Detection of self-biting behavior of mink by loop-mediated isothermal amplification (LAMP) and sequence-characterized amplified regions (SCAR).

Self-biting disease occurs in most farmed fur animals in the world. The mechanism and rapid detection method of this disease has not been reported. We applied bulked sergeant analysis (BSA) in combination with RAPD method to analyze a molecular genetic marker linked with self-biting trait in mink group. The molecular marker was converted into SCAR and loop-mediated isothermal amplification (LAMP) marker for rapid detection of this disease. A single RAPD marker A10 amplified a specific band of 1000bp in self-biting minks. The sequences of the bands exhibited 73% similarity to the Canis Brucella. SCAR and LAMP marker were designed for the specific fragment of RAPD marker A10 and validated in 30 self-biting minks and 30 healthy minks. c2 test showed difference (p0.05) with SCAR and significant difference (p0.01) with LAMP in the detection rate between the two groups, but LAMP method was more accurate than SCAR method. This indicated that LAMP can be used as a positive marker to detect self-biting disease in minks.

Bioinformatic analysis based on the complete coding region of the MSTN gene within and among different species.

Myostatin, encoded by the MSTN gene (previously GDF8), is a member of the transforming growth factor-ß superfamily, which normally acts to limit skeletal muscle mass by regulating the number and growth of muscle fibers. In this study, a total of 84 myostatin gene sequences with known complete coding regions (CDS) and corresponding amino acid sequences were analyzed from 17 species, and differentiation within and among species was studied using comparative genomics and bioinformatics. Characteristics of the nucleotide and amino acid sequences were also predicted. The results indicated that a total of 569 polymorphic sites, including 53 singleton variable sites and 516 parsimony informative sites, which could be sorted into 44 haplotypes, were detected from 17 species. Observed genetic diversity was higher among species than within species, and Vulpes lagopus was more polymorphic than other species. There was clear differentiation of the myostatin gene among species and the reconstructed phylogenetic tree was consistent with the NCBI taxonomy. The myostatin gene was 375-aa long in most species, except for Mus musculus (376 aa) and Danio rerio (373 aa). The amino acid sequences of myostatin were deemed hydrophilic, and had theoretical pI values of <7.0, mostly due to the acidic polypeptide. The instability index of the myostatin protein was 40.48-51.63, indicating that the polypeptide is not stable. The G+C content of the CDS nucleotide sequence in different species was 40.60-51.69%. The predicted promoter region of the Ovis aries myostatin gene was 150-220 bp upstream of the start codon.

Correlation analysis between single nucleotide polymorphism of DRD1 gene and stereotyped behavior of blue fox.

This study was performed to investigate the correlation between stereotyped behavior of the blue fox and single nucleotide polymorphisms (SNPs) of the DRD1 gene. We choose the DRD1 gene as a major gene for investigating the correlation of gene polymorphism and self-biting disease by means of direct sequencing. Part of the DRD1 gene exon of the blue fox was cloned; the length of the whole sequence was 864 bp. Four SNPs were detected and analyzed by the chi-square analysis; the results showed that the gene polymorphism of T206C in the DRD1 gene had a significant correlation with self-biting (P < 0.01). Therefore, marker-assistant selection on self-biting of blue foxes using these SNPs can be applied to select healthy individuals.

Quantitative candidate gene association studies of metabolic traits in Han Chinese type 2 diabetes patients.

