AVIAN POXVIRUS INFECTION IN A FLAMINGO (PHOENICOPTERUS RUBER) OF THE LISBON ZOO.

Avian poxviruses (APV) are very large viruses spread worldwide in a variety of hosts. They are responsible for a disease usually referred to as pox, mainly characterized by nodular lesions on feather-free regions of the body. On May 2010, a young American flamingo (Phoenicopterus ruber) of the Lisbon Zoo (Portugal) developed a nodular lesion suggestive of poxvirus infection on its right foot. Avipoxvirus was isolated from the lesion and a fragment of the P4b-encoding gene was amplified by polymerase chain reaction. The nucleotide sequence of the amplicon was determined and analyzed. A close relationship (100% identity) was observed between the flamingo poxvirus and isolates from great bustard (Hungary 2005), house sparrow (Morocco 2009), MacQueen's bustard (Morocco 2011), and Houbara bustard (Morocco 2010 and 2011), suggesting interspecies transmission as a possible source of infection. To strengthen the investigation, the 5' and 3' ends of genes cnpv186 and cnpv 187, respectively, were also analyzed. The cnpv186-187 fragment exhibited 100% identity with MacQueen's bustard and Houbara bustard isolates, both from Morocco 2011. Phylogenetic analyses based in both fragments grouped the flamingo isolate consistently within clade B2 of canarypox. However, the phylogenetic relationships among the different representatives of avian poxviruses were more comprehensive in the tree based on the concatenated coding sequences of the cnpv186-187 fragment, rather than on the P4b-coding gene. The clearer displacement and distribution of the isolates regarding their host species in this last tree suggests the potential usefulness of this genomic region to refine avian poxvirus classification.

Molecular and biochemical characterization of caffeine synthase and purine alkaloid concentration in guarana fruit.

Guarana seeds have the highest caffeine concentration among plants accumulating purine alkaloids, but in contrast with coffee and tea, practically nothing is known about caffeine metabolism in this Amazonian plant. In this study, the levels of purine alkaloids in tissues of five guarana cultivars were determined. Theobromine was the main alkaloid that accumulated in leaves, stems, inflorescences and pericarps of fruit, while caffeine accumulated in the seeds and reached levels from 3.3% to 5.8%. In all tissues analysed, the alkaloid concentration, whether theobromine or caffeine, was higher in young/immature tissues, then decreasing with plant development/maturation. Caffeine synthase activity was highest in seeds of immature fruit. A nucleotide sequence (PcCS) was assembled with sequences retrieved from the EST database REALGENE using sequences of caffeine synthase from coffee and tea, whose expression was also highest in seeds from immature fruit. The PcCS has 1083bp and the protein sequence has greater similarity and identity with the caffeine synthase from cocoa (BTS1) and tea (TCS1). A recombinant PcCS allowed functional characterization of the enzyme as a bifunctional CS, able to catalyse the methylation of 7-methylxanthine to theobromine (3,7-dimethylxanthine), and theobromine to caffeine (1,3,7-trimethylxanthine), respectively. Among several substrates tested, PcCS showed higher affinity for theobromine, differing from all other caffeine synthases described so far, which have higher affinity for paraxanthine. When compared to previous knowledge on the protein structure of coffee caffeine synthase, the unique substrate affinity of PcCS is probably explained by the amino acid residues found in the active site of the predicted protein.

Guarana: revisiting a highly caffeinated plant from the Amazon.

ETHNOPHARMACOLOGICAL RELEVANCE: Guarana (Paullinia cupana Kunth var. sorbilis (Mart.) Ducke) has been traditionally consumed by indigenous communities of the Amazon region. It is valued mainly for its stimulant property because of its high content of caffeine, which can be up to 6% in the seeds. AIM OF THE REVIEW: The purpose of this review is to revisit this typically Brazilian plant, addressing economic considerations, the chemical makeup of the seeds and pharmacological properties so far investigated. RESULTS: Guarana is primarily produced in the Brazilian states of Amazonas and Bahia, and approximately 70% of the production is used by the industry of soft and energy drinks. The other 30% becomes guarana powder for direct consumption in capsules or dilution in water, or it serves as a raw material for the pharmaceutical and cosmetics industries. In addition to its stimulant property, guarana has other therapeutic properties, which have aroused the interest of the scientific community. CONCLUSION: This review shows that other guarana properties may be explored and how scarce are the studies regarding agronomic, plant pathology, physiology and breeding. So far, caffeine has been the main reason to study guarana and still will lead the researches because the demand for this alkaloid by food and pharmaceutical industry, and a strongly growing market related with beauty products. However, guarana has other components and there is great interest in studies designed to elucidate the effects of guarana's bioactive components and their potential pharmacological applications. Significant part of the guarana production in Brazil still comes from Indians tribes in the Amazon State, and any improvement in this plant, in any aspect, may propitiate a positive economic impact in their lives.