Molecular Identification of a New Member of the Clover Proliferation Phytoplasma Group (16SrVI) Associated with Centaurea Solstitialis Virescence in Italy

In the United States, yellow starthistle (Centaurea solstitialis) is an annual invasive weed with Mediterranean origins. Malformed plants displaying witches' broom, fasciations, abortion of buds and flower virescence symptoms were observed in central Italy. Attempts to transmit the causal agent from the natural yellow starthistle host to periwinkle by grafting, resulted in typical symptoms of a phytoplasma, i.e. yellowing and shortening of internodes. The detection of phytoplasmas was obtained from both symptomatic yellow starthistle and periwinkle by the specific amplification of their 16S-23S rRNA genes. PCR amplification of extracted DNA from symptomatic plant samples gave a product of expected size. Asymptomatic plants did not give positive results. An amplicon obtained by direct PCR with universal primers P1/P7 was cloned and sequenced. The homology search using CLUSTALW program showed more than 99% similarity with Illinois elm yellows (ILEY) phytoplasma from Illinois (United States) and 97% with Brinjal little leaf (BLL) phytoplasma from India. Digestion of the nested-PCR products with restriction enzymes led to restriction fragment length polymorphism patterns referable to those described for phytoplasmas belonging to the clover proliferation (16S-VI) group. Since this is a previously undescribed disease, the name Centaurea solstitialis virescence has been tentatively assigned to it. This is a new phytoplasma with closest relationships to ILEY and BLL, but distinguishable from them on the basis of 16S rDNA homology, the different associated plant hosts and their geographical origin.     

A new standard area diagram set for assessment of severity of soybean rust improves accuracy of estimates and optimizes resource use

Soybean rust (SBR), caused by Phakopsora pachyrhizi, is the most important yield-damaging fungal disease of soybean due to severe reduction in healthy leaf area and acceleration of leaf fall. In experimental research, SBR severity is estimated visually aided/trained by a standard area diagram (SAD) developed and validated during the mid-2000s (Old SAD). In this study, we propose a new SAD set for SBR with six true-colour diagrams following linear increments (c. 15% increments) amended with four additional diagrams at low (<10%) severities, totalling 10 diagrams (0.2%, 1%, 3%, 5%, 10%, 25%, 40%, 55%, 70%, and 84%). For evaluation, 37 raters were split into two groups. Each assessed severity in a 50-image sample (0.25%–84%), first unaided and then using either the Old SAD or the New SAD. Accuracy, precision, and reliability of estimates improved significantly relative to unaided estimates only when aided by the New SAD (accuracy >0.95). Low precision (<0.78) and a trend of underestimation with an increase in severity were the main issues with the Old SAD, which did not differ from unaided estimates. Simulation to evaluate the impact of the errors by different methods on hypothesis tests, showed that the new SAD was more powerful for detecting the smallest difference in mean control (e.g., 70% vs. 65% disease reduction) than the Old SAD; the latter required a 2-fold increase in sample size to achieve the same power. There is a need to improve some SADs, taking advantage of new knowledge and technology to increase accuracy of the estimates, and to optimize both resource use efficiency and management decisions.     

Enzyme Bioprospection of Marine-Derived Actinobacteria from the Chilean Coast and New Insight in the Mechanism of Keratin Degradation in Streptomyces sp. G11C

