Stability of Citrus tristeza virus protective isolate 'Pêra IAC' according to SSCP analysis of old and new lines of three sweet orange varieties

Clonal cleaning, followed by pre-immunization with protective complexes of Citrus tristeza virus(CTV), allowed the commercial cultivation of Pêra sweet orange, a variety that has great importance for Brazilian citriculture but is sensitive to the virus. The use of mild protective isolates in other citrus varieties, even those more tolerant to CTV, can also be of interest to prevent the spread of severe isolates. The aim of this study was to characterize, by means of SSCP (Single Strand Conformational Polymorphism) analysis of the coat protein gene, CTV isolates present in plants of the sweet orange cultivars Pêra, Hamlin and Valencia propagated from four budwood sources: 1) old lines, 2) nucellar lines, 3) shoot-tip-grafted lines, and 4) shoot-tip-grafted lines pre-immunized with the mild CTV protective isolate 'PIAC'. We also evaluated the correlation of the obtained SSCP patterns to stem pitting intensity, tree vigor and fruit yield. SSCP results showed low genetic diversity among the isolates present in different trees of the same variety and same budwood source and, in some cases, in different budwood sources and varieties. Considering tristeza symptoms, lower intensity was noted for plants of new, shoot-tip-grafted and pre-immunized shoot-tip-grafted lines, compared to old lines of the three varieties. The observed SSCP patterns and symptomatology suggested that more severe CTV complexes infect the plants of old lines of all three varieties. The protective complex stability was observed in the SSCP patterns of CTV isolates of some shoot-tip-grafted and pre-immunized clones. It was concluded that the changes detected in other electrophoretic profiles of this treatment did not cause loss of the protective capacity of CTV isolate 'PIAC' inoculated in the pre-immunization.

Germination of Caesalpinia leiostachya (benth.) ducke seeds classified by size and shape

Caesalpinia leiostachya (Benth.) Ducke (Caesalpiniaceae) is a Brazilian legumenous tree whose seeds present different sizes and shapes, with dormancy imposed by a waterproof coat. In order to improve that quality of lot, seeds were classified by width in sieves of circular meshes (13, 14, 15, 16 and 17) and manually separated by shape in elongate, round and angular. Masses of 1000 seeds were determined and the thickness of each width and shape seedclass was measured, and coat permeability of the seeds classified by the width was investigated. Seeds were scarified in concentrated sulphuric acid and submitted to the germination test. Both final percentage and speed of germination index were evaluated. The classification of the seeds improved the quality of the lot, and the coat permeability was not affected by seed width. Elongated-flat seeds are of low physiological quality and should be discarded. Masses of 1000 seeds, percentage final and speed of germination increased with the width of the seeds. Both elongated and angular seeds had similar germinative behavior, and round seeds are of greater thickness and of superior physiological quality.

Benefits of automated acclimatization during the pre-milking phase of lactating girolando cows

The aim of this paper was to evaluate the automated acclimatization effects during pre-milking of cows on thermal conditioning, physiology, milk production and cost-benefit of the automated adiabatic evaporative cooling system (AECS). The treatments 20; 30; 40 min and control consisted of exposure time of pre-milking cows to the automated AECS. Sixteen cows were used with an average daily milk yield of 19 kg, distributed in a 4 x 4 Latin square design. The Tukey's test (P<0.05) was used to compare the means. The environmental variables, dry bulb temperature (DBT, ºC) and relative humidity (RH, %), were recorded every minute, which allowed the determination of the system efficiency through the Temperature and Humidity Index (THI). The respiratory rate (RR), rectal temperature (RT) and temperature of the coat (TC) were measured before and after the acclimatization. The 40 min treatment kept the environmental variables and the comfort indexes within recommended limits. The physiological variables (RR, RT and TC) were lower in the 40 min treatment and reflected positively on milk production, which increased 3.66% compared to the control treatment. The system was profitable, having a 43 days return on investment and a monthly revenue increase of R$ 1,992.67.

