Classification of differentiating oocytes during ovarian cycle in the giant freshwater prawn, Macrobrachium rosenbergii de man

Based on the light microscopic observations of cells' sizes, chromatin patterns, amount of lipid droplets and yolk granules, the female germ cells could be classified into four different phases, which include 1) oogonia (Oog), 2) primary oocytes (pOc), 3) secondary oocytes (sOc), and 4) mature oocyte (mOc). Oog are small oval-shaped cells with irregular-shaped nuclei sizing 4–6 μm in diameter. They rest on the connective tissue germinal cord at the tip of each ovarian pouch (lobule). Oogonia increase their number through mitotic division, and the daughter cells move into ovarian pouch where they undergo first meiotic division to become primary oocytes, which have various steps of 1st meiotic prophase accumulating at the innermost zone of the ovarian pouch. The primary oocytes are small oval-shaped cells (8.5–10 μm in diameter) with large nuclei containing chromatin in various states of condensation that finally transform into chromatids. Their nuclei are surrounded by thin rim of faint blue-stained cytoplasm. The secondary oocytes derived from 2nd meiosis and comprise five steps: Oc1 and Oc2, classified as previtellogenic oocytes, Oc3 and Oc4, classified as vitellogenic oocytes, and mature oocyte (mOc) The zones of ovarian pouch are defined based on the accumulation of various steps of developing oocytes, namely, oogenic, previtellogenic, vitellogenic and mature zones, respectively. The ovarian cycle is divided into five stages based on the number and types of oocytes present in each stage. Stage 0 and I are spawn and spent stages. Stage II and III are proliferative and premature stages, while stage IV is mature stage. During ovarian stage I, each ovarian pouch contains primarily oogonia, primary oocytes, Oc1 and a few Oc2. In stage II, the pouch contains mainly Oc2 and Oc3, while in stage III the predominant cells are Oc4. Mature oocytes appear synchronously, in stage IV. The ovulating mature oocytes pass through the thin disrupted wall of ovarian pouch into subcapsular space, that leads into the oviduct situated on the ventro-lateral side of the ovarian lobe. At spawning, the ovarian pouches break down and only connective sheaths and hemolymph sinuses remain. The germinal cords and islets of oogonia remain in the central area of stage 0 ovary. The ovarian capsule, including the muscular layer, becomes attenuated as the ovary progresses from stage 0 to IV. The hemolymph vessels become highly convoluted in the central area of the ovary, and they branch radially into smaller hemolymph sinuses around each oogenic pouch.     

Alcohol diluent provides the optimal formulation for calcium chloride non-surgical sterilization in dogs

Background

Surgical castration is widely used to sterilize male dogs, but has significant impacts on time to perform the operation, recovery of the animals as well as cost, which can limit population control programs. Previous research has shown intratesticular injection of calcium chloride dihydrate (CaCl₂) in saline to be a promising alternative to surgery. However, long-term azoospermia was not maintained at dosages low enough to avoid side effects. In the search for an optimized formulation, the current investigation is the first study on long-term sterilization effects of intratesticular injection of CaCl₂ in either lidocaine solution or alcohol in dogs. CaCl₂ at 20% concentration in lidocaine solution or alcohol was administered via intratesticular injection to groups of 21 dogs each. The treated animals were examined at 2, 6, and 12 months for sperm production, blood levels of testosterone, and side effects; at time zero and 12 months for testicular size and semen volume. The experimentally treated animals were compared to a control group receiving saline injection only.

Results

Testicles of dogs treated with CaCl₂ in either diluent significantly decreased in size. After administration of CaCl₂ in lidocaine solution, sterility was achieved for at least 12 months in 75% of treated dogs. However, optimal long-term contraceptive effectiveness was achieved with CaCl₂ in alcohol, which resulted in azoospermia over the 12-month study period. Testosterone levels significantly decreased following treatment with CaCl₂, and sexual activity disappeared. Although testosterone returned to baseline levels by 12 months for the group treated with CaCl₂ in lidocaine, dogs injected with CaCl₂ in alcohol had a 63.6% drop in testosterone level, which remained at the low end of physiological range throughout the study. No adverse effects were noted.

