Characterization of runs of homozygosity islands in American mink using whole-genome sequencing data.

The genome-wide analysis of runs of homozygosity (ROH) islands can be an effective strategy for identifying shared variants within a population and uncovering important genomic regions related to complex traits. The current study performed ROH analysis to characterize the genome-wide patterns of homozygosity, identify ROH islands and annotated genes within these candidate regions using whole-genome sequencing data from 100 American mink (Neogale vison). After sequence processing, variants were called using GATK and Samtools pipelines. Subsequent to quality control, 8,373,854 bi-allelic variants identified by both pipelines remained for further analysis. A total of 34,652 ROH segments were identified in all individuals, among which shorter segments (0.3-1 Mb) were abundant throughout the genome, approximately accounting for 84.39% of all ROH. Within these segments, we identified 63 ROH islands housing 156 annotated genes. The genes located in ROH islands were associated with fur quality (EDNRA, FGF2, FOXA2 and SLC24A4), body size/weight (MYLK4, PRIM2, FABP2, EYS and PHF3), immune capacity (IL2, IL21, PTP4A1, SEMA4C, JAK2, CCNA2 and TNIP3) and reproduction (ADAD1, KHDRBS2, INSL6, PGRMC2 and HSPA4L). Furthermore, Gene Ontology and KEGG pathway enrichment analyses revealed 56 and 9 significant terms (FDR-corrected p-value < 0.05), respectively, among which cGMP-PKG signalling pathway, regulation of actin cytoskeleton, and calcium signalling pathway were highlighted due to their functional roles in growth and fur characteristics. This is the first study to present ROH islands in American mink. The candidate genes from ROH islands and functional enrichment analysis suggest possible signatures of selection in response to the mink breeding targets, such as increased body length, reproductive performance and fur quality. These findings contribute to our understanding of genetic characteristics, and provide complementary information to assist with implementation of breeding strategies for genetic improvement in American mink.

Effect of Vitamin A Supplementation on Growth Performance, Serum Biochemical Parameters, Intestinal Immunity Response and Gut Microbiota in American Mink (Neovison vison).

This experiment investigated the effect of vitamin A supplementation on growth, serum biochemical parameters, jejunum morphology and the microbial community in male growing-furring mink. Thirty healthy male mink were randomly assigned to three treatment groups, with 10 mink per group. Each mink was housed in an individual cage. The mink in the three groups were fed diets supplemented with vitamin A acetate at dosages of 0 (CON), 20,000 (LVitA) and 1,280,000 IU/kg (HVitA) of basal diet. A 7-day pretest period preceded a formal test period of 45 days. The results show that 20,000 IU/kg vitamin A increased the ADG, serum T-AOC and GSH-Px activities, villus height and villus height/crypt depth ratio (p < 0.05). The mRNA expression levels of IL-22, Occludin and ZO-1 in the jejunum of mink were significantly higher in the LVitA group than those in the CON and HVitA groups (p < 0.05). Vitamin A supplementation increased the diversity of jejunum bacteria, decreased the ratio of Firmicutes to Bacteroidetes and increased the relative abundance of Akkermansia, uncultured bacterium f Muribaculaceae, Allobaculum, Lachnospiraceae NK4A136 group, Rummeliibacillus and Parasutterella. The comparison of potential functions also showed enrichment of glycan biosynthesis and metabolism, transport and catabolism pathways in the vitamin A supplementation groups compared with the CON group. In conclusion, these results indicate that dietary vitamin A supplementation could mediate host growth by improving intestinal development, immunity and the relative abundance of the intestinal microbiota.

Dietary supplementation of Ascophylum nodosum improved kidney function of mink challenged with Aleutian mink disease virus.

