Characterization of fluoroquinolone resistance in Escherichia coli strains from ruminants.

Thirty-seven fluoroquinolone-resistant Escherichia coli strains from ruminants (according to Clinical and Laboratory Standards Institute guidelines) were screened by molecular methods for mutations in the quinolone resistance-determining region (QRDR) of the gyrA and parC genes and for the presence of the qnrA gene. One of the strains studied was an enterohemorrhagic E. coli (EHEC) strain potentially pathogenic for humans. Three E. coli strains resistant to enrofloxacin (minimal inhibitory concentration [MIC] = 2 microg/ml) but not to ciprofloxacin (MIC = 1 microg/ml) presented single mutations in the gyrA and parC genes, while 34 strains resistant to both fluoroquinolones presented double and single mutations in gyrA and parC, respectively (31 strains), or double mutations in gyrA and parC (3 strains). The EHEC strain presented a double amino acid substitution in the GyrA protein (Ser-83-->Leu and Asp-87-->Gly) and a double amino acid substitution in the ParC protein (Gly-78-->Cys and Ser-80-->Arg), one of which has not been previously described. The present study shows that most of the mutations in the QRDR of the gyrA and parC genes of fluoroquinolone-resistant E. coli strains from ruminants are the same as those seen in E. coli strains from other animal species and humans and that there are no differences in mutation patterns in the QRDR of E. coli strains from healthy ruminants and those with diarrhea. No strains carried qnrA, which indicates that this gene does not play an important role in the selection of fluoroquinolone-resistant E. coli strains from ruminants.     

Evaluation of phenolic compounds and antioxidant activity of blueberries and modelization by artificial neural networks

ABSTRACT

The study aimed at evaluating the influence of different production conditions, conservation, and extraction procedures on the total phenolic compounds and antioxidant activity of blueberries by DPPH and ABTS methods. The production factors considered were origin, altitude of the farm location, and age of the bushes. The conservation conditions considered were freezing as opposed to the fresh product. The extraction procedures included two different solvents and two orders of extraction. The data analysis was carried out by training artificial neural networks to model the data and extract information from the model.

The results obtained revealed that the type of extract and the order of extraction influenced the concentrations of phenolic compounds as well as the antioxidant activity of the different samples studied. Also the origin of the farms from where the blueberries were harvested significantly influence those properties, showing that the blueberries from Oliveira do Hospital had less phenolic compounds and lower antioxidant activity. Also older bushes at higher altitudes seem to produce berries richer in these properties. Regarding conservation, no influence was observed for phenols but a slight influence could be detected for antioxidant activity.     

RABBIT HEMORRHAGIC DISEASE VIRUSES DETECTED IN PET RABBITS IN A COMMERCIAL LABORATORY IN EUROPE

Rabbit hemorrhagic disease (RHD) is an important cause of disease and mortality in wild and domestic European rabbits (Oryctolagus cuniculus) throughout the world. Testing for 2 distinct RHD virus types (RHDV/RHDVa and RHDV2) was carried out on samples collected from 684 rabbits submitted from veterinary practices and private owners throughout Europe between January 2015 and June 2017. Four (0.6%) were positive for RHDV/RHDVa and 257 (37.4%) were positive for RHDV2. RHDV/RHDVa was detected in individual samples from Germany and the Netherlands, while RHDV2 was found in animals from Germany, Great Britain, Luxembourg, The Netherlands, Spain, Switzerland, Poland, Belgium, Austria, Sweden, and Finland.     

