浏阳黑山羊瘤胃上皮细胞周期分布、增殖和凋亡特点

本研究旨在建立浏阳黑山羊瘤胃上皮细胞的体外培养模型,并对其周期分布、增殖和凋亡特点进行研究。试验采集60日龄浏阳黑山羊的瘤胃上皮组织,应用0.25%胰蛋白酶+0.02%乙二胺四乙酸(EDTA)消化法对山羊瘤胃上皮组织进行消化,得到单个的山羊瘤胃上皮原代细胞进行体外培养。通过倒置显微镜对原代和传代培养阶段细胞形态进行观察,采用细胞计数法检测细胞的生长活性,应用细胞免疫组化学方法对传代细胞进行鉴定,并用流式细胞术检测山羊瘤胃上皮传代细胞周期分布情况和凋亡比率。结果显示:1)经0.25%胰蛋白酶+0.02%EDTA消化获得的山羊瘤胃上皮原代细胞,培养1 d开始贴壁生长,2 d开始生长较快(对数期),呈典型的"波峰"状生长,3~4 d生长最为迅速,7 d生长速度平稳(平台期)。2)经细胞免疫组化学方法的鉴定,细胞胞浆为黄褐色,即细胞角蛋白19呈阳性表达。3)膜联蛋白-V/碘化丙啶联合染色显示,随着培养时间的延长,细胞凋亡比率显著增加(P0.01)。结果表明,通过0.25%胰蛋白酶+0.02%EDTA的消化方法成功得到了浏阳黑山羊瘤胃上皮细胞,可为今后研究反刍动物瘤胃相关机制与功能提供模型。     

The impact of fish and drought on frog breeding in temporary waters in Zimbabwe

The breeding of frogs in four ponds near Harare, Zimbabwe, was investigated during a wet rainy season (2000/01) and a dry one (2001/02). During 2000/01 eight and nine species bred in two ponds in abandoned gravel pits that never contained fish, but only four species bred in these in 2001/02 and the relative abundance was reduced by about 50%. Pond 3 was a small dam that filled after the rains and was invaded by fish once it overflowed. Five species bred in it in 2000/01, but breeding activity was curtailed once it was invaded by catfish, Clarias gariepinus, some of which were found with frog remains in their stomachs. The pond did not overflow in the following season, and was therefore not invaded by fish; although only four species bred in it their relative abundance was considerably higher. Pond 4 was a permanent pond that always contained fish. Only three species bred in it and the relative abundance of tadpoles was always low. Species belonging to the Ranidae seemed to be most severely affected by drought and this may explain the absence of some species, such as the African bullfrog, Pyxicephalus adspersus, that was once abundant around Harare.     

Sex Identification of the Black Swan (Cygnus atratus) using the Locus-specific PCR and Implications for its Reproduction

Over the last 4-5 years the small captive population of black swans (Cygnus atratus) has consistently failed to reproduce at the Chengdu Research Base of Giant Panda Breeding. The probable cause was hypothesized to be an abnormal sex distribution of the population. The black swan is an example of a sexually monomorphic species. The locus-specific polymerase chain reaction (PCR) approach based on the chromo-helicase-DNA-binding 1 (CHD1) gene, was adopted for the sex determination of the black swans. For this purpose, F1, F2 and R primers were designed using the primerselect software for amplification of the CHD1 gene region. DNA agarose gel electrophoresis showed that the female control displayed two bands, whereas only a single band was found in the male control. Sequence analyses of all seven unknown sex black swans demonstrated the sex-specific DNA band for female. Therefore, it was inferred that all the individuals of the black swan population are females, which has resulted in unfertilized eggs and reproduction failure. This method can be extended to the sexing of other monomorphic avian species and will assist in the design of breeding projects.     

