Expression of verocytotoxic Escherichia coli antigens in tobacco seeds and evaluation of gut immunity after oral administration in mouse model

Verocytotoxic Escherichia (E.) coli strains are responsible for swine oedema disease, which is an enterotoxaemia that causes economic losses in the pig industry. The production of a vaccine for oral administration in transgenic seeds could be an efficient system to stimulate local immunity. This study was conducted to transform tobacco plants for the seed-specific expression of antigenic proteins from a porcine verocytotoxic E. coli strain. Parameters related to an immunological response and possible adverse effects on the oral administration of obtained tobacco seeds were evaluated in a mouse model. Tobacco was transformed via Agrobacteium tumefaciens with chimeric constructs containing structural parts of the major subunit FedA of the F18 adhesive fimbriae and VT2e B-subunit genes under control of a seed specific GLOB promoter. We showed that the foreign Vt2e-B and F18 genes were stably accumulated in storage tissue by the immunostaining method. In addition, Balb-C mice receiving transgenic tobacco seeds via the oral route showed a significant increase in IgA-positive plasma cell presence in tunica propria when compared to the control group with no observed adverse effects. Our findings encourage future studies focusing on swine for evaluation of the protective effects of transformed tobacco seeds against E. coli infection.     

Anatomic distribution and electrophysiologic properties of accessory atrioventricular pathways in dogs

OBJECTIVE: To evaluate the anatomic distribution and electrophysiologic properties of accessory pathways (APs) in dogs. DESIGN: Case series. ANIMALS: 10 dogs with tachyarrhythmias associated with an AP. PROCEDURES: Each dog underwent electrophysiologic testing to determine the inducibility of documented and undocumented arrhythmias and to identify location, conduction properties, and antegrade and retrograde effective refractory periods of the APs. Radiofrequency catheter ablation was then performed. RESULTS: 15 APs were identified; 7 dogs each had a single AP, and 3 had multiple APs. Fourteen of the 15 APs were right-sided (6 right free wall, 4 posteroseptal, 3 midseptal, and 1 anteroseptal), and 1 was left-sided (left free wall). All APs conducted in an all-or-none fashion. Unidirectional retrograde conduction was observed in 11 APs, and bidirectional conduction was observed in 4. All documented tachyarrhythmias could be induced during electrophysiologic testing; atrial fibrillation was also inducible in 2 dogs. Mean +/- SD cycle duration of orthodromic atrioventricular reciprocating tachycardia was 215.80 +/- 44.87 milliseconds. Mean shortest R-R interval during atrial fibrillation was 247.33 +/- 83.17 milliseconds. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that in dogs, most APs are right-sided, had unidirectional retrograde conduction, and are associated with various arrhythmias, including orthodromic atrioventricular reciprocating tachycardia and atrial fibrillation without evidence of pre-excitation.     

Mesenteric vascular thrombosis associated with disseminated abdominal visceral hemangiosarcoma in a cat

An adult castrated male cat was evaluated because of a 4 day history of lethargy and partial anorexia. Physical examination revealed abdominal pain with a palpable fluid wave. Cytologic and biochemical analyses of peritoneal effusion were suggestive of septic peritonitis. On surgical exploration of the abdomen, the mesenteric vessels had no palpable pulses and they contained gross thromboses. The intestines were white with no visible peristalsis. Necropsy findings included disseminated, poorly differentiated hemangiosarcoma throughout the abdomen. Mesenteric arterioles contained fibrin thrombi. To the author's knowledge, no previous reports exist of complete mesenteric vascular thrombosis associated with disseminated abdominal visceral hemangiosarcoma in a cat.     

Soil organic matter content and chemical composition under two rotation management systems in a Mediterranean climate

The crop rotation system in organic farming is a determinant factor to accumulate and preserve soil organic matter (SOM), and in depth knowledge on its effects is still lacking. Tillage intensity in particular is crucial to maintain soil aggregates and protect SOM from degradation. The evolution of SOM was tested in two adjacent fields under two different rotation cropping systems (low-intensity tillage and high-intensity tillage), and the effect of a further cultivation of legume in both fields was evaluated using ¹³carbon (C)-nuclear magnetic resonance (NMR) and elemental analysis of samples isolated through combined aggregate size and density fractionation. The two adjacent fields had been managed using the following organic farming methods for 13 seasons since 1998: i) alfalfa-based, with nitrogen (N) enrichment and low-frequency tillage with alfalfa (Medicago sativa) (9 seasons), winter wheat (Triticum durum) (3 seasons), and broad bean (Vicia faba) (1 season) and ii) cereal-based, with N depletion and annual tillage with barley (Hordeum vulgare) (7 seasons), sunflower (Helianthus annuus) (2 seasons), broad bean (Vicia faba) (3 seasons), and bare fallow (1 season). Soil sampling was carried out at the end of the 13-year rotation (T0, November 2011) and after winter wheat and chickpea cultivation in both fields over two subsequent years (T1, July 2013). Bulk organic C was significantly higher in the alfalfa-based system than in the cereal-based system at both T0 and T1, with SOM occluded in soil aggregates and associated with mineral particles. In terms of the macroaggregates heavy fraction at T0, the alfalfa-based field contained twice the organic C of that in the cereal-based field, as well as three times the organic C in the occluded particulate organic matter (POM). The occluded POM (oPOM) had a lower aryl/O-alkyl C ratio in the alfalfa-based system than in the cereal-based system, suggesting that oPOM undergoes a lower degree of decomposition during low-intensity management. The aryl/O-alkyl C ratios of the macro-and microaggregate oPOM decreased from T0 to T1 in the cereal-based system, suggesting increased protection of these fractions by soil aggregates. Thus, including legumes in crop rotation appears to positively affect the accumulation of SOM associated with mineral particles and within soil aggregates.     

