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.     

Comment on "The incidence of fire in Amazonian forests with implications for REDD"

Arag?o and Shimabukuro (Reports, 4 June 2010, p. 1275) reported that fires increase in agricultural frontiers even as deforestation decreases and concluded that these fires lead to unaccounted carbon emissions under the United Nations climate treaty's tropical deforestation and forest degradation component. Emissions from post-deforestation management activities are, in fact, included in these estimates--but burning of standing forests is not.     

Predicting mature cork biomass with t years of growth from one measurement taken at any other age

Cork is a natural product that is extracted from the outer bark of the cork oak tree. According to Portuguese legislation, the interval between two consecutive cork extractions on the same tree must be equal or greater than 9 years. Although the majority of the cork oak stands are debarked at the end of this period, this rotation may not be the optimum in many cases. The existing models for cork weight prediction can only be used for trees debarked at a 9 years or, in one model, at a 10-years rotation period, since the data used for its development was limited to these growth periods. The development of a method that allows for the prediction of the mature cork biomass with t years of growth, based in one measurement taken at any other age, was the main objective of this work. The method is based on the knowledge that the density of the cork tissue is constant between the inner and outer cork rings, being significantly different from density of the cork back. It can be implemented using two different equations that were developed during this work: a model to estimate cork biomass with 9 years of age and a model to estimate the cork back weight proportion at 9 years of age. For the first model, four different alternative models were developed, considering different levels of information collected during forest inventory. The model to estimate the cork back weight proportion leads to the biomass of cork tissue. Cork biomass at t years is obtained by decreasing or increasing the biomass of cork tissue according to the difference in cork thickness between t and 9 years of growth. The proposed method was evaluated by comparing the observed and estimated values of cork biomass from an independent data set with corks with 9, 10 and 11 years of age. The results showed similar precision for corks with 9, 10 or 11 years of age. As expected the precision of the predictions increases when the model to estimate cork biomass with 9 years of age uses more information. The presented method should be an important tool for cork oak stand management, for the prediction of the evolution of carbon stocks in cork oak stands, and will allow analyzing the impact in cork biomass production of decreasing or extending the interval between two consecutive cork extractions.     

Molecular identification of a 16SrIII-B phytoplasma associated with cassava witches’ broom disease

Since its arrival in the British Isles in 1845 Phytophthora infestans has remained the most destructive pathogen of potato. In the ensuing period, the British and Irish P. infestans populations have undergone major displacements following the immigration of novel strains. Here we report the re-emergence of the Ib mitochondrial DNA haplotype in the British and Irish P. infestans populations associated with the 6_A1 genotype. Historically associated with the previously panglobally distributed clonal lineage US-1, the Ib haplotype has not been detected (with the exception of a single isolate in the mid 1990s) in the British or Irish P. infestans populations since the early 1980s. The 6_A1 isolates analysed possessed mtDNA Ib, but were otherwise quite unlike US-1, having the Pep allozyme genotype 96/96 and novel RG57 and SSR fingerprints. These genetic characteristics strongly suggest that the appearance of the 6_A1 genotype in these populations has resulted from migration (possibly after a recombination event elsewhere). This study highlights the advantages of utilising a range of different markers in pathogen monitoring.     

Photocatalytic Degradation of Phenol by Thermal Titanium Dioxide Thin Layer Electrodes

Electrochemical processes in industrial effluents have been studied as a means to obtain higher efficiency in wastewater treatment. Heterogeneous photocatalysis appears as a low-cost alternative through the use of lower wattage lamps and thermal TiO₂ films. Photocatalysis became a clean process for water treatment due to hydroxyl radicals generated on semiconductor surface. Such radicals are able to degrade several organic compounds. This study used different electrodes and analytical methods for degradation of phenol molecules to reduce treatment costs, improve efficiency, and identify compounds formed during the decomposition of phenolic molecules. Thermal growth of TiO₂ film was observed on the titanium electrode in rutile form. Application of an electrical potential on the Ti/TiO₂ working electrode increases efficiency in reducing concentration of phenol after photocatalytic treatment. Still, high energy radiation (UVC) showed best degradation rates in photolytic process. Different compounds formed during the degradation of phenol were also identified in the UVC?CPE treatment.     

Antimicrobial and Antimycobacterial Activity of Cyclostellettamine Alkaloids from Sponge Pachychalina sp.

Cyclostellettamines A – F (1 – 6) isolated from the sponge Pachychalina sp. and cyclostellettamines G - I, K and L (7 – 11) obtained by synthesis were evaluated in bioassays of antimicrobial activity against susceptible and antibiotic-resistant Staphylococcus aureus, Pseudomonas aeruginosa and antibiotic-susceptible Escherichia coli and Candida albicans, as well as in antimycobacterial activity against Mycobacterium tuberculosis H37Rv bioassays. The results obtained indicated that cyclostellettamines display different antimicrobial activity depending on the alkyl-chain size, suggesting that, if a mechanism-of action is implied, it is dependent on the distance between the two pyridinium moieties of cyclostellettamines.     