Recent genome-wide association studies have identified many loci associated with type 2 diabetes mellitus (T2DM), hyperuricemia, and obesity in various ethnic populations. However, quantitative traits have been less well investigated in Han Chinese T2DM populations. We investigated the association between candidate gene single nucleotide polymorphisms (SNPs) and metabolic syndrome-related quantitative traits in Han Chinese T2DM subjects. Unrelated Han Chinese T2DM patients (1975) were recruited. Eighty-six SNPs were genotyped and tested for association with quantitative traits including lipid profiles, blood pressure, body mass index (BMI), serum uric acid (SUA), glycated hemoglobin (HbA1c), plasma glucose [fasting plasma glucose (FPG)], plasma glucose 120 min post-OGTT (P2PG; OGTT = oral glucose tolerance test), and insulin resistance-related traits. We found that CAMTA1, ABI2, VHL, KAT2B, PKHD1, ESR1, TOX, SLC30A8, SFI1, and MYH9 polymorphisms were associated with HbA1c, FPG, and/or P2PG; GCK, HHEX, TCF7L2, KCNQ1, and TBX5 polymorphisms were associated with insulin resistance-related traits; ABCG2, SLC2A9, and PKHD1 polymorphisms were associated with SUA; CAMTA1, VHL, KAT2B, PON1, NUB1, SLITRK5, SMAD3, FTO, FANCA, and PCSK2 polymorphisms were associated with blood lipid traits; CAMTA1, SPAG16, TOX, KCNQ1, ACACB, and MYH9 polymorphisms were associated with blood pressure; and UBE2E3, SPAG16, SLC2A9, CDKAL1, CDKN2A/B, TCF7L2, SMAD3, and PNPLA3 polymorphisms were associated with BMI (all P values <0.05). Some of the candidate genes were associated with metabolic and anthropometric traits in T2DM in Han Chinese. Although none of these associations reached genome-wide significance (P < 5 x 10(-8)), genes and loci identified in this study are worthy of further replication and investigation.

Effects of dietary protein levels on digestibility of nutrients and growth rate in young female mink (Mustela vison).

This study investigated digestibilities of nutrients and feed efficiency in female mink at the different dietary protein levels during the mink growth period. Effects of dietary protein on growth performance of minks were also measured. Sixty 45-day-old healthy female minks were randomly assigned to 6 treatment groups with 10 animals in each group. Animals were fed diets varying in protein levels: 28% (Group I), 30% (Group II), 32% (Group III), 34% (Group IV), 36% (Group V) and 38% (Group VI), respectively. The digestibilities of key nutrients were determined on Day 14 after initiating the experiment and the last 3 days. From the beginning of the study, body weight and feed intake were weighed and recorded every other week in order to calculate the average daily bodyweight gain and the feed efficiency. The trial had demonstrated that nitrogen intake was greatly significantly different, which was affected by dietary protein levels (p < 0.001). Growth performance of minks was impaired when dietary protein level was at 28%. When dietary protein level was at 34%, minks had the best daily gains, feed efficiency, and digestibilities of some key nutrients.

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.

Effects of dietary fat levels on nutrient digestibility and production performance of growing-furring blue foxes (Alopex lagopus).

The objective of this study was to determine whether nutrient digestibility and production performance of growing-furring blue foxes (Alopex lagopus) are affected by different dietary fat levels. Sixty-four young animals were randomly assigned to four groups (A, B, C, D) provided with diets containing approximately 12%, 26%, 40%, 54% fat in the dry matter respectively. When dietary fat level was increased, the apparent digestibility of main nutrients except for crude carbohydrates, and gross energy were improved (p < 0.0001). The amount of nitrogen excreted was reduced and the biological value of protein was enhanced (p < 0.0001). Over the experimental phase, the efficiency of metabolizable energy (ME) used for gain in group B (26% fat content) was higher than that in other groups. When diets with 12-26% fat content were fed, there was an increasing tendency in skin size. But it had negative effects on skin size and fur quality when the amount of fat content was over 40%. In conclusion, the experiment showed that dietary fat could significantly improve some nutrient utilization and significantly reduce feed/gain ratio as a main energy source. The most preferable fur quality and efficiency of ME used for gain were obtained when diet contained 26% fat level in growing-furring period.

Effects of diets with different protein and DL-methionine levels on the growth performance and N-balance of growing minks.

