Reduction and Growth Inhibition of Listeria monocytogenes by Use of Anti-Listerial Nisin, P100 Phages and Buffered Dry Vinegar Fermentates in Standard and Sodium-Reduced Cold-Smoked Salmon.

Cold-smoked salmon are ready-to-eat products that may support the growth of pathogenic Listeria monocytogenes during their long shelf-life. Consumption of such contaminated products can cause fatal listeriosis infections. Another challenge and potential risk associated with CS salmon is their high levels of sodium salt. Excess dietary intake is associated with serious health complications. In the present study, anti-listerial bacteriocin (nisin), P100 bacteriophages (Phageguard L, PGL) and fermentates (Verdad N6, P-NDV) were evaluated as commercial bio-preservation strategies for increased control of L. monocytogenes in standard (with NaCl) and sodium-reduced (NaCl partially replaced with KCl) CS salmon. Treatments of CS salmon with nisin (1 ppm) and PGL (5 × 107 pfu/cm2) separately yielded significant initial reductions in L. monocytogenes (up to 0.7 log) compared to untreated samples. Enhanced additive reductions were achieved through the combined treatments of nisin and PGL. Fermentates in the CS salmon inhibited the growth of Listeria but did not lead to its eradication. The lowest levels of L. monocytogenes during storage were observed in nisin- and PGL-treated CS salmon containing preservative fermentates and stored at 4 °C, while enhanced growth was observed during storage at an abusive temperature of 8 °C. Evaluation of industry-processed standard and sodium-replaced CS salmon confirmed significant effects with up to 1.7 log reductions in L. monocytogenes levels after 34 days of storage of PGL- and nisin-treated CS salmon-containing fermentates. No differences in total aerobic plate counts were observed between treated (PGL and nisin) or non-treated standard and sodium-reduced CS salmon at the end of storage. The microbiota was dominated by Photobacterium, but with a shift showing dominance of Lactococcus spp. and Vagococcus spp. in fermentate-containing samples. Similar and robust reductions in L. monocytogenes can be achieved in both standard and sodium-replaced CS salmon using the bio-preservation strategies of nisin, PGL and fermentates under various and relevant processing and storage conditions.

Decision tree scoring system to guide selection for consumer preference in sweetpotato breeding trials.

BACKGROUND: Previously, a lexicon and protocol for quantitative descriptive analysis (QDA) was established for the Uganda sweetpotato breeding program. The implication of QDA scores for priority sensory attributes on consumer preference should be determined to interpret results efficiently and make decisions effectively. The present study aimed to develop a gender-responsive decision tree to obtain an overall sweetpotato eating quality score to facilitate demand-led targeted breeding selection. It focused on Kamuli and Hoima districts (Uganda) and uses pre-lease advanced clones ('NKB3', 'NKB105', 'NKB135', 'D11' and 'D20'), released varieties ('NASPOT 8' and 'NAROSPOT 1') and landraces ('Muwulu-Aduduma', 'Umbrella'). RESULTS: Including boiled sweetpotato sensory characteristics, namely mealy, sweet taste, sweetpotato smell, firm and not fibrous, in breeding design would benefit end-users, especially women given their role in varietal selection, food preparation and marketing. 'D20', 'NASPOT 8' and 'NAROSPOT 1' were most liked in both districts. 'NKB3' and 'D11' were the least liked in Hoima, whereas 'Muwulu-Aduduma' was the least liked in Kamuli. There was a positive correlation between color and overall liking (r2 = 0.8) and consumers liked the color (average rating ≥ 6 on a nine-point hedonic scale) of all genotypes. Threshold values (average rating on 11-point scales) for consumer acceptability were identified (sweet taste = 6, sweetpotato aroma and flavor = 6, firmness = 3, and mealiness = 4). A regression decision tree tool was created to calculate an eating quality selection index when screening lines in breeding programs using the values. CONCLUSION: Decision trees that include consumer needs and gender considerations would facilitate demand-led breeding and make varietal selection in sweetpotato breeding programs more effective. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Mapping the Kitchen Microbiota in Five European Countries Reveals a Set of Core Bacteria across Countries, Kitchen Surfaces, and Cleaning Utensils.