Absorption of carbohydrate-derived nutrients in sows as influenced by types and contents of dietary fiber

The current investigation was undertaken to study the absorption and plasma concentration of carbohydrate-derived nutrients [glucose, short-chain fatty acids (SCFA), and lactate] and the apparent insulin production in sows fed diets containing contrasting types and contents of dietary fiber. Six sows were fed 3 experimental diets, low fiber (LF; 177 g of dietary fiber and 44 g of soluble fiber/kg of DM), high soluble fiber (HF-S; 429 g of dietary fiber and 111 g of soluble fiber/kg of DM), and high insoluble fiber (HF-I; 455 g of dietary fiber and 74 g of soluble fiber/kg of DM), in a repeated crossover design. Variations in dietary concentration and solubility of dietary fiber were obtained by substituting starch-rich wheat and barley in the LF diet with dietary fiber-rich co-products (sugar beet pulp, potato pulp, pectin residue, brewers spent grain, pea hulls, and seed residue, which have distinct physicochemical properties). The main carbohydrate component of the LF diet was starch and nonstarch polysaccharides (cellulose and noncellulosic polysaccharides) for the 2 high dietary fiber diets. Consumption of the LF diet resulted in increased and rapid glucose absorption at 0 to 4 h postfeeding. With the HF-I diet, the glucose absorption pattern was similar but at a decreased rate, whereas it was decreased and delayed with the HF-S diet (diet, P < 0.001; time, P < 0.001). These differences were also reflected in the insulin response. The quantitative absorption of SCFA at 0 to 10 h postfeeding was greater when feeding the HF-S diet compared with the LF diet (P < 0.001) and intermediate when feeding the HF-I diet (P < 0.001). The study showed that feeding the high dietary fiber diets resulted in a increased and more uniform uptake of SCFA than when feeding the LF control. Moreover, the HF-S diet reduced diurnal variation in glucose and insulin concentrations.     

Analysis of Dynamics in Cd, Fe, and Pb in Particulate Matter by using the Fisher–Shannon Method

The Fisher–Shannon information plane (FS), defined by the Fisher information measure and the Shannon entropy power, is proposed to investigate the complex dynamics of the concentration of three heavy metals (Cd, Fe, and Pb) in four different fractions of particulate matter data, recorded in Tito, a small industrial area of southern Italy. In the FS plane, the three metals are aggregated in three different clusters, characterized by different degrees of order. This result is related to different sources of the three metals.     

Resistance to the cabbage root fly, <Emphasis Type="Italic">Delia radicum</Emphasis> (Diptera,Anthomyiidae), of turnip varieties (<Emphasis Type="Italic">Brassica rapa</Emphasis> subsp. <Emphasis Type="Italic">rapa</Emphasis>)

The cabbage root fly Delia radicum L. (Diptera: Anthomyiidae) is one of the major pests of many Brassica crops in the temperate areas of Europe and North America. At present, turnip (B. rapa ssp. rapa L.) varieties resistant to the pest does not exist. With the aim to fill this gap, a no-choice tolerance test of 56 accessions among turnips, turnip tops and turnip greens was performed under controlled conditions by introducing D. radicum eggs. Plant survival, leaf and root conditions, pupae number and weight significantly varied among plant accessions. Ten putatively resistant and ten susceptible accessions (control group) were selected from this first screening, transplanted in the field and exposed to natural infestation to detect antibiosis and antixenosis mechanisms. Both in the laboratory and in the field, pupae number significantly varied within accessions and between resistant and susceptible group, although pupal weight did not, indicating the absence of antibiosis effect on this early stage. In the field, the number of galleries was significantly lower in the resistant group in comparison with the control. Resistant accessions had smaller size, and a smaller, white and mostly buried root. Within the resistant and susceptible accessions, larger plants harboured more pupae, however purple roots were those most preferred, and the hosted pupae weighed most. Three accessions from the resistant group (MBGBR0178, MBGBR0570 and MBGBR0371) stand out for resistance to D. radicum possibly through antixenosis mechanisms.     

Characterization of stable humic–enzyme complexes of different soil ecosystems through analytical isoelectric focussing technique (IEF)

Conventional parameters (total organic carbon, total nitrogen, humic acids and fulvic acids) frequently associated with soil quality were measured and compared with the amount and activity of β-glucosidases associated with the humic–enzyme complex. Soil quality may be defined as the maintenance of an “enzymatically active humic carbon pool” which may express soil resilience and ecological functions. The combination of ultrafiltration (UF) and isoelectric focussing (IEF) permitted the extraction and purification of the humic-bound β-glucosidases. Even though humic-β-glucosidases represent only a small part of soil humic matter either in terms of total soil β-glucosidase activity (3–21%) or as a proportion of the humic-C content (4–8%), they proved to be discriminating parameters (significative differences at P < 0.05) for defining soil biochemical quality. Specifically, on the basis of the amount of β-glucosidase activity in the humic fraction >10⁴ Da (IEF peaks in the pH range 3.5–4.5), the soils were distributed in a decreasing order of quality: undisturbed meadow > tree plantation > 60-year vineyard > agricultural soil > 1-year vineyard.