Conclusions

A single, bilateral intratesticular injection of 20% CaCl₂ in 95% ethanol was a reliable method for induction of sterilization in 18–28 kg male dogs in this study. The approach showed long-term efficacy and reduced sexual behavior. This chemical method of sterilization might provide an effective, efficient alternative to surgical castration that can have positive impacts on dog welfare.     

日本沼虾饲料最适蛋白质、脂肪含量及能量蛋白比的研究

本文研究了日本沼虾(Macrobrachium nipponense)饲料的最适蛋白质含量、脂肪含量及能量蛋白比。试验用饲料由酪蛋白、糊精、混合油、复合维生素和无机盐混合物等组成。本试验用的日本沼虾体重为 1.77±0.23克,试验期间水温为 21—25℃。用增重率、饲料系数和肝胰脏α-淀粉酶活力等作为评价指标。试验结果表明:日本沼虾配合饲料适宜蛋白质含量为 36.8—42.27％,适宜脂肪含量为6—12％。当配合饲料的蛋内质含量在适宜范围内,饲料能量蛋白比(C/P)为8千卡/克蛋白左右,每公斤配合饲料的总能量为3006—3561千卡是最为适宜的。当饲料总能量在一定范围内,随着饲料碳水化合物含量的增加,肝胰脏α-淀粉酶活力增强。     

青虾“红体病”组织病理学观察

用连续切片的方法对患有红体病的青虾进行组织病理学研究，结果表明患病青虾鳃部、肌肉组织和肝胰腺部位病症比较明显，不同患病阶段组织变形差异显著。鳃片细胞在患病初期表现为胞质收缩，后期则细胞膨胀破裂。细胞内物质外溢，细胞边缘界限不完整；肌肉组织在后期出现肌纤维束之间间隙，且部分肌纤维有断裂的现象；中期肝小叶间隔出现空隙和腺细胞变形，在后期则表现为小叶溃烂。造成青虾因红体病死亡的原因，初期主要是因为鳃片的组织病变导致呼吸困难，进而造成青虾生理异常；后期主要是因为肝胰脏的溃烂造成消化功能全面丧失。     

Serotonin induces ovarian maturation in giant freshwater prawn broodstock, Macrobrachium rosenbergii de Man

This study investigated the effects of serotonin (5-hydroxytryptamine or 5HT) on ovarian development in Macrobrachium rosenbergii de Man. Adult female prawns at the ovarian stage I (spent) were injected with 5HT at 1, 5, 10, 20 and 50 μg g^− 1 body weight (BW) intramuscularly on days 0, 5 and 10, and sacrificed on day 15. The doses as related to the effect could be categorized into three levels: low (1 and 5 μg g^− 1 BW of 5HT), medium (10 and 20 μg g^− 1 BW of 5HT) and high (50 μg g^− 1 BW of 5HT). The low-dose, especially at 1 μg g^− 1 BW, caused prawns to exhibit a significant increase in ovarian index (ovarian weight/body weight × 100) (5.79 ± 0.09%) as compared to the control (1.49%). The ovaries of most of these prawns could develop to stage IV (mature) and contained synchronously mature oocytes while most of the control ovaries remained at stage I and II (proliferative), and contained only oogonia to previtellogenic (Oc1, Oc2) and early vitellogenic oocytes (Oc3). The medium- and high-dose treated prawns exhibited ovaries that could reach stages III and IV and contained various types of oocytes of different maturity. Pretreatment with 5HT receptor antagonist, cyproheptadine (CYP), at 10 μg g^− 1 BW before 5HT injection significantly suppressed the effect of 5HT. Intramuscular injection of the 5HT-primed thoracic ganglion culture medium into CYP-pretreated prawns resulted in the increase of ovarian index about 5–6 times more than in the control, and in the groups injected with 5HT-primed media from muscle strip, eyestalk and brain. The ovaries of most prawn could develop up to stage IV and contained synchronously developed vitellogenic (Oc4) and mature oocytes (Oc5). These findings suggest that 5HT indirectly induces ovarian development and oocytes maturation in M. rosenbergii, probably via a putative ovarian stimulating factor released from the thoracic ganglia.     

