Characterisation of mecA gene negative Staphylococcus aureus isolated from bovine mastitis milk from Northern Germany.

Staphylococcus aureus (S. aureus) is an important causative agent of contagious intermammary infections in dairy cattle. S. aureus is also considered as an important foodborne pathogen and cause of food poisoning cases and outbreaks worldwide. In order to understand the molecular ecology of S. aureus, the present study compared phenotypic and genotypic characteristics of 70 S. aureus isolates from bovine mastitis milk samples collected during the period from August 2001 to March 2014 in different regions of Northern Germany. The S. aureus isolates were characterised phenotypically, as well as genotypically for their genetic diversity using multi-locus sequence typing (MLST), spa typing and the presence of virulence genes encoding 16 staphylococcal enterotoxins (sea-selu), toxic shock syndrome toxin (tst), thermonuclease (nuc), clumping factor (clfA and clfB), coagulase (coa) and the methicillin resistance gene mecA. A total of 16 sequence types were grouped into eight clonal complexes (CCs), and 17 spa types were identified. These included six novel sequence types and one novel spa type. The majority of bovine mastitis milk-associated sequence types belonged to the clonal complex CC5, CC97, CC133, and CC151 and showed closely related genotypes or lineages with sequence types of human origin. The genotype CC133 (ST133-t1403) was predominant, constituting 27.1% of the isolates. In addition, the S. aureus isolates displayed nine different enterotoxigenic profiles. All S. aureus were methicillin-susceptible (MSSA). The current study provides new information on phenotypic and genotypic traits of S. aureus isolates from bovine mastitis. The comparison of characteristics of isolates from the present study originating from mastitis milk showed similarities with human isolates. This might help to better understand the distribution of S. aureus in the one health context.

Assessment of pasteurisation of edible insects using enzymatic tests (activity of alkaline phosphatase and lactoperoxidase) applied in dairy products.

Industrialising edible insects goes along with quality control and hazard analysis and critical control points. One of those steps is assessing heat treatment. For the present contribution, the potential of enzymatic heat assessment tests used in the dairy industry (alkaline phosphatase and lactoperoxidase) to detect heat treatment in several insect species ( Acheta domesticus, Gryllus assimilis, Gryllus bimaculatus, Locusta migratoria, Schistocerca gregaria, Chilecomadia moorei, Galleria mellonella, Bombyx mori, Pachnoda marginata, Tenebrio molitor, Zophobas atratus, Apis mellifera, and Hermetia illucens) was evaluated. Insect material was homogenised, diluted, and the enzymatic tests (Lactognost®, Peroxtesmo®) were carried with these liquids as if they were milk. All species but C. moorei, B. mori, P. marginata, and A. mellifera showed alkaline phosphatase activity in raw samples and none in heated (10 min at 100 ℃) ones, while only G. mellonella, T. molitor, and Z. atratus reacted accordingly with lactoperoxidase. In trial 2 focusing only on alkaline phosphatase activity, inactivation of the enzyme after 5, 10, and 15 min of heating occurred species specific within a range of 60-86 ℃, i.e. within ordinary pasteurisation schemes. Thus and for the time being, heat treatment in many edible insect species can be assessed using alkaline phosphatase activity test kits. In contrast to milk samples, positive results may display bluish or greenish colours, and the time until a reliable reading is possible is extended to 1-1.5 h (24 h in the case of Gryllidae).

Microbiology and foodborne pathogens in honey.

Honey has been considered a relatively safe foodstuff due to its compositional properties, with infant botulism caused by Clostridium botulinum being the most prominent health risk associated with it. Our review is focused on the honey microflora along the food chain and evaluates the pathogenic potential of those microorganisms found in honey. This product may contain a great variety of bacteria and, particularly, fungi that eventually entered the food chain at an early stage (e.g., via pollen). For many of these microorganisms, opportunistic infections in humans have been recorded (e.g., infections by Staphylococcus spp., Citrobacter spp., Escherichia coli, Hafnia alvei, Aspergillus spp., Fusarium spp., Trichoderma spp., Chaetomium spp.), although direct infections via honey were not registered.

Development and validation of a loop mediated isothermal amplification (LAMP) assay for the detection of Staphylococcus aureus in bovine mastitis milk samples.

Staphylococcus (S.) aureus is one of the most important animal pathogens causing bovine mastitis. Also, it is a major human pathogen that may produce a variety of toxins which cause staphylococcal food poisoning. In the present study a LAMP assay based on gene nuc to identify S. aureus was developed and validated. The specificity of the LAMP assay was confirmed by using 70 S. aureus isolates and 21 non-S. aureus strains. The optimal temperature-time combination to amplify gene nuc successfully was 65 °C and 30 min. The analytical sensitivity of the developed LAMP assay was 0.26 pg of S. aureus DNA per reaction. The limit of detection evaluated with milk spiked with S. aureus was 9 × 102 CFU mL-1. The final results of this assay were available within less than 2 h. The present study showed that the LAMP assay based on gene nuc appeared to be rapid and simple, and could also be used to identify S. aureus isolates from mastitis milk of dairy cows.

Food Safety Hazards and Microbiological Zoonoses in European Meat Imports Detected in Border Inspection in the Period 2008-2013.

