Multiply-primed rolling-circle amplification (MPRCA) of PCV2 genomes: Applications on detection, sequencing and virus isolation

Multiply-primed rolling-circle amplification (MPRCA) was used to amplify porcine circovirus type 2 (PCV2) genomes isolated from tissues of pigs with signs of post-weaning multisystemic wasting syndrome (PMWS). Two of the amplified PCV2 genomes were cloned in prokaryotic plasmids and sequenced. Both were nearly identical (1767 nt) except for one silent substitution in the region coding for the capsid protein (ORF2). In addition, they showed high nucleotide sequence similarity with PCV2 isolates from others countries (93–99%). To investigate whether the MPRCA amplified PCV2 genomes could be used to produce infectious virus, the cloned genomes were isolated from the plasmids, recircularized and used for transfection in PK-15 cells. This procedure led to the production of infectious virus to titres up to 10^5.55 TCID₅₀/mL. It was concluded that MPRCA is a useful tool to amplify PCV2 genomes aiming at sequencing and virus isolation strategies, where particularly useful is the fact that it allows straightforward construction of PCV2 infectious clones from amplified genomes. However, it was less sensitive than PCR for diagnostic purposes.     

Six Edible Wild Fruits as Potential Antioxidant Additives or Nutritional Supplements

Total antioxidant activity of six non-cultivated but traditionally collected fruits from the south of Europe was assessed by measuring their ability to reduce the hydroxyl radical ( OH^· ) \left( {{\hbox{O}}{{\hbox{H}}^\bullet }} \right) and hydrogen peroxide (H₂O₂), and their Trolox equivalent antioxidant capacity (TEAC). This antioxidant activity was compared with that shown by the synthetic antioxidants BHA (E-320), BHT (E-321) and propyl gallate (E-310). Total phenolics, ascorbic acid and the carotenoid content of the fruits were also analyzed. All fruits showed a high ability to scavenge the OH^· {\hbox{O}}{{\hbox{H}}^\bullet } radical, ranging from 60.61% to 81.04% inhibition for Rosa canina and Crataegus monogyna, respectively. The H₂O₂ scavenging capacity and the TEAC value varied widely, ranging between 3.63% and 87.26% inhibition of H₂O₂ and between 0.47 and 416.64 mM trolox g⁻¹ FW for Sorbus domestica and Rosa canina, respectively. The antioxidant activity of fruits was higher than that of the synthetic additives analyzed, except in the TEAC assay. The phenolic and carotenoid content of R. canina was much higher than that of the other fruits analyzed and its ascorbic acid concentration was also high, reflecting its higher efficacy towards ABTS ^·- {\hbox{ABT}}{{\hbox{S}}^{ \bullet - }} (TEAC assay) and H₂O₂ species_. In spite of these associations, the correlation coefficients between total antioxidant activity and the antioxidant compounds analyzed were not very significant; only phenolics and carotenoids showed a marginal correlation with the TEAC assay. The results support the possible use of R. canina as natural antioxidant to replace the synthetic additives, as well as their use in the production of functional foods with a high antioxidant activity.     

Effects of Sewage on the Chemical Composition of Piracicaba River,Brazil

Water samples were collected from 7 locations along major rivers of Piracicaba River basin for 22 months. The 4 upstream points represent non-polluted sites and the 3 downriver points represent polluted sites. Due to sewage input, concentrations of dissolved organic carbon (DOC), dissolved inorganic carbon (DIC), major conservative cations and anions increased significantly in the polluted sites. The major increases were observed for sodium, chloride and sulfate. Nitrate was an exception for this pattern, with similar concentrations between polluted and non-polluted sites. The probable cause was reduction of nitrate to ammonium in the polluted sites, where anoxic conditions prevail. Most of the variables had an inverse correlation with water discharge, especially in the polluted points. The sewage load was diluted by precipitation and surface waters.     

