Isolation and characterization of Pseudomonas aeruginosa from sheep with fleece rot in northern and middle Jordan

A total of 162 sheep fleece samples were collected from 2315 sheep clinically examined for evidence of dermatitis. The sheep belonged to 32 flocks raised in northern and middle Jordan. Eighty-three samples showed no obvious abnormalities, whereas the remainder showed exudation (79 samples), fleece discoloration (18) and fleece roughness (40) and abscesses (7). Seventeen Pseudomonas aeruginosa isolates were obtained from these samples. Antibacterial resistance in vitro was common; resistance to tetracycline, amoxycillin, erythromycin and cotrimoxazole was shown by > 90% of the isolates. Resistance to norfloxacin (29.4% of isolates), ciprofloxacin (17.6%) and amikacin (17.6%) was also demonstrated. Fourteen isolates were serum resistant when assessed after 1-3 h incubation in sheep and calf sera, and their count increased by 2-2.9 and 2.5-3.5 respectively.     

Selection response to fleece weight,wool characteristics,and heritability estimates in yearling Romney sheep

The juvenile live weights, yearling fleece weight and wool characteristics of 2987 Romney progeny of 114 sires born over 9 breeding seasons in a fleece-weight-selected and a random control line were analysed. The high fleece weight-selected flock performance was higher (P < 0.001) for weaning weight and spring weight (6.9 and 8.4%), higher (P < 0.001) for greasy and clean fleece weight (23.8 and 24.3%), and also higher (P < 0.001) for FD by 1 μm, staple length by 6 mm, and wool yellowness by 0.3 unit than random control yearling sheep. Heritability estimates for weaning and spring live weight, greasy fleece weight, clean fleece weight, yield, fibre diameter, staple length, staple strength, loose wool bulk, brightness and yellowness were 0.15, 0.51, 0.35, 0.36, 0.40, 0.57, 0.41, 0.24, 0.46, 0.12 and 0.14 respectively. The heritability estimates are within the range of the long wool sheep breeds studied previously. The high selection differential achieved in the initial screening commercial flocks however facilitated to approach a selection response peak in a shorter selection duration and thus resulted in a non-significant genetic gain in greasy fleece weight when averaged over the 9 years of selection.     

竹鼠绿脓杆菌的分离和鉴定

对广西某养殖户送检的病竹鼠进行病理剖检及细菌分离鉴定,根据发病临床症状,病理变化,病原分离鉴定及致病性试验结果,确诊为绿脓杆菌感染。对分离菌进行药敏试验,结果显示所分离到的绿脓杆菌细菌对氧氟沙星、诺氟沙星、环丙沙星、庆大霉素、阿米卡星和阿奇霉素敏感;对头孢他啶、头孢西丁、利福平和阿莫西林耐药。     

Observations on blood leucocytes and lymphocyte subsets in sheep infected with bovine leukaemia virus: a progressive study

Haematological parameters and reactivity of lymphocyte antigens to monoclonal antibodies were studied over a 10-month period in sheep experimentally infected with bovine leukaemia virus (BLV). BLV-inoculated animals seroconverted within 1 month and showed a significant lymphocytosis 2-6 weeks after infection. Control animals inoculated with BLV-free lymphocytes showed a stronger and more immediate neutrophil response than those inoculated with BLV-positive lymphocytes. One month after infection, BLV-inoculated sheep showed a relative increase of cells bearing antigens T4, T6, T8 and T19, and 10 months into the trial, MHC II lymphocytes increased, T6 remained elevated, but T4 helper cells were significantly decreased in number. Lymphoma tissue showed the presence of T8 cells, and lymph nodes from seroconverted sheep had areas of concentrated T4 staining cells. These results demonstrate responses in cellular immune mechanisms to infection with BLV.     

