Hb, HCT and MCV values in SPF rare minnow and significantly higher MCV values of female CV rare minnow than those in male individuals.

Author contribution

The authors’ contributions were as follows: LS designed the research. LS, NQ and MH finished this experiment. LS analyzed the data and wrote the paper. CX, XX and JW provided language help. All authors reviewed and approved the final manuscript.

Funding

This research was supported by a grant from the National Key Technology R & D Program (2011BAI15B01-41) and the Hubei Key Technology R&D Program (2015BCE098).

Data availability

Data of the present article are available under request.

Code availability

Not applicable

Ethics approval

All the experimental procedures were performed in accordance with the Committee on the Ethics of Animal Experiments of Institute of Hydrobiology, Chinese Academy of Science (Permit Number 20180516).

Consent to participate

All authors agree to participate in this study.

Consent for publication

All authors agree to participate in the publication of this article.

Conflict of interest

The authors declare no competing interests

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No competing interests reported.

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Hematologic and serum biochemical characteristics of rare minnow (Gobiocypris rarus) in two different culture environments

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