Biopohjaiset polymeerit dispersiopäällystyksessä

Työn aiheena oli tehdä muotoiltavissa oleva rasvankestävä pakkauskartonki. Polymeeridispersiopäällystettyjä pakkauskartonkeja käytetään kertakäyttöisissä tuotteissa kuten vuoissa, kupeissa ja lautasissa. Tuotteiden on kestettävä rasvaa niiden lopputarkoituksen mukaisesti. Pakkausala jatkaa vuosittaista kasvuaan vauhdilla. Uusia kierrätettäviä biopohjaisia ja luonnolle ystävällisempiä pakkaustuotteita on kehitettävä kasvun tyydyttämiseksi. Biopohjaiset pakkaustuotteet ovat mahdollisia ratkaisuja ympäristöön kohdistuvien ongelmien vähentämiseksi ja öljypohjaisten raaka-aineiden korvaajiksi. Diplomityön teoriaosuudessa keskityttiin dispersiopäällystyksessä käytettyihin biopolymeereihin ja niiden toimivuuteen suojaavina kalvoina. Teoriaosuudessa käsiteltiin myös rasvankestoa ja rasvankestävien tuotteiden materiaaliominaisuuksia. Kirjallisuuden perusteella havaittiin luonnonpolymeerien alhaisen kuiva-ainepitoisuuden ja korkean päällystemäärätarpeen muodostamat haasteet dispersiopäällystyksessä. Pohjakartongin karheudella ja tiiveydellä sekä suojaavan polymeerin kalvon rakenteella huomattiin olevan suuri merkitys rasvankeston saavuttamiseksi. Kokeellinen osa jakautui kolmeen osakokonaisuuteen: laboratoriokokeisiin, esipilotointiin ja varsinaiseen pilot-koeajoon. Esikokeiden perusteella suojaavat kalvot, joiden raaka-aineena käytettiin biopohjaisia dispersiopolymeerejä, antoivat riittäviä rasvankesto-ominaisuuksia, mutta eivät kestäneet konvertointia. Pienellä synteettisten polymeerien lisäyksellä pystyttiin parantamaan päällystettyjen kartonkien rasvankestoa sekä konvertoitavuutta. Pilot-koeajonäytteiden testaustulokset tukivat esikokeissa tehtyjä havaintoja. Tämän työn perusteella kohtuullinen lisäysmäärä synteettistä polymeeriä voi parantaa merkittävästi biopohjaisen suojaavan kalvon antamaa rasvankestoa sekä päällystetyn kartongin konvertoitavuutta.

Comparison of two commercial kits and two extraction methods for fecal glucocorticoid analysis in ocelots (Leopardus pardalis) submitted to ACTH challenge

The ocelot (Leopardus pardalis) is included in list of wild felid species protected by CITES and is part of conservation strategies that necessarily involve the use of assisted reproduction techniques, which requires practical and minimally invasive techniques of high reproducibility that permit the study of animal reproductive physiology. The objective of this study was to compare and validate two commercial assays: ImmuChem Double Antibody Corticosterone 125I RIA from ICN Biomedicals, Costa Mesa, CA, USA; and Coat-a-Count Cortisol 125I RIA from DPC, Los Angeles, CA, USA, for assessment of fecal glucocorticoid metabolites in ocelots submitted to ACTH (adrenocorticotropic hormone) challenge. Fecal samples were collected from five ocelots kept at the Brazilian Center of Neotropical Felines, Associação Mata Ciliar, São Paulo, Brazil, and one of the animals was chosen as a negative control. The experiment was conducted over a period of 9 days. On day 0, a total dose of 100 IU ACTH was administered intramuscularly. Immediately after collection the samples were stored at 20C in labeled plastic bags. The hormone metabolites were subsequently extracted and assayed using the two commercial kits. Previously it was performed a trial with the DPC kit to check the best extraction method for hormones metabolites. Data were analyzed with the SAS program for Windows V8 and reported as means ± SEM. The Schwarzenberger extraction method was slightly better when compared with the Wasser extraction method (103,334.56 ± 19,010.37ng/g of wet feces and 59,223.61 ± 12,725.36ng/g of wet feces respectively; P=0,0657). The ICN kit detected an increase in glucocorticoid metabolite concentrations in a more reliable manner. Metabolite concentrations (ng/g wet feces) on day 0 and day 1 were 66,956.28 ± 36,786.93 and 92,991.19 ± 28,555.63 for the DPC kit, and 205,483.32 ± 83,811.32 and 814,578.75 ± 292,150.47 for the ICN kit, respectively. The limit of detection for the ICN kit was 7.7 ng/mL for 100% B/Bo (25ng/mL for 88%B/Bo) and for the DPC kit it was 0.2ug/dL for 90.95% B/Bo (1ug/dL for 81.27% B/Bo). In conclusion it was confirmed that the Schwarzenberger extraction method and the ICN kit are superior for extracting and measuring fecal glucocorticoid metabolites in ocelot fecal samples.