Analysis of world strains of Anaplasma marginale using major surface protein 1a repeat sequences

Anaplasma marginale is a tick-borne pathogen of cattle that causes the disease bovine anaplasmosis worldwide. Major surface proteins (MSPs) are involved in host-pathogen and tick-pathogen interactions and have been used as markers for the genetic characterization of A. marginale strains and phylogenetic studies. MSP1a is involved in the adhesion and transmission of A. marginale by ticks and varies among geographic strains in the number and sequence of amino-terminal tandem repeats. The aim of this study was to characterize the genetic diversity of A. marginale strains collected from countries in North and South America, Europe, Asia, Africa and Australia, inclusive of all continents. In this study, we characterized 131 strains of A. marginale using 79 MSP1a repeat sequences. These results corroborated the genetic heterogeneity of A. marginale strains in endemic regions worldwide. The phylogenetic analyses of MSP1a repeat sequences did not result in clusters according to the geographic origin of A. marginale strains but provided phylogeographic information. Seventy-eight percent of the MSP1a repeat sequences were present in strains from a single geographic region. Strong (> or =80%) support was found for clusters containing sequences from Italian, Spanish, Chinese, Argentinean and South American strains. The phylogenetic analyses of MSP1a repeat sequences suggested tick-pathogen co-evolution and provided evidence of multiple introductions of A. marginale strains from various geographic locations worldwide. These results contribute to the understanding of the genetic diversity and evolution of A. marginale and tick-pathogen interactions.     

Purification and characterization of a 45-kDa concealed antigen from the midgut membranes of Ornithodoros erraticus that induces lethal anti-tick immune responses in pigs

Ornithodoros erraticus is an argasid tick that can transmit severe diseases such as human relapsing fever and African swine fever. In the search for a vaccine against this parasite, a crude extract of tick midgut membranes (GME) was obtained that in pigs and mice induced a protective response able to kill up to 80% of the nymphs in the first 72 h post-feeding and to reduce the fecundity of females by more than 50%. To identify the protective antigens, the GME was subjected to successive biochemical fractionations and the resulting simpler protein fractions were inoculated in pigs. A 45-kDa antigen, the so-called Oe45, was detected, purified and demonstrated to be responsible for the protection induced by the GME. Oe45 seems to be a membrane protein that is presumably expressed on the luminal membrane of midgut epithelial cells. Oe45 consists of at least two differently charged bands (cationic and neutral), which show antigenic cross-reactivity. The possibility that these bands might be different isoforms of the same protein is discussed. Although Oe45 is constitutively expressed at low levels throughout the trophogonic cycle, its expression is up-regulated by the ingestion of blood, as suggested by the higher levels observed between 6 and 72 h post-feeding.     

In vivo long-term effects of retinoic acid exposure in utero on induced hyperplastic epidermal foci in murine skin

Adult Naval Medical Research Institute (NMRI) mice, after prenatal exposure to retinoic acid (RA), were treated with a standard two-stage skin carcinogenesis regime to characterize hyperplastic epidermal foci that precede the appearance of cutaneous papillomas, and to investigate the in vivo long-term action of RA on adult mouse skin treated with DMBA (7,12 dimethyl benz[a]anthracene) and TPA (12-O-tetradecanoylphorbol 13-acetate). The results demonstrate that RA administered to pregnant mice had a long-term inhibitory action on the cell differentiation and development of hyperplastic lesions occurring prior to cancer on the adult skin of their offspring as well as a stimulatory effect on cell proliferation of these hyperplastic lesions.     

Spongiform neurodegenerative disease in a Persian kitten

A congenital encephalopathy with spongiform degeneration and prominent neuronal apoptosis was observed in a 4-month-old Persian male cat with a history of depressed mental status and ataxia. On clinical examination, signs included right head tilt, ventroflexion of the head and neck, and tetraparesis. Histological examination of the central nervous system revealed multifocal, bilateral and symmetrical vacuolar degeneration of the neuropil, mainly involving the cerebellar and vestibular nuclei area, the caudal colliculi, the mesencephalic nuclei, the tegmental area and the deeper layer of the cerebral cortex. Accumulation of phosphorylated neurofilaments was detected in neuronal perikarya of the deep cortical layers, hippocampus and thalamus. Numerous pyknotic and apoptotic neurons were also observed in the cerebral cortex. These neuropathological changes differ from those observed in previous reports of spongiform degeneration of the grey matter in cats and were suggestive of a congenital neurodegenerative disease.     

