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A biotechnological process for treatment and recycling poultry feathers as a feed ingredient.

A strain of Kocuria rosea with keratinolytic capacity was cultured aerobically on submerged feathers to obtain a fermented feather meal (FFM). This FFM enriched with cells of K. rosea mainly contains crude protein (71%). The pepsin digestibility of the fermented product (88%) was similar to the value of the commercial feather meal and more than 70% greater that untreated feathers. The bacterial biomass improved the content of amino acids lysine (3.46%), histidine (0.94%) and methionine (0.69%). Additionally, the amino acid availability tested by in vivo assay was greater than commercial feather meal. The microbial cells also supplied carotenoid pigments to FFM (68 ppm). These results suggest that feather meal enriched with K. rosea may be useful in animal feeding as protein and pigment source.

Amino Acids↗

Efficacy of cellular vaccines and genetic adjuvants against bacterial kidney disease in chinook salmon (Oncorhynchus tshawytscha).

DNA adjuvants and whole bacterial cell vaccines against bacterial kidney disease (BKD) were tested in juvenile chinook salmon. Whole cell vaccines of either a nonpathogenic Arthrobacter spp. or an attenuated Renibacterium salmoninarum strain provided limited prophylactic protection against acute intraperitoneal challenge with virulent R. salmoninarum, and the addition of either synthetic oligodeoxynucleotides or purified R. salmoninarum genomic DNA as adjuvants did not increase protection. However, a combination of both whole cell vaccines significantly increased survival among fish naturally infected with R. salmoninarum, and the surviving fish treated with the combination vaccine exhibited reduced levels of bacterial antigens in the kidney. This is the first demonstration of a potential therapeutic effect of a whole cell vaccine against BKD.

Actinomycetales Infections↗

Genomic and transcriptomic characterization of genes expressed at 20 MPa by the marine actinobacterium Kocuria flava.

A marine hydrocarbonoclastic actinobacterium Kocuria flava IOS11 was isolated from 3500 m deep-sea water of the Indian Ocean. The isolate efficiently degraded phenanthrene (250 mg/L) achieving 82 and 98% of degradation at 0.1 MPa and 20 MPa, respectively within a period of 5 days. Whole genome, transcriptomee and metabolomic analysis elucidated its phenanthrene biodegradation efficiency under in situ deep-sea conditions. The genome sequence comprises 3.47 Mb distributed across 88 scaffolds with a high GC content of 74.30%. The genome analysis encoded 3126 genes including 3052 protein coding sequences with functional annotation identifying a broad array of genes associated with PAHs degradation, environmental stress adaptation, biosurfactant and siderophore synthesis. Transcriptome profiling under 0.1 and 20 MPa conditions with phenanthrene as a sole carbon source revealed enhanced expression of hydrocarbon degrading genes, transporters, biosurfactant associated enzymes and stress responsive genes including integrases, DNA repair protein Rad, alanine ligase, heat and cold shock proteins under high pressure conditions underscoring the deep-sea adaptation capabilities of the strain. The degradation pathway of phenanthrene was proposed through integrated genome, transcriptome and metabolomic analysis. These studies provided K. flava IOS11 as a metabolically versatile and pressure adapted bacterium with promising potential for bioremediation application in extreme marine environment.

Transcriptome↗

Detection of Renibacterium salmoninarum in tissue samples by sequence capture and fluorescent PCR based on the 16S rRNA gene.

The 16S rRNA genes from eight isolates of Renibacterium salmoninarum with different origins and dates of isolation were sequenced to evaluate the possibility to construct a diagnostic PCR system with target sites within this gene. The sequences were found to be identical but for one single position in one of the isolates, and two regions with an adequate number of nucleotide differences as compared to closely related species were identified. Species-specific fluorescent PCR primers complementary to these regions were constructed as well as oligonucleotides for DNA preparation by sequence capture. A mimic molecule was constructed to be used as an internal control. The PCR was specific and allowed the detection of DNA equivalent to 1-10 R. salmoninarum genomes per reaction. The DNA preparation with sequence capture and analysis by PCR with a mimic was found to be a reliable method for analysis of kidneys from fish with BKD. The amount of PCR inhibiting substances present in the tissue was reduced, and the relevant DNA was concentrated in the capture step. Furthermore, the use of the mimic molecule in the system assured that false negative results could be identified.

