Characteristics of nucleotides and amino acids in cDNA sequence of xyloglucan endotransglycosylase cloned from <Emphasis Type="Italic">Anthocephalus chinensis</Emphasis>

Xyloglucan endotransglycosylase is an identified relaxation factor with functions of easing and extending of plant cell walls.Its activities are directly related to plant growth and elongation of organisms.Anthocephalus chinensis is a tall and fast growing evergreen tree species.We cloned the full cDNA sequence of AcEXT genes which is abundantly expressed in the cambium of A.chinensis.The sequence analysis of nucleotides and amino acids revealed the presence of a 1396 bp full cDNA sequence,including a 960 bp complete open reading frame(ORF) encoding a 320 amino acid protein.The deduced amino acid sequence of AcXET was homologous to the other known XET proteins and contained the conserved EIDFE catalytic site which was specific to all the XETs.Our data should serve as a foundation for further insight into AcXET gene molecular mechanisms during wood formation and cell wall engineering of woody plants.     

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.     

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.     

A simple and efficient method for isolating small RNAs from different plant species

Background  

Small RNAs emerged over the last decade as key regulators in diverse biological processes in eukaryotic organisms. To identify and study small RNAs, good and efficient protocols are necessary to isolate them, which sometimes may be challenging due to the composition of specific tissues of certain plant species. Here we describe a simple and efficient method to isolate small RNAs from different plant species.     

Spatial structure of shell polychromatism in <Emphasis Type="Italic">Cepaea hortensis</Emphasis> in relation to a gradient of landscape fragmentation in Western France

Because of their highly polymorphic shell patterns, Cepaea land snails have been the subject of numerous studies in ecological genetics. Here, we investigated the spatial structure of polychromatism in Cepaea hortensis in agricultural landscapes with zones from low to high hedgerow densities. Our main purpose was to search for a relationship between landscape composition and spatial structuring of chromatism. Despite significant spatial heterogeneity in the three landscapes sampled, only the high hedgerow density landscape showed a significant spatial structuring of shell polymorphism. In order to understand this result, an investigation of daily movement patterns in relation to habitat form was carried out on a mark-release experiment under semi-artificial conditions. This experiment revealed a strong influence of a linear corridor on snail dispersal. In the field, spatial heterogeneity of shell polymorphism, related to the effects of genetic drift, was shaped by restricted dispersal in narrow corridors. In the more enclosed one, i.e. where hedgerow density was the highest, the significant spatial structure we detected involved a balance between local genetic drift and environmentally mediated gene flow. This isolation-by-distance pattern resulted from direct gene exchange through fields between neighbouring populations. When applying landscape distances based on hedgerow length, no significant spatial correlation with polychromatism was found. In the more fragmented sites, random genetic drift seemed to be the prevailing force and, at the scale of the whole sampled area, selective pressures potentially interfere with these genetic drift-dispersal events.     

Pharmacokinetic disposition of triclabendazole in cattle and sheep; discrimination of the order and the rate of the absorption process of its active metabolite triclabendazole sulfoxide

A comparative pharmacokinetic study was conducted to determine the order and the rate of absorption of triclabendazole (TCBZ) in cattle and sheep. A commercial suspension of TCBZ (Biofasiolex, Biogénesis S.A., Argentina) was administered at a dose rate of 10 mg/kg by the oral route to six Holstein female calves and six Corriedale female sheep. The plasma concentration profiles of the metabolites triclabendazole sulfoxide (TCBZ-SO) and triclabendazole sulfone (TCBZ-SO₂) were analysed by means of the non-compartmental method. The order of the absorption process of the active metabolite, TCBZ-SO, was determined by construction of curves of cumulative absorbed fraction of the drug by means of the Wagner-Nelson method. The appearance of TCBZ-SO in plasma of cattle and sheep resembles the entry of a constant quantity of drug into the organism per unit time. This is explained by the reservoir effect of the rumen, which acts as a biological slow-release system for TCBZ-SO and its precursor TCBZ to the posterior digestive tract where they are absorbed. The plasma concentration profiles of TCBZ-SO in both species were well described by a one-compartment open model with zero-order process of absorption and first-order process of elimination. The values of AUC_0-∞ and C _max of TCBZ-SO did not differ between species, while other kinetic parameters except for λ _z had higher values in calves than in sheep. In the case of TCBZ-SO₂, t _max was the only parameter that did not differ between species, while other kinetic parameters except for λ _z had higher values in calves than in sheep.     