Mikrohuokoisen pinnoitteen kehittäminen suodatinkankaan päälle

Mikrohuokoisia membraaneja käytetään monissa suodatussovelluksissa kuten vedenpuhdistuksessa. Tämän työn tavoite oli pinnoittaa suodatinkankaita mikrohuokoisella membraanilla. Membraanimateriaalin ja suodatuskankaan yhdistelmällä tavoiteltiin komposiittimateriaalia, jolla on membraanin selektiivinen erotuskyky ja kankaan mekaaniset ominaisuudet. Tämän työn kirjallisuusosa käsittelee membraanin valmistusta faasi-inversiomenetelmällä, pinnoitteeksi soveltuvia membraanimateriaaleja sekä pinnoitteen lujittamista. Kokeellisessa osassa suodatuskankaalle kehitettiin sopivaa pinnoitusmenetelmää. Pinnoitemateriaaleina käytettiin akryylidispersioainetta, polyamidia ja polysulfonia. Tuloksista huomattiin, ettei akryylidispersioaine soveltunut pinnoitemateriaaliksi. Kangasta onnistuttiin pinnoittamaan polyamidilla ja pinnoitusolosuhteita kontrolloimalla saatiin aikaiseksi pinnoite, joka pysyi kankaassa kiinni ja jonka rakenne koostui noin 12 μm huokosista. Polymeeriliuoksen pitoisuuden huomattiin vaikuttavan pinnoitteen rakenteeseen, mutta siihen jäi edelleen suuria avoimia huokosia. Suuret yksittäiset huokoset laskivat kuplapistepainetta ja tekivät huokoskokojakaumasta leveämmän. Polyamidipitoisuudella 14 ja 16 m–% pinnoite oli tunkeutunut kankaan sisään ja sen rakenne oli huokoinen, mutta pintakerrokseen muodostunut tiivis ihokerros tukki materiaalia. Tulosten perusteella kankaiden pinnoittaminen membraanikerroksella on mahdollista mutta pinnoitusprosessia täytyy edelleen kehittää, jotta päästään pienempään huokoskokoon ja kapeampaan huokoskokojakaumaan pinnoitteessa.

Blade and rod coating of low solid dispersions

Työn aiheena oli tehdä ohut barrierkalvo terä- tai sauvapäällystys menetelmällä. Erilaisissa elintarvikepakkauksissa käytetään hyviä barrier-ominaisuuksia omaavia ohuita päällysteitä. Elintarvikepakkauksen tehtävä on suojata pakattua tuotetta ympäristöltä, mahdollistaa helppo kuljetus ja säilytys sekä antaa tarvittavat tiedot tuotteesta tuotteen käsittelijöille ja loppukäyttäjille. Diplomityön teoriaosuudessa keskityttiin barrierpäällystykseen, eri päällystysmenetelmiin, niiden erityisvaatimuksiin ja ominaisuuksiin. Teoriaosuudessa käsiteltiin myös vaadittavia barrier-ominaisuuksia ja haasteita niiden saavuttamisessa. Kirjallisuuden perusteella haasteiksi nousivat helposti muodostuvat mikroreiät. Kokeellinen osa jakautui kahteen osakokonaisuuteen: laboratoriokokeisiin ja pilot-koeajoon. Laboratoriokokeita tehtiin ennen pilot-ajoa, jotta pilot-koeajoon voitiin valita parhaat päällystereseptit. Pilot-koeajonäytteiden päällystemäärät osoittautuivat liian pieniksi ja siksi laboratoriossa tehtiin jatkotutkimuksia riittävän päällystemäärän saavuttamiseksi. Tämän työnperusteella pohjakartongin ominaisuuksilla, erityisesti karheudella, on merkittävä vaikutus päällystyksen onnistumisessa ja yksinkertaisilla resepteillä ja päällystysmenetelmillä ei saada tarpeeksi laadukasta kalvoa.

