养猪生产中后代性别的预选——发展现状和应用前景

动物精子的性别可通过流式精子分选仪和DNA标识进行鉴定,而利用鉴定性别的精子(即性控精子)并借助人工授精技术或其它授精技术产生的后代,在过去5年中估计已多达30000个(其中大多数是母牛)。有关文献资料证明,能有效地区分X精子和Y精子的唯一标记物是精子染色体中的DNA。众所周知,目前世界各地采用的方法是Beltsville精子分选技术,该技术根据X精子群和Y精子群中DNA相对含量上的差异,用荧光染液(Hoechst33342染液)标记精子,随后利用流式细胞仪分选经荧光标记的精子,从而达到分离精子的目的。目前,X精子或Y精子正常的生产速度是每小时1500万个,该项技术已在家畜、实验动物和动物园动物中应用,如果将该技术应用在人上,在预选后代性别比例上可达90%～95%的成功率。因动物品种不同,性控精子在动物体内的授精部位也不相同。常规的人工授精技术、宫内授精技术、输卵管内授精技术、用于胚胎移植的体外受精(in-vitro fertilization,IVF)技术或子宫角深部授精技术均能有效地使动物怀孕,至于利用哪种技术进行性控精子的精则取决于动物品种。尽管所有动物都能获得高纯度的分选精子,但是在实际生产中利用低剂量精子还难以让母猪怀孕。子宫角深部授精技术每次授精0.5～1.00亿个性控精子已能产生可喜的效果:利用特制的输精管,输入常规人工授精所需精子量的五十分之一的性控精子,足以使动物怀孕。性控精子通过常规的授精技术能够被猪成功利用前,还需重新设计输精管,同时输精的次数和每次授精时精子数量也需作进一步的研究。分选精子的低温保存技术已被牛人工授精普遍采用。尽管已能利用冷冻的性别分选精子生产小猪,但是性别分选精子经冷冻和解冻处理后在常规生产中的应用还未达到最理想的效果。本文将讨论猪精子性别分选的最新研究成果及其发展趋势,并重点探讨将性别分选技术应用于养猪生产中必须对其进行必要的技术开发。     

Ultrastructural and cytochemical studies in normal Japanese quail (Coturnix coturnix japonica) eosinophils and in those from birds with experimentally induced eosinophilia

Normal eosinophil development in the Japanese quail (Coturnix coturnix japonica) was similar to that described in the fowl and the duck, with granulogenesis occurring in the Golgi apparatus. The characteristic lipid droplets were small in the immature eosinophils, and after staining specifically for lipid, small moieties were also traced to the Golgi apparatus. In mature eosinophils the lipid droplets measured between 1.0 and 1.5 micron in diameter and they were surrounded by profiles of smooth endoplasmic reticulum. Eosinophilia was difficult to induce in quails; injections of either horse serum or bovine serum albumin (BSA)/aluminium hydroxide produced a poor response. In some quails in which eosinophilia was produced, however, eosinophil granules showed many crescentic and vacuolated forms. The lipid droplets in the activated eosinophils were fused in many cells to form large intracellular aggregates of lipid. Quail eosinophils, which hitherto have been regarded as peroxidase-negative, had strong activity in the lipid droplets of cells from stimulated birds. It is postulated that this peroxidase-positive reaction may represent a form of ceroid or lipofuscin pigment resulting from lipid peroxidation. Acid phosphatase and trimetaphosphatase reactions were reduced in many activated cells, with a large proportion of granules being non-reactive. The results of dietary manipulations in quails appear to suggest that in stressful situations the eosinophil metabolism is altered and there is a reduction in the number of lipid droplets in the cell.     

Raw mung beans as a protein source for bred gilts

A study involving 546 crossbred gilts from six seasons was conducted to evaluate raw mung beans as a partial replacement for soybean meal in diets for gilts during gestation. Gilts were randomly allotted to either a control sorghum grain-soybean meal diet or a diet in which a portion of the soybean meal was replaced with mung beans. In the first three seasons, gilts were fed diets in which the protein supplement was totally soybean meal or 89% mung beans (high level) and 11% soybean meal. In the last three seasons the level of mung beans in the supplemental protein was reduced to 61% mung beans with 39% soybean meal (moderate level). Feeding the high level of mung beans decreased (P less than .05) weight gain during gestation and reduced (P less than .05) weight loss during lactation compared with gilts fed the control diet or the moderate level of mung beans. Little difference was noted in litter size at birth, but litter size at 21 d for gilts fed moderate levels of mung beans was less (P less than .05) than for gilts fed the control diet or the high level of mung beans. Little difference was noted in survival rate to 21 or 42 d or individual and litter weights at birth and 21 d. Pig and litter weights at 42 d, however were reduced in gilts fed the high level of mung beans (P less than .05 and P less than .10, respectively) compared with the control diet.(ABSTRACT TRUNCATED AT 250 WORDS)     

Fine structural and cytochemical studies of eosinophils from fowls and ducks with eosinophilia

The ultrastructure and cytochemistry of eosinophils from adult fowl and ducks with either spontaneous or experimentally induced eosinophilia were examined. The results showed that a high proportion of the eosinophils in the peripheral blood of eosinophilic birds had ultrastructural features different from those of normal eosinophils. In both species, there was a reduction in cell size. Fowl eosinophil granules showed similar morphological changes to those seen in the quail with many crescentic and vacuolated forms being present. In eosinophilic ducks, the crystalline interna of the specific granules were often fragmented or were either partially or completely lysed. Cytochemically, peroxidase activity in both species was generally unaltered in abnormal eosinophils compared with those from normal birds. This correlates with the findings in man. However, amounts of acid phosphatase and trimetaphosphatase were reduced in many cells, with a large proportion of granules being non-reactive. The latter observation corresponds with that in quail but differs from man, in which stimulated eosinophils have increased enzymatic activity.     