鸡源绿脓杆菌的分离鉴定及药敏试验

为确诊引起秦皇岛某鸡场细菌性疾病的主要病原菌,分析其耐药情况,为临床用药和治疗提供依据,本试验对送检的病死鸡肝脏中分离得到的4株菌进行形态特征观察、生化鉴定、免疫荧光染色观察、细菌16S rRNA基因PCR扩增与测序分析及药敏试验.结果显示,4株分离菌均为革兰氏阴性、带绿色荧光的短杆菌,PCR扩增出的特异性条带与预期大小相吻合,确定为绿脓杆菌.药敏试验结果显示,分离株对庆大霉素、阿米卡星、大观霉素、头孢他啶、头孢曲松、左氧氟沙星、恩诺沙星、氟苯尼考、多黏菌素B和磷霉素高度敏感,对四环素和强力霉素中度敏感,而对新霉素、头孢噻肟、头孢噻吩、阿莫西林、氨苄西林、红霉素、复方新诺明和磺胺甲氧异唑耐药.提示,该分离菌对常用抗生素耐药情况严重,养殖场应根据药敏试验结果选择敏感药物进行合理用药.     

Isolation,Identification and Drug Susceptibility Test of Pseudomonas aeruginosa in Chickens

In order to detect the main bacterial pathogens and analyze the drug resistance for providing therapeutical guidance for drug administration,4 stains were isolated from death chickens of a breeding chicken farm in Qinhuangdao area.All 4 isolates were identified by morphological characteristics,biochemical test,immunofluorescent staining,amplification of 16S rRNA genes by PCR,sequence analysis and drug susceptibility test.The results showed that all 4 isolates were gram-negative,bacillus brevis with green fluorescent,and the sequence of 16S rRNA gene of isolates were analyzed and test results showed that specific amplified band was consistent with the expected size.The isolated bacteria were identified to be Pseudomonas aeruginosa.Results of drug susceptibility test indicated that the bacterial isolates were highly sensitive to gentamycin,amikacin,spectinomycin,ceftazidine,ceftriaxone,levofloxacin,enrofloxacin,florfenicol,polymyxin B and fosfomycin,intermidiate sensitive to tetracycline and doxycyclin,resistant to neomycin,cefotaxime,cephalothin,amoxicillin,ampicillin,erythromycin,trimethoprim-sulfamethoxazole and oxygen sulfamethoxazole isoxazole.The results suggested that the resistances of Pseudomonas aeruginosa isolates to commonly used antibiotics were serious.So antibiotics should be reasonably selected based on the result of drug susceptibility test in this farm.     

中药对柔嫩艾美尔球虫感染鸡外周血T细胞亚群的影响

为研究中药对柔嫩艾美尔球虫感染鸡免疫水平的影响,本实验采用流式细胞技术检测柔嫩艾美尔球虫感染鸡外周血CD4+T细胞亚群,以及CD8+T细胞亚群数目的动态变化。结果表明,接种球虫后,感染给中药组鸡CD4+、CD8+T细胞亚群数目及百分比(CD4+T/CD8+T)均高于感染不给药组,9d时,感染给中药组最高,分别为9.77和7.53,与感染不给药组差异显著(p0.05)。感染球虫组CD4+T/CD8+T比值均低于不感染组,感染给中药组在接种后第6d和第9d均高于感染不给药组。感染给中药组、不感染给中药组和不感染不给药组CD4+T/CD8+T平均值均在2-1之间,分别为1.11、1.33和1.25,而感染不给药组仅为0.93。     

Peripheral lymphocyte subsets as a prognostic indicator of mortality and morbidity in healthy dogs

To evaluate the relationship among immune status and increased morbidity and mortality, peripheral blood lymphocytes (CD3(+), CD4(+), CD8(+) and CD21(+) cells) from 32 healthy dogs over 8 years of age were analyzed. Twenty-five of the 32 dogs were followed-up for 3 years after the analysis; and 14 dogs were found to be diseased, and nine dogs died. There was no notable difference between the ages of the dogs that died compared with the ones that survived. The relative percentage of CD4(+) and the CD4(+):CD8(+) ratio decreased notably in dogs falling ill compared with healthy dogs. The relative percentage of CD3(+) lymphocytes showed a notable decrease in dogs that died within 3 years in comparison with dogs that survived. In a discriminant analysis of morbidity and mortality, most patients were correctly classified as diseased or not and surviving or dead, respectively. These results indicate that the immunophenotypes of peripheral blood lymphocytes in older dogs offer promise as parameters for evaluating mortality and morbidity.     

