BoLA class I polymorphism and in vitro immune response to M. bovis antigens

SUMMARY: From a sample of 119 Friesian calves, serologically typed for BoLA class I, 47 subjects were chosen expressing 9 different MHC types (A6, A6.9, A10, A11, A14, A15, A30, W16, M103) with the same age and reared in the same farm conditions. The animals were s.c. injected with a water in oil suspension of killed M. bovis and the treatment was repeated two days later. Before the treatment and 21 days later, calves were bled and on PBM (peripheral blood mononuclear leucocytes) were performed the following tests: 1. Lymphocyte Stimulation with bovine and avian PPDs (Purified protein derivative of Mycobacterium bovis and Mycobacterium avium, respectively). 2. Phagocytic activity towards M. bovis. 3. Class II molecules expression on cell surface. 4. Percentage of leucocyte populations and subpopulations. In the in vitro Lymphocyte Stimulation test, all the animals and classes were responders. Animals bearing A10 BoLA class I presented c.p.m. (counts per minute) and index values higher than the other cattle; these values were significantly positively related both to bovine and avian PPDs (P < .01). By variance analysis A14 BoLA type showed a slight positive significant correlation with more efficient phagocytic activity. BoLA class I type did not seem to significantly affect percentage of class II positive cells and leucocyte percentages on PBM. ZUSAMMENFASSUNG: Der BoLA Klasse I Polymorphismus und in vitro immunologische Antwort gegen die Antigene von M. bovis Aus einer Stichprobe von 119 für BoLA Klasse I serologisch typisierten Friesian K?lber, wurden 47 Subjekte ausgew?hlt, die 9 verschiedene MHC Typen ausdrückten (A6, A6.9, A10, A11, A14, A15, A30, W16, M103). Alle waren gleich alt und in gleichen Haltungsbedingungen. Die Tiere wurden mit einer Wasser-in-Ol Suspension abget?teter M. bovis subkutan injiziert und die Behandlung nach zwei Tagen wiederholt. Vor und 21 Tage nach Behandlung wurden die folgenden Tests ausgeführt: 1. Lymphozyten-Stimulationstest mit bovinen und Geflügel PPDs. 2. Phagozyten Aktivit?t gegen M. bovis. 3. Zeil-Oberfl?chen, Expression der Klasse II Moleküle. 4. Anteile der Lymphozyten Populationen und Subpopulationen. Im in vitro Lymphozyten-Stimulationstest waren alle Tiere und Klassen responder. Tiere mit A10 BoLA I zeigten h?here c.p.m. und Indexwerte als die anderen; diese Werte waren in signifikant positiver Beziehung mit der PPD von M. bovis und auch mit M. avium (P < .01). BoLA Typ A14 zeigte leicht signifikant positive Korrelation mit wirksamerer Phagozyten Aktivit?t. BoLA Klasse I Typ scheint nicht den Prozentsatz der positiven Zellen der Klasse II und der Leukozyten der PBM signifikant zu beeinflussen. RESUMEN: Polimorfismo de BoLA clase I y immunidad a los antigenos del M. bovis Se escojeron 47 novillos dentro de un grupo de 119 animales que segun analisis previamente hecha tenian BoLA de clase I. Estos 47 novillos fueron escojidos de manera que tuvieran 9 distintos tipos de MHC (A6, A6.9, A10, A11, A14, A15, A30, W16, M103), la misma edad, las mismas condiciones de cria. Estos animales fueron inoculados subcutaneo con M. bovis matados en una suspension oleosa y la misma inoculacion fue repetida una secunda vez despues de dos dias. Por cada animal se tomaron muestras de sangre antes y 21 dias despues de la inoculacion de arriba. Las muestras de sangre fueron pruebaoas con: 1. Stimulacion Lymhocitaria con PPD bovina y avicola. 2. Actividad phagocitaria a M. bovis. 3. Expresion sobre la superficie celular de moleculas de clase II. 4. Porcentaje de poblaciones y de subpob-laciones de leucocitos. Todos los animales y todos los tipos de MHC dieron respuestas positivas en las pruebas de Stimulacion Lymphocitaria. Los animales que tenian la BoLA A10 presentaron valores de c.p.m. y indices mas altos de los demas animales. Estos valores se encontraron significativamente y positivamente relacionados sea a la PPD bovina que a la PPD avicola. Por medio de la analisis de varianzas se encontro que el tipo BoLA A14 muestraba una correlacion significativa y algo positiva con una mejor actividad fagocitaria. Los tipos de clase BoLA I no parecieron que influenzaran de manera appreciable el porcentaje de positividad por la clase II y el porcentaje de leucocitos en la sangre PBM.     

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。     

Tangential Chains of Neuroblasts in the Subpial Layer of the Adult Rabbit Cerebellum

Veterinary Research Communications -     

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.     

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.     

Protective Effect of <Emphasis Type="Italic">Juglans regia</Emphasis> L. Walnut Extract Against Oxidative DNA Damage

Walnuts (Juglans regia L.) are relevant components of the Mediterranean diet providing important macronutrients, micronutrients and other bioactive constituents including unsaturated fatty acids, proteins, fiber, vitamins, minerals, phytosterols and polyphenols. Although the walnut beneficial effects in human health are widely recognized by a lot of epidemiologic studies very little is known regarding its effect on damaged DNA. The aim of the present study was to investigate the effect of Juglans regia L. ethanolic extract from kernel on the induction of DNA strand breaks by thiol/Fe³⁺/O₂ mixed function oxidase, tert-butyl hydroperoxide or UVC radiations in acellular and cellular models. Plasmid DNA cleavage and fast Halo assay were used to monitor oxidative damage to DNA. Both approaches showed protection of oxidatively injured DNA. These results agree with a lot of scientific proofs which recommend walnut as dietary adjunct in health promotion and prevention as well as in treatment of lifestyle-related oxidative diseases.