Reaction phenotyping of vinblastine metabolism in dogs

Vinblastine is a vinca alkaloid used either as a single agent or in combination therapy for the treatment of canine mast cell tumours and lymphomas. The objective of this study was to determine which isoform of cytochrome P450 enzyme is responsible for the majority of vinblastine metabolism in dogs. A panel of eight recombinant canine cytochrome P450 enzymes (CYP1A1, CYP1A2, CYP3A12, CYP3A26, CYP2B11, CYP2C41, CYP2C21 and CYP2D15) were incubated in vitro with vinblastine. Findings were confirmed by the use of canine polyclonal antibodies of cytochrome P450 enzymes (CYP1A1, CYP3A12, CYP2B11 and CYP2C21) that were pre‐incubated with individual and pooled hepatic microsomes that were purified from canine liver. Substrate depletion was observed in the presence of recombinant CYP3A12, whereas depletion did not substantially occur when microsomes were pre‐incubated with polyclonal antibodies against CYP3A12. These findings confirmed that CYP3A12 is the major cytochrome P450 isoform responsible for the metabolism of vinblastine in dogs.     

Implications of hypoxia tolerance for wetland refugia use in Lake Nabugabo,Uganda

Aquatic hypoxia can affect predator‐prey interactions by altering the success rate of the predator and/or the vulnerability of prey. For example, in the Lake Victoria basin of East Africa, native prey exploit hypoxic wetlands as refugia from predation by introduced Nile perch (Lates niloticus). Here, it is predicted that species exploitation of wetlands depends on their hypoxia tolerance relative to the heterogeneity of wetland hypoxia. In this study, we compared the hypoxia tolerance of four fish taxa that differ in their use of hypoxic wetlands in Lake Nabugabo, Uganda: the cichlid Pseudocrenilabrus multicolor victoriae that inhabits the dense swamp interior; and three taxa that inhabit wetland ecotones including Nile tilapia (Oreochromis niloticus), L. niloticus and juvenile endemic haplochromine cichlids. We characterised hypoxia tolerance by exposing fish to progressive hypoxia and quantifying variation in aquatic surface respiration (ASR). The effect of body size on tolerance was explored in L. niloticus by quantifying ASR behaviour across a range of size classes. ASR behaviour was also compared between O. niloticus groups from wetland versus open‐water habitats to detect habitat‐associated intraspecific variation. The most tolerant taxon was the swamp specialist P. multicolor, indicated by its low ASR thresholds and small percentage of fish using ASR during the final sample interval. The other three taxa did not differ in ASR behaviour, and no differences were detected between O. niloticus groups. Body size effects were present for L. niloticus suggesting a lower tolerance to hypoxia in larger‐bodied individuals, thus limiting their ability to penetrate wetlands.     

The influence of different management systems and age on intestinal morphology, immune cell numbers and mucin production from goblet cells in post-weaning pigs

At time of weaning, the immune system in piglets is not fully mature resulting in reduced growth and increased mortality. Early-weaned pigs transported to a segregated early weaning (SEW) facility have enhanced performance and gut development compared to conventional (CONV) pigs which may be due, in part, to decreased pathogen challenge. To gain further insight into SEW enhanced performance and gut development, gut samples from pigs weaned at 19+/-2 days were assessed during the post-weaning (PW) period. The numbers of cells expressing CD2, CD4, CD8, and CD172 (the 74-22-15 (SWC3) antibody is now known to be specific for CD172), MHC class II, and CD25 were quantified using immunohistochemistry. Additionally, samples of duodenum, jejunum, and ileum were evaluated for the production of neutral, acidic, and sulfuric mucins from goblet cells and morphological measurements were also made. No effects due to the management systems alone were observed for any of the parameters. However, there were interactive effects of age/time post-weaning and management system on the immune cells as well as on the mucin secreting goblet cells. There were no differences in gut morphology between SEW and CONV reared pigs.     

Germany/Australia Index of Sperm Sex Sortability in Elephants and Rhinoceros

Flow cytometric sexing of spermatozoa followed by application in artificial insemination or in vitro fertilization provides a unique opportunity to predetermine the sex of offspring and might enhance the conservation management of endangered species in captivity such as the elephant and rhinoceros. To obtain an indication of the sortability of spermatozoa from these species, the relative DNA differences between X and Y chromosome bearing spermatozoa (fresh, frozen thawed, epididymal) from three rhinoceros species [white ( Ceratotherium simum ), black ( Diceros bicornis ), Indian ( Rhinoceros unicornis )] and both elephant species, the Asian and the African elephant ( Elephas maximus, Loxodonta Africana ), were determined through separation of spermatozoa into X and Y chromosome bearing populations, using a modified high speed flow cytometer. The head profile areas of spermatozoa from all five species were measured using light microscopy. By multiplying the relative DNA differences and the head profile areas, the sperm sorting indices were calculated to be 47, 48 and 51 for white, black and Indian rhinoceros respectively. The calculated sorting index for the Asian elephant was 66. In the African elephant, we determined the highest sorting index of 76. These results indicate the practicability of flow cytometric sex sorting of spermatozoa from the tested rhinoceros species and both elephant species. The lower sorting indices in rhinos indicate that sex sorting of spermatozoa from the rhinoceros will be more challenging than in elephants.