A PCR-based assay for discriminating Cervus and Rangifer (Cervidae) antlers with mitochondrial DNA polymorphisms

This study describes a method for discriminating Rangifer antlers from true Cervus antlers using agarose gel electrophoresis, capillary electrophoresis, quantitative real-time PCR, and allelic discrimination. Specific primers labeled with fluorescent tags were designed to amplify fragments from the mitochondrial D-loop genes for various Cervus subspecies and Rangifer tarandus differentially. A 466-bp fragment that was observed for both Cervus and Rangifer antlers served as a positive control, while a 270-bp fragment was specifically amplified only from Rangifer antlers. Allelic discrimination was used to differentiate between Cervus and Rangifer antlers, based on the amplification of specific alleles for both types of antlers. These PCR-based assays can be used for forensic and quantitative analyses of Cervus and Rangifer antlers in a single step, without having to obtain any sequence information. In addition, multiple PCR-based assays are more accurate and reproducible than a single assay for species-specific analysis and are especially useful in this study for the identification of original Cervus deer products from fraudulent Rangifer antlers.     

Flow cytometric assessment of ethylene glycol monoethyl ether on spermatogenesis in rats

The effects of ethylene glycol monoethyl ether (EGEE) on testicular cell populations in rats were investigated by a flow cytometric method. Rats were administered by gavage with EGEE at the various doses of 0 (saline alone), 100, 200, 400, and 800 mg/kg body weight/day for 4 weeks. The treatment of EGEE caused decreases in the weight of testis and epididymis and in the number of testicular cells. Histopathologically, exfoliation of germ cells into the tubular lumen was observed at the doses of above 200 mg/kg. The treatment of EGEE at the dose of 400 mg/kg caused moderate testicular degeneration. A significant depletion of haploid cells and a disproportionate ratio of diploid and tetraploid cells were observed as determined by flow cytometric analysis. These results indicate that the toxic effect of EGEE on the male reproductive system may be strongly associated with the disproportion of testicular germ cells.     

Development of a multiplex PCR assay to detect Edwardsiella tarda,Streptococcus parauberis,and Streptococcus iniae in olive flounder (Paralichthys olivaceus)

A multiplex PCR protocol was established to simultaneously detect major bacterial pathogens in olive flounder (Paralichthys olivaceus) including Edwardsiella (E.) tarda, Streptococcus (S.) parauberis, and S. iniae. The PCR assay was able to detect 0.01 ng of E. tarda, 0.1 ng of S. parauberis, and 1 ng of S. iniae genomic DNA. Furthermore, this technique was found to have high specificity when tested with related bacterial species. This method represents a cheaper, faster, and reliable alternative for identifying major bacterial pathogens in olive flounder, the most important farmed fish in Korea.     

Cell proliferation and neuroblast differentiation in the dentate gyrus of high-fat diet-fed mice are increased after rosiglitazone treatment

In this study, we determined how rosiglitazone (RSG) differentially affected hippocampal neurogenesis in mice fed a low-fat diet (LFD) or high-fat diet (HFD; 60% fat). LFD and HFD were given to the mice for 8 weeks. Four weeks after initiating the LFD and HFD feeding, vehicle or RSG was administered orally once a day to both groups of mice. We measured cell proliferation and neuroblast differentiation in the subgranular zone of the dentate gyrus using Ki67 and doublecortin (DCX), respectively, as markers. In addition, we monitored the effects of RSG on the levels of DCX and brain-derived neurotrophic factor (BDNF) in hippocampal homogenates. At 8 weeks after the LFD feeding, the numbers of Ki67- and DCX-positive cells as well as hippocampal levels of DCX and BDNF were significantly decreased in the RSG-treated group compared to the vehicle-treated animals. In contrast, the numbers of Ki67- and DCX-positive cells along with hippocampal levels of DCX and BDNF in the HFD fed mice were significantly increased in the RSG-treated mice compared to the vehicle-treated group. Our data demonstrate that RSG can modulate the levels of BDNF, which could play a pivotal role in cell proliferation and neuroblast differentiation in the hippocampal dentate gyrus.     

