Silver nanoparticles preplant bulb soaking affects tulip production

Nanomaterials and particularly silver nanoparticles (AgNPs) have increased great interest in many fields of science. The element may affect plant growth and development but so far research studies have been scarce and their results disparate. Tulip is one of the most important ornamental plants and its production technology is constantly being improved. The aim of this study was to investigate the effects of AgNPs on growth and yield attributes of tulip cv. ‘Pink Impression’. Before planting, the tulip ‘Pink Impression’ bulbs were soaked in the following concentrations of AgNPs: 25, 50, 100 and 150?mg?L^-1. Control bulbs were soaked in deionized water. The plants were forced in a greenhouse for two growing seasons. The tulips treated with 100?mg?L^?1 AgNPs flowered earlier, had longer cut-flower stem, larger petals, greater stem diameter and cut-flower fresh weight. Moreover, at this concentration of AgNPs, they showed increased leaf greenness index (SPAD), stomatal conductance, root fresh weight, root length and produced daughter bulbs of the greatest weight. The study outcomes indicate that AgNPs may be used as plant growth promoters in horticulture.     

Otolith elemental characteristics of whitefish (Coregonus lavaretus) from brackish waters of the Gulf of Bothnia,Baltic Sea

To investigate whether European whitefish (Coregonus lavaretus) groups in the brackish (≤7%) Gulf of Bothnia (Baltic Sea) express specific otolith elemental characteristics, concentrations of elements (Ca, Ba, Sr, Zn, Mn, Fe) in whole otoliths were studied by inductively coupled plasma‐optical emission spectrometry. Whitefish (N = 67) were sampled from six different latitudinal sites; four in the sea and two in rivers along the west coast of Finland, and from a fresh water lake for comparison. The concentration of several elements showed significant differences among sampling sites. Otolith Ba concentrations were higher at the northern sampling sites compared to the southern ones, thereby showing negative association to capture‐habitat salinity. In contrast, otolith Sr and Zn concentrations were lower at the northern sampling sites, thereby associating positively to capture‐habitat salinity. Otolith Mn concentrations did not associate with latitude or salinity but markedly varied between sampling sites. Elevated Fe concentrations occurred in fish otoliths from the two northernmost sites. Ca concentrations were at the same levels in otoliths from all whitefish samples. Otoliths of whitefish from the lake had very high Ba and low Sr concentrations compared to concentrations in otoliths from whitefish in the sea and rivers. Our results show that otolith elemental characteristics of whitefish vary considerably at various geographic scales showing high potential for population identification using elemental fingerprints. Analysis of otolith elemental characteristics may therefore provide an efficient tool for the identification of main areas of residence, spawning locations, and migration routes, thus contributing towards the sustainable management of whitefish stocks.     

Intestinal metabolism of weaned piglets fed a typical United States or European diet with or without supplementation of tributyrin and lactitol

The aim of the study was to investigate the effects of supplementation of a microencapsulated blend of tributyrin and lactitol (TL) to a standard European (EU) diet without antibiotic growth promoters on intestinal metabolism and mucosa development of weaned piglets and to compare it with a standard US diet containing animal proteins, zinc oxide, copper sulfate, and carbadox. Ninety piglets weaned at 21 d were divided into 3 dietary groups consisting of 5 replicates each: 1) US diet supplemented with 55 mg/kg of carbadox, and 2.5% each of plasma proteins and spray-dried blood cells in the first phase, 3,055 mg/kg of Zn in the first and second phases, and 180 mg/kg of Cu in the third phase; 2) EU diet based on vegetable proteins and no antibiotics; and 3) the same EU diet supplemented with 3,000 mg/kg of microencapsulated TL. The study was divided into 3 phases: 0 to 7, 8 to 21, and 22 to 35 d. On d 7, 21, and 35, animals were weighed, and feed consumption and efficiency were determined. On d 14 and 35, one pig per pen was killed, and the intestinal contents and mucosa from the proximal, middle, distal jejunum and the ileum were sampled. Intestinal wall sections were fixed for histological analysis, and intestinal content was used for VFA, ammonia, and polyamine analysis. Throughout the study (d 0 to 35), the US diet had greater ADG and ADFI than the EU diet (P < 0.05). The EU diet supplemented with TL tended to have 11% greater ADG (P = 0.17). Feeding the EU diet caused a reduction in proximal and middle jejunum villi length by 10% (P < 0.05) and an increase in crypt size in proximal jejunum (P < 0.05) compared with the US diet, probably due to an increased rate of cell loss and crypt cell production. The TL supplementation resulted in longer villi along the jejunum and less deep crypts in the proximal jejunum (+15.9 and -8.9%, respectively; P < 0.05) than the unsupplemented EU diet. The TL diet increased the concentrations of cadaverine and putrescine in the small intestine (P < 0.05) and seemed to increase cadaverine, histamine, putrescine, and spermine in the large intestine by 1.5- to 10-fold compared with the US or EU diet. In conclusion, although the US diet had a greater effect on growth performance and mucosal trophic status than the EU diets, the supplementation with slowly released TL seemed to be an effective tool to partially overcome the adverse effects of vegetable protein diets.     