鲤鱼种间、种内杂交中杂种优势强度表现的某些规律的初步研究

从1986年至1990年于我所松浦试验场,用黑龙江野鲤、荷包红鲤、兴国红鲤、红镜鲤、散鳞镜鲤、德国镜鲤和柏氏鲤进行了种间、种内14个组合的杂交,系统观察了不同组合亲本和杂种F_1在鳞被、体色、体型、生长、成活率、抗寒力、抗病力和起捕率等8个性状的遗传传递规律及其杂种优势强度,结果是.(1)鲤鱼种内异源杂交几乎所有的杂交组合的杂种F_1都表现出生长优势和对环境抗性的增强,其大小随亲本的亲缘远近和鳞被、体色、体型等性状间的差异而不同。一般品系间杂交,杂种生长的优势率在10％左右;品种间杂交,单交种的生长优势率在15—30％,三杂交种的生长优势率在30％以上。在抗寒、抗病、成活率等抗性上,表现出受其中一个亲本的影响,且有明显的母本效应,并可遗传传递给杂种后代。鳞被、体色、体型的基因效应,对杂种的生长有明显的促进作用。(2)鲤鱼种间杂交几乎不表现杂种优势,其特点:是柏氏鲤和黑龙江野鲤两个野生种间杂交,成活率、抗寒力强烈的表现不亲和性;而柏氏鲤与家养种间的杂交,表现一定的亲和性。亲本和杂种F_1的鳞被、体色仍表现显隐性关系,而体型、抗寒、抗病、成活率、起捕率等受柏氏鲤种质的强烈影响。获得抗寒力的杂种,在第二代时抗寒力明显消失,说明基因转移和整合的不稳定性。用柏氏鲤与散鳞镜鲤杂交的杂种F_1与另一抗寒很强的杂种F_1(黑龙江野鲤♀×红镜鲤♂)的双杂交种F_1,抗寒明显增强。     

罗氏沼虾繁殖习性的研究

本文对罗氏沼虾的繁殖习性进行了较为详细地描述。性成熟的雌虾必须经过生殖蜕皮方可交配、产卵；受精卵粘附于腹肢刚毛上；在水温２４℃左右的条件下，幼体孵出约需２７天；性成熟雌虾能多次产卵。     

白洋淀日本沼虾血细胞的初步研究

对取自华北白洋淀种群的日本沼虾(Macrobrachium nipponense)血细胞进行显微形态观察、分类、测量和计数。依据细胞的形态,胞质中颗粒的数量和密度以及细胞的核质比把日本沼虾血细胞分为透明细胞(hyaline cell,HC)、半颗粒细胞(semigranular cell,SGC)和颗粒细胞(granular cell,GC)3类。血细胞大小依次为颗粒细胞>半颗粒细胞>透明细胞。血细胞密度为(1.13±0.50)×106个/mL,其中GC占48.5%,SGC占31.8%,HC占19.7%。并对我国日本沼虾的华南种群、华中种群及白洋淀所在地华北种群的血细胞大小和数量进行了比较。     

罗氏沼虾浙江养殖群体与缅甸自然群体遗传差异的RAPD分析

利用随机扩增多态DNA(RAPD)技术，对浙江省罗氏沼虾养殖群体和新引进的缅甸自然群体的遗传差异进行了比较分析，以期从分子水平了解罗氏沼虾的种群遗传多样性背景及与引种的关系。采用经筛选的22个10bp的随机引物对罗氏沼虾两群体各20尾进行群体RAPD分析。22个引物共检测到139个位点。养殖群体的多态位点比例为30．22％，群体平均杂合度为0．2646，Shannon多样性指数为0．0780，群体内各个体之间的遗传共享度为0．9353，遗传距离为0．0647；自然群体的多态位点比例为33．81％，群体的平均杂合度和Shannon多样性指数分别为0．2888和0．0940，群体内各个体之间的遗传共享度为0．9201，遗传距离为0．0799；两群体之间的遗传距离为0．1845。自然群体的遗传多样性水平比养殖群体要高。利用随机引物S9和S52，在罗氏沼虾两群体间扩增出2条稳定、明显的群体间特异性标记带。     

罗氏沼虾早苗的中间培育及成虾高产养殖

介绍了在塑料棚控温土地内对罗氏沼虾早繁苗进行放养前中间培育及池塘成虾高产养殖的具体过程和方法，探讨了罗氏沼虾早苗中间培育和成虾养殖的一些技术问题。