Generation and characterisation of Tn5-tagged <Emphasis Type="Italic">Xanthomonas oryzae</Emphasis> pv. <Emphasis Type="Italic">oryzae</Emphasis> mutants that overcome <Emphasis Type="Italic">Xa23</Emphasis>-mediated resistance to bacterial blight of rice

Xanthomonas oryzae pv. oryzae causes bacterial blight of rice. Xa23, a bacterial blight resistance gene identified originally in wild rice, Oryza rufipogon, is dominant and resistant to all X. oryzae pv. oryzae field isolates tested. The corresponding avirulence gene avrXa23 is unknown. Here we report the generation of a random insertion mutant library of X. oryzae pv. oryzae strain PXO99 using a Tn5-derived transposon tagging system, and identification of mutant strains that are virulent on CBB23, a near-isogenic rice line containing Xa23. A total of 24,192 Tn5 inserted clones was screened on CBB23 by leaf-cutting inoculation and at least eight of them caused lesions on CBB23 comparable to those on JG30, the susceptible recurrent parent of CBB23. Polymerase chain reaction and Southern blot analysis showed that all the eight mutants, designated as P99M1, P99M2, P99M3, P99M4, P99M5, P99M6, P99M7 and P99M8, have a single Tn5-insertion in their genomes. The flanking DNA sequences of the Tn5-insertion sites were isolated by PCR-walking and sequenced. Bioinformatic analysis of the flanking sequences, by aligning them with the whole genome sequences of X. oryzae pv. oryzae strains PXO99, KACC10331 and MAFF311018 through NCBI, revealed that the Tn5-insertions disrupted genes that encode TAL effector AvrBs3/PthA, ISXo1 transposase, Type II secretion system protein-like protein or outer membrane protein, glycogen synthase, cytochrome C5 and conserved hypothetical protein. Further identification of these mutants will facilitate the molecular cloning of avirulence gene avrXa23. The authors C.-L. Wang, A.-B. Xu contributed equally to this work; Y. Gao and Y.-L. Fan contributed equally to this work.     

Doxycycline treatment for <Emphasis Type="Italic">Dirofilaria immitis</Emphasis> in dogs: impact on <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> and <Emphasis Type="Italic">Enterococcus</Emphasis> antimicrobial resistance

Doxycycline is an antibiotic that, in addition to the classic antibacterial use, is also prescribed to fight parasitic diseases, like heartworm disease in dogs. Despite the concern that the overuse of this antibiotic may decrease susceptibility of clinically important bacteria, the consequences of the prolonged doxycycline therapy in heartworm-infected dogs have never been studied before. We have analyzed the impact of this therapy on Staphylococcus aureus and Enterococcus antimicrobial resistance. In this study, 17 heartworm-infected dogs (10 that had completed the doxycycline treatment and 7 dogs that had not yet begun) were included. Twenty-four isolates of Staphylococcus aureus were obtained from two locations of each dog. After treatment, 73.3% of isolates were resistant to at least one antibiotic but only 22.2% of isolates before treatment. Most of doxycycline resistant isolates were obtained from dogs that have received treatment. Erythromycin resistance or intermediate susceptibility was detected in 45.6% of isolates, most of them from dogs after treatment. For Enterococci, 48 isolates were obtained from fecal samples (25 before treatment and 23 after treatment). Before treatment, 32% of isolates were resistant at least to one antibiotic while after, this data increase up to 65%. Comparing isolates before and after treatment, a clear increase in resistance to doxycycline (12% against 21.74%) and erythromycin (20% against 39.13%) was observed. Although the present work is a preliminary research, the results encourages the development of further studies to determinate the effect of prolonged doxycycline therapy on antimicrobial resistance.     

The occurrence of postharvest gray-mold rot of sweet persimmon caused by <Emphasis Type="Italic">Botrytis cinerea</Emphasis> in Korea

In this study, we identified the causative agent of postharvest gray-mold rot in sweet persimmon fruit that were collected from Gangneung, Gangwon Province, Korea in October 2016. Symptoms included extensive growth of mycelia on post harvested fruit. The fungus was isolated from infected fruit and cultured on potato dextrose agar (PDA). For identification of the fungus, we examined morphology characteristics and rDNA sequencing analysis of the fungus and confirmed its pathogenicity according to Koch’s postulates. The results of morphological examinations, pathogenicity tests, 5.8S rDNA sequences of the internal transcribed spacer regions (ITS1 and ITS4) and the five nuclear protein-coding genes G3PDH, HSP60, RPB2, MS547 and TUB revealed that the causal agent of postharvest gray-mold rot on sweet persimmon fruit in Korea was Botrytis cinerea.