Multi#ex—CAPS技术及其在番茄遗传鉴定中的应用

 ：应用Multiplex—PCR和CAPS(Cleaved amplified polymorphic sequence)原理研究建立了Mutiplex—CAPS技术。该技术包括：(1)在PCR扩增反应体系加入2对、3对或多对核苷酸引物，经30～40个扩增循环后得到每对引物的特异扩增片段；(2)应用同一种限制性内切酶消化这些不同引物经扩增后获得的特异片段；(3)琼脂糖凝胶电泳溴化乙锭染色检测酶切片段长度的多态性。利用该技术对番茄(Lycopersicon es—cdentum)TA517及其近等基因系的分析表明，Multiplex—CAPS能同时检测2个、3个或多个CAPS标记的多态性，其效果等同于每个CAPS标记的多态性的叠加，重复性好，分析效率高，降低成本数倍。     

Long‐Term Tillage and Cropping System Effects on Chemical and Biochemical Characteristics of Soil Organic Matter in a Mediterranean Semiarid Environment

Several studies have reported how tillage and cropping systems affect quantity, quality, and distribution of soil organic matter (SOM) along the profile. However, the effect of soil management on the chemical structure of SOM and on its hydrophobic and hydrophilic components has been little investigated. In this work, the long‐term (19 years) effects of two cropping systems (wheat monoculture and wheat/faba bean rotation) and three tillage managements (conventional, reduced, and no tillage) on some chemical characteristics of SOM and their relationships with labile carbon (C) pools were evaluated. Soil samples were taken from the topsoil (0–15 cm) of a Chromic Haploxerert (central Sicily, Italy). After 19 years of different tillage and cropping systems management, total organic C significantly differed among treatments with the labile organic C pools showing the greater amount in no till and in wheat/faba bean plots. Hydrophobic and hydrophilic components of SOM, determined by diffuse reflectance infrared Fourier transform spectroscopy, were mainly affected by cropping system, whereas aromatic components of SOM by tillage. Soil organic matter components and characteristics showed significant correlations with the soil biochemical parameters, confirming the expected synergism between chemical and biochemical properties. This study demonstrated that (i) no tillage and crop rotation improve the chemical and biochemical properties of SOM of Vertisols under semiarid environment; and (ii) tillage management and cropping systems have affected, after 19 years, more the chemical and biochemical properties of SOM than its quantity. Copyright © 2014 John Wiley & Sons, Ltd.     

Laboratory tests for diagnosing and monitoring canine leishmaniasis

Although several reviews on canine leishmaniasis have been published, none thoroughly described clinicopathologic abnormalities and their clinical usefulness. The aim of this review was to provide information concerning current diagnostic tests relevant for clinical pathologists and from a practical perspective. Specifically, in canine leishmaniasis, nonregenerative normocytic normochromic anemia, thrombocytopenia, or leukogram changes may be present. Clinical chemistry and urinalysis may indicate renal dysfunction (azotemia, decreased urine specific gravity, proteinuria) and an inflammatory/immune response (increased acute phase proteins [APP] or α₂‐ and/or γ‐globulins). Although a potential gammopathy is usually polyclonal, it may also appear oligo‐ or monoclonal, especially in dogs coinfected by other vector‐borne pathogens. When lesions are accessible to fine‐needle aspiration (lymphoadenomegaly, nodular lesions, joint swelling), cytology is strongly advised, as the presence of Leishmania amastigotes in a pattern of pyogranulomatous inflammation or lymphoplasmacytic hyperplasia is diagnostic. If the cytologic pattern is inconclusive, the parasite should be identified by histology/immunohistochemistry or PCR on surgical biopsies. Alternatively, cytology and PCR may be performed on bone marrow samples where amastigotes, along with erythroid hypoplasia, myeloid hyperplasia, plasmacytosis, or secondary dysmyelopoiesis can be observed. Dogs with overt leishmaniasis generally have high antibody titers, while low titers predominate in immunologically resistant infected dogs or in exposed dogs with no parasite confirmation. Quantitative serology is recommended in clinically suspect dogs as high‐titer antibodies titers may confirm the clinical diagnosis. In confirmed and treated dogs, renal function and inflammatory/immune response variables should be periodically monitored.     

Tolerance of benzalkonium chloride, formalin, malachite green, and potassium permanganate in goldfish and zebrafish

OBJECTIVE: To determine tolerance of goldfish and zebrafish to benzalkonium chloride, formalin, malachite green, and potassium permanganate. DESIGN: Tolerance study. ANIMALS: Adult goldfish (Carassius auratus) and zebrafish (Danio rerio). PROCEDURES: Groups of fish (n = 10/group) were exposed to each disinfectant at the therapeutic dosage; at 0.25, 0.5, 3, and 5 times the concentration used for the therapeutic dosage; and at the concentration used for the therapeutic dosage but for 3 or 5 times the recommended exposure time. RESULTS: In both species, exposure to malachite green at the therapeutic dosage resulted in toxic effects, including death. Exposure to formalin at the therapeutic dosage resulted in toxic effects in goldfish, but not zebrafish, and exposure to potassium permanganate resulted in toxic effects in zebrafish, but not goldfish. On the basis of the ratio of therapeutic dosage to median lethal dosage, in goldfish, formalin was more toxic than benzalkonium chloride, which was more toxic than malachite green, which was more toxic than potassium permanganate. In zebrafish, potassium permanganate was more toxic than formalin and benzalkonium chloride, which were approximately equally toxic and more toxic than malachite green. Extending treatment time increased the toxicity of potassium permanganate in zebrafish and the toxicity of formalin and malachite green in goldfish, but did not alter the toxicity of the other disinfectants. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that there was no consistency between zebrafish and goldfish in their tolerance to disinfectants, and that therapeutic dosages reported in the literature for these disinfectants were not always safe.     