Marine actinobacteria are viewed as a promising source of enzymes with potential technological applications. They contribute to the turnover of complex biopolymers, such as pectin, lignocellulose, chitin, and keratin, being able to secrete a wide variety of extracellular enzymes. Among these, keratinases are a valuable alternative for recycling keratin-rich waste, which is generated in large quantities by the poultry industry. In this work, we explored the biocatalytic potential of 75 marine-derived actinobacterial strains, focusing mainly on the search for keratinases. A major part of the strains secreted industrially important enzymes, such as proteases, lipases, cellulases, amylases, and keratinases. Among these, we identified two streptomycete strains that presented great potential for recycling keratin wastes—Streptomyces sp. CHA1 and Streptomyces sp. G11C. Substrate concentration, incubation temperature, and, to a lesser extent, inoculum size were found to be important parameters that influenced the production of keratinolytic enzymes in both strains. In addition, proteomic analysis of culture broths from Streptomyces sp. G11C on turkey feathers showed a high abundance and diversity of peptidases, belonging mainly to the serine and metallo-superfamilies. Two proteases from families S08 and M06 were highly expressed. These results contributed to elucidate the mechanism of keratin degradation mediated by streptomycetes.     

Detection of bacterial contamination and DNA quantification in stored blood units in 2 veterinary hospital blood banks

Blood transfusions in veterinary medicine have become increasingly more common and are now an integral part of lifesaving and advanced treatment in small and large animals. Important risks associated with transfusion of blood products include the transmission of various infectious diseases. Several guidelines suggest what infectious agents to screen for in canine and feline transfusion medicine. However, while the risk of bacterial contamination of blood products during storage and administration has not been documented in veterinary medicine, it has emerged as a cause of morbidity and mortality in human transfusion medicine. Clinical experience shows that the majority of blood component bacterial contaminations are caused by only a few species. Unlike other types of bacteria, psychrotolerant species like Pseudomonas spp. and Serratia spp. can proliferate during the storage of blood units at 4°C from a very low titer at the time of blood collection to a clinically significant level (> 10⁵ CFU/mL) causing clinical sepsis resulting from red blood cell concentrate transfusions in human medicine. The purpose of this report was to describe the detection and quantification procedures applied in 4 cases of bacterial contamination of canine and feline blood units, which suggest the need for further investigations to optimize patients’ safety in veterinary transfusion medicine.     

Tolerance of benzalkonium chloride, formalin, malachite green, and potassium permanganate in goldfish and zebrafish

OBJECTIVE: To determine tolerance of goldfish and zebrafish to benzalkonium chloride, formalin, malachite green, and potassium permanganate. DESIGN: Tolerance study. ANIMALS: Adult goldfish (Carassius auratus) and zebrafish (Danio rerio). PROCEDURES: Groups of fish (n = 10/group) were exposed to each disinfectant at the therapeutic dosage; at 0.25, 0.5, 3, and 5 times the concentration used for the therapeutic dosage; and at the concentration used for the therapeutic dosage but for 3 or 5 times the recommended exposure time. RESULTS: In both species, exposure to malachite green at the therapeutic dosage resulted in toxic effects, including death. Exposure to formalin at the therapeutic dosage resulted in toxic effects in goldfish, but not zebrafish, and exposure to potassium permanganate resulted in toxic effects in zebrafish, but not goldfish. On the basis of the ratio of therapeutic dosage to median lethal dosage, in goldfish, formalin was more toxic than benzalkonium chloride, which was more toxic than malachite green, which was more toxic than potassium permanganate. In zebrafish, potassium permanganate was more toxic than formalin and benzalkonium chloride, which were approximately equally toxic and more toxic than malachite green. Extending treatment time increased the toxicity of potassium permanganate in zebrafish and the toxicity of formalin and malachite green in goldfish, but did not alter the toxicity of the other disinfectants. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that there was no consistency between zebrafish and goldfish in their tolerance to disinfectants, and that therapeutic dosages reported in the literature for these disinfectants were not always safe.     