Microbiological contaminations and other food safety hazards are omnipresent within the European Union (EU) and a considerable risk for consumers, particularly in imported meat and meat products. The number of rejections at external EU borders has been increasing in recent years. Official authorities in each member state are therefore obliged to notify border rejections of food and animal feed due to a direct or indirect risk to human or animal health. This study explored the trends and temporal and spatial distribution of notifications on food safety hazards between January 2008 and December 2013 with a special emphasis on microbiological zoonoses in meat and meat products including poultry at border checks resulting from the rapid alert system for food and feed (RASFF). Results indicated that border rejection notifications are increasing exponentially, frequently due to Salmonella in poultry and shiga-toxin-producing E. coli in meat and meat products.

Similarities and differences among fluid milk products: traditionally produced, extended shelf life and ultrahigh-temperature processed.

Extended shelf life milk is a relatively new kind of fluid milk, generally manufactured by high-temperature treatment and/or micro-filtration. Being advertised as 'pasteurized milk with an extended shelf life', its flavour, compositional quality and labelling was questioned. Extended shelf life (high-temperature treatment), pasteurized ('traditionally produced') and ultrahigh-temperature milk were, therefore, compared at the beginning and end of shelf life. In triangle tests, panellists distinguished clearly between all products. High-temperature treatment milk's flavour was closer to ultrahigh-temperature and traditionally produced milk in the beginning and at the end of shelf life, respectively. Physicochemically and bacteriologically, all three types could be distinguished. Since 'extended shelf life' comprises many process varieties (each affecting flavour differently), consumer information and appropriate package labelling beyond 'long-lasting' is necessary, e.g. by mentioning the heat treatment applied.

In vitro effects of very low levels of aflatoxin B1 on free radicals production and bactericidal activity of bovine blood neutrophils.

As one of the most potent and hazardous feed/food-originated mycotoxins, aflatoxin (AF) B1 is regarded as a potent immunosuppressor in dairy cows. Neutrophils (PMN), as key effector cells against pathogens, have a high potential to kill engulfed microbes. To investigate the in vitro effects of very low doses of AFB1 on blood PMN functions, we examined the effects of biologically relevant concentrations of AFB1 on the phagocytosis and non-phagocytosis dependent luminol, representative of mainly intracellular free radicals, and isoluminol, representative of mainly extracellular free radicals, chemiluminescence (CL), necrosis and apoptosis of PMN. Isolated blood PMN from healthy dairy cows (n=12) were exposed to 0, 0.01, 0.05 and 0.5 ng/ml of AFB1 for 0.5 and 18 h depending on the assay. Further, blood PMN of healthy dairy cows (n=8) were exposed to 0.5 ng/ml of AFB1 for 3h and myeloperoxidase (MPO) activity, superoxide anion (O2⁻) production, phagocytosis and killing activities against Staphylococcus (S.) aureus and Escherichia (E.) coli, were examined. Though the effect of extremely low doses of AFB1 were less pronounced, at 0.5 ng/ml the production of free radicals was greatly enhanced, especially extracellularly. In contrast to isoluminol CL, the AFB1-treated PMN showed a remarkably impaired phagocytosis-depended luminol CL. PMN necrosis and apoptosis were not affected by AFB1. MPO activity, O2⁻ production, phagocytosis rates and killing of E. coli and S. aureus by AFB1-treated PMN were significantly lower than those of non-treated ones. Our results show the extracellularly pro-oxidant and antiphagocytic properties of very low doses of AFB1 for bovine PMN. The scope of the suppressive effects of the in vitro AFB1 levels on cellular innate immune functions should be considered for high yielding dairy cows.

Inverse halftoning using binary permutation filters.

The problem of reconstructing a continuous-tone image given its ordered dithered halftone or its error-diffused halftone image is considered. We develop a modular class of nonlinear filters that can reconstruct the continuous-tone information preserving image details and edges that provide important visual cues. The proposed nonlinear reconstruction algorithms, denoted as binary permutation filters, are based on the space and rank orderings of the halftone samples provided by the multiset permutation of the "on" pixels in a halftone observation window. For a given window size, we obtain a wide range of filters by varying the amount of space-rank ordering information utilized in the estimate. For image reconstructions from ordered dithered halftones, we develop periodically space-varying filters that can account for the periodical nature of the underlying screening process. A class of suboptimal but simpler space-invariant reconstruction filters are also proposed and tested. Constrained LMS type algorithms are employed for the design of reconstruction filters that minimize the reconstruction mean squared error. We present simulations showing that binary permutation filters are modular, robust to image source characteristics, and that they produce high visual quality image reconstruction.

The effect of split virus influenza vaccination on theophylline pharmacokinetics.

The effects of split virus influenza vaccine on the pharmacokinetics of theophylline and the in vivo and in vitro induction of interferon were studied in 23 volunteers. No effect on the total body clearance, half-life, or mean residence time of theophylline was found as a result of influenza virus vaccine. Additionally, no interferon activity was detected in vivo for as long as 24 h after vaccination. Finally, no production of interferon occurred in tonsil or peripheral lymphocyte cultures inoculated with split virus vaccine. We conclude that split virus influenza vaccine does not affect theophylline pharmacokinetics. In addition, we find no evidence that split virus influenza vaccine functions as an interferon inducer in humans.