Determination of anthocyanins from camu-camu (Myrciaria dubia) by HPLC-PDA, HPLC-MS, and NMR

Camu-camu [Myrciaria dubia (HBK) McVaugh] is a small fruit native to the Amazonian rain forest. Its anthocyanin profile has now been investigated for the first time. Fruits from two different regions of the S?o Paulo state, Brazil, were analyzed. The major anthocyanins were isolated by high-speed countercurrent chromatography. HPLC-PDA, HPLC-MS/MS, and 1H NMR were used to confirm the identity of the main anthocyanins of camu-camu. Cyanidin-3-glucoside was identified as the major pigment in the fruits from both regions, representing 89.5% in the fruits produced in Iguape and 88.0% in those from Mirandópolis, followed by the delphinidin-3-glucoside, ranging between 4.2 and 5.1%, respectively. Higher total anthocyanin contents were detected in the fruits from Iguape (54.0 +/- 25.9 mg/100 g) compared to those from Mirandópolis (30.3 +/- 6.8 mg/100 g), most likely because of the lower temperatures in the Iguape region.     

Headspace solid phase microextraction (SPME) analysis of flavor compounds in wines. Effect of the matrix volatile composition in the relative response factors in a wine model.

The application of headspace solid phase microextraction (SPME) for flavor analysis has been studied. Headspace SPME sampling was tested for nine common wine flavor compounds in 10% (v/v) aqueous ethanol: linalool, nerol, geraniol, 3-methyl-1-butanol, hexanol, 2-phenylethanol, ethyl hexanoate, ethyl octanoate, and ethyl decanoate. The chemical groups (monoterpenoids, aliphatic and aromatic alcohols, and esters) showed specific behavior in SPME analysis. SPME sampling parameters were optimized for these components. Relative response factors (RRFs), which establish the relationship between the concentration of the compound in the matrix liquid solution and the GC peak area, were estimated for all compounds. Log(10)(RRF) varied from 0 (3-methyl-1-butanol) to 3 (ethyl decanoate), according to their molecular weight. Quantification by SPME was shown to be highly dependent on the matrix composition; the compounds with higher RRF were the less affected. As a consequence, the data obtained with this methodology should be used taking into consideration these limitations, as shown in the analysis of four monovarietal Bairrada white wines (Arinto, Bical, Cerceal, and Maria Gomes).     

Cytogenetical and histological studies in testis of Tayassu tajacu (Cateto), Tayassu pecari (Queixada) and a natural interspecific hybrid

The karyotype, the histological structure of the testis and the synaptonemal complex (SC) of the mammalian species Tayassu tajacu , Tayassu pecari and of an interspecific male hybrid captured in nature were analysed. The specimens of T. tajacu (2n=30) and T. pecari (2n=26) exhibited seminiferous tubules with germ cells in all spermatogenesis stages. In the SC studies both species had a regular structure, easily identified in the autosomes and in the sex chromosomes. The hybrid (2n=28) had seminiferous tubules with almost all germinal cells in the spermatogonium stage and only a few cells in the primary spermatocyte stage. Gross abnormalities in SC were observed. A few lateral elements showed regular or partially regular synapsis, other lateral elements were synapsed as multivalents, and most axial elements remained unsynapsed. The results suggest that the karyotypes of the parental species have sufficient differentiation to avoid chromosome synapsis. Alternatively, the hypothesis of the existence of genetic incompatibilities between the parental species is discussed.     

Check-list for scientific names of common parasitic fungi

This list is a continuation of Series 1a (Neth. J. Pl. Path. 78, Suppl. 1, 1972), an account of the nomenclature of common parasitic fungi on trees and shrubs as used in official publications of the Netherlands Society of Plant Pathology and the Netherlands Ministry of Agriculture and Fisheries.     

Check-list for scientific names of common parasitic fungi. Series 2a: Fungi on field crops: beet and potato; caraway,flax and oilseed poppy

This list gives a first account of the nomenclature of common parasitic fungi on field crops as used in official publications of the Netherlands Society of Plant Pathology and the Netherlands Ministry of Agriculture and Fisheries.     

Mitochondrial damage as an early event of monensin-induced cell injury in cultured fibroblasts L929

The present study was designed to identify, submicroscopically, the primary organelle or target structure for monensin in cultured murine fibroblasts L929. In addition, the effect of the drug on cell size and surface membranes of the cells were analysed; cellular proliferation, collagen secretion, and necrosis and apoptosis were re-evaluated. At the lowest concentration of monensin the foremost ultrastructural alteration occurred in the mitochondria, characterized by increased matrix density with disorganized and less distinct crystae. Incubation with monensin at higher concentrations resulted in severe mitochondrial damage and marked dilatation of the Golgi apparatus and rough endoplasmic reticulum cisternae. Fibroblasts exposed to higher concentrations of monensin were enlarged with decreased number of filopodia and hollows in the surface membrane. Moreover, monensin inhibited the cell proliferation, increased immunohistochemical positiveness for collagen type I in a dose-dependent manner, and, at high concentrations, caused cell necrosis whereas apoptosis was not induced. Taken together, these results show that monensin induces early mitochondrial damage, possibly causing an energy deficit that led to inhibition of fibroblasts proliferation and accumulation of collagen causing dilatation of Golgi apparatus and rough endoplasmic reticulum. Moreover, the mitochondrial damage would also explain the monensin-induced necrosis.     