Chokeberry polyphenols reduce prooxidative influence of oxidized fats in rat diets

The aim of the study was to estimate nutritional and physiological consequences of combined supplementation of rat casein diets with a strongly oxidized mixture of fats and extract of polyphenols from chokeberry. The experiment was conducted on 80 young Wistar rats divided into 8 groups. The animals were fed diets supplemented with 8% of fresh or oxidized fats and 0.4, 0.8 and 1.2% of polyphenol extract together. Addition of the oxidized fats caused significant reduction in animal growth without a decrease in nitrogen digestibility. On the other hand, the extract of polyphenols suppressed nitrogen utilization; animal growth was however similar to the control group. Analysis of the variance showed negative influence of the oxidized fats on cholesterol and triacyloglycerol concentration in the blood of the experimental animals. Increased activity of both aminotransferases in the blood of rats fed diets containing the oxidized fats indicated on liver malfunction. There was no effect of the experimental factors on the antioxidant status of the blood, but a dose-dependent reduced content of malonylodialdehyde in the blood of rats fed diets supplemented with the extract of polyphenols suggested that the extract had a preventive effect against prooxidative activity of the oxidized fats.     

Surgical treatment of hypospadias. Techniques and results in six dogs

Various types of hypospadias and methods of surgical treatment of this anomaly in 6 dogs was described in this article. Methods of surgical treatment were set up individually for each case and depended on the type of hypospadias and coexisting anomalies. Shortening of the penis was performed in two dogs, amputation in three cases and an urethral reconstruction in one patient. Castration was performed in three cases. Persistent frenulum of the prepuce was found in two dogs. Additionally three patients were submitted to the plastic surgery of the prepuce anomalies. In one dog, the loose and free falling part of the prepuce was suspended and attached to the abdominal integument according to the original own method. In all dogs, uncomplicated wound healing was observed. Surgical procedures resulted in improved cosmesis with reducing clinical signs of hypospadias and other coexistent anomalies.     

Protothecosis as a cause of chronic diarrhoea in a dog

The clinical form of the protothecosis in animals is most commonly observed in countries with a warm and moist climate, only a few reports describing cases of this infection in cooler areas of the word exist. In the case of large bowel infection in dogs, organisms colonise the lamina propria and submucosa causing severe necrotizing ulcerative or haemorrhagic enterocolitis. In this report the intestinal form of protothecosis in 1.5-year-old, male, mongrel dog with chronic hemorrhagic diarrhoea is described. History revealed that the dog spent some time in the countryside and afterwards diarrhoea with fresh blood appeared. The results of morphological and biochemical blood analysis were normal and stool examination did not reveal the presence of parasites. Treatment with anti-inflammatory doses of prednisone, metronidazole and enrofloxacin followed by sulphasalazine resulted in a short period of improvement, but was followed by deep deterioration of animal status. Because of the relapse diagnostic laparotomy was performed and tissue samples of the colon and jejunum were obtained for histopathology. On the basis of the clinical signs, exploratory laparotomy findings and histopathology the diagnosis of canine intestinal prototecosis was made and medical treatment was recommended.     

The effect of copper nanoparticles and copper (II) salt on redox reactions and epigenetic changes in a rat model

The aim of the study was to evaluate the effects of a diet containing different levels of Cu in two different chemical forms (carbonate and nanoparticles) on redox reactions and epigenetic changes in a rat model. For 4 weeks, five experimental groups (eight rats in each) were fed diets with two dosages of added Cu (standard—6.5 mg/kg or half of the standard dosage—3.25 mg/kg, and as a negative control no additional Cu in the mineral mixture) in two forms (standard—CuCO₃ and copper nanoparticles). Addition of Cu nanoparticles resulted in higher Cp (ceruloplasmin) activity and LOOH (lipid peroxides) and MDA (malondialdehyde) content, as well as decrease the CAT (catalase) activity and level of PC (protein carbonyl), 3‐NT (3‐nitrotyrosine), 8‐OHdG (8‐hydroxydeoxyguanosine), GSH + GSSG (total glutathione) and DNA methylation. Reducing the dose of copper resulted in a decrease in the level of LOOH and GSH + GSSG as well as CAT activity, but increased the level of PC and methylated DNA. Based on these evidence, we concluded that addition of copper nanoparticles in the diet reduces protein oxidation and nitration as well as DNA oxidation and methylation. Lowering the level of Cu in the diet increases the oxidation of proteins and DNA methylation.     