Actinomycetales Infections↗

Gamma irradiation and cellular damage in Kocuria rosea: investigation by one- and two-dimensional infrared spectroscopy.

Fourier transform infrared (FT-IR) spectroscopy was used to investigate the radiation-induced effects on Kocuria rosea. Bacterial suspensions at the stationary phase were exposed to increasing doses of gamma radiation. In the region 1350-840cm(-1), assigned to phosphodiester backbone, nucleic acids, and sugar rings, the radical damaging effects were dose-dependent, with the first threshold at 2.75kGy and the second at 13.75kGy inducing more striking spectral variations. Postirradiation reincubation did not significantly affect the biomolecular response, except in the spectral range 1100-1000cm(-1). These observations suggest the occurrence of new phylogenetic characteristics for K. rosea following irradiation. Moreover, two-dimensional analysis was used to highlight correlated evolutions of molecular species as radical aggression increased. The results point to an evolutionary scheme during the time course of irradiation. Thus, one- and two-dimensional IR analyses are convenient means of investigating the metabolic events following oxidative stress generated by either chemical or physical agents.

Cell Membrane↗

The structural identifiability and parameter estimation of a multispecies model for the transmission of mastitis in dairy cows.

A structural identifiability analysis is performed on a mathematical model for the coupled transmission of two classes of pathogen. The pathogens, classified as major and minor, are aetiological agents of mastitis in dairy cows that interact directly and via the immunological reaction in their hosts. Parameter estimates are available from experimental data for all but four of the parameters in the model. Data from a longitudinal study of infection are used to estimate these unknown parameters. A novel approach and application of structural identifiability analysis is combined in this paper with the estimation of cross-protection parameters using epidemiological data.

Animals↗

Degradation characteristics of a dibenzofuran-degrader Terrabacter sp. strain DBF63 toward chlorinated dioxins in soil.

To obtain basic information towards applying a dibenzofuran (DF)-degrader Terrabacter sp. strain DBF63 to bioremediate dioxin-contaminated soil, we investigated the degradative potential of strain DBF63 for either chlorinated or polychlorinated dibenzo-p-dioxins and dibenzofurans (ClxDD/ClxDF) in soil. In the soil slurry system with a soil to water ratio of 1:5 (w/v), the DF-grown DBF63 cells degraded 90% of 1 ppm 2,8-Cl2DF, whereas only 40% of 1 ppm 2,3-Cl2DD during the 7-day incubation. The degradation rates of 2-CIDF, 2-ClDD, 2,8-Cl2DF and 2,3-Cl2DF by strain DBF63 in the soil slurry system (5-day incubation) were approximately 89%, 65%, 78% and 32%, respectively. These results suggest that strain DBF63 was able to degrade mono- to dichlorinated dibenzofurans more effectively than mono- to dichlorinated dibenzo-p-dioxins. Using the same soil slurry system, we performed a preliminary bioremediation experiment using the actual dioxin-contaminated soil at an incineration site. We found that approximately 10% of tetra- to hexa-chlorinated congeners was decreased by a single inoculation with DBF63 cells within a 7-day incubation.

Biodegradation, Environmental↗

The gram positive cocci.

Recent changes in taxonomy of the gram positive cocci are discussed. Views on these changes and practical methods of differentiating the staphylococci, micrococci, streptococci, and aerococci are presented. Simplified schemes, using acceptable clinical laboratory techniques, are presented that either differentiate or categorize the pathologically important gram positive coccal species.

Bacteria↗

Micrococcal urinary-tract infections in young women.