Environmental impact of razor clam harvesting using salt in Ria Formosa lagoon (Southern Portugal) and subsequent recovery of associated benthic communities

• 1. Razor clams are found in different habitats ranging from sheltered systems (e.g. lagoons and estuaries) to open coasts. They are distributed worldwide and comprise a small number of species of high economic value. Depending on the specific habitat and species exploited, different mechanical and hand‐harvesting techniques are employed. While the environmental effects resulting from mechanized methods have been described by several authors, the impacts caused by traditional hand‐harvesting methods remain unknown. Therefore, a study was undertaken in Ria Formosa lagoon (South of Portugal) addressing the environmental effects resulting from harvesting Solen marginatus with salt.
• 2. No significant impact on the sediment was found; the main effect was an increase in salinity after covering the area with salt, which decreased rapidly with the flood tide and after a few hours had returned to pre‐harvesting levels.
• 3. No effects on benthic communities were observed, with similar fluctuation patterns recorded in control and experimental areas, the observed differences being attributed to the natural variability of benthic populations.
• 4. Based on the results obtained in the present study, a razor clam fishery using salt in intertidal areas can be considered environmentally ‘friendly’.

Copyright © 2008 John Wiley & Sons, Ltd.     

Broodstock conditioning of the Portuguese oyster (Crassostrea angulata,Lamarck, 1819): influence of different diets

The Portuguese oyster Crassostrea angulata shows great potential in oyster farming. The conservation of pure populations of this species is important for production diversification and biodiversity preservation. In this way, the zootechnological development for seed hatchery production is extremely important. Broodstock conditioning is a key step in the process of rearing bivalves in a hatchery. Many factors regulate the reproductive cycle, being food one of the most important ones. To evaluate the effect of different diets on C. angulata reproductive performance, broodstock were conditioned with different food regimes formulated fundamentally by flagellates (Diet 1 – Pavlova lutheri and Isochrysis galbana clone T‐ISO; Diet 2 – P. lutheri, T‐ISO and Skeletonema costatum) and constituted fundamentally by diatoms (Diet 3 – S. costatum and Chaetoceros calcitrans; Diet 4 – P. lutheri, S. costatum and C. calcitrans). During conditioning, samples of oysters were collected to evaluate condition index, gonadal development and biochemical composition. At the end of the conditioning period, oysters were induced to spawn to evaluate reproductive output (fecundity, fertilization rate and D‐larvae development). The diets had an impact on the gametogenesis process, energy storage and reproductive output performance, being the best results those obtained in broodstock fed with the diatoms‐predominant diets. However, those fed with diets majority flagellates had an unsuccessful performance. Holistic approaches incorporating all results in this study reveal and reinforce the idea that the diatom species used presented the nutritional requirements to C. angulata broodstock, being essential in the conditioning phase.     

Surface Glucan Structures in Aeromonas spp.

Aeromonas spp. are generally found in aquatic environments, although they have also been isolated from both fresh and processed food. These Gram-negative, rod-shaped bacteria are mostly infective to poikilothermic animals, although they are also considered opportunistic pathogens of both aquatic and terrestrial homeotherms, and some species have been associated with gastrointestinal and extraintestinal septicemic infections in humans. Among the different pathogenic factors associated with virulence, several cell-surface glucans have been shown to contribute to colonization and survival of Aeromonas pathogenic strains, in different hosts. Lipopolysaccharide (LPS), capsule and α-glucan structures, for instance, have been shown to play important roles in bacterial–host interactions related to pathogenesis, such as adherence, biofilm formation, or immune evasion. In addition, glycosylation of both polar and lateral flagella has been shown to be mandatory for flagella production and motility in different Aeromonas strains, and has also been associated with increased bacterial adhesion, biofilm formation, and induction of the host proinflammatory response. The main aspects of these structures are covered in this review.