BACKGROUND: Feed additives which can ease the negative effects of infection by the Aleutian mink disease virus (AMDV) are of interest to mink farmers. The effects of kelp meal (Ascophylum nodosum) supplementation on immune response, virus replication and blood parameters of mink inoculated with AMDV were assessed. AMDV-free black mink (n = 75) were intranasally inoculated with a local strain of AMDV and fed a commercial pellet supplemented with kelp meal at the rates of 1.5% or 0.75% of the feed or were kept as controls (no kelp) for 451 days. Blood was collected on days 0 (pre-inoculation), 31, 56, 99, 155, 366 and 451 post-inoculation (dpi). RESULTS: No significant difference was observed among the treatments for the proportion of animals positive for antibodies against the virus measured by the counter-immunoelectrophoresis (CIEP), viremia measured by PCR, antibody titer measured by quantitative ELISA, total serum protein measured by a refractometer or elevated levels of gamma globulin measured by iodine agglutination test at the sampling occasions. At the termination of the experiment on 451 dpi, there were no differences among treatments for antibody titer measured by CIEP, total serum protein, albumin, globulins, albumin:globulin ratio, alkaline phosphatase, gamma-glutamyl transferase, and proportions of PCR positive spleen, lymph node or bone marrow samples, but blood urea nitrogen and creatine levels were significantly lower in the 1.5% kelp supplemented group than in the controls. CONCLUSION: Kelp supplementation improved kidney function of mink infected with AMDV with no effect on liver function, immune response to infection by AMDV or virus replication.

Effects of dietary vitamin E supplementation on the reproductive performance of yearling female mink (Neovison vison) fed wet fish-based feed.

This study was conducted to investigate the effects of dietary vitamin E (VE) supplementation on the reproductive performance of female mink, preweaning growth performance of their kits, and antioxidative status and immune functions of dams and kits. Yearling mink dams (n = 180) were randomly assigned to five treatment groups (n = 36). The dietary treatments included a basal diet supplemented with VE at 0 (control), 80, 160, 320, or 640 mg/kg DM. Compared with the control, the addition of 160-320 mg/kg VE decreased (P = 0.0362) the percentage of pre-weaning mortality of mink kits. At weaning, the average body weight was greater (P = 0.0408) in kits fed 320 mg/kg supplemental VE. In mink dams, the addition of 80-320 mg/kg VE increased (P = 0.0125) serum SOD. Supplementation of 320 or 640 mg/kg VE decreased (P = 0.0260) serum reactive oxygen species (ROS). Furthermore, feeding a large dose of VE (640 mg/kg diet) resulted in increased (P = 0.0245) serum α-tocopherol. In mink kits, the addition of 320 or 640 mg/kg VE increased serum α-tocopherol (P = 0.0207) and IgG (P = 0.0464). Supplementation of 640 mg/kg VE decreased (P = 0.0471) serum ROS. The present results indicate that VE supplementation improved the antioxidative status of mink dams and enhanced the immune functions, decreased pre-weaning mortality and enhanced weaning weight of their kits. Taken together, the effective VE supplementation was 320 mg/kg of diet for yearling female mink.

Commonly Used Vomiting Animal Models / 中国实验方剂学杂志

Vomiting is a common clinical symptom. Long-term severe vomiting could seriously reduce the life quality of patients, so it is necessary to be intervened with antiemetic drugs. Traditional Chinese medicine has a long history in the treatment of vomiting with remarkable a curative effect. It has the advantages of multi-component, multi-target and multi-system synergistic antagonism. However, due to the active substance and unclear mechanism, it is urgent to adopt an internationally recognized vomiting model system to evaluate the antiemetic characteristics, elucidate the vomiting mechanism, and provide reference for better clinical application. Therefore, this paper systematically introduces several vomiting animal models that are widely used at home and abroad. According to the authors' own experimental experience, this paper focuses on the rat and mice pica models for reference of relevant researchers. Specifically, ferrets are an internationally recognized ideal vomiting animal model, and the golden standard for evaluating the effects of antiemetic drugs, suncus murinus is the smallest mammalian vomiting model. Rodents have no vomiting reflexes, but studies have shown that its pica behavior is equivalent to the vomiting behavior of other species. Because the easy availability and operation, the model has been promoted and applied in mainland China. Guizhou mini-pig model is a self-developed medium-sized mammalian vomiting model with a similar anatomical structure and vomiting characteristics to human, and worthy of popularization and application. In conclusion, different vomiting models have their own characteristics that need to be optimized according to the purpose of experiments and samples.