The distribution of external taste buds in flower fish (Pseudophoxinus antalyae)

Purpose of this study was to determine the distribution of external taste buds of flower fish (Pseudophoxinus antalyae) on different body locations (lip, lateral, ventral and dorsal areas, dorsal-pelvic-pectoral-anal fins). In this species, it was found that the density of taste buds was greater on lip epithelium than that of lateral (between pectoral-anal, anal-caudal fins), ventral (between pelvic and anal fins) and dorsal (close to the head) locations. It was observed that the shape of taste bud changed depending upon the epithelium thickness and it was found that some of these structures protruded towards to the exterior of epithelium.     

贵州中部的鹭鸟群巢

2006年5月,对贵州中部的鹭鸟群巢进行了调查,共记录到11个繁殖群、1587巢,种类包括小白鹭(Egretta garzetta)、牛背鹭(Bubulcus ibis)、池鹭(Ardeola bacchus)、夜鹭(Nycticorax nycticorax)和苍鹭(Ardea cinema).最大的繁殖群在黄平风云崖和赤水宴府.种类以小白鹭为优势种,其巢数量占总巢数的49.7%;而苍鹭最少,仪占0.1%.所有鹭鸟巢均只发现于贵州中、北部,而南部的雷公山地区没有调查到.鹭鸟群巢在贵州中部的分布明显受到农田种植模式(冬天种植小麦和油菜,夏天种植水稻)的影响,繁殖巢主要分布于稻作区.但在贵州南部的主要稻作区却没有发现鹭鸟繁殖群巢,其原因尚待调查.对贵州黄平和赤水这两个较大鹭鸟群巢所在地需重点加以保护.     

台湾泥鳅死亡致病菌的分离鉴定及药敏试验

为确定捕捞转场后造成台湾泥鳅（Paramisgurnus dabryanus ssp.Taiwan）死亡的病原,本试验对台湾泥鳅进行病理解剖、病原分离。对分离菌进行形态学观察、生理生化特性测定及16S rDNA基因测序鉴定,并进行人工感染试验、药敏试验。结果显示,从泥鳅肝脏、脾脏、肾脏及肠道组织中分离到形态一致的优势菌株,经纯化保存,命名为XJC。菌株XJC葡萄糖发酵、D-葡萄糖、鸟氨酸、苯丙氨酸及尿素利用阳性,其他所测生化指标均为阴性,与摩氏摩根菌（Morganella morganii）一致;16S rDNA基因序列与摩氏摩根菌相似性达99.6%,被鉴定为摩氏摩根菌。感染试验结果表明,菌株XJC对台湾泥鳅具有致病性,被感染的泥鳅出现了肠道充血典型病理变化。从发病泥鳅体内分离到与菌株XJC一致的菌株,可以判定菌株XJC是泥鳅病原菌。药敏试验结果表明,菌株XJC对氨基糖苷类、喹诺酮类及第三代头孢类药物表现出敏感,对四环素、青霉素、红霉素、多肽类、呋喃唑酮、洁霉素及第一、第二代头孢类药物均等表现耐药,生产中可选用氨基糖苷类、喹诺酮类药物进行摩氏摩根菌疾病的防治。本试验结果可为台湾泥鳅摩氏摩根菌病诊断和防治提供参考。     

Detection of European bat lyssavirus 2 (EBLV-2) in a Daubenton's bat (Myotis daubentonii) from Magdeburg, Germany

In Europe bat rabies in Daubenton's bats (Myotisdaubentonii) and in Pond bats (Myotis dasycneme) caused by the European bat lyssavirus 2 (EBLV-2) has been confirmed in less than 20 cases to date. Here we report the second encounter of this virus species in Germany. A Daubenton's bat found grounded in the zoological garden in Magdeburg died shortly after. In the frame of a retrospective study the bat carcass was eventually transferred to the national reference laboratory for rabies at the Friedrich-Loeffler-Institute for rabies diagnosis. Lyssavirus was isolated and characterized as EBLV-2.     