Induced plant defences in biological control of arthropod pests: a double‐edged sword

Biological control is an important ecosystem service delivered by natural enemies. Together with breeding for plant defence, it constitutes one of the most promising alternatives to pesticides for controlling herbivores in sustainable crop production. Especially induced plant defences may be promising targets in plant breeding for resistance against arthropod pests. Because they are activated upon herbivore damage, costs are only incurred when defence is needed. Moreover, they can be more specific than constitutive defences. Nevertheless, inducible defence traits that are harming plant pest organisms may interfere with biological control agents, such as predators and parasitoids. Despite the vast fundamental knowledge on plant defence mechanisms and their effects on natural enemies, our understanding of the feasibility of combining biological control with induced plant defence in practice is relatively poor. In this review, we focus on arthropod pest control and present the most important features of biological control with natural enemies and of induced plant defence. Furthermore, we show potential synergies and conflicts among them and, finally, identify gaps and list opportunities for their combined use in crop protection. We suggest that breeders should focus on inducible resistance traits that are compatible with the natural enemies of arthropod pests, specifically traits that help communities of natural enemies to build up. © 2017 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

The risk of salmonellae shedding by dogs fed Salmonella-contaminated commercial raw food diets

Twenty-eight research dogs were enrolled to determine the prevalence of salmonellae shedding after consumption of 1 Salmonella-contaminated commercial raw food diet meal. Sixteen dogs were exposed to Salmonella-contaminated commercial raw food diets and 12 to Salmonella-free commercial raw food diets. Seven of the exposed dogs shed salmonellae 1-7 days after consumption of Salmonella-contaminated raw food diets. None of the dogs fed Salmonella-free diets shed salmonellae. No clinical signs were observed in either group. Five of the 7 dogs shed the same serotypes as those recovered from food samples used for feeding. Results showed the same serotypes and antimicrobial resistance pattern in 2 of the 7 shedders. Dogs fed Salmonella-contaminated raw food diets can shed salmonellae and may, therefore, be a source of environmental contamination potentially leading to human or animal illness.     

Current knowledge about the processes of luteolysis and maternal recognition of pregnancy in camelids

Camelids have many unique reproductive features that considerably differ from those of other domestic species. Females are induced ovulators with subsequent development of a corpus luteum (CL) with a short lifespan. Plasma progesterone concentration starts to increase on day 4, peaks on day 8–9 and, in non-pregnant animals, basal concentration is reached around day 10–11 post-induction of ovulation. Luteolytic pulses of prostaglandin F_2α (PGF_2α) are firstly detected on day 7 or 8 (approximately on day 5–6 after ovulation), with maximal luteolytic peaks observed between days 9 and 11 post-mating, in coincidence with a high endometrial expression of cyclooxygenase 2, a limiting enzyme in prostaglandins synthesis. Unlike other species, oxytocin seems not to be involved in the luteolytic process in these species. The CL is the main source of progesterone secretion, and its function is required to support pregnancy. Despite constant research efforts, aspects of reproduction and maternal recognition of pregnancy in camelids remain not fully understood. A transient decrease and subsequent recovery in plasma progesterone concentration are observed after day 9 post-mating in pregnant animals in association with a pulsatile release of PGF_2α and a transitory decrease in CL vascularization. Thus, embryo recognition should occur between days 8 and 12 post-mating. In camels, conceptus tissues exhibit aromatizing activity with the capacity to synthesize large amounts of oestradiol. Similarly, llama blastocysts secrete oestradiol-17β during the preimplantation stage, with a higher production during the elongation period. An increase in the endometrial expression of oestrogen receptor α is also observed on day 12 post-mating. All these evidences suggest that oestrogen could be the signal released by the embryo at the time of its recognition in camelids. Besides, nearly 98% of pregnancies are carried out in the left horn. A decrease in the endometrial expression of mucin 1 and 16 genes has been reported, suggesting that these changes are crucial for successful embryo implantation; however, no differences have been observed between horns. Thus, maternal recognition of pregnancy in camelids is a particularly complex process that must occur in a concise time to allow the rescue of the CL and embryo survival.     

Effects of training on potassium homeostasis during exercise and skeletal muscle Na+,K(+)-ATPase concentration in young adult and middle-aged Dutch Warmblood horses