Distribution of α-tocopherol stereoisomers in mink (Mustela vison) organs varies with the amount of all-rac-α-tocopheryl acetate in the diet.

Synthetic α-tocopherol has eight isomeric configurations including four 2R (RSS, RRS, RSR, RRR) and four 2S (SRR, SSR, SRS, SSS). Only the RRR stereoisomer is naturally synthesised by plants. A ratio of 1·36:1 in biopotency of RRR-α-tocopheryl acetate to all-rac-α-tocopheryl acetate is generally accepted; however, studies indicate that neither biopotency of α-tocopherol stereoisomers nor bioavailability between them is constant, but depend on dose, time, animal species and organs. A total of forty growing young male mink were, after weaning, assigned one of the following treatments for 90 d: no α-tocopherol in diet (ALFA_0), 40 mg/kg RRR-α-tocopheryl acetate (NAT_40), 40 mg/kg all-rac-α-tocopheryl acetate (SYN_40) and 80 mg/kg feed all-rac-α-tocopheryl acetate (SYN_80). Mink were euthanised in CO2 and blood was collected by heart puncture. Mink were pelted and liver, heart, lungs, brain and abdominal fat were collected for α-tocopherol stereoisomer analysis. The proportion of RRR-α-tocopherol decreased in all organs and plasma with increasing amount of synthetic α-tocopherol stereoisomers in the diet (P≤0·05), whereas the proportion of all synthetic α-tocopherol stereoisomers increased with increasing amount of synthetic α-tocopherol stereoisomers in the diet (P≤0·05). The proportion of α-tocopherol stereoisomers in plasma, brain, heart, lungs and abdominal fat showed the following order: RRR>RRS, RSR, RSS>Σ2S, regardless of α-tocopherol supplement. The liver had the highest proportion of Σ2S stereoisomers, and lowest proportion of RRR-α-tocopherol. In conclusion, distribution of α-tocopherol stereoisomers differs with dose and form of α-tocopherol supplementation. The results did also reveal the liver's role as the major organ for accumulation of Σ2S α-tocopherol stereoisomers.

Effects of maternal treatment with ß-hydroxy-ß-metylbutyrate and 2-oxoglutaric acid on femur development in offspring of minks of the standard dark brown type.

The aim of the study was to evaluate the effect of the diet, mother type and sex of the offspring on the mechanical and geometric parameters of long bones as well as bone tissue density in minks. Primiparous and multiparous dams were supplemented with ß-hydroxy ß-methylbutyrate (a metabolite of leucine, at the daily dosage of 0.02 g/kg of body weight) and/or 2-oxoglutaric acid (a precursor of glutamine, at the daily dosage of 0.4 g/kg of body weight) during gestation. The diet did not influence bone tissue density and the length of the humerus. An increase in the length of the femur was noted in male offspring delivered by multiparous dams. The diet resulted in an increase in the weight of the humerus in males from multiparous dams and a decrease in offspring from primiparous dams. Heavier femora were noted in male offspring delivered by both types of dams. The maximum elastic strength of the humerus was higher in the offspring delivered by multiparous than primiparous dams, irrespective of the offspring sex. The diet resulted in reduction in the ultimate strength of the femur in the male offspring delivered by primiparous dams. Only females born by multiparous dams, irrespective of the diet, showed a significant increase in the cross-sectional area of the humerus, while a significant decline was noted in males delivered by multiparous dams and in all the offspring delivered by primiparous dams. An increase in the cross-sectional area of the femur was noted in the offspring delivered by multiparous dams, while reduction was observed in the offspring delivered by primiparous dams. These results have shown for the first time that the presence of ß-hydroxy-ß-methylbutyrate or 2-oxoglutaric acid in the diet of pregnant primiparous or multiparous dams unambiguously affects the geometry and mechanical properties of offspring's long bones.

Effects of Different Sources and Levels of Zinc on Growth Performance, Nutrient Digestibility, and Fur Quality of Growing-Furring Male Mink (Mustela vison).