Morphological and histochemical characterization of the mucosa of the digestive tract in Engraulis anchoita

The histomorphological aspects as well as the histochemical content and distribution of glycoproteins (GPs) in the mucosa of the digestive tract of the anchovy Engraulis anchoita were studied. The buccopharyngeal cavity is lined by a squamous stratified epithelium with mucous superficial cells; the oesophagus shows two zones, cranial with a squamous stratified epithelium with mucous superficial cells and caudal with a columnar secretory epithelium. Finally, the stomach presents both the cranial and pyloric portion lined with a simple columnar epithelium. Tubular branched glands, formed by a single type of glandular cell, located along the stomach, are more numerous in the cranial portion. The GPs were identified with (1) oxidizable vicinal diols; (2) sialic acids and some of their chain variants, C7 or C9; (3) sialic acid residues with O-acyl substitution at C7 or C8; (4) carboxyl groups and (5) sulphate groups. Histochemical tests showed that the buccopharyngeal cavity presented the largest amount of the different types of mucosubstances. Epithelial secretory cells were found in the oesophagus, which synthesized a large quantity of sialosulphoglycoproteins likely to be related to a protective role. The surface epithelium of the stomach synthesizes and secretes acid and neutral GPs, probably related to the movement of fluids and to the absorption of easily digested substrates, respectively. Although great differences exist between different species, in E. anchoita as in other fish species, the wall of the digestive tract is composed of the four layers classically described for vertebrates. The GPs secreted by the epithelial cells are suggested to be important for the protection and inhibition of microorganisms. In addition, they are involved in enzymatic digestion of food, absorptive functions and lubrication of the alimentary tract.     

Distributive and phagocytic characteristics of hepatic macrophages in five cetaceans belonging to Delphinidae and Ziphiidae

Details of morphology and distribution of hepatic macrophages in cetaceans were investigated using the immunohistochemistry with an antibody (SRA-E5) generated against human macrophage scavenger receptor antigen. Liver samples were obtained from five species of cetaceans (Baird's beaked whales, short-finned pilot whales, Risso's dolphins, bottlenose dolphins, and pantropical spotted dolphins). Except for two species of whales, the number of SRA-E5-positive Kupffer cells was greatest in the perivenous zone (zone 3), followed by the mid-zonal (zone 2) and periportal (zone 1) zones; this distribution pattern was different from that in cattle examined here and previously reported rodents with the highest number in zone 1. The frequency of Kupffer cell in each of zones was significantly different among species, and interestingly, the total mean of the Kupffer cell number in three zones increased as the body-length of species was small. In cetaceans, Kupffer cells in zone 1 appeared larger and more stellate in shape, whereas those in zone 3 were smaller and rounder. All cetaceans but Baird's beaked whales had the black pigment-containing Kupffer cells, with the greatest number in zone 3, and macrophages with the similar pigments were also seen in the hepatic intermediate septa, indicating an active phagocytosis. Most of the black pigments were considered to be lipofuscin and such pigments were not seen in the bovine livers. These results indicate that cetacean hepatic macrophages show differences in the distribution and phagocytosis among hepatic lobular zones, or between cetacean species and terrestrial animals.     

Seroprevalence of tularemia in wild bears and hares in Japan

Tularemia is a zoonotic disease caused by Francisella tularensis. The distribution of the pathogen in Japan has not been studied well. In this study, seroprevalence of tularemia among wild black bears and hares in Japan was determined. Blood samples collected from 431 Japanese black bears (Ursus thibetanus japonicus) and 293 Japanese hares (Lepus brachurus) between 1998 and 2009 were examined for antibodies against F. tularensis by micro-agglutination test (MA) or enzyme-linked immunosorbent assay. By subsequent confirmatory tests using western blot (WB) and indirect immunofluorescence assay (IFA), eight sera from Japanese black bears were definitely shown to be seropositive. All of these eight bears were residents of the northeastern part of main-island of Japan, where human tularemia had been reported. On the other hand, no seropositive Japanese hares were found. These results suggest that Japanese black bears can serve as sentinel for tularemia surveillance and may help understand the distribution of F. tularensis throughout the country. This is the first report on detection of antibody to F. tularensis in black bears of Japan.     