Mechanical injury and fungal infection induce acquired resistance in Norway spruce

Norway spruce trees (Picea abies (L.) Karst.) pretreated by wounding and fungal infection showed highly enhanced resistance to a subsequent challenge inoculation with the pathogenic bluestain fungus Ceratocystis polonica (Siem.) C. Moreau. This is the first time the effectiveness of the constitutive and inducible defenses has been shown to depend on prior wounding and infection in conifers, although such acquired resistance has previously been found in several angiosperms. Trees that were pretreated with a combination of 12 bark wounds (1.6 x 10 cm), four fungal inoculations and four sterile inoculations 1-15 days before mass inoculation with C. polonica at 400 inoculations per square meter over a 0.8 m stem section had significantly shorter necroses in the phloem, less bluestained sapwood, and less dead cambium than untreated control trees. Pretreatment with four fungal or sterile inoculations alone did not lead to enhanced resistance. Pretreatment by bark wounding alone seemed to provide an intermediate degree of resistance compared to bark wounding, fungal inoculations and sterile inoculations combined. All trees had a marked increase in the number of resin ducts in the year of inoculation compared with previous years, suggesting that formation of traumatic resin ducts play an important role in the development and maintainance of enhanced resistance.     

Mapping landscapes: Integrating GIS and social science methods to model human-nature relationships in Southern Cameroon

Participatory mapping and GIS are both necessary to model the interactions between humans and their environment. A case study from the forest margin in the Congo Basin demonstrates how data from participatory community mapping and other social science methods can be prepared for quantitative modelling. This approach bridged the gap between spatial modelling data and social decision-making in space by elaborating a geographically consistent social representation of the landscape and giving a geographical base to the connection between land use, its cultural representation, and its social management. This was achieved through an iterative process of GIS cartography, using feedback from village informants and field checking, to transpose the spatial references from participatory mapping sketches into reliable geographic locations. As well as demonstrating the utility of such data for modelling, this work clarified the distribution of land rights among the six main owner-clans spread through the eight hamlets in the watershed. The ‘basin’ of spatial resources and its relation to the rules of land use and natural resource management were defined for each clan. Land-use systems at the forest-agriculture interface in the study area proved to be complex, strongly driven by social rules and influenced by history and settlement strategies. These social and historical aspects established the framework within which communities make current decisions and interventions. The authors thank the people of Akok for their collaboration and hospitality. Village technician Michel Engueng and village contact Adolphe Ze facilitated and assisted this work. Our thanks also to Julie Mbazo’o and Remy Assoumou, both members of the ‘social side’ working group of the FLORES modelling team. Christopher Legg’s contribution to the discussion of this work through his constant reviewing of the CamFlores model is gratefully recognized. Finally, thanks to the whole CIFOR-ACM team for numerous scientific and methodological inputs. The Italian Foreign Ministry funded the work of Valentina Robiglio (APO-GIS specialist) through an APO contract with IITA. The European Union (Tropical Forestry budget line DG VIII) contributed to the CIFOR’s research program on the ‘Adaptive Collaborative Management of Forests’ (ACM) in Africa, and to the Alternative to Slash and Burn (ASB) program in Cameroon on ‘Environmental Services and Rural Livelihoods’ through the World Agroforestry Centre.     

Anaesthetic management for balloon dilation of cor triatriatum dexter in a dog

A three-month-old female Rottweiler puppy was referred for intravascular correction of a previously identified cor triatriatum dexter. Echocardiography confirmed the presence of a hyperechoic membrane that divided the right atrium into a cranial and caudal chamber. A foramen in this membrane allowed the blood to flow from the caudal to the cranial chamber. Balloon dilation of the defect under transthoracic echocardiographic guidance was scheduled for the following day. The dog was premedicated with 0.5 μg/kg sufentanil and 0.2 mg/kg midazolam administered intravenously. General anaesthesia was induced with 2 mg/kg propofol and maintained with inhaled isoflurane in oxygen; at the same time, a constant rate infusion of 0.5 μg/kg/h sufentanil was administered by means of an infusion pump. Uneventful ventricular and supraventricular tachyarrhythmias developed during the placement of catheters and balloon dilation. At the end of procedure, when the guide wire and balloon catheter were removed, normal sinus rhythm was observed. To the authors’ knowledge, no previous reports have described the anaesthetic management of a balloon dilation procedure for cor triatriatum dexter in dogs.