A rapid MTT colorimetric assay to assess the proliferative index of two Indian strains of Theileria annulata

A study was undertaken to compare the proliferative index of macroschizont-infected lymphoblastoid cells of two Indian strains [Izatnagar (IZT) and Parbhani (PBN)] of Theileria annulata by an in vitro MTT [3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide], colorimetric assay. Culture conditions were standardized to define the optimal cell concentration in 96-well microculture plates to yield nearly 100% living cells for measurement of the metabolized formazan activity. A cell concentration of 1.5x10(5) cells/ml was found to be optimal for effective discrimination of the parasite strains. On the basis of conversion of MTT by the actively proliferating lymphoblastoid cells, the PBN strain of T. annulata stimulated a 2.5-fold increase in formazan activity in comparison to the IZT strain. The in vitro MTT assay was found to be a simple and convenient method for assessing the cell activation rate and growth, obviating the need for radioactive material for the assay. The results of the proliferation assay are discussed in relation to previously documented information on the biological characteristics of this important pathogen of cattle.     

脱脂乳发酵体系中乳酸菌的高效离子交换色谱分析

应用高效离子交换色谱对脱脂乳发酵体系中的嗜热链球菌和保加利亚乳杆菌进行色谱表征。结果表明，单菌培养过程中保加利亚乳杆菌菌体形态的变化在其相应色谱行为上得到了体现，二者的色谱峰有比较好的稳定性。两种菌的峰面积与菌数之间呈现出良好的线性关系，可决系数R^2分别为0．9957和0．9916。因此本方法的建立，可用于酸奶发酵过程中乳酸菌的分析与表征，有望为乳酸菌混合物的研究提供一种快捷的新方法。     

Factors associated with failure in breeding soundness examination of Western USA rams

Breeding-soundness examination (BSE) and eradication of Brucella ovis infection in rams are critical components of flock-health programs. The aims of this retrospective, cross-sectional study were to describe the results of BSE in a large sample of rams in the Western USA and to determine the association between BSE outcome and the semen collection method (penis manually extended vs. retained in the preputial cavity), ram body-condition score (BCS), the presence of ulcerative posthitis, and the size of the flock of origin. We evaluated the first BSE in a given year for rams from Colorado, Wyoming, and Utah, USA, from 2000 through 2007. Breeding-soundness examination consisted of physical examination, scrotal circumference and BCS measurement, semen collection by electroejaculation, and microscopic examination of semen motility, morphology, and leukocyte concentration. We assigned a reason for failure to each failed BSE and used multivariable logistic and Poisson regressions to measure associations between ram and flock variables and the risk or reason for failure on BSE. A non-random, owner-selected subset of rams was tested for antibodies to B. ovis by serum indirect ELISA (iELISA). The Rogan-Gladen corrected B. ovis seroprevalence was measured. Of the 14,667 BSEs performed on 11,804 rams, 29.0% were classified as "failed;" the most common reason for failure was substandard semen parameters (43.8%). Breeding-soundness examinations were more likely to have been categorized as failure for inflammatory causes when performed on rams from medium-sized flocks (OR 1.6; 95% CI 1.1, 2.3) and large flocks (OR 1.4; 95% CI 1.0, 1.9) (P=0.02), suggesting that larger flocks are at higher risk of contagious diseases. The adjusted seroprevalence of B. ovis antibodies among tested rams in this study was 10.0%. Of 233 rams seropositive to B. ovis, 125 (53.6%) were subclinical, a finding that supports the importance of this test in ram BSE. We found that emaciation in rams was associated with an increased risk of BSE failure from substandard semen parameters (P<0.001), but ulcerative posthitis and the semen collection method were not (P=0.09 and 0.34, respectively). However, collection of semen with the penis retained in the preputial cavity resulted in greater odds of leukospermia relative to semen collection with the penis extended (OR 4.1; 95% CI 2.9, 5.9; P<0.001), presumably from contamination of the semen sample with preputial leukocytes. For ram BSE, therefore, semen collection with the penis manually extended from the sheath is recommended to limit leukocyte contamination of the sample.     

利用质量控制图监控不同实验条件对布鲁氏菌试管凝集实验结果的影响

运用建立的质量控制图,监控不同实验条件对布鲁氏菌试管凝集实验结果可能造成的影响。在每次日常检测时对标准阳性血清进行试管凝集实验并使用浊度仪测量其上清液的光密度(OD值),在收集足够有效数据后,绘制当前条件下的质量控制图。通过模拟日常实验条件下孵育温度、时间等容易产生偏离的条件,记录不同实验条件下标准阳性血清试管凝集实验后上清液的OD值,绘制控制用控制图。运用绘制的质量控制图对不同实验条件进行监控,发现孵育温度和孵育时间均对实验结果产生较大影响,从而导致失控。采用质量控制图对布鲁氏菌试管凝集实验进行监控,可提高质量监控的有效性、科学性。严格控制实验条件是保证实验结果的重要前提。     

Association of double-stranded RNA with whitefly-mediated silvering in squash

A disorder of unknown aetiology appearing as silvering on squash (Cucurbita pepo) leaves has become prevalent in south Florida. Typical symptoms, observed after 2 weeks of exposure to adults of Bemisia tabaci , the sweet potato whitefly (SPWF) include vein clearing followed by silvering of whole leaves. Fruits from affected plants are blanched or streaked longitudinally. The silverleaf syndrome was not mechanically transmissible when crude extracts of symptomatic leaves were used as inoculum. Two sizes of double-stranded RNA (dsRNA) of c. 4.6 and 4.2 kilobase pairs (kbp) were consistently observed in leaf extracts of caged, whitefly-infested, symptomatic plants. In contrast, caged plants not exposed to whiteflies showed no symptoms and contained no detectable dsRN A. In addition, squash plants exposed to SPWF colonies from California neither developed silverleaf symptoms nor contained any dsRNA. Double-stranded RNA could also be readily detected in whitefly-infested yellow summer squash and zucchini from the field. However, asymptomatic eggplant and watermelon infested with SPWF from nearby fields did not contain detectable dsRNA. Dot-spot hybridization assays using an RNA-specific probe detected homologous sequences in both adults and nymphs of SPWF that induced silverleaf symptoms but not in adults and nymphs of SPWF that could not induce silverleaf symptoms. These results, along with the non-endogenous nature of the detected dsRNA suggest that the dsRNA is or is associated with the causal agent of whitefly-mediated leaf silvering in squash.     

