Characterization of the goldfish fecal microflora by the fluorescent in situ hybridization method

ABSTRACT:   Bacterial populations in goldfish feces were characterized by the fluorescent in situ hybridization (FISH) method. A total of nine different group-specific rRNA-targeted oligonucleotide probes were used. Approximately half of the microbial cells (57.8 ± 16.7%) were detected with a probe EUB338 and found to be bacteria. The microbial cells in 33–35 of the 35 samples from five specimens strongly hybridized with probes ALF1b, BET42a and GAM42a, suggesting that goldfish intestinal bacteria are mainly composed of α, β and γ-subclasses of Proteobacteria. The fact that a probe AER66 reacted with 25.6 ± 14.2% of the total microbial cells in all 35 samples, demonstrated that genus Aeromonas was the dominant species in the goldfish intestines. Genus Bacteroides including Bacteroides type A detected with a probe BAC303 was observed in 15 of 35 samples while other taxonomic groups determined with HGC69a, CF39a and P72 were detected in 6–11 of 35 samples. These results strongly suggest that Bacteroides shows the greatest daily fluctuation and interindividual variation in the intestines of goldfish. Moreover, the FISH method was proven to be useful for rapid enumeration of taxonomic groups of fish intestinal bacteria.     

不同品系西方蜜蜂基因组DNA多态性图谱构建及杂交分析

本研究应用随机引物K（5'-CGGCCCCTGT-3'）对不同品系西蜂的基因组DNA扩增，获得了能区分不同品系西蜂的多态性DNA图谱，从多态性图谱中选取一个共同片段K1680bp，标记成探针，探针K1680bp与不同品系西蜂的基因组DNA匀有杂交，从而证明片段K1680bp是西蜂蜂种的共性，为西蜂的鉴定提供了直接依据。     

犬贾第虫包囊排出规律的研究

对自然感染贾第虫的比格犬进行粪检调查 ,结果表明犬贾第虫包囊排出呈间歇性 ,其间歇期为 7～ 8d ,排囊后 1 2～ 1 4d达到排囊高峰期 ,峰期持续 2～ 3d,排囊持续时间为 2 5～ 2 7d。用 2只幼犬进行免疫抑制接种犬贾第虫 ,接种后第 7天出现排包囊 ,排囊后 6～ 7d达到排囊高峰期 ,峰期持续 4～ 5d。受染动物出现腹泻 ,体重减轻     

荧光原位杂交检测野生六出花(Alstroemeria aurea) 染色体命运*

 利用从野生六出花(Alstroemeria aurea) 中分离的一个种特异性串联重复序列(A001-I) 作荧光原位杂交的探针, 检测野生六出花染色体在其一系列杂交后代中的命运。试验结果表明, 除最长的一对染色体外, 该方法能够检测到在其一系列杂种后代中其余7 对染色体是否存在。     

兔防御素cDNA表达质粒的构建及其表达

将兔防御素（MCP－1）cDNA插入真核表达载体pcDNA3的EcorRⅠ和XbaⅠ酶切位点之间，构建了兔MCP－1cDNA的真核表达质粒pcDEF。通过脂质体转染，使兔MCP－1 cDNA在COS－7细胞中表达，在转染60、84、108h后，提取总RNA。采用RT－PCR，在288bp的位置扩增出1条特异性带；RNA斑点印迹杂交表明，兔MCP－1 cDNA在60、84、108h均有表达。     

寿光鸡杂交试验

用寿光鸡与黑乌骨鸡、斗鸡和AA鸡进行杂交试验，结果表明：①寿光鸡与黑乌骨鸡杂交，其F2代体增重介于两品种之间，跖长比寿光鸡降低17．2％；羽型多表现为扁羽，约15％为丝状羽；皮肤为白色；均为5趾。②鲁西斗鸡与寿光鸡杂交F3代与寿光鸡相比，体增重降低6．3％，料重比增加5．9％，胸宽增加5．3％。③寿光鸡与AA鸡杂交，F1代增重速度比寿光鸡本身提高85．2％，料重比降低14．3％。④白乌骨鸡比黑乌骨鸡增长快，F1代比黑乌骨鸡100日龄增重提高24％，比白乌骨鸡本身增重快9．1％。     