Modulatory effects of chitosan adipate on the T and B lymphocyte subsets in mice

This study examined the subsets of T lymphocytes in the thymus, spleen and mesenteric lymph nodes as well as the subsets of B lymphocytes in the spleen and mesenteric lymph nodes in mice administered chitosan adipate (20 mg/kg) intraperitoneally once or four times at 24 h intervals. The results showed that chitosan adipate decreased the percentage of immature CD4⁺CD8⁺ thymic T cells and increased the percentage of mature CD4⁺ and CD8⁺ thymocytes. The most significant stimulating effect was observed after four injections. A single exposure to chitosan adipate increased the percentage of CD4⁺ mesenteric lymph node cells, but four injections of the drug increased the percentage of CD4⁺ and CD8⁺ mesenteric lymph node cells. Chitosan adipate had no effect on the subset of splenic T cells. In contrast, chitosan adipate administered either once or four times increased the percentage of CD19⁺ splenocytes but had no effect on the percentage of CD19⁺ mesenteric lymph node cells. Overall, chitosan adipate induces the maturation and differentiation of thymocytes, and regulates the number of B splenic cells and lymph node T cells irrespective of the number of doses.     

种蛋孵化死胚及孵化厅环境样本中铜绿假单胞菌的分离鉴定及其耐药性分析

本研究旨在了解种鸡场中种蛋孵化死胚和周围环境中感染或污染铜绿假单胞菌(Pseudomonas aerugino-sa,PA)的比率及分离株的耐药性.从广东省的5个规模化种鸡场采集孵化厅死胚及其周围环境样本进行PA的分离鉴定,采用K-B纸片扩散法分析其抗菌药物敏感性,并通过常规PCR检测对应的耐药基因.结果 显示,在采集...     

小熊猫源铜绿假单胞菌耐药性和耐药基因的研究

为了鉴定小熊猫(Ailurus fulgens)肺源致病菌并研究其耐药性和耐药基因,从死亡小熊猫肺部组织中无菌分离培养致病菌,进行革兰氏染色及16S rDNA测序鉴定。采用纸片扩散法(K-B法)进行细菌药敏试验和小鼠致病性试验,应用PCR法检测分离株的28种β-内酰胺酶相关基因、6种氨基糖苷类修饰酶基因(AMEs)、外膜蛋白D2基因(oprD2)、磺胺耐药基因(qacE△1-sul1)、3种整合子基因(intⅠ1、2、3)和主动外排泵调节基因(mexR)等共40种耐药基因。肺组织样品中分离培养的1株菌鉴定为铜绿假单胞菌,革兰氏染色阴性,药敏试验对氯霉素敏感,对β-内酰胺类、头孢类、氨基糖甙类、大环内酯类、硝基咪唑类、喹诺酮类和利福霉素等均具有耐药性,小鼠表现与小熊猫相似的症状而死亡。5种耐药基因TEM、aac(3)-Ⅱ、ant(3″)-Ⅰ、qacE△1-sul1、intⅠ1为阳性,其他35种为阴性。该小熊猫肺源铜绿假单胞菌具有较强的致病性和多药耐药性,其耐药机制与5种耐药基因有关。     

绵羊妊娠期外周血T细胞及其亚群的观察

应用酸性α-萘酯酶(ANAE)组化反应标记绵羊妊娠期外周血T细胞,与间情期相比,T细胞总数和辅助性T细胞(Th)无明显变化(P>0.05),而抑制性T细胞(Ts)明显增加(P<0.05).间情期和妊娠期Th/Ts分别为4.6和2.6.结果提示,妊娠期羊可能通过增加Ts细胞来压低母体免疫水平.     