First identification of reciprocal translocations in Polish pigs

SUMMARY: Two different reciprocal translocations, rcp (8;14) (p21;q25) and rcp (7;13) (q13;q46), were described for the first time in Poland in the Polish Landrace and Duroc pigs. Observation of synaptonemal complexes showed the presence of a quadrivalent composed of translocational chromosomes and their homologues. Individuals carrying translocations were characterized by normal external appearance and correct semen parameters. Analysis of fertility made on the basis of litter size demonstrated that fertility decreased by 25% in the carrier of translocation rcp (8;14) and by 52% in the carrier of translocation rcp (7;13). ZUSAMMENFASSUNG: Zum erste mal festgestelte gegenseitige Translokationen bei Schweine in Polen Bein Polnischen Landras und Duroc Schweinen wurden zum erstenmal in Polen, zwei verschiedene reciproke Translokationen-rcp (8;14) (p21;q25) und rcp (7;13) (q13;q46)-festgestellt. Anhand der Beobachtungen des Synaptonemal Complex wurde die Anwesenheit eines Tetravalent nachgewiesen, welches aus Translokationschromosomen und seinen Homologen zusammengesetzt ist. Die Translokationstiere hatten normale Exterieur und Spermaparameter. Aufgrund der Fortpflanzungsergebmissen wurden verminderte Fruchtbarkeit bei Translokationstieren rcp (8;14)-um 25% und bei Translokationstieren rcp (7;13)-um 52% festgestellt. RéSUMé: Les translocations réciproques identifiées pour la prèmière fois chez les porcs levés en Pologne Chez les porcs des races Landrace et Duroc on a constaté, pour la première fois en Pologne deux translocations réciproques différentes-rcp (8;14) (p21;q25) ainsi que rcp (7;13) (q13;q46). L'observation des complexes synaptonémals a manifesté la présence du quadrivalent composé de chromosomes de translocation et de leurs homologes. Les animaux-les porteurs de la translocation ont se caractérisé par l'extérieur normal et par les paramètres réguliers du sperme. L'analyse de la fertilité expérimenté selon de la taille des portées a démontré la réduction de la fertilité pour le porteur de la translocation rcp (8;14) à 25% cependant pour les porteurs de la translocation rcp (7;13) à 52%.     

A simple and efficient CRISPR/Cas9 platform for induction of single and multiple,heritable mutations in barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.)

Background

Genome editing of monocot plants can be accomplished by using the components of the CRISPR/Cas9 (clustered regularly interspaced short palindromic repeat/CRISPR associated Cas9) technology specifically optimized for these types of plants. Here, we present the development of RNA-guided Cas9 system for simplex and multiplex genome editing in barley.

Results

We developed a set of customizable RNA-guided Cas9 binary vectors and sgRNA modules for simplex and multiplex editing in barley. To facilitate the design of RNA-guided Cas9 constructs, the pBract derived binary vectors were adapted to Gateway cloning and only one restriction enzyme was required for construction of the sgRNA. We designed a synthetic, codon optimized Cas9 gene containing the N terminal SV40 nuclear localization signal and the UBQ10 Arabidopsis 1st intron. Two different sgRNAs were constructed for simplex editing and one polycistronic tRNA-gRNA construct (PTG) for multiplex editing using an endogenous tRNA processing system. The RNA-guided Cas9 constructs were validated in transgenic barley plants produced by Agrobacterium-mediated transformation. The highest mutation rate was observed in simplex editing of the cytokinin oxidase/dehydrogenase HvCKX1 gene, where mutations at the hvckx1 locus were detected in 88% of the screened T₀ plants. We also proved the efficacy of the PTG construct in the multiplex editing of two CKX genes by obtaining 9 plants (21% of all edited plants) with mutations induced in both HvCKX1 and HvCKX3. Analysis of the T₁ lines revealed that mutations in the HvCKX1 gene were transmitted to the next generation of plants. Among 220 screened T₁ plants we identified 85 heterozygous and 28 homozygous mutants, most of them bearing frameshift mutations in the HvCKX1 gene. We also observed independent segregation of mutations and the Cas9-sgRNA T-DNA insert in several T₁ plants. Moreover, the knockout mutations of the Nud gene generated phenotype mutants with naked grains, and the phenotypic changes were identifiable in T₀ plants.

Conclusions

We demonstrated the effectiveness of an optimized RNA-guided Cas9 system that can be used for generating homozygous knockout mutants in the progeny of transgenic barely plants. This is also the first report of successful multiplex editing in barley using a tRNA processing system.