SEED DORMANCY BREAKING TREATMENTS FORAFRICAN PURSLANE (Zaleya pentandra)

ABSTRACT Understanding the mechanisms involved in releasing seed dormancy is crucial for effective plant management and renewal of species in the arid zone. Zaleya pentandra is an emerging invasive weed of the arid areas of Pakistan. We investigated the effects of different dormancy breaking treatments on the germination of Z. pentandra seeds. Seeds were treated with hot water (by placing them in boiling water for 5, 15, 30, 60, 90, 120, and 150 min), dry heat (by placing them in a preheated oven at 70 oC for 1, 2, and 4 hours; at 70 oC for 1, 2, 3, and 4 days, and at 200 oC for 5, 10, 15, 30, and 45 min) and stratification (by placing them at 2-5 ºC in a refrigerator for 5, 10, 30, and 60 min; for 3, 6, and 12 hours, and for 1, 2, 4, 8, 15, and 30 days). Seeds also were soaked in thiourea ([(NH2)2CS] (0, 2,500, 5,000, 7,500, and 10,000 mg L-1 for 24 h at 30 oC) and in KNO3 (0, 10,000, 20,000, 30,000, 40,000, 50,000, and 60,000 mg L-1 for 24 h at 30 oC). Additionally, seeds were scarified with HCl (for 3, 6, 9, 12, 15, 18, and 21 h), HNO3 (for 3, 6, 9, 12, 15, 18, and 21 h), and H2SO4 (for 20, 40, 60, 80, 100, and 120 min at 30 oC) and also mechanically scarified with sandpaper. Zaleya pentandra seeds showed typical signs of hard seed coat dormancy. Mechanical scarification and acid treatments promoted seed germination to a varying degree. Seed scarification with HNO3 for 12 to 18 h as well as with HCl for 12 h and 15 h was efficient in breaking dormancy of Z. pentandra seeds, providing germination up to 92.5%. Seed scarification with H2SO4 from 20 to 120 min showed little effect, whereas hot water, dry heat, stratification and various concentrations of thiourea and KNO3 were ineffective in breaking Z. pentandra seed dormancy.

Estrutura do tegumento seminal de quatro espécies de Leguminosae (Caesalpinioideae), ocorrentes numa área de caatinga (PE ­ Brasil)

Neste estudo observou-se que os tegumentos seminais de Caesalpinia ferrea, C. pyramidalis, Senna martiana e S. spectabilis var. excelsa estão compostos de um estrato de macrosclereídes, revestido por cutícula e subcutícula, seguido por uma camada de osteosclereídes, parênquima esclerificado e camada interna também de osteosclereídes. A impermeabilidade dos tegumentos seminais, possivelmente, se restringe à região das macroesclereídes.

Possibility to add a barrier layer to a fibre tray package in the moulding stage

The main objective of the present study was to analyze the best approach on how to coat paperboard trays at the pressing stage. The coating gives the paperboard enhanced barrier and mechanical properties. The whole process chain of the barrier coating development was studied in the research. The methodology applied includes obtaining the optimum temperature at which good adhesion and bonding is formed between paperboard and skin film. Evaluation of mechanical properties after the coatings; such as cracking, curling and barrier properties was performed.

Morphology and anatomy of the developing fruit of Maclura tinctoria, Moraceae

(Morphology and anatomy of the developing fruit of Maclura tinctoria, Moraceae). Maclura tinctoria (L.) D. Don ex Steudel was selected for the present study due of its economic and medicinal importance. The purpose of this investigation is to present a detailed description of the fruit development, specially by: (a) defining the fruit type presented by the species, and (b) characterizing the seed type of the species based upon the presence or not of mechanical tissue on the seed-coat. The fruit originates from the subglobose female inflorescence which consists of small unipistillate flowers with superior ovary, unilocular and uniovular apical placentation. The mature fruit is multiple, constituted of small drupes. The ovule is ana-campylotropous, suspended, bitegmic and crassinucellate. The mature seed is flattened, slightly ovated, cream colored, with unspecialized membrane coat with thin-walled cells more or less crushed. The seed has parenchymatic endosperm with lipophilic content. The embryo is straight, with two cotyledons of the same size. Ontogenetic studies reveal that the fruits are infrutescences. The fleshy edible part is derived from the perigone and inflorescence axis. The drupes consist of a single pyrene of macrosclereids.

Viability and vigor of jamun (Syzygium cumini) seeds

Jamun (Syzygium cumini L. Skeels) (Black plum, Damson plum) fruits weigh between 2-5 g at maturity. Fresh seeds represented 20-80% of the total fruit weight; the seed coat and cotyledons contributed 6% and 94% to the total seed weight respectively, while the weight of the embryonic axis was insignificant. Only the embryonic axis stained with Tetrazolium, not the cotyledons. The seeds are polyembryonic with up to four embryos, of which at most three germinate. Decoated seeds germinated faster than coated seeds under nursery conditions, with high significant germination percentages, dry matter production rates and vigor indices. The lack of staining of the cotyledon by tetrazolium was probably due to the presence of an impermeable layer. Decoating seeds for faster germination is recommended.