The objective of this study was to investigate the effects of different sources and levels of zinc (Zn) on growth performance, nutrient digestibility, serum biochemical parameters, and fur quality in growing-furring male mink. Animals in the control group were fed a basal diet with no Zn supplementation. Mink in the other nine treatments were fed the basal diet supplemented with Zn from either grade Zn sulfate (ZnSO4·7H2O), Zn glycinate (ZnGly), or Zn pectin oligosaccharides (ZnPOS) at concentrations of either 100, 300, or 900 mg Zn/kg dry matter. One hundred and fifty healthy 15-week-old male mink were randomly allocated to ten dietary treatments (n = 15/group) for a 60-day trial from mid-September to pelting in December. Mink in the Zn-POS groups had higher average daily gain than those in the control group (P < 0.05). Zn source slightly improved the feed/gain (P = 0.097). N retention was increased by Zn addition (P < 0.05). Mink supplemented with dietary Zn had higher (P < 0.05) pancreas Zn level than the control group. Fur length was greater (P < 0.05) in ZnGly and ZnPOS groups compared with the control. In addition, fur length and fur density increased (linear, P < 0.05) with Zn supplementation in the diet. In conclusion, our data show that dietary Zn addition improves growth performance by increasing nitrogen retention and fat digestibility in growing-furring mink and Z-POS is equally bioavailable to mink compared to ZnGly.

Therapeutic effect of Pseudomonas aeruginosa phage YH30 on mink hemorrhagic pneumonia.

Hemorrhagic pneumonia caused by Pseudomonas aeruginosa remains one of the most costly infectious diseases among farmed mink and commonly leads to large economic losses during mink production. The objective of this study was to investigate the potential of using phages as a therapy against hemorrhagic pneumonia in mink. A broad-host-range phage from the Podoviridae family, YH30, was isolated using the mink-originating P. aeruginosa (serotype G) D7 strain as a host. The genome of YH30 was 72,192bp (54.92% G+C), contained 86 open reading frames and lacked regions encoding known virulence factors, integration-related proteins or antibiotic resistance determinants. These characteristics make YH30 eligible for use in phage therapy. The results of a curative treatment experiment demonstrated that a single intranasal administration of YH30 was sufficient to cure hemorrhagic pneumonia in mink. The mean colony count of P. aeruginosa in the blood and lung of YH30-protected mink was less than 10(3) CFU/mL (g) within 24h of bacterial challenge and ultimately became undetectable, whereas that in unprotected mink reached more than 10(8) CFU/mL (g). Additionally, YH30 dramatically improved the pathological manifestations of lung injury in mink with hemorrhagic pneumonia. Our work demonstrates the potential of phages to treat P. aeruginosa-caused hemorrhagic pneumonia in mink.

Interactions between retinol, α-tocopherol and cholecalciferol need consideration in diets for farmed mink (Mustela vison).

A sufficient but balanced vitamin supplementation is a prerequisite for a satisfactory growth pattern and an effective immune system in mink and all other species. The fat-soluble vitamins are very sensitive to over- or under-supply because they interact with each other with respect to dose-response and chemical form. The purpose of the present study was to investigate the effect of increasing the amount of retinol in combination with RRR-α-tocopherol or all-rac-α-tocopherol in the feed given to growing mink on their retinol, cholecalciferol and α-tocopherol concentrations in plasma and selected organs. The results showed that the mink met their retinol requirements from the basal diet, but there were no negative effects of supplying various amounts of retinol on their plasma α-tocopherol concentrations. On the other hand, the study showed that the cholecalciferol status in plasma, assessed as the 25-hydroxycholecalciferol concentration, was low when retinol was supplemented in the feed at high levels. In addition, supplementation with RRR-α-tocopherol in the feed negatively affected the plasma concentration of 25-hydroxycholecalciferol compared with supplementation with all-rac-α-tocopherol. In general, female mink had higher concentrations of fat-soluble vitamins in plasma than male mink.

Uptake of selenium and mercury by captive mink: Results of a controlled feeding experiment.