Pseudomonas syringae pv. coryli, the Causal Agent of Bacterial Twig Dieback of Corylus avellana

ABSTRACT Thirty-eight bacterial strains isolated from hazelnut (Corylus avellana) cv. Tonda Gentile delle Langhe showing a twig dieback in Piedmont and Sardinia, Italy, were studied by a polyphasic approach. All strains were assessed by fatty acids analysis and repetitive sequence-based polymerase chain reaction (PCR) fingerprinting using BOX and ERIC primer sets. Representative strains also were assessed by sequencing the 16S rDNA and hrpL genes, determining the presence of the syrB gene, testing their biochemical and nutritional characteristics, and determining their pathogenicity to hazelnut and other plants species or plant organs. Moreover, they were compared with reference strains of other phytopathogenic pseudomonads. The strains from hazelnut belong to Pseudomonas syringae (sensu latu), LOPAT group Ia. Both fatty acids and repetitive-sequence-based PCR clearly discriminate such strains from other Pseudomonas spp., including P. avellanae and other P. syringae pathovars as well as P. syringae pv. syringae strains from hazelnut. Also, the sequencing of 16S rDNA and hrpL genes differentiated them from P. avellanae and from P. syringae pv. syringae. They did not possess the syrB gene. Some nutritional tests also differentiated them from related P. syringae pathovars. Upon artificial inoculation, these strains incited severe twig diebacks only on hazelnut. Our results justify the creation of a new pathovar because the strains from hazelnut constitute a homogeneous group and a discrete phenon. The name of P. syringae pv. coryli is proposed and criteria for routine identification are presented.     

Aerobic bacterial flora of the nasal cavity in Gulf of California sea lion (Zalophus californianus) pups

Nasal swab samples from clinically healthy California sea lions pups (Zalophus californianus) from six different reproductive rookeries in the Gulf of California were collected to determine the type and frequency of the representative aerobic bacterial microflora of their nasal mucosa. A total of 114 samples were examined and 100 bacterial isolates were identified and typified by microbiological and biochemical standard tests. Fifty four isolates corresponded to Gram positive bacteria (54%) and 46 isolates to Gram negative bacteria (46%). Fifteen bacterial genera were identified, including Micrococcus, Arcanobacterium, Corynebacterium, Moraxella, Neisseria, Escherichia, Kurthia, Acinetobacter, Staphylococcus, Brevibacillus, Bacillus, Klebsiella, Stenotrophomonas, Pseudomonas and Aeromonas. The most frequently isolated genera were Moraxella (24%), Micrococcus (18%), and Corynebacterium (15%). These results show the presence in the nasal cavity of sea lions of several microorganisms. Although considered part of the normal microflora, they may also be opportunistic pathogens for their hosts and may act as a potential natural sentinel of environmental changes.     

Higher incidence of Malassezia pachydermatis in the eyes of dogs with corneal ulcer than in healthy dogs

Malassezia pachydermatis is usually associated with otitis and dermatitis in dogs but it can also cause diseases in other species, including humans. In a human neonatal intensive care unit, M. pachydermatis was isolated from an infant's ocular discharge. Therefore, the aim of this study was to ascertain the presence of Malassezia spp. and its possible consequences in dogs' eyes. This research included 19 dogs with unilateral or bilateral corneal ulcers and 60 healthy dogs. A total of 158 clinical specimens from both the groups were obtained from the conjunctival sac of each eye by a calibrated platinum loop. The samples were placed on Dixon and blood agar, incubated at 35 degrees C, and examined daily for 15 days. Then, the strains were subcultured on Sabouraud agar. Of 22 clinical specimens collected from the eyes with corneal ulcers, five cultures (23%) were positive for M. pachydermatis. Of 16 samples collected from the contralateral healthy eye, cultures were positive in three samples (19%). Three animals had unilateral corneal ulcer and positive cultures for M. pachydermatis in both the eyes. Two dogs had unilateral corneal ulcer and positive cultures for M. pachydermatis in the same eye. However, from the 120 samples of 60 healthy dogs, only four clinical specimens (3%) had positive cultures for M. pachydermatis. The findings of M. pachydermatis, in a considerable percentage of clinical specimens from dogs with corneal ulcer, suggest its possible role at least as an aggravating factor in the pathophysiology of this disease.     

Antibacterial activity of coffee extracts and selected coffee chemical compounds against enterobacteria

The in vitro antimicrobial activity of commercial coffee extracts and chemical compounds was investigated on nine strains of enterobacteria. The antimicrobial activity investigated by the disc diffusion method was observed in both the extracts and tested chemical compounds. Even though pH, color, and the contents of trigonelline, caffeine, and chlorogenic acids differed significantly among the coffee extracts, no significant differences were observed in their antimicrobial activity. Caffeic acid and trigonelline showed similar inhibitory effect against the growth of the microorganisms. Caffeine, chlorogenic acid, and protocatechuic acid showed particularly strong effect against Serratia marcescens and Enterobacter cloacae. The IC(50) and IC(90) for the compounds determined by the microtiter plate method indicated that trigonelline, caffeine, and protocatechuic acids are potential natural antimicrobial agents against Salmonella enterica. The concentrations of caffeine found in coffee extracts are enough to warrant 50% of the antimicrobial effect against S. enterica, which is relevant to human safety.