Study of the effects of ß-myrcene on rat fertility and general reproductive performance

ß-Myrcene (MYR) is a monoterpene found in the oils of a variety of aromatic plants including lemongrass, verbena, hop, bay, and others. MYR and essential oils containing this terpenoid compound are used in cosmetics, household products, and as flavoring food additives. This study was undertaken to investigate the effects of MYR on fertility and general reproductive performance in the rat. MYR (0, 100, 300 and 500 mg/kg) in peanut oil was given by gavage to male Wistar rats (15 per dose group) for 91 days prior to mating and during the mating period, as well as to females (45 per dose group) continuously for 21 days before mating, during mating and pregnancy, and throughout the period of lactation up to postnatal day 21. On day 21 of pregnancy one-third of the females of each group were submitted to cesarean section. Resorption, implantation, as well as dead and live fetuses were counted. All fetuses were examined for external malformations, weighed, and cleared and stained with Alizarin Red S for skeleton evaluation. The remaining dams were allowed to give birth to their offspring. The progeny was examined at birth and subsequently up to postnatal day 21. Mortality, weight gain and physical signs of postnatal development were evaluated. Except for an increase in liver and kidney weights, no other sign of toxicity was noted in male and female rats exposed to MYR. MYR did not affect the mating index (proportion of females impregnated by males) or the pregnancy index (ratio of pregnant to sperm-positive females). No sign of maternal toxicity and no increase in externally visible malformations were observed at any dose level. Only at the highest dose tested (500 mg/kg) did MYR induce an increase in the resorption rate and a higher frequency of fetal skeleton anomalies. No adverse effect of MYR on postnatal weight gain was noted but days of appearance of primary coat, incisor eruption and eye opening were slightly delayed in the exposed offspring. On the basis of the data presented in this paper the no-observed-adverse-effect level (NOAEL) for toxic effects on fertility and general reproductive performance can be set at 300 mg of ß-myrcene/kg body weight by the oral route.

The leaves of green plants as well as a cyanobacterium, a red alga, and fungi contain insulin-like antigens

We report the detection of insulin-like antigens in a large range of species utilizing a modified ELISA plate assay and Western blotting. We tested the leaves or aerial parts of species of Rhodophyta (red alga), Bryophyta (mosses), Psilophyta (whisk ferns), Lycopodophyta (club mosses), Sphenopsida (horsetails), gymnosperms, and angiosperms, including monocots and dicots. We also studied species of fungi and a cyanobacterium, Spirulina maxima. The wide distribution of insulin-like antigens, which in some cases present the same electrophoretic mobility as bovine insulin, together with results recently published by us on the amino acid sequence of an insulin isolated from the seed coat of jack bean (Canavalia ensiformis) and from the developing fruits of cowpea (Vigna unguiculata), suggests that pathways depending on this hormone have been conserved through evolution.

Heterofucans from Dictyota menstrualis have anticoagulant activity

Fucan is a term used to denote a family of sulfated L-fucose-rich polysaccharides which are present in the extracellular matrix of brown seaweed and in the egg jelly coat of sea urchins. Plant fucans have several biological activities, including anticoagulant and antithrombotic, related to the structural and chemical composition of polysaccharides. We have extracted sulfated polysaccharides from the brown seaweed Dictyota menstrualis by proteolytic digestion, followed by separation into 5 fractions by sequential acetone precipitation. Gel electrophoresis using 0.05 M 1,3-diaminopropane-acetate buffer, pH 9.0, stained with 0.1% toluidine blue, showed the presence of sulfated polysaccharides in all fractions. The chemical analyses demonstrated that all fractions are composed mainly of fucose, xylose, galactose, uronic acid, and sulfate. The anticoagulant activity of these heterofucans was determined by activated partial thromboplastin time (APTT) using citrate normal human plasma. Only the fucans F1.0v and F1.5v showed anticoagulant activity. To prolong the coagulation time to double the baseline value in the APTT, the required concentration of fucan F1.0v (20 µg/ml) was only 4.88-fold higher than that of the low molecular weight heparin Clexane® (4.1 µg/ml), whereas 80 µg/ml fucan 1.5 was needed to obtain the same effect. For both fucans this effect was abolished by desulfation. These polymers are composed of fucose, xylose, uronic acid, galactose, and sulfate at molar ratios of 1.0:0.8:0.7:0.8:0.4 and 1.0:0.3:0.4:1.5:1.3, respectively. This is the fist report indicating the presence of a heterofucan with higher anticoagulant activity from brown seaweed.