Tummaruk P, Tienthai P
Department of Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn University, Bangkok, Thailand.
Reprod Domest Anim. 2010 Apr;45(2):208-13. doi: 10.1111/j.1439-0531.2008.01205.x. Epub 2008 Oct 13.
The aim of this study was to investigate the number of spermatozoa in the crypts of the utero-tubal junction (UTJ) and the oviduct of sows approximately 24 h after intrauterine insemination (IUI) and deep intrauterine insemination (DIUI) and compared with that of conventional artificial insemination (AI). Fifteen crossbred Landrace x Yorkshire (LY) multiparous sows were used in the experiment. Transrectal ultrasonography was performed every 4 h to examine the time of ovulation in relation to oestrous behaviour. The sows were inseminated with a single dose of diluted fresh semen by the AI (n = 5), IUI (n = 5) and DIUI (n = 5) at approximately 6-8 h prior to the expected time of ovulation, during the second oestrus after weaning. The sperm dose contained 3000 x 10(6) spermatozoa in 100 ml for AI, 1,000 x 10(6) spermatozoa in 50 ml for IUI and 150 x 10(6) spermatozoa in 5 ml for DIUI. The sows were anaesthetized and ovario-hysterectomized approximately 24 h after insemination. The oviducts and the proximal part of the uterine horns (1 cm) on each side of the reproductive tracts were collected. The section was divided into four parts, i.e. UTJ, caudal isthmus, cranial isthmus and ampulla. The spermatozoa in the lumen in each part were flushed several times with phosphate buffer solution. After flushing, the UTJ and all parts of the oviducts were immersed in a 10% neutral buffered formalin solution. The UTJ and each part of the oviducts were cut into four equal parts and embedded in a paraffin block. The tissue sections were transversely sectioned to a thickness of 5 mum. Every fifth serial section was mounted and stained with haematoxylin and eosin. The total number of spermatozoa from 32 sections in each parts of the tissue (16 sections from the left side and 16 sections from the right side) was determined under light microscope. The results reveal that most of the spermatozoa in the histological section were located in groups in the epithelial crypts. The means of the total number of spermatozoa in the sperm reservoir (UTJ and caudal isthmus) were 2296, 729 and 22 cells in AI, IUI and DIUI groups, respectively (p < 0.01). The spermatozoa were found on both sides of the sperm reservoir in all sows in the AI and the IUI groups. For the DIUI group, spermatozoa were not found on any side of the sperm reservoir in three out of five sows, found in unilateral side of the sperm reservoir in one sow and found in both sides of the sperm reservoir in one sow. No spermatozoa were found in the cranial isthmus, while only one spermatozoon was found in the ampulla part of a sow in the IUI group. In conclusion, DIUI resulted in a significantly lower number of spermatozoa in the sperm reservoir approximately 24 h after insemination compared with AI and IUI. Spermatozoa could be obtained from both sides of the sperm reservoir after AI and IUI but in one out of five sows inseminated by DIUI.
本研究旨在调查经子宫内输精（IUI）和深部子宫内输精（DIUI）后约24小时，母猪子宫输卵管连接处（UTJ）隐窝和输卵管中的精子数量，并与传统人工授精（AI）进行比较。实验选用了15头长白猪与约克夏猪杂交的经产母猪。每4小时进行一次经直肠超声检查，以确定排卵时间与发情行为的关系。在断奶后的第二个发情期，于预计排卵时间前约6 - 8小时，分别通过AI（n = 5）、IUI（n = 5）和DIUI（n = 5）给母猪单次输精稀释后的新鲜精液。AI组的精子剂量为100 ml含3000×10⁶个精子，IUI组为50 ml含1000×10⁶个精子，DIUI组为5 ml含150×10⁶个精子。输精后约24小时，将母猪麻醉并进行卵巢子宫切除术。收集生殖道两侧的输卵管和子宫角近端（1 cm）。将该部位分为四个部分，即UTJ、峡部尾端、峡部头端和壶腹部。用磷酸盐缓冲溶液对各部分管腔内的精子冲洗数次。冲洗后，将UTJ和输卵管各部分浸入10%中性缓冲福尔马林溶液中。将UTJ和输卵管各部分切成四等份并包埋在石蜡块中。组织切片横向切成5μm厚。每隔五张连续切片进行裱片，并用苏木精和伊红染色。在光学显微镜下确定组织各部分32个切片（左侧16个切片，右侧16个切片）中的精子总数。结果显示，组织学切片中的大多数精子成组位于上皮隐窝中。AI组、IUI组和DIUI组精子储存部位（UTJ和峡部尾端）的精子总数平均值分别为2296、729和22个（p < 0.01）。AI组和IUI组的所有母猪在精子储存部位两侧均发现有精子。DIUI组中，五头母猪中有三头在精子储存部位两侧均未发现精子，一头母猪在精子储存部位单侧发现精子，一头母猪在精子储存部位两侧均发现精子。峡部头端未发现精子，IUI组的一头母猪在壶腹部仅发现一个精子。总之，与AI和IUI相比，DIUI导致输精后约24小时精子储存部位的精子数量显著减少。AI和IUI后精子储存部位两侧均可发现精子，但DIUI输精的母猪中有五分之一仅一侧发现精子。