Anti-obesity activity of diglyceride containing conjugated linoleic acid in C57BL/6J ob/ob mice

This study was to investigate the anti-obesity effects of diglyceride (DG)-conjugated linoleic acid (CLA) containing 22% CLA as fatty acids in C57BL/6J ob/ob male mice. There were four experimental groups including vehicle control, DG, CLA, and DG-CLA. The test solutions of 750 mg/kg dose were orally administered to the mice everyday for 5 weeks. CLA treatments significantly decreased mean body weight in the obese mice throughout the experimental period compared to the control (p < 0.01). All test solutions significantly decreased the levels of triglyceride, glucose and free fatty acids in the serum compared with control (p < 0.05). The levels of total cholesterol were also significantly reduced in DG and DG-CLA groups compared with the control group (p < 0.05). CLA significantly decreased weights of renal and epididymal fats compared with the control (p < 0.05). DG and DG-CLA also significantly decreased the epididymal fat weights compared with the control (p < 0.05). A remarkable decrease in the number of lipid droplets and fat globules was observed in the livers of mice treated with DG, CLA, and DG-CLA compared to control. Treatments of DG and CLA actually increased the expression of peroxisome proliferator-activated receptor gamma. These results suggest that DG-CLA containing 22% CLA have a respectable anti-obesity effect by controlling serum lipids and fat metabolism.     

Effect of ACTH on plasma corticosterone and cortisol in eagles and condors

The effect of ACTH on plasma corticosterone and cortisol was determined in 12 eagles (Haliaeetus leucocephalus) and in 6 Andean condors (Vultur gryphus). In all raptors, the concentration of plasma corticosterone was substantially greater than that of cortisol. After ACTH administration, the eagles had a marked increase (P less than 0.001) in plasma corticosterone concentrations, but not in plasma cortisol. Administration of saline solution did not induce increased plasma corticosterone concentrations in the eagles. The condors had a smaller increase (P less than 0.002) in plasma corticosterone concentrations after ACTH administration, as compared with that of the eagles. However, administration of saline solution in 2 condors resulted in an increase in corticosterone similar to the increase after ACTH administration. In the condor, a stress-related release of endogenous ACTH may have an effect similar to that induced by exogenously administered ACTH. Plasma cortisol concentrations did not increase significantly after administration of ACTH or saline solution in either raptor species.     

Lumbar spondylectomy in the subhuman primate.

Removal of the 2nd or 3rd lumbar vertebrae (or both) has been accomplished in the subhuman primate (Macaca mulatta). Variations between this animal and the dog in posture, vertebral column anatomy, and spinal cord blood supply made no apparent difference in the results when compared with those in previous experiments. All macaques were able to clinb and to use their hind legs in a normal manner within 3 days after surgical operation. Once hair had regrown over the surgical incision, persons not familiar with the animals were not able to identify the animals that had undergone surgical operation. If a single vertebra was removed, use of spinous process and a single vertebral body plate was sufficient to stabilize the vertebral column. Except for 1 rhesus macaque whose spinous process plates were removed 20 days after the operation, vertebral columns of macaques (n = 3) that underwent single spondylectomy healed in nearly normal alignment. With the removal of 2 lumbar vertebrae, the previously described stabilization procedure was not sufficient to prevent kyphotic deformity of the vertebral column. Spinous process plates from 1 macaque were removed after 64 days. However, all these macaques (n = 3) could climb and use their legs as well as others in the colony.     

Prevalence of Campylobacter spp isolated from the intestinal tract of pigs raised in an integrated swine production system

OBJECTIVE: To enumerate the prevalence of Campylobacter isolates in the intestinal tract of market-weight swine raised in an integrated swine operation in Texas. SAMPLE POPULATION: Samples of cecal contents were collected from 595 pigs (mean body weight, 110 kg [242 lb]) at time of slaughter. Pigs were off-spring of Yorkshire-Landrace sows and Duroc or Hampshire boars. Pigs originated from 4 farrow-to-finish farms. PROCEDURE: During a 9-month period, visits were made to a slaughter plant to remove cecal contents from market-weight hogs. Samples were obtained from 50 pigs/visit from designated farms so that samples were obtained 3 times from pigs of each of 4 farms. Isolation of Campylobacter spp was accomplished by use of enrichment broth and restrictive media, using microaerophilic conditions. RESULTS: Campylobacter spp were isolated from 70 to 100% of the pigs, depending on the farm and the date the samples were collected. Campylobacter coli was isolated from 20 to 100% (mean, 60%) of samples, and C jejuni was isolated from 0 to 76% (mean, 31%) of samples. Campylobacter lari was isolated from 2 pigs. Concentrations of C coli or C jejuni ranged from 10(3) to 10(7) colony-forming units/g of cecal content. CONCLUSIONS AND CLINICAL RELEVANCE: Campylobacter coli generally is accepted as a common inhabitant of the intestinal tract of swine. However, analysis of results of this study suggests that a relatively high prevalence of C jejuni may be found in pigs raised on specific farms.