OBJECTIVE: To investigate the effects of moderate short-term training on K+ regulation in plasma and erythrocytes during exercise and on skeletal muscle Na+,K(+)-ATPase concentration in young adult and middle-aged horses. ANIMALS: Four 4- to 6-year-old and four 10- to 16-year-old Dutch Warmblood horses. PROCEDURE: The horses underwent a 6-minute exercise trial before and after 12 days of training. Skeletal muscle Na+,K(+)-ATPase concentration was analyzed in gluteus medius and semitendinosus muscle specimens before and after the 12-day training period. Blood samples were collected before and immediately after the trials and at 3, 5, 7, and 10 minutes after cessation of exercise for assessment of several hematologic variables and analysis of plasma and whole-blood K+ concentrations. RESULTS: After training, Na+,K(+)-ATPase concentration in the gluteus medius, but not semitendinosus, muscle of middle-aged horses increased (32%), compared with pretraining values; this did not affect the degree of hyperkalemia that developed during exercise. The development of hyperkalemia during exercise in young adult horses was blunted (albeit not significantly) without any change in the concentration of Na+,K(+)-ATPase in either of the muscles. After training, the erythrocyte K+ concentration increased (7% to 10%) significantly in both groups of horses but did not change during the exercise trials. CONCLUSIONS AND CLINICAL RELEVANCE: In horses, the activation of skeletal muscle Na+,K(+)-ATPase during exercise is likely to decrease with age. Training appears to result in an increase in Na+,K(+)-ATPase activity in skeletal muscle with subsequent upregulation of Na+,K(+)-ATPase concentration if the existing Na+,K(+)-ATPase capacity cannot meet requirements.     

COMPUTED TOMOGRAPHIC FINDINGS IN 57 CATS WITH PRIMARY PULMONARY NEOPLASIA

Primary pulmonary neoplasia is relatively uncommon in cats and generally has a poor prognosis. In this multicenter, retrospective study of 57 cats with pulmonary neoplasia, the most frequent presenting signs were anorexia/inappetence (39%) and cough (37%). The pulmonary tumors were considered to be incidental findings in 9% cats. In computed tomographic (CT) images, primary pulmonary tumors appeared as a pulmonary mass in 55 (96%) cats and as a disseminated pulmonary lesion without a defined mass in two (4%) cats. Most pulmonary tumors were in the caudal lobes, with 28 (49%) in the right caudal lobe and 17 (30%) in the left caudal lobe. CT features associated with pulmonary tumors included mass in contact with visceral pleura (96%), irregular margins (83%), well‐defined borders (79%), bronchial compression (74%), gas‐containing cavities (63%), foci of mineral attenuation (56%), and bronchial invasion (19%). The mean (range) maximal dimension of the pulmonary masses was 3.5 cm (1.1–11.5 cm). Additional foci of pulmonary disease compatible with metastasis were observed in 53% cats. Pleural fluid was evident in 30% cats and pulmonary thrombosis in 12% cats. The histologic diagnoses were 47 (82%) adenocarcinomas, six (11%) tumors of bronchial origin, three (5%) adenosquamous cell carcinomas, and one (2%) squamous cell carcinoma. In this series, adenocarcinoma was the predominant tumor type, but shared many features with less common tumor types. No associations were identified between tumor type and CT features. Prevalence of suspected intrapulmonary metastasis was higher than in previous radiographic studies of cats with lung tumors.     

Management of downy mildew of sweet basil (Ocimum basilicum) caused by Peronospora belbahrii by means of resistance inducers, fungicides, biocontrol agents and natural products

To control downy mildew of sweet basil (Ocimum basilicum L.), incited by Peronospora belbahrii Thines, a number of compounds were tested in 2011 and 2012 under glasshouse conditions. These included copper-based fungicides, biocontrol agents, and compounds previously reported to induce resistance in plants to pathogens. Results were compared with those provided by fungicides registered for downy mildew control in Italy. The tested compounds were used alone or applied in rotation in spray programs. In all trials, the greatest reduction in disease incidence and severity was found with treatments that included metalaxyl-M?+?copper hydroxide, a mineral fertilizer ‘Alexin’, mandipropanid, and azoxystrobin. The glucohumates activator complex and acibenzolar-S-methyl also provided significant disease control (P?<?0.05). The mineral fertilizer Alexin, the glucohumates activator complex and acibenzolar-S significantly reduced disease incidence and severity 20 days after the last treatment compared with the untreated control (P?<?0.05). Among the copper-based products, the greatest reductions in disease incidence and severity were provided by copper hydroxide with terpenic alcohols and copper oxychloride?+?copper hydroxide. The mineral fertilizer Kendal and prohexadione-Ca, as well as mustard oil, partially reduced disease incidence and severity compared with the untreated control (P?<?0.05), whereas the biocontrol agent Bacillus subtilis QST 713 and thyme oil extract were not effective in two out of three trials. When different combinations of various products used in rotation were tested, effective control was found either using a rotation of fungicides with compounds that can induce resistance, as well as by using rotation with different resistance-inducing compounds on their own.