Serologic survey for Toxoplasma gondii in wild animals in Florida

Blood samples were collected for serum separation from 114 species of wild animals (25 species of mammals, 82 species of birds, and 7 species of reptiles) in Florida. Each of the 3,471 samples was tested for antibodies to Toxoplasma gondii, using the indirect hemagglutination test. The highest prevalences of T gondii antibodies were 19% in armadillos (Dasypus novemcinctus), 18% in raccoons (Procyon lotor), 13% in black rats (Rattus rattus), and 11% in opossums (Didelphis marsupialis). Antibody prevalences were significantly higher in male than in female raccoons (P less than 0.05) and in adult than in nonadult raccoons and opossums (P less than 0.005). A high proportion of seropositive animals was found in three other mammalian species: 4 of 4 black bears (Ursus americanus), 2 of 3 bobcats (Lynx rufus), and 2 of 8 Norway rats (Rattus norvegicus) tested. Antibodies were found in 8 of the 1,279 avian serums; they were not found in any of the 13 reptilian serums tested. There were no significant geographic variations in antibody prevalence in any species.     

Genetic variation of chicken MC1R gene in different plumage colour populations

1. Genetic variation in the chicken MC1R gene was analysed through bioinformatic methods after polymerase chain reaction (PCR) amplification and sequencing of the coding region of MC1R gene from 5 different populations with 4 plumage colours (black, grey plumage with black spots, yellow plumage with black spots, red). 2. A total of 11 novel variations were detected in Hebei chickens, of which 8 were non-synonymous. Allele distribution analysis showed that the wild-type e(+) (Brown Leghorn) allele was mainly found in Hy-Line Brown and Lohmann Brown, the dominant Extended black E(AY220304) allele was mainly found in Hebei chicken with black plumage, whereas the Buttercup (e(bc)) allele was rare. 3. Nucleotide diversity (π) within each colour strain of Hebei chickens (0·0047-0·0052) was significantly greater than that of Hy-Line Brown (0·0024) or Lohmann Brown (0·0043). 4. The results indicate that there is abundant polymorphism in the MC1R gene, especially in Hebei chicken, which was associated with its rich plumage colour diversity.     

Comparison of black and white light for collecting Culicoides imicola and other livestock-associated Culicoides species in South Africa

Comparison of the effectiveness of 8W fluorescent black and white light sources, in two 4x4 Latin squares (16 replicates) designs under South African conditions, showed black light to be up to three time more effective in collecting Culicoides imicola Kieffer (Diptera, Ceratopogonidae) and other South African Culicoides species. Four Culicoides species, which were collected in low numbers with black light, were not collected in traps equipped with the white light source. No significant difference was found in the parous rate of the C. imicola populations as determined by the two light sources. The study highlighted the superiority of black light as a preferred collection method for C. imicola, considered to be the most widespread and abundant vector of livestock orbiviruses. The results underline the need to develop and adopt standard techniques for measuring the variables of vectorial capacity.     

安哥拉山羊与建昌黑山羊及其杂种后代的DNA指纹分析

以 (CA/GATA/TCC) 5探针 ,HinfI酶切 ,研究安哥拉山羊和建昌黑山羊及安哥拉山羊与建昌黑山羊级进杂交的F2 、F3的DNA指纹图谱。结果 ,在供试山羊中 ,个体平均检出 1 8.2± 0 .4条谱带 ,安哥拉山羊、F2 、F3的鉴别机率分别为 1 .38× 1 0 - 12 ,1 .1 8× 1 0 - 13,0 .50× 1 0 - 10 ;群体内相似系数安哥拉山羊为0 .4 539,F2 为 0 .4 0 77,F3为 0 .61 1 1 ;亲子鉴定的父权概率W =0 .9889;安哥拉山羊与建昌黑山羊比较 ,2 .3kb和 8.6kb谱带为安哥拉山羊的特异谱带     