Typical and atypical manifestations of Anaplasma phagocytophilum infection in dogs

Eighteen clinically ill dogs, naturally infected with Anaplasma phagocytophilum, were examined at a veterinary practice in Baxter, Minnesota. A clinical examination, complete blood cell count, enzyme- linked immunosorbent assay (ELISA) for A phagocytophilum, Borrelia burgdorferi, and Ehrlichia canis antibodies and Dirofilaria immitis antigen, and a polymerase chain reaction test for A phagocytophilum DNA were obtained for all dogs. Physical examination findings included fever, arthropathy, lymphadenopathy, epistaxis, acute gastritis, cervical hyperpathia, and central nervous system dysfunction. Complete blood cell count abnormalities included thrombocytopenia, morulae in neutrophils, anemia, leukopenia, eosinopenia, lymphopenia, and monocytosis. Seroreactivity to A phagocytophilum was found in 61%, B burgdorferi antibodies in 17%, and D immitis antigen in 5% of the dogs. Fever, arthropathy, neurologic dysfunction, and epistaxis are clinical syndromes that can be associated with A phagocytophilum infection. Treatment with doxycycline resulted in rapid resolution of clinical signs in all dogs.     

Performance and meat quality traits of beef heifers fed with two levels of concentrate and ruminally undegradable protein

The effects of two levels of concentrate and ruminally undegradable protein (RUP) on performance, intake, digestibility, carcass characteristics, meat quality traits, and commercial cuts yield were assessed. Twenty crossbred heifers (240 kg average body weight) were used. At the beginning of the trial, four animals were slaughtered as reference group and the 16 remaining animals were randomly assigned to four treatments, in a 2 × 2 factorial design: two levels of concentrate (40% and 80%, dry matter (DM) basis) and two levels of RUP (48.79% and 27.19% of CP). At the end of the trial, all the animals were slaughtered. There was no interaction (P > 0.05) between concentrate and RUP levels. Dry matter intake and nutrients digestibility was not affected (P > 0.05) by RUP level. Heifers fed the highest RUP level had greater (P < 0.05) average daily gain (ADG). Concentrate level did not affect (P > 0.05) DMI and ADG. Heifers fed diets with 80% concentrate had greater intake of TDN and EE, and lower intake of NDF (P < 0.05). The digestibilities of all nutrients, except the NDF, were greater (P < 0.05) for the 80% concentrate diets. The RUP levels did not affect (P > 0.05) the carcass characteristics and carcass gain composition. Heifers fed 80% concentrate diets had larger (P < 0.05) longissimus muscle area and lower gain of bones (P < 0.05). Concentrate level did not affect (P > 0.05) the composition of carcass gain. There was no effect (P > 0.05) of RUP and concentrate levels on meat quality traits and commercial cut yields.     

Genetic analysis of Anisakis typica (Nematoda: Anisakidae) from cetaceans of the northeast coast of Brazil: new data on its definitive hosts

Prevalence and seasonal variations of helminth infections and their association with morbidity parameters were studied in traditionally reared Cambodian cattle. Four villages in two provinces of West Cambodia were visited on monthly intervals over a period of 11 months, during which 2391 animals were faecal and blood sampled for parasitological and haematological examinations. The body condition score (BCS), faecal consistency (diarrhoea score, DS), colour of the ocular conjunctivae (FAMACHA(?)) and packed cell volume were determined for each individual animal. The overall proportion of samples that was positive for gastrointestinal nematodes was 52%, 44% and 37% in calves (from 1 to 6 months), young animals (6 to 24 months) and adults (over 24 months), respectively, while geometric mean faecal egg counts (FECs) for each of these age categories were 125, 66 and 15 eggs per gram, respectively. Six genera of strongyles were found in the faecal cultures, i.e. in descending order of occurrence, Cooperia, Oesophagostomum, Haemonchus, Trichostrongylus, Mecistocirrus and Bunostomum. The prevalences of Fasciola and Paramphistomum, estimated by coprological examination, varied between 5-20% and 45-95%, respectively. Logistic mixed models were used to investigate associations of morbidity markers with the presence of parasite infection. A low BCS was associated with gastrointestinal nematode and liver fluke infections, and soft faecal consistency with Paramphistomum infections. However, other factors such as nutritional deficiencies and intercurrent diseases are likely to enhance the effects of parasites and should therefore be considered when using these morbidity parameters as indicators of parasitism.