Divergent selection for fleece weight in French Angora rabbits: Non-genetic effects, genetic parameters and response to selection

In order to explore genetic variability of wool production and other quantitative traits, an 8-cohort divergent selection experiment for total fleece weight (TFW) was carried out in French Angora rabbits. Studies were made on the wool production of 669 female rabbits born between 1994 and 2001 and having produced wool from the third to 12th harvests. The aim of the selection experiment was to obtain two divergent lines (low and high) on total fleece weight. The studied traits included total fleece weight, weight of the two qualities of wool (WAJ1 and WAW1), homogeneity (HOM), live body weight at ages of 4 (LW4), 8 (LW8), 12 (LW12), 16 (LW16), and 20 (LW20) weeks and then 9 weeks before each harvest (9LW). A preliminary analysis of non-genetic factors was done with the GLM procedure. The genetic parameters and genetic trends were analysed using a BLUP animal model. Heritability estimates for TFW, WAJ1, WAW1, HOM, LW4, LW8, LW12, LW16, LW20 and 9LW were 0.38, 0.30, 0.10, 0.06, 0.30, 0.09, 0.14, 0.32, 0.39 and 0.45, respectively. Genetic and phenotypic correlations between TFW and WAJ1 were high (0.98 ± 0.01 and 0.89 ± 0.01, respectively). There was a low genetic correlation between TFW and 9LW (0.26 ± 0.12). After eight cohorts of selection, the divergence between the lines was approximately three genetic standard deviations. Selection for total fleece weight had a generally beneficial effect on fleece quality.     

Adhesion molecules and lymphocyte subsets in milk and blood of periparturient Holstein cows.

Migration of leukocytes into the mammary gland is an essential element of resistance to infection which is likely influenced by expression of adhesion molecules. The contribution of subsets to mammary gland resistance remains unclear. Mononuclear cells from milk and blood of dairy cows were examined for variation in CD4+, CD8+, and WC1+ (Workshop Cluster 1; marker for gammadelta T cells) lymphocyte phenotypes and expression of LFA-1 and L-selectin at several time points during the periparturient period and at Week 16 of lactation. Proportions of CD4+ T cells were higher (p < or = 10.05) in blood than milk at all times between Week 0 and Week 16 relative to calving; the inverse was true of CD8+ cells. Expression of L-selectin was lower (p < or = 0.05) on CD4+ cells and higher on CD8+ cells from milk. The WC1+ subset was more frequent in blood than in milk except at calving when the opposite was true. After calving, proportions of L-selectin+ WC1+ cells decreased steadily to Week 16. Expression of LFA-1 was examined on mononuclear cell populations and found to be lower on milk cells and did not vary over time. We conclude that proportions of T cells subsets differ significantly between blood and milk, particularly around calving. Corresponding variations in L-selectin expression may indicate a role for this molecule in regulating the movement of CD8+ and WC1+ T cells into the bovine mammary gland.     

外周血中CD4+、CD8+T、CD4+CD25+Treg在绵羊接种布鲁氏菌M5-90弱毒疫苗后的动态变化

为研究绵羊接种布鲁氏菌弱毒M5-90株后外周血中CD4+、CD8+T、CD4+CD25+Treg细胞的动态变化规律,本研究选择11只健康绵羊,每10 d免疫一次,共免疫3次,分别在免疫前、免疫后10d、20 d、30 d利用流式细胞术检测外周血中CD4+、CD8+T、CD4+CD25+Treg淋巴细胞亚群.在免疫后的第20 d,CD4+T、CD8+T细胞百分含量达到最高水平(P＜0.05)后均缓慢下降;在第10d,CD4+CD25+Treg细胞缓慢升高,至20 d、30 d均显著升高(p＜0.05);在布鲁氏菌M5-90疫苗免疫应答过程中CD4+CD25+Treg细胞参与了机体的免疫反应调控,对CD4+T、CD8+T淋巴细胞的比例进行调节,并且维持CD4+/CD8+比值稳定,起到平衡Th1/Th2细胞间反应的作用.     