This study was performed to evaluate the effects of diets with different protein and dl-methionine (Met) levels on nitrogen (N) retention, nutrient digestibility, growth performance, and some blood parameters in growing minks. Eighty healthy male minks were selected and randomly divided into five groups with different types of diet. The dietary protein levels, expressed as percentage of dry matter (DM), were 36% (HP) and 28% (LP), corresponding to average 363g and 295g protein/kg DM, respectively. LP was supplemented with Met (0.3%, 0.6% and 0.9% DM); the codes were LP+M1, LP+M2 and LP+M3, respectively. From July to middle of September, the average daily gain and feed: gain ratio (F/G) of the minks that received the diet with 0.6% Met added to the low protein diet was better than feeding HP and other groups. Fecal N and Urinary N of group LP+M2 were the lowest one, in contrast, the daily retention of N was the highest one. Digestibility of DM and CP were not affected by different diets, but digestibility of fat declined with dietary protein level decreasing. Serum urea nitrogen (SUN) was affected by different protein and Met levels. Considering all factors the best performance could be observed offering LP+M2, the prime level of Met was 13.87 g/kg DM in dietary, and 258.5 g digestible protein kg(-1) DM was enough for mink in growing period. Furthermore, addition of Met in low protein diets for minks would be beneficial in terms of reduced feed expenses and lower nitrogen emissions to the environment.

Genetic evidence for domestic raccoon dog rabies caused by Arctic-like rabies virus in Inner Mongolia, China.

An outbreak of rabies affected domestic raccoon dogs on an animal farm in Inner Mongolia, China in 2007. A study was conducted to characterize the aetiological agent and clarify the origin of the rabies virus. Brain tissues were obtained from five rabid raccoon dogs. Viral nucleoprotein antigen was detected in the brain tissues and five rabies viruses were isolated from these rabid animals. Phylogenetic analysis of the N and G gene sequences showed that these isolates were closely related to Arctic-like rabies viruses isolated from the far-eastern region of Russia and South Korea, but distinct from the rabies viruses that are widely distributed in endemic areas in China. Epidemiological data suggested that the likely source of infection was from one wild raccoon dog that was captured and placed in the same type of pen used for domestic raccoon dogs.

Selective and lithography-independent fabrication of 20 nm nano-gap electrodes and nano-channels for nanoelectrofluidics applications.

A new method has been developed to selectively fabricate nano-gap electrodes and nano-channels by conventional lithography. Based on a sacrificial spacer process, we have successfully obtained sub-100-nm nano-gap electrodes and nano-channels and further reduced the dimensions to 20 nm by shrinking the sacrificial spacer size. Our method shows good selectivity between nano-gap electrodes and nano-channels due to different sacrificial spacer etch conditions. There is no length limit for the nano-gap electrode and the nano-channel. The method reported in this paper also allows for wafer scale fabrication, high throughput, low cost, and good compatibility with modern semiconductor technology.

Odontoblastic Exosomes Attenuate Apoptosis in Neighboring Cells.

Each odontoblast is tightly linked to other odontoblasts. They form a line of defense and are capable of withstanding external stimuli, particularly the inflammation caused by caries. Thus, we investigated exosomes derived from odontoblasts as an intercellular mechanism by which inflamed odontoblasts are protected from apoptosis. CD63, an exosome marker, was expressed at high levels in caries-affected regions of the dental pulp. We conducted an ex vivo experiment by applying different concentrations of lipopolysaccharide (LPS) to the odontoblast-like cells (mineralization was induced in stem cells derived from the apical papilla). Odontoblast-like cells treated with a high concentration of LPS (20 µg/mL LPS, severely affected) exhibited an accelerated release of exosomes, which attenuated the LPS-induced cell apoptosis of odontoblast-like cells treated with a low concentration of LPS (1 µg/mL LPS, mildly affected). Next, we blocked exosome uptake with chlorpromazine, and the rescue effect vanished. Based on our findings, severely inflamed odontoblasts attenuate the apoptosis of mildly inflamed neighboring cells through an exosome-mediated intercellular signaling pathway.

Comparative studies on testis, epididymis and serum hormone concentrations in foxes, and hybrids during the pre-breeding period.