Estimating evapotranspiration in the Padova Botanical Garden

In 1996, intensive building operations near the botanical garden of Padova, the oldest botanical garden in the world, altered a long-established equilibrium between groundwater and plants and threatened the lives of some of them. To avoid water stress, an advanced irrigation system was installed. For design purposes, better knowledge of the water cycle and the monthly average evapotranspiration (ET) in the area was needed. Due to the complex canopy stand of the site, ET was estimated using the water balance method, integrating mathematical models with the Arc/Info Geographical Information System. The water balance was estimated in 1997 and 1998, and results were used to derive an empirical mean crop coefficient of the botanical garden, to simulate the long-term water requirements using the product of reference ET and the apparent crop coefficient to estimate ET from the garden. Two types of hydrological behaviour were identified: one in the central area of the garden, where reduced ground cover diminishes ET and increases runoff and percolation. In the external area, the ET was higher because of the presence of many trees. The empirical mean monthly crop coefficient ranged between 0.56 and 0.83, indicating that ET in the entire area is always less than grass reference ET.     

In vitro formation of metabolic-intermediate cytochrome P450 complexes in rabbit liver microsomes by tiamulin and various macrolides

Tiamulin and a number of macrolides were evaluated as to their ability in forming metabolic-intermediate (MI) complexes with cytochrome P450 in liver microsomes from rabbits bred for meat production. Complex formation, which occurred only in preparations where the expression of P450 3A was increased as the result of rifampicin pre-treatment and with different kinetics, was in the order tiamulin > erythromycin > TAO approximately roxithromycin approximately tylosin and did not take place with tilmicosin and spiramycin. Most of the tested compounds underwent an oxidative N-dealkylation and a good relationship could be found between the rate of N-dealkylase activity in induced preparations and the aptitude in generating MI complexes. Although the results from in vitro studies should be interpreted with caution, it is suggested that the potential for in vivo drug interactions also exists in the rabbit for tiamulin and for four out of the six tested macrolides.     

Discrimination of ‘San Marzano’ accessions: A comparison of minisatellite, CAPS and SSR markers in relation to morphological traits

‘San Marzano’ (SM) is one of the most widely known tomato (Solanum lycopersicum L.) cultivars, and is a classic example of a local variety with a premium value. Unfortunately, the original cultivated form is underrepresented in the Protected Denomination of Origin (PDO) area because of the incidence of contaminant and phenotypically similar genotypes. Our aim was to examine the ability of three DNA marker systems (minisatellite, cleaved amplified polymorphic sequence (CAPS) and simple sequence repeat (SSR)) to reveal the genetic diversity of tomato accessions that were, based on a morphological analysis, very similar. The data indicate that both minisatellites and SSRs can be used to genetically distinguish the analysed materials. Furthermore, these two marker systems depict relationships consistent with the hierarchal pattern obtained by the morphological data. As locally cultivated tomato accessions are often characterised by some degree of genetic variability, our results will be valuable in facilitating the purification, management and breeding of tomato germplasms. The differences between the marker systems employed are also discussed in relation to their usefulness in the agro-food chain.     

Closed reduction and surgical fixation of elbow joint luxation in an adult mare and in a 3-month-old foal using knotless bone anchors and FiberTape

This report describes the successful surgical stabilisation of elbow joint luxation using knotless bone anchors and FiberTape to repair the medial collateral ligament in an 11-year-old mare (Case 1) and both medial and lateral collateral ligaments in a 3-month-old foal (Case 2). Radiography (both cases) and computed tomography (Case 2) were used to diagnose subluxation (Case 1) and complete luxation (Case 2) with suspected rupture of both collateral ligaments. Case 1 was supported in a rescue sling for 7 weeks postoperatively to prevent recumbency. Recovery was good, and the mare was sound at the walk at the time of discharge 8 weeks after surgery and 1 year later at the follow-up visit. Case 2 was discharged 3 weeks after surgery and was sound at the trot when re-evaluated 2 and 6 months postoperatively. Although elbow luxation accompanied by torn collateral ligaments is rare in horses, successful outcomes have been described. In the present case report, joint stabilisation and a successful outcome were achieved, for the first time, with closed reduction and an open approach for placement of knotless bone anchors and FiberTape. Limitations of our study included inability to directly assess the integrity of the implants during healing and relying mainly on clinical signs to assess healing.     

Investigation of biofilm production and icaA and icaD genes in Staphylococcus aureus isolated from heifers and cows with mastitis

Biofilm formation and antimicrobial resistance of Staphylococcus aureus are important virulence factors in cases of mastitis in dairy cows. However, few studies have investigated mastitis strains isolated from heifers. Within this context, the objective of the present study was to investigate biofilm formation on Congo red agar, the presence of the icaA and icaD genes by polymerase chain reaction (PCR), and the percentage of in vitro antimicrobial resistance of 110 S. aureus isolates from mammary gland secretions of heifers and cows with mastitis. PCR detected the icaA and icaD genes in 98% and 100% of isolates, respectively. However, only 55.5% of all isolates produced a biofilm on Congo red agar. Antimicrobial susceptibility testing revealed that 47.0% of isolates from heifers and 70.4% of isolates from cows were resistant to at least one of the antimicrobial agents tested. Resistance to penicillin and/or ampicillin was the most frequent (44.5%). These results indicate the need to implement prophylactic and control measures of mastitis for heifers. Heifers and cows can carry resistant strains with the capacity of biofilm production, a fact representing a threat to public health and animal well‐being and generating losses to dairy farmers.