Captive, juvenile, ranch-bred, male mink (Neovison vison) were fed diets containing various concentrations of methyl-mercury (MeHg) and selenium (Se) for a period of 13 weeks and then sacrificed to determine total Hg levels in fur, blood, brain, liver and kidneys and total Se concentrations in brain tissue. As MeHg concentrations in the diet increased, concentrations of total Hg in the tissues also increased with the highest level occurring in the fur > liver = kidney > brain > blood. Concentrations of Hg in the fur were correlated (r(2) > 0.97) with liver, kidney, blood and brain concentrations. The addition of Se to the mink diet did not appear to affect most tissue concentrations of total Hg nor did it affect the partitioning of Hg between the liver:blood, kidney:blood and brain:blood; however, partitioning of Hg between fur and blood was apparently affected.

Therapeutic effect of the YH6 phage in a murine hemorrhagic pneumonia model.

The treatment, in farmed mink, of hemorrhagic pneumonia caused by multidrug-resistant Pseudomonas aeruginosa strains has become increasingly difficult. This study investigated the potential use of phages as a therapy against hemorrhagic pneumonia caused by P. aeruginosa in a murine hemorrhagic pneumonia model. An N4-like phage designated YH6 was isolated using P. aeruginosa strain D9. YH6 is a virulent phage with efficient and broad host lytic activity against P. aeruginosa. No bacterial virulence- or lysogenesis-related ORF is present in the YH6 genome, making it eligible for use in phage therapy. In our murine experiments, a single intranasal administration of YH6 (2 × 10(7) PFU) 2 h after D9 intranasal injections at double minimum lethal dose was sufficient to protect mice against hemorrhagic pneumonia. The bacterial load in the lungs of YH6-protected mice was less than 10(3) CFU/g within 24 h after challenge and ultimately became undetectable, whereas the amount of bacteria in the lung tissue derived from unprotected mice was more than 10(8) CFU/g within 24 h after challenge. In view of its protective efficacy in this murine hemorrhagic pneumonia model, YH6 may serve as an alternative treatment strategy for infections caused by multidrug-resistant P. aeruginosa.

Effects of dietary copper on organ indexes, tissular Cu, Zn and Fe deposition and fur quality of growing-furring male mink (Mustela vison).

The objectives of this study were to study the effects of different levels of dietary copper on organ indexes, tissular Cu, Zn and Fe deposition and fur quality of mink in the growing-furring periods. One hundred and five standard dark male mink were randomly assigned to seven groups with the following dietary treatments: basal diet with no supplemental Cu (Control); basal diet supplemented with either 6, 12, 24, 48, 96 and 192 mg/kg Cu from copper sulphate, respectively. The colour intensity scores displayed a linear trend (P = 0.057). The spleen Cu concentrations responded in a linear (P < 0.05) fashion with increasing level of Cu, but copper supplementation did not affect speen concentrations of Fe or Zn. Supplemental dose of Cu linearly increased (P < 0.05) liver Cu and Fe concentrations but did not alter (P > 0.10) liver Zn. Our results indicate that Cu plays an important role in the pigmentation in growing-furring mink, and supplemental dietary Cu in growing-furring mink improve hair colour, and copper has limited effects on liver mineral deposition.

Copper bioavailability, blood parameters, and nutrient balance in mink.