Silver fox and blue fox belong to different genera, and the hybrid males are reproductively sterile. In the present study, there was a comparison of testicular and epididymal morphology and serum hormone concentrations among silver foxes, blue foxes, and the hybrids during the pre-breeding period, using 20 male silver foxes, 20 male blue foxes, 15 male HSBs (silver fox male × blue fox female hybrids) and 15 male HBSs (blue fox male × silver fox female hybrids), respectively. Hybrids had a smaller diameter of seminiferous tubules than pure-species males, and testes of hybrid males did not differ in mean size and relative weight from pure-species males. There were many Sertoli cells and spermatogenic cells in silver foxes and blue foxes, while numbers of spermatogonia and primary spermatocytes were less with no secondary spermatocytes in the hybrids. Furthermore, mean serum testosterone and estradiol concentrations in the hybrids were less, and FSH, LH, and PRL were greater than that in silver foxes and blue foxes (P < 0.05), suggesting that lesser concentrations of testosterone and estradiol and greater concentrations of FSH, LH and prolactin can inhibit the completion of spermatogenesis during the pre-breeding period. The results indicate that fox hybrid sterility may result from failures at the early stages of spermatogenesis.

Effects of sources and concentrations of zinc on growth performance, nutrient digestibility, and fur quality of growing-furring female mink (Mustela vison).

A completely randomized 3 × 3 + 1 factorial experiment was conducted to evaluate the effects of sources and concentrations of Zn on growth performance, nutrient digestibility, serum biochemical endpoints, and fur quality in growing-furring female black mink. One hundred fifty healthy 15-wk-old female mink were randomly allocated to 10 dietary treatments ( = 15/group) for a 60-d trial. Animals in the control group were fed a basal diet, which consisted of mainly corn, soybean oil, meat and bone meal, and fish meal, with no Zn supplementation. Mink in the other 9 treatments were fed the basal diet supplemented with Zn from either zinc sulfate (ZnSO), zinc glycinate (ZnGly), or Zn pectin oligosaccharides (ZnPOS) at concentrations of either 100, 300, or 900 mg Zn/kg DM. The results showed that mink in the ZnPOS groups had higher ADG than those in the ZnSO groups (main effect, < 0.05). The addition of Zn reduced the G:F ( < 0.05). In addition, CP and crude fat digestibility were linearly increased with Zn supplementation ( < 0.05) and N retention tended to increase with Zn addition ( = 0.08). Dietary Zn supplementation increased the concentration of serum albumin and activity of alkaline phosphatase ( < 0.05). There was a linear effect of dietary Zn on the concentration of tibia Zn and pancreatic Zn ( < 0.05). For fur quality characteristics, the fur density and hair color of mink were improved by dietary Zn concentration ( < 0.05). Compared with ZnSO (100%), relative bioavailability values of ZnGly were 115 and 118%, based on tibia and pancreatic Zn, respectively, and relative bioavailability values of ZnPOS were 152 and 142%, respectively. In conclusion, this study demonstrates that Zn supplementation can promote growth and increase nutrient digestibility and fur quality and that ZnPOS is more bioavailable than ZnSO and ZnGly in growing-furring female mink.

Comparative studies on testicular and epididymal morphology, and serum hormone concentrations in foxes and the hybrids during the breeding season.

The silver fox and the blue fox belong to different genera, and the hybrid males are fully or partially sterile. In the present study, the objective was to evaluate the causes of hybrid male sterility, and therefore analyze the differences in testicular, and epididymal morphology and serum hormone concentrations among silver foxes, blue foxes, and the hybrids during the breeding season. Samples were collected from 20 male silver foxes, 20 male blue foxes, 15 male HSBs (silver fox female × blue fox male hybrids) and 14 male HBSs (blue fox male × silver fox female hybrids), respectively. Seminal evaluation showed large numbers of sperm present in the semen of blue foxes and silver foxes, but no sperm present in the hybrids. Mean testicular volume and the diameter of seminiferous tubules in silver foxes and blue foxes were greater than in the hybrids; and there were many Sertoli cells, spermatogenic cells, and sperm in silver foxes and blue foxes, while spermatogenic cells decreased with no sperm in the hybrids. Mean serum LH and prolactin concentrations in silver foxes and blue foxes were less and testosterone was greater than in the hybrids (P<0.05). The results indicate that germ cell meioses in the hybrids were arrested at the prophase stage of meiosis, and that lesser concentrations of testosterone and greater concentrations of LH and prolactin can inhibit the completion of spermatogenesis.