Meiotic and Isozymic Analyses of Tall Fescue × Giant Fescue Hybrids and Amphiploids1

The meiotic behavior of three tall fescue (Festuca arundinacea, 2n = 6x = 42) genotypes, giant fescue (F. gigantea, 2n = 6x = 42), and their reciprocal F₁ hybrids and C₁, amphiploids was evaluated to determine the parental genomic relationships. Isozyme banding patterns were used to confirm the parental identity of the hybrids and amphiploids. At meta-phase I, the parents had predominantly bivalent pairing. The hybrids had an average of 9.51 I, 16.02 II, 0.12 III, 0.02 IV, and the amphiploids had 2.17 I, 38.82 II, 0.60 III, 0.58 IV, 0.01 V—VIII. The prevalence of bivalent pairing in both hybrids and amphiploids suggested a homoeologous relationship between the six genomes, with four of the six being more closely related. Bivalent pairing in the amphiploids indicated genetic regulation of chromosome pairing. Zymograms were obtained for acid phosphatase (ACPH), alcohol dehydrogenase (ADH), glutamate oxaloacetate transaminase (GOT), malate dehydrogenase (MDH), 6-phosphogluconate dehydrogenase (6-PGD) and phosphoglucoisomerase (PGI). The three tall fescue and giant fescue parents had different zymograms for ACPH, MDH, 6-PGD and PGI; thus, the tall fescue parents of the hybrids and amphiploids could be determined based on the banding patterns of these four enzymes. Phenotypes were determined for ACPH-1, PGI-2 and 6-PGD-1. ACPH-1 may be used to follow the introgression of giant fescue chromatin into a certain tall fescue genotype.     

陶赛特羊与青海半细毛羊杂交试验

试验在高寒牧区天然草地放牧条件下 ,测定陶×半细杂交一代羊出生到 4月龄和 6月龄的生长发育指标 ,结果表明 :在青海高原严酷的生态环境条件下 ,杂交一代羔羊生长发育快于青海半细毛羊 ,4月龄活重为 2 2 .73kg ,6月龄活重为 2 7.96kg ,分别比青海半细毛羔羊提高 3 .0 2kg、3 .64kg(P <0 .0 1)。体长、体高、胸围、胸宽、管围和尻宽显著高于青海半细毛羔羊 (P <0 .0 1)     

Identification of marine algicidal Flavobacterium sp. 5 N-3 using multiple probes and whole-cell hybridization

ABSTRACT: We designed five 16S rRNA-targeted oligonucleotide probes (Sp probes) specific for Flavobacterium sp. 5 N-3, which inhibits the growth of red tide phytoplankton Gymnodinium mikimotoi (Dinophyceae). These probes were evaluated by whole-cell hybridization against 5 N-3 cells incubated under laboratory conditions. The fluorescence signal from the cell detected with Sp probe mix5, a mixture of the five probes, was 8.4-fold higher than that obtained with only one Sp probe (Sp01RF). The signal obtained by this method was strong enough to recognize 5 N-3 cells and count them under the epifluorescence microscope, while the signal was often undetectable when a single probe was used. Fluorescence intensities of cells at stationary phases and of 'starved' cells in sterile seawater using Sp probe mix5 were low but still sufficient for enumeration. These Sp probes did not hybridize with 11 strains from the Cytophaga/Flavobacteria/Bacteroides phylum and did react with strain 5 N-3 following whole-cell hybridization. These results show that 5 N-3 cells cultivated under our laboratory conditions can be detected by whole-cell hybridization with the five designed probes. These data also suggest that this technique may be useful for detection of an algicidal bacterium 5 N-3 in the natural environment.