Histological Characteristics of the Tracheobronchial Tree of the Least Shrew (Cryptotis Parva)

The least shrew (Cryptotis parva) is a small vomit‐competent insectivorous species which has recently been introduced as an emesis animal model in the laboratory. In this study, the respiratory system of the least shrew was examined and compared with the well‐established larger species routinely used in the laboratory. Five least shrews (4–5 g body weight, 45–60 days old) were used. Standard histological procedures were followed for light microscopic examination. The lining epithelium of the trachea was found to be pseudostratified ciliated columnar (PSCC). Three types of cells were easily identified, basal and ciliated as well as few goblet cells interspersed among the ciliated cells and they were not clearly recognizable. A few tracheal seromucous glands were located at the free end of the C‐shaped cartilaginous rings. The cartilaginous rings are replaced by smooth muscle cells before the bronchi enter into the lung. The lining epithelium of tracheobronchial tree gradually changes into simple cuboidal epithelium that lacks goblet cells. However, the division of the tracheobronchial tree is similar to other mammalian species. On the other hand, the principal bronchus lacks cartilaginous plaques as it becomes intrapulmonary bronchus. The wall of the bronchi is supported by thick layers of spirally arranged smooth muscles. Two types of cells were readily recognizable: basal and ciliated cells, with rarely observed goblet cells. In addition, the PSCC epithelium changes into simple cuboidal much earlier in the bronchial division relative to other species.     

饲料蛋白质水平对台湾泥鳅生长性能、形体指标和体成分的影响

本试验旨在探索饲料蛋白质水平对台湾泥鳅生长性能、形体指标及体成分的影响。选用初始体重为(8.57±0.35)g的台湾泥鳅720尾,随机分成4组,每组3个重复,每个重复60尾。4组台湾泥鳅投喂蛋白质水平分别为25%、30%、35%和40%的试验饲料,养殖时间为60d。结果表明:随着饲料蛋白质水平的增加,台湾泥鳅末重和增重率均呈先升高后趋于稳定的变化趋势,且均在饲料蛋白质水平为35%时达到最大。饲料系数随着饲料蛋白质水平的升高呈先降低后趋于稳定的变化趋势,且在饲料蛋白质水平为40%时最低。饲料蛋白质水平从35%增加到40%,末重、增重率和饲料系数无显著变化(P0.05)。基于增重率和饲料系数的折线模拟结果表明,台湾泥鳅达到最大增重率的饲料蛋白质水平为34.57%,饲料系数最低的饲料蛋白质水平为35.47%。随着饲料蛋白质水平的升高,蛋白质效率、蛋白质沉积率和成活率均呈先升高后降低的变化趋势。通过二次多项式回归分析可知,台湾泥鳅蛋白质效率和蛋白质沉积率最高时的饲料蛋白质水平分别为33.61%和34.68%。随着饲料蛋白质水平的升高,台湾泥鳅脏体比和肝体比均呈先降低后趋于稳定的变化趋势,鱼体粗蛋白质含量呈先升高后趋于稳定的变化趋势,鱼体粗脂肪含量呈先降低后趋于稳定的变化趋势。结果提示,适量增加饲料蛋白质水平可提高鱼体生长速度,增加鱼体可食用部分比例,提高蛋白质效率和蛋白质沉积率,但饲料蛋白质水平过高,会降低蛋白质沉积率和蛋白质效率。在本试验条件下,综合考虑生长性能指标和蛋白质利用情况,台湾泥鳅适宜饲料蛋白质水平为33.61%~35.47%。     

Corneal vascularization in the Florida manatee (Trichechus manatus latirostris) and three-dimensional reconstruction of vessels