In a prospective study in young women, novobiocin-resistant subgroup-3 micrococci were the second commonest cause, after Escherichia coli; of acute urinary infections. Proteus mirabilis was the only other causative organism. Symptoms, pyuria, or possible aetiological factors were the same in micrococcal and coliform infections. The infecting micrococcus "biotype" was only rarely found among the normal flora of the genitourinary tract of young women, though other micrococci and staphylococci were commonly present. Evidently, the infecting micrococci are selectively pathogenic in the urinary tract. Micrococcal infections, like coliform infections; commonly followed sexual intercourse, but there was no evidence that the micrococci were sexually transmitted. The infecting biotype was rarely found in the male urethra or prepuce.

Drug Resistance, Microbial↗

Detection of bacterial pathogens in environmental samples using DNA microarrays.

Polymerase chain reaction (PCR) is an important tool for pathogen detection, but historically, it has not been possible to accurately identify PCR products without sequencing, Southern blots, or dot-blots. Microarrays can be coupled with PCR where they serve as a set of parallel dot-blots to enhance product detection and identification. Microarrays are composed of many discretely located probes on a solid substrate such as glass. Each probe is composed of a sequence that is complimentary to a pathogen-specific gene sequence. PCR is used to amplify one or more genes and the products are then hybridized to the array to identify species-specific polymorphism within one or more genes. We illustrate this type of array using 16S rDNA probes suitable for distinguishing between several salmonid pathogens. We also describe the use of microarrays for direct detection of either RNA or DNA without the aid of PCR, although the sensitivity of these systems currently limits their application for pathogen detection. Finally, microarrays can also be used to "fingerprint" bacterial isolates and they can be used to identify diagnostic markers suitable for developing new PCR-based detection assays. We illustrate this type of array for subtyping an important food-borne pathogen, Listeria monocytogenes.

Actinomycetales Infections↗

Evolution of microbial populations during traditional Feta cheese manufacture and ripening.

In three different dairies (A, B and C) located in Peloponess region (Southern Greece), traditional Feta cheese trials took place February to March using mixtures of sheep's and goat's milk. Only small variations in the evolution of microbial groups were observed during the whole ripening period. The main groups, such as thermophilic cocci, mesophilic lactococci, thermophilic lactobacilli, nonstarter lactic acid bacteria (NSLAB), presumptive Leuconostoc, enterococci and micrococci, reached their highest levels during the first 16 days, and then declined approximately 1-2 log units until the end of ripening. The remaining groups investigated, comprising yeasts, coliforms and Escherichia coli, were highest at day 4. The yeasts remained constant, while coliforms and E. coli decreased sharply and were not detectable after 120 days of ripening. A number of 146 isolates (dairy A) taken from all stages of the manufacturing and ripening process were purified and studied. Lactobacillus plantarum (58/146) and isolates of related species Lactobacillus pentosus and Lactobacillus paraplantarum (16/146) were the most common microorganisms found during cheese ripening. Streptococcus thermophilus (23/146) and Lactobacillus delbrueckii subsp. bulgaricus (20/146) were detected in high levels up to 20 days, and then gradually reduced. Enterococcus faecium (29/146) was found in all manufacturing and ripening stages.

Animals↗

Effect of nitrate and nitrite curing salts on microbial changes and sensory quality of non-fermented sausages.

The effects of nitrate and nitrite curing salts on microbial changes and sensory quality of non-fermented sausages of small diameter were investigated. During pre-ripening (day 5), levels of lactic acid bacteria and yeasts were slightly higher in nitrite-made sausages than in those made with nitrate. In contrast, nitrite discouraged the growth of psychrotrophs as occurs in fermented sausages. By the end of ripening (day 26), levels of microorganisms were similar in both batches of sausages except for psychrotrophs being higher in those made with nitrite. Nitrate-made sausages showed higher aroma and taste intensity.

Animals↗

Brining time effect on physicochemical and microbiological parameters in Idiazábal cheese.

