青花菜不同器官中4-甲基亚磺酰丁基硫苷及萝卜硫素含量分析

 【目的】青花菜的抗癌功能主要与4-甲基亚磺酰丁基硫苷的降解产物萝卜硫素有关,通过测定不同青花菜材料花球、茎和叶片中两者的含量并分析两者含量间的相关性,为青花菜高营养品质育种提供理论依据。【方法】采用反相高效液相色谱法,对24个不同青花菜材料花球、茎和叶片中4-甲基亚磺酰丁基硫苷含量,及其中的10份材料花球、茎和叶片中萝卜硫素含量进行测定和分析。【结果】青花菜不同基因型间以及不同器官间4-甲基亚磺酰丁基硫苷和萝卜硫素含量差异均极显著,4-甲基亚磺酰丁基硫苷和萝卜硫素含量均为花球含量最高,其次为茎,叶片含量最低;不同材料花球中4-甲基亚磺酰丁基硫苷平均含量分别为茎和叶片的4.4和13.97倍,萝卜硫素平均含量分别为茎和叶片的4和8.7倍,而有的基因型材料茎和叶片中4-甲基亚磺酰丁基硫苷的含量较高,约为花球中的50%;参试的10个不同基因型材料的花球、茎和叶片中的萝卜硫素含量与其前体4-甲基亚磺酰丁基硫苷含量间相关系数分别为0.9886**,0.9994**,0.9935**。【结论】不同青花菜材料间不同器官中4-甲基亚磺酰丁基硫苷含量存在显著差异;在青花菜萝卜硫素开发应用中,可选择其前体4-甲基亚磺酰丁基硫苷作为筛选指标,并可筛选茎和叶中4-甲基亚磺酰丁基硫苷含量较高的材料作为提取萝卜硫素的原料。     

Methyl‐β‐Cyclodextrin Improves Sperm Capacitation Status Assessed by Flow Cytometry Analysis and Zona Pellucida‐Binding Ability of Frozen/Thawed Bovine Spermatozoa

Mammalian sperm undergo a series of biochemical transformations in the female reproductive tract that are collectively known as capacitation. Cyclodextrins added to the sperm culture medium have been described to induce in vitro sperm capacitation, enabling its use in protein‐free media. However, the additive capacitating effect of methyl‐β‐cyclodextrin (MβCD) in the medium containing bovine serum albumin (BSA) is unknown in the bovine species. In this study, we evaluated the effects of incubating frozen–thawed bovine spermatozoa in a BSA‐containing medium supplemented with MβCD on different sperm quality and functional parameters. Sperm viability decreased with the addition of MβCD in a dose‐dependent manner (p < 0.05), and DNA damage could be observed but only with the highest concentration of MβCD. However, pre‐incubation of spermatozoa in MβCD‐supplemented medium improved the capacitation status as assessed by the increase in plasma membrane fluidity, intracellular calcium concentration, induced acrosome reactivity and zona pellucida (ZP)‐binding ability (p < 0.05). Thus, we conclude that MβCD supplementation is able to enhance the capacitation status of frozen–thawed bovine spermatozoa cultured in capacitation medium containing BSA and could result in a valid strategy for its application on artificial reproductive technologies such as in vitro fertilization or intracytoplasmic sperm injection.     

Linking macrodetritivore distribution to desiccation resistance in small forest fragments embedded in agricultural landscapes in Europe

Purpose

Most of the agricultural landscape in Europe, and elsewhere, consists of mosaics with scattered fragments of semi-natural habitat like small forest fragments. Mutual interactions between forest fragments and agricultural areas influence ecosystem processes such as nutrient cycling, a process strongly mediated by the macrodetritivore community, which is however, poorly studied. We investigated macrodetritivore distribution patterns at local and landscape-level and used a key functional trait (desiccation resistance) to gain mechanistic insights of the putative drivers.

Methods

Macrodetritivores were sampled in forest edges-centres of 224 European forest fragments across 14 landscapes opposing in land use intensity. We used a multilevel analysis of variance to assess the relative contribution of different spatial scales in explaining activity-density and Shannon-diversity of woodlice and millipedes, together with a model-based analysis of the multivariate activity-density data testing the effect on species composition. Secondly, we tested if desiccation resistance of macrodetritivores varied across communities at different spatial scales using linear mixed effect models.

Results

Forest edge-centre and landscape use intensity determined activity-density and community composition of macrodetritivores in forest fragments, while fragment characteristics like size and continuity were relatively unimportant. Forest edges and higher intensity landscapes supported higher activity-density of macrodetritivores and determined species composition. Forest edges sustained woodlouse communities dominated by more drought tolerant species.

Conclusions

Landscape use intensity and forest edges are main drivers in macrodetritivore distribution in forest fragments with desiccation resistance a good predictor of macrodetritivore distribution. Key functional traits can help us to predict changes in community structure in changing landscapes.