The cornea of the Florida manatee is unique and unusual in its anatomy in that blood vessels have been found throughout. In all other animal species, this is considered a pathological condition impeding vision, and is usually caused by injury or trauma. The purpose of this study was to more clearly describe corneal vascularization by examining the architecture through three-dimensional reconstruction in order to find possible patterns in size, distribution, and location of blood vessels relative to gender, age, location, and season. Twenty-six eyes from 22 individuals were prepared for histologic examination and subsequent three-dimensional reconstruction. Every specimen examined had blood vessels in the cornea, comprising an average of 0.3% of total surface density (volume) of the cornea. No differences were found between individuals based on gender, age, and season. Environmental influences were not a significant factor either, which was not originally anticipated. The presence of vessels at the level of the anterior epithelium was surprising and it appeared that the vascularization was directed more anteriorly than was originally thought. The presence of blood vessels in a prenatal eye was also found. In all the eyes examined, no signs of injury or trauma could be observed. The presence of blood vessels appears to minimally impair vision based on their low density, size, and location. The association of vessels with the anterior epithelium and development of vessels within the fetus point to an evolutionary adaptation possibly due to the manatee's unique ability to move between water bodies.     

山羊咽扁桃体和咽鼓管扁桃体的组织结构观察

选取健康10月龄奶山羊10头,断头宰杀后取咽扁桃体和咽鼓管扁桃体,应用组织学光镜和电镜制片技术研究咽扁桃体和咽鼓管扁桃体的显微和亚显微组织结构.结果表明:山羊咽扁桃体和咽鼓管扁桃体的黏膜上皮主要由2～3层多边形上皮细胞组成,部分区域只有单层扁平细胞,相邻上皮细胞间空隙很大,上皮细胞表面有丰富的微绒毛.上皮细胞之间和黏膜上皮下方固有层内有大量淋巴细胞浸润.扁桃体的实质部分由数个次级淋巴小结和弥散淋巴组织构成,弥散淋巴组织中有大量分布的淋巴管和毛细血管后微静脉.此外,在紧贴黏膜上皮细胞下方的固有层和淋巴滤泡中可观察到少量的树突状细胞.结果提示山羊的咽扁桃体和咽鼓管扁桃体可作为鼻腔免疫的主要诱导位点和效应部位.     

Clinical evaluation of the CA530-VET hematology analyzer for use in veterinary practice

BACKGROUND: The CA530-VET is a completely automated impedance cell hematology analyzer, which yields a 16-parameter blood count including a 3-part leukocyte differential. OBJECTIVES: The aim of this study was to examine the operational potential of the CA530-VET and its value for use in veterinary practice. METHODS: The analyzer was tested for blood carry-over, precision, and accuracy. Comparison methods included the CELL-DYN 3500, microhematocrit centrifugation, manual platelet (PLT) counting for feline and equine species, and a 100-cell manual WBC differential. Blood samples for comparison of the methods were obtained from 242 dogs, 166 cats, and 144 horses. RESULTS: The carry-over ratio (K) was 0.28% for RBC, 0.59% for PLT, 0.32% for WBC, and 0.18% for hemoglobin (HGB) concentration. Coefficients of variation (CVs) for within-batch precision and duplicate measurement of blood samples were clearly within the required limits, except for duplicate platelet counts in cats (8.7%) and horses (9.5%). The WBC count was in excellent agreement for dogs and horses and RBC count was in excellent agreement for horses. The accuracy of feline WBC counts was not acceptable, with the exception of values at the high end of the range. RBC counts in dogs and cats, and HGB concentration and MCV in all 3 species were sufficiently accurate. The CA530-VET HCT results were in excellent agreement with microhematocrit results in horses but exceeded the maximum allowed inaccuracy for cats and dogs. In all species, PLT counts established mechanically and manually were not in adequate agreement. Large differences were found between the CA530-VET and the manual differential percentage for lymphocytes and "mid-sized cells" (monocytes and basophilic granulocytes). CONCLUSIONS: The CA530-VET can be considered useful for routine canine, feline, and equine blood cell analyses. It should not be considered accurate, however, for PLT counts, feline total WBC counts in the subnormal and normal range, and leukocyte differentials, except for granulocytes.