Pharmacokinetics of Enrofloxacin and Its Metabolite Ciprofloxacin after Intramuscular Administration of Enrofloxacin in Goats

The pharmacokinetics of enrofloxacin and its active metabolite ciprofloxacin were investigated in goats after a single intramuscular administration of enrofloxacin at 2.5 mg/kg body weight. The plasma concentrations of enrofloxacin and ciprofloxacin were determined simultaneously by a HPLC method. The peak concentrations (C _max) of enrofloxacin (1.13 g/ml) and ciprofloxacin (0.24 g/ml) were observed at 0.8 and 1.2 h, respectively. The elimination half-life (t _1/2), volume of distribution (V _d(area)), total body clearance (Cl_B) and mean residence time (MRT) of enrofloxacin were 0.74 h, 1.42 L/kg, 1329 ml/h per kg and 1.54 h, respectively. The t _1/2, area under the plasma concentration–time curve (AUC) and the MRT of ciprofloxacin were 1.38 h, 0.74 g h/ml and 2.73 h, respectively. The metabolic conversion of enrofloxacin to ciprofloxacin was appreciable (36%) and the sum of the plasma concentrations of enrofloxacin and ciprofloxacin was maintained at or above 0.1 g/ml for up to 4 h. Enrofloxacin appears to be useful for the treatment of goat diseases associated with pathogens sensitive to this drug.     

Effect of practical diets with different protein levels on the performance of Farfantepenaeus paulensis juveniles nursed in a zero exchange suspended microbial flocs intensive system

Farfantepenaeus paulensis juveniles (72 ± 24 mg), were reared in a suspended microbial flocs system and fed practical diets containing increasing amounts of crude protein (250, 300, 350, 400 and 450 g kg^?1 CP). Development of microbial flocs was promoted by high aeration rates and fertilization with wheat bran and molasses. Flocs were composed of detritus in the form of flocculated matter colonized by heterotrophic bacteria, cocoid and filamentous cyanobacteria, flagellate and ciliate protozoa and rotifers. Proximate composition analysis of the suspended microbial floc showed CP levels of 304 g kg^?1. After 45 days, mean shrimp survival were above 89%, with no significant differences between treatments. Shrimp fed diets with 350 g kg^?1 or higher CP content achieved significant higher (P < 0.05) final weight (0.66–0.68 g), weight gain (0.58–0.61 g) and instantaneous growth rate (0.049–0.050), with feed conversion rates (2.17–2.30) significantly lower (P < 0.05). Results show that, when rearing is carried out in a suspended microbial flocs system, dietary CP levels can be kept at 350 g kg^?1. Furthermore, results confirm that microbial‐based systems allow shrimp culture without compromising the surrounding environment and shows the possible reduction of production costs and fish meal dependence.     

Molecular genetic mapping of quantitative trait loci associated with loaf volume in hexaploid wheat (Triticum aestivum)

Major efforts in wheat research are being made to improve the yield and quality of wheat. Loaf volume (Lv) is the main quality parameter deciding the bread making potential of wheat. To genetically dissect quantitative trait loci (QTLs) for Lv, a Recombinant Inbred Line (RIL) population (F₈) was developed from a cross between two Indian wheat varieties “HI 977” and “HD 2329”. A total of 914 SSR and 100 ISSR primers were used for molecular analysis and the genetic map comprising 19 chromosomes was constructed with 202 SSR markers and 2 HMW glutenin subunit loci: Glu-B1 and Glu-D1. The phenotypic data were collected from six environments including three different agro-climatic zones for 2 consecutive years. Dissection of Lv through AMMI model revealed significant G×E variance for the trait. QTL analysis was performed using composite interval mapping. A total of 30 QTLs for Lv were detected and significant QTLs were identified on 6B and 6D chromosomes; 1B, 1D, 2A, 3A, 5B and 5D also contributed genetically to Lv. Association between 6B and 6D QTLs and variable expression of gliadins on group 6 chromosomes were discussed. QTLs detected in this study were compared with other QTL analysis in wheat.     

Campylobacter

Species within the genus, Campylobacter, have emerged over the last three decades as significant clinical pathogens, particularly of human public health concern, where the majority of acute bacterial enteritis in the Western world is due to these organisms. Of particular concern are the species, C. jejuni and C. coli, which are responsible for most of these gastrointestinal-related infections. Although these organisms have already emerged as causative agents of zoonoses, several aspects of their epidemiology and pathophysiology are only beginning to emerge. Trends in increasing antibiotic resistance are beginning to emerge with oral antibiotics, which may be the drug of choice for when it is necessary to intervene chemotherapeutically. This review wishes to examine (i) emerging clinical aspects of the disease, such as Guillain Barre syndrome (GBS), (ii) the association between these organisms and poultry as a natural host, (iii) environmental aspects of Campylobacter epidemiology, (iv) the emergence of atypical campylobacters (v) emerging trends in antibiotic resistance, (vi) adoption of modern methods for the detection of campylobacters.