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.

Cloning and characterization of the cDNA sequences of two venom peptides from Chinese scorpion Buthus martensii Karsch (BmK).

From a cDNA library made from venom glands of Chinese scorpions of Buthus martensii Karsch, full-length cDNAs encoding precursors of two venom peptides have been isolated using a cDNA probe synthesized by polymerase chain reaction. Sequence analysis of the cDNAs revealed that one encoded precursor was 85 amino acid residues long including a signal peptide of 19 residues and a mature peptide (named BmK T) of 66 residues, and another encoded precursor was 84 residues long containing the same length signal peptide and a mature peptide (BmK M4 isoform, named BmK M4') of 64 residues. The analysis of amino acid sequence similarity indicated that the BmK T was homologous with both mammalian and insect toxins from BmK scorpion or other scorpions, and the BmK M4' was highly homologous with the members of the mammalian neurotoxin family of BmK, having two point mutations in amino acid residue sequence compared to BmK M4, a natural toxin from BmK.

Surgical treatment of 32 cases of long-term atopic keratoconjunctivitis using the amniotic membrane.

PURPOSE: To evaluate the use of surgical treatment with amniotic membrane for long-term atopic keratoconjunctivitis. Damaged corneas were repaired with various techniques: amniotic membrane transplantations, amniotic membrane coverings, amniotic membrane fillings (AMFs), and amniotic membrane inlay fillings, the latter of which were combined with glycerol-preserved corneal transplants. METHODS: This retrospective study was conducted on 37 eyes belonging to 37 patients with atopic keratoconjunctivitis. Thirty-two patients were classified into four groups according to surgical technique. Five patients undergoing medical management served as controls. Surgical outcome was measured by recovery time and long-term visual improvement. RESULTS: In all surgical eyes, integrity of ocular tissues was effectively restored and symptoms were reduced at 24.4 ± 13 days post recovery. Mean best-corrected visual acuity improved from 0.6 ± 0.2 to 0.198 ± 0.16 logarithm of the minimum angle of resolution (P<0.001). There were no intraoperative or postoperative complications, with the exception of two recurring cases, both controlled by medication. Recovery time of the control groups lasted 52 ± 16 days. In controls, mean best-corrected visual acuity improved from 0.74 ± 0.15 to 0.54 ± 0.29 logarithm of the minimum angle of resolution (P ≤ 0.05). The vision improvement has significant difference for surgical treatment vs medical. (Mann-Whitney U-test, U = 119, P < 0.05, one tailed).Vision improvements remained stable during a mean follow-up period of 21.7 ± 3.8 months. CONCLUSION: Patients suffering from severe chronic atopic keratoconjunctivitis and its complications can benefit from suitable surgical treatments: transplants, covers, fillings, or corneal graft surgeries supplemented with AMFs.

[Estimate the safety of baculovirus insecticides: the history and the status Quo].

This article reviews the evaluation of security of baculovirus used as a pesticide. Since 1970s, the scholars have done a lot of experiments with kinds of baculovirus to test the security of a large number kinds of living things even our human beings. Almost all experiments proved that baculovirus is secure, but some experiments came to different conclusions. These gave rise to great debate twice when they were published, but these conclusions have been proved to be wrong with later test by other scientists or the author himself. Since 90s baculovirus have been used a great deal as the vector to express the foreign gene. Some of them reported the expression in mammalian cells, which brought the suspicious of the baculovirus safety. This article made an analysis and a conclusion about them. Also, this article laid emphasis on the security of recombination baculovirus with the results of the security experiment of self-made recombination baculovirus pesticide. In the last analysis, this article draws a conclusion that the baculovirus and recombinant baculovirus insecticides are secure.