     

生态型农业观光园规划设计

通过阐述园区规划设计原则及理论根据,结合实例分析了生态型农业观光园的设计内容,有利于更好的研究与开发农业观光园项目。     

Analgesic effect of pre-operative etodolac and butorphanol administration in dogs undergoing ovariohysterectomy

The analgesic, bleeding, and renal effects of dogs pre‐medicated with etodolac with and without butorphanol were evaluated. Twenty‐four 1‐year‐old healthy dogs, weighing 19 ± 3 kg (mean ± SD) were randomly assigned to four treatment groups (n = 6): control (C), etodolac (E), butorphanol (B), and etodolac with butorphanol (EB). Etodolac (12–14 mg kg^?1 PO) was given 1 hour before propofol induction and isoflurane maintenance anesthesia. Butorphanol (0.4 mg kg^?1 IV) was given immediately following endotracheal intubation. Control dogs received only propofol (8 mg kg^?1 to effect) and isoflurane anesthesia. All dogs were mechanically ventilated to maintain Pe ′CO₂ between 35 and 45 mm Hg (4.7–6.0 kPa). Lactated Ringer's solution was given at 10 mL kg^?1 hour^?1 during anesthesia. Plasma cortisol concentrations were assessed 1 day prior to surgery (baseline), immediately prior to anesthesia induction, and every 30 minutes until 5 hours following extubation, and 1 day after surgery. Total duration of anesthesia was 50 minutes and total surgery duration was 30 minutes. Isoflurane concentration area under the curve (AUC) over time during the anesthesia was compared among treatment groups. Buccal mucosal bleeding time (BMBT) was assessed 1 day before E administration and during surgery. Urine GGT to urine creatinine ratio, BUN, and plasma creatinine were taken daily from 1 day before to 3 days after surgery. Behavioral pain scores (numerical rating scale) were assessed by two observers blinded to the treatment during the 5‐hour recovery period at 30 minute intervals until 3 hours, and again at 5 hours after extubation. All data were analyzed using anova . Multiple comparisons were performed if the anova was significant. Alpha value was set at 0.05. Plasma cortisol concentrations significantly increased from time of extubation in all the treatment groups. They did not return to the baseline until 5, 2.5, 1.5, and 1.5 hours after extubation in the C, B, E, and EB groups, respectively. Isoflurane AUC was not significantly different among treatment groups. Dogs treated with EB had significantly less behavioral pain than all other groups throughout the 5‐hour recovery period. No significant difference was found between treatment groups or within treatment groups over time in BMBT, or any renal variables. This study demonstrated that (i) pre‐operative administration of E provides profound analgesia during the post‐operative period without renal or bleeding side‐effects in dogs undergoing OHE; and (ii) a combination of butorphanol–etodolac provides the best analgesic effect during the post‐operative period based on the behavioral pain score.     

Genetic Description of Factor XI Deficiency in Holstein Semen in Western Japan

Factor XI deficiency was detected in Holstein cows and mummified foetuses in Japan; however, no report is available about the occurrence of Factor XI deficiency in Holstein semen in Japan. Five hundred cows in twelve dairy farms in Hiroshima Prefecture, Japan were under the study. Genomic DNA was extracted from the cows using a commercial DNA kits and screened to Factor XI mutation. Based on the information of the carrier cows found in the cattle population, four Holstein bulls were analysed for Factor XI mutation. DNA was extracted from bull's semen using phenol chloroform method. Extracted genomic DNA of the bull's semen was typed for Factor XI using specific polymerase chain reaction (PCR) primers. The resultant PCR was sequenced using big dye terminator sequencing method. The pedigree of the bulls was investigated. Furthermore, the inheritance of Factor XI mutation to next generation was estimated. Out of the 500 cows, five were heterozygous to Factor XI. Moreover, out of the four bulls, one was found to carry the mutation of Factor XI; it was also a complex vertebral malformation (CVM) carrier. In DNA sequencing, the insertion mutation of 76 bp of poly-adenine that characterizes the Factor XI deficiency was detected in the carrier bull as well as the carrier cows. Pedigree analysis of the carrier bull revealed that his father and mother ID were 2247419A and 14189172A, respectively, that originated from USA Holstein. Out of six daughter cows born to the carrier bull, one cow (16.6%) inherited Factor XI mutation, while three of them (50.0%) inherited CVM mutation. Autosomal recessive genes that affect cow's reproduction have a particular concern to dairy industry. To our knowledge this is the first report of Factor XI mutation in Holstein semen in Japan.     

Cervical Ripening and Parturition in Cows are Driven by a Cascade of Pro-Inflammatory Cytokines

The final stages of cervical ripening and parturition resemble an inflammatory process. Although the role of cytokines in both spontaneous and experimentally induced parturitions has been described in several small laboratory animals and humans, the involvement of pro‐inflammatory and regulatory cytokines in physiologic parturition in cows has not been determined. In this study, the cytokine expression profiles were assessed in bovine cervical tissue at several stages of pregnancy and at parturition. Serial biopsy samples of the cervix were obtained from 10 cows on day 185 and day 275 of pregnancy (which was on average 5.4 days before parturition) and at parturition. Messenger RNA expression levels of interleukin (IL)‐1β, IL‐6, IL‐8, IL‐10 and tumour necrosis factor (TNF)α were determined using real‐time polymerase chain reaction and the number of neutrophils and eosinophils was estimated by Luna and Sirius Red staining. At parturition, IL‐8 expression had increased 430‐fold (p < 0.001) when compared with that of the day 185 of pregnancy, large numbers of neutrophils had invaded the cervix while eosinophils remained scarce, IL‐1β had increased eightfold (p < 0.05) and IL‐6 had not changed significantly. Additionally, IL‐10 was increased by 10‐fold (p < 0.001) and TNFα decreased by 57% (p < 0.05) when compared with that of the day 185 of pregnancy. The large increase in expression of IL‐8, enabling the influx of neutrophils, is indicative of its important role in the final stage of cervical ripening and at parturition. As previous studies have shown that neutrophils excrete matrix metalloproteinases (MMP), this might contribute to softening of the cervix. In contrast, the only slightly increased levels of IL‐1, steady concentrations of IL‐6 and decreased TNFα, the potential consequences of increased IL‐10 expression, indicate that final cervical of cows in ripening at term parturition is an inflammatory process influenced by regulatory cytokines.