Physicochemical and microbiological parameters were compared for three brining times (12, 24 and 36 h) for fresh, young, semihard and hard Idiazabal cheese. Longer brining time produced higher salt, dry matter and salt-moisture ratio and lower water activity values for all types of cheese according to ripening time, while non-significant changes were observed for pH. In fresh cheese (1-15 days ripening), non-significant differences for microbiological counts in relation to brining time were observed, except for moulds. In young and hard cheeses, Lactobacillus and Leuconostoc showed lower counts with longer brining times. In contrast, Micrococaceae, yeast and moulds were stimulated by higher salt content in matured cheeses. In addition. this work has proved that there are lower water activity values and lower microbiological counts in longer-matured Idiazabal cheeses. For the different brining and ripening times, positive correlations were observed among most of the microbial groups studied, but a different behavior was established for Enterococcus, Clostridium tyrobutyricum, yeast and moulds.

Cheese↗

Infochemicals in mosquito host selection: human skin microflora and Plasmodium parasites.

The interaction between the African malaria vector Anopheles gambiae and its human host has traditionally been viewed within a bitrophic context, considering only the human and the mosquito. Recently, the influence of the Plasmodium parasite on the interaction has been recognized, because it affects the physiology and/or behaviour of humans and mosquitoes. However, studies on odour-mediated host-seeking behaviour of An. gambiae and other Diptera have provided evidence that a fourth group of organisms should be taken into consideration. Human skin microflora play a role in the production of odorous compounds that might function as kairomones for mosquitoes. Here, Marieta Braks, Rob Anderson and Bart Knols introduce the role of human microflora into the process of odour-mediated host selection and review the interaction in a multipartite context so as to identify research avenues that will enhance our limited knowledge of this aspect of malaria transmission.

Animals↗

Spectroscopic studies on poly(ethylene glycol)-lysozyme interactions.

In the present paper, different spectroscopic methods were applied to evaluate conformational changes of hen egg-white lysozyme (HEWL) in various solvents and in the presence of poly(ethylene glycol) (PEG). In citrate (0.007M, pH=6), or in Tris (0.1M, pH=7.4), no conformational change of the protein was measured across the range of concentrations tested. In addition, HEWL in ultra-pure water revealed no irreversible conformational change and no activity loss, at least at low concentrations (< or =0.2mg/ml). Whereas PEG can induce a reorganization of water molecules, no change of the secondary and tertiary protein conformations was observed in the presence of PEG. In addition, in the presence of PEG of various molecular weights, no change of enzymatic activity of the HEWL was observed across the range of concentrations tested.

Circular Dichroism↗

Arthrobacter, Brachybacterium and Planococcus isolates identified from antarctic sea ice brine. Description of Planococcus mcmeekinii, sp. nov.

Three facultative psychrophilic Gram-positive bacterial strains were isolated from brine samples from the sea ice community in Antarctica. All strains were coccoid to rod-shaped and exhibited broad salinity and temperature ranges for growth. The three strains were subjected to 16S rDNA sequencing and subsequent phylogenetic analysis. All possess unique 16S rDNA sequences indicating they are new, previously unreported organisms. Phylogenetic analyses coupled with phenotypic characterization indicated that one of the strains is most closely related to the low mol% G + C genus Planococcus for which a new species, P. mcmeekinii, is proposed. The two other strains are members of the high mol% G + C Gram-positive bacteria and most closely related to the genera Arthrobacter and Brachybacterium. This study reports the first phylogenetic evidence that Gram-positive bacteria reside in the marine sea ice brine.

Actinomycetales↗

Identification of oxacillin-susceptible and oxacillin-resistant Staphylococcus aureus using commercial latex agglutination tests.

Four hundred thirteen Staphylococcus sp. were identified by Staphaurex, Staphaurex Plus, and BACTiStaph kits using tube coagulase as reference. Among 222 coagulase-positive isolates, 56 were oxacillin-resistant Staphylococcus aureus. All tests were accurate in distinguishing between coagulase-positive and -negative staphylococci with sensitivities and specificities > or = 97% and only nine discrepancies.

Agglutination Tests↗