A 3 × 3 + 1 factorial experiment was conducted based on a completely randomized design to evaluate the effects of different sources of copper on plasma metabolites, nutrient digestibility, relative copper bioavailability, and retention of some minerals in male mink. Animals in the control group were fed a basal diet, which mainly consisted of corn, fish meal, meat and bone meal, and soybean oil, with no copper supplementation. Mink in the other 9 treatments were fed the basal diet supplemented with Cu from reagent-grade copper sulfate (CuSO4), tribasic copper chloride (TBCC), or copper methionine (CuMet). Copper concentrations of the experimental diets were 50, 100, and 150 mg Cu/kg DM. Blood samples were collected via the toe clip at the end of study (d 42) to determine blood hematology and blood metabolites. A metabolism trial of 4 d was conducted during the last week of experimental feeding. There was a linear (P < 0.01) effect of dose of Cu on plasma Cu concentrations, ceruloplasmin concentration, and Cu-Zn superoxide dismutase activity. A linear response to Cu dose was noted for fat (P < 0.05) digestibility. Supplemental dose of Cu linearly increased (P < 0.05) liver Cu and decreased (P < 0.05) liver Zn level but did not alter liver Fe. The concentration of liver Cu of the mink fed with TBCC and CuMet diets was greater (P < 0.05) than that fed CuSO4. Compared with CuSO4 (100%), relative bioavailability values of TBCC were 104 and 104%, based on serum ceruloplasmin and liver copper, respectively, and relative bioavailability values of CuMet were 130 and 111%. CuMet and TBCC are more bioavailable than CuSO4. In conclusion, the relative bioavailability of CuMet obtained in this study was greater than that of CuSO4 and TBCC. Dose of Cu had an important effect on the regulating ceruloplasmin concentration, Cu-Zn superoxide dismutase activity, and the digestion of dietary fat in mink.

Effects of dietary copper on elemental balance, plasma minerals and serum biochemical parameters of growing-furring male mink (Mustela vison).

The objectives of this study were to study the effects of different levels of dietary copper on copper and zinc balance, plasma minerals and serum biochemical parameters of mink in the growing-furring periods. One hundred and five standard dark male mink were randomly assigned to seven groups with the following dietary treatments: basal diet with no supplemental Cu (Control); basal diet supplemented with either 6, 12, 24, 48, 96, or 192 mg/kg Cu from copper sulfate, respectively. The average daily gain (ADG) linearly (P = 0.0026, P = 0.0006) responded to increasing levels of Cu; maximal growth was seen in the Cu24 group. Feed efficiency tended to improve with the increase of dietary copper level (linear P = 0.0010, quad, P = 0.0011). Fecal copper, urinary copper, retention copper responded in a linear (P < 0.05) fashion with increasing level of Cu. The effect of level of Cu was linear (P < 0.001) for plasma Cu concentration. The serum glutamic-oxalacetic transaminase (GOT) and glutamic-pyruvic transaminase (GPT) activities were increased linearly (P < 0.05) with dose of Cu, but serum total protein (TP) and albumin (ALB) concentrations decreased linearly (P < 0.05) as dietary copper levels increased. Effect of level of Cu was linear (P < 0.001) for serum ceruloplasmin (CER) concentration or Cu-Zn superoxide dismutase (Cu-Zn SOD) activity. Supplemental dose of Cu linearly decreased serum triglyceride (TG) (P = 0.011) and total cholesterol (TC) (P = 0.007). Our results indicated that the activity of Cu-dependent enzymes was enhanced by increasing dietary Cu concentration and that supplementation of Cu in the diet of mink could alter the plasma lipid profile and copper concentration.

Effects of different dietary protein levels and DL-methionine supplementation on hair growth and pelt quality in mink (Neovision vision).

The effect of different dietary protein levels and DL-methionine (Met) supplementation on hair growth and the resulting pelt quality in mink was studied. Four groups of male mink were fed with four isocaloric diets containing 32% (P32), 24% (P24), 16% (P16) or P24+Met (0.8%) crude protein of dry matter (DM) from September to December. Skin biopsies were taken at the pelting. Histological techniques and computer-assisted light microscopy were used to determine the ratio of activity (ROA) of under hairs and guard hairs respectively. The results showed that when the dietary protein level reduced from 32% to 16%, body length, number and diameter of under hairs and guard hairs of minks declined, and pelt length and pelt weight of minks decreased significantly (p < 0.05). These parameters were similar between P32 and P24 with Met supplementation (p > 0.05). The hair follicle density of the winter coat was not influenced by the dietary protein levels and Met supplementation (p > 0.05). Low-protein diets content led to a reduction of hair follicle developing to next phase. It was documented that 24% crude protein of DM with Met supplementation during growing-furring period was sufficient for minks to express their genetic capacity to develop hair follicles and achieve the prime fur characteristics. Overall this study demonstrated that hair growth and hair properties in pelts are very dependent on the dietary protein and Met supply in the growing-furring period of minks.