水貂源铜绿假单胞菌的分离鉴定、耐药性及毒力基因检测

为查明河北某水貂养殖场水貂死亡原因,本研究进行了病理组织学检查、细菌分离培养、细菌形态学观察、生化鉴定、16S rDNA鉴定、药敏试验、耐药基因和毒力基因检测、动物回归试验等。结果显示,共分离到3株铜绿假单胞菌,16S rDNA序列与GenBank中铜绿假单胞菌相应序列相似性达到100%,且3株分离菌均能使大鼠死亡。药敏结果表明,分离菌对左氧氟沙星、庆大霉素、阿米卡星、多黏菌素B敏感,对β-内酰胺类、氨基糖苷类、大环内酯类、喹诺酮类、四环素类等抗生素耐药。分离菌携带3种超广谱β-内酰胺酶基因（bla_CTX-M1、bla_OXA-2、bla_OXA-10）、3种碳青霉烯酶基因（bla_VIM-1、bla_SPM-1、bla_KPC-1）及吸附相关基因、T1SS-T3SS分泌系统、氧化应激相关基因、群体行为调控基因、磷脂酶相关的17种毒力基因。本研究结果为水貂绿脓杆菌病的临床诊断和治疗提供了参考依据。     

Differential cytokine expression in T cell subsets from bovine peripheral blood

Although distinct cytokine expression in T cell subsets is well understood in mice and humans, limited information is available on bovine T cell subsets. In the present study, we analyzed the mRNA expression of 10 kinds of cytokines and CD25 expression in CD4⁺, CD8⁺, WC1⁺ and WC1^-γδ T cell subsets in bovine peripheral blood by Concanavalin A (Con A) stimulation. CD25 expression was significantly increased in CD4⁺, CD8⁺ and WC1⁺γδ T cells, but not in WC1^-γδ T cells by Con A stimulation. In CD4⁺ T cells, the mRNAs of Interleukin (IL)-2, IL-6, IL-10, interferon (IFN)-γ, tumor necrosis factor (TNF)-α, TNF-β and transforming growth factor (TGF)-β were expressed in control cultures, and IL-3, IL-4 and granulocyte-macrophage colony stimulating factor (GM-CSF) were newly expressed when the cells were stimulated with Con A. CD8⁺ T cells expressed the mRNAs of IL-6, TNF-α, TNF-β and TGF-β in control cultures, and newly expressed those of IL-2, IFN-γ and GM-CSF, but did not express those of IL-3, IL-4 or IL-10 after Con A stimulation. The cytokine expression profile of WC1⁺γδ T cells was similar to that of CD8⁺ T cells. However, WC1^-γδ T cells did not express any cytokine mRNA except TGF-â mRNA. These results will contribute to elucidate the participation of T cell subsets in immune responses against infectious disease in cattle.     

Age-related changes in leukocytes and T cell subsets in peripheral blood of Japanese Black cattle

It is well known that the immune system changes with age during development and maturation in Holstein cattle. But age-related changes in leukocytes and T cell subsets in peripheral blood of Japanese Black cattle still remain unclear. The aim of the present study was to investigate comparative changes of leukocytes (granulocytes, monocytes, B cells and T cells) and T cell subsets (CD4⁺, CD8⁺, γδ, CD8⁺γδ and WC1⁺γδ T cells) in Japanese Black cattle aged 0.5, 1, 2, 6, 18 and 36–41 (adult) months on flow cytometry using specific monoclonal antibodies for the cell surface markers. T cell proportion was approximately 40% in 2-month-old cattle and decreased to 20.6% in adults. In contrast, B cell proportion significantly increased from 7.4% to 28.2% with age. In T cell subsets the percentage of CD4⁺ T cells significantly increased from 40.5% to 60%, but that of WC1⁺γδ T cell subset significantly decreased with age. The percentages of CD8⁺ and CD8⁺γδ T cells did not change. The present study details the proportional changes in leukocyte and T cell subsets with age in the peripheral blood of Japanese Black cattle and these findings are similar to those described for Holstein cattle.     

兔绿脓假单胞菌性肺炎的诊断与防治

本文报道近年来我省某试验养兔场兔群中流行一种生前以呼吸困难、口鼻出现血样分泌物症状,死后剖检以肺组织变性坏死为特征的疫情,经临床观察、实验诊断、病理剖检和病理组织学检查、流行病学调查及动物致病性实验研究,鉴定为绿脓假单胞菌(Ps.aeruginosa)引起的肺炎,该病特点传染快、死亡率高。通过流行病学调查,探索该病发病流行规律,为防治本病提供科学依据。