The residential kitchen is often heavily colonized by microbes originating from different sources, including food and human contact. Although a few studies have reported the bacterial composition in cleaning utensils and surface samples there is limited knowledge of the bacterial diversity across different sample types, households, and countries. As part of a large European study, we have identified the microbiota of 302 samples from cleaning utensils (sponges and cloths), kitchen surfaces (sinks, cutting boards, countertops, tap handles, and a pooled sample of other handles) in 74 households across 5 countries (France, Hungary, Norway, Portugal, and Romania). In total, 31 bacterial phyla were identified, with Proteobacteria, Firmicutes, Bacteroidota, and Actinobacteria being the most abundant. Despite large variations in households with respect to kitchen standards, kitchen practices, cleaning regimes, and diet and considerable differences in bacterial diversity between samples, eight bacterial genera/families commonly associated with environmental sources were identified in most samples and defined as a core microbiota: Acinetobacter, Pseudomonas, Enhydrobacter, Enterobacteriaceae, Psychrobacter, Chryseobacterium, Bacillus, and Staphylococcus. These genera/families were also among the bacteria with the highest relative abundance across all samples, in addition to Yersiniaceae, Kocuria, Pantoea, and Streptococcus. Taxa associated with potential pathogens and fecal indicators were low in abundance but broadly distributed throughout the households. The microbial composition of surface samples indicated that the microbial composition on kitchen surfaces is more characteristic for the particular country than the object type, while the microbiota of cleaning utensils was similar across countries but differed between types (sponge or cloth). IMPORTANCE There is limited knowledge of the characteristics, differences, and similarities of the bacterial composition in residential kitchens. Here, we report the microbiota of cleaning utensils (sponges and cloths) and five different surface samples in 74 households across five European countries. In addition to increasing the knowledge of the kitchen microbiota from many geographical areas, this study identified a core microbiota in European residential kitchens despite large variations in kitchen practices and kitchen design and standards across countries and households.

Levels of nitrate, nitrite and nitrosamines in model sausages during heat treatment and in vitro digestion - The impact of adding nitrite and spinach (Spinacia oleracea L.).

Nitrite derivatives react with endogenous precursors forming N-nitrosamines associated with development of colorectal cancer. The present study aims to investigate the formation of N-nitrosamines in sausage during processing and in vitro gastrointestinal digestion after adding sodium nitrite and/or spinach emulsion. The INFOGEST digestion protocol was used to simulate the oral, gastric, and small intestinal phases of digestion, and sodium nitrite was added in the oral phase to mimic the input of nitrite from saliva as it has shown to affect the endogenous formation of N-nitrosamines. The results show that the addition of spinach emulsion, in spite of it being a source of nitrate, did not affect the nitrite content in either batter, sausage, or roasted sausage. The levels of N-nitrosamines increased with the added amount of sodium nitrite, and further formation of some volatile N-nitrosamines was observed during roasting and in vitro digestion. In general, N-nitrosamine levels in the intestinal phase followed the same trend as in the undigested products. The results further indicate that nitrite present in saliva may cause a significant increase in N-nitrosamine levels in the gastrointestinal tract and that bioactive components in spinach may protect against the formation of volatile N-nitrosamines both during roasting and digestion.

Microbial Safety and Sensory Analyses of Cold-Smoked Salmon Produced with Sodium-Reduced Mineral Salts and Organic Acid Salts.

Cold-smoked (CS) salmon contains high levels of sodium salts, and excess dietary sodium intake is associated with an array of health complications. CS salmon may also represent a food safety risk due to possible presence and growth of the foodborne pathogen Listeria monocytogenes which may cause fatal human infections. Here we determine how reformulated CS salmon using commercial sodium-reduced salt replacers containing KCl (e.g., Nutek, Smart Salt, SOLO-LITE) and acetate-based preservative salts (Provian K, proviant NDV) affect sensory properties, quality, and microbial safety. Initial sensory screening of sodium-reduced CS salmon was followed by L. monocytogenes growth analyses in selected variants of reformulated CS salmon, and finally by analyses of CS salmon variants produced in an industrial smokehouse. Projective mapping indicated overall minor sensory changes in sodium-replaced samples compared with a conventional product with NaCl. Growth of L. monocytogenes was temperature-dependent (4 °C vs. 8 °C storage) with similar growth in sodium-reduced and conventional CS salmon. The addition of 0.9% of the preservative salts Provian K or Provian NDV gave up to 4 log lower L. monocytogenes counts in both sodium-reduced and conventional cold-smoked salmon after 29 days of chilled storage. No changes in pH (range 6.20−6.33), aw levels (range 0.960−0.973), or weight yield (96.8 ± 0.2%) were evident in CS salmon with salt replacers or Provian preservative salts. Analyses of CS salmon produced with selected mineral salt and preservative salt combinations in an industrial salmon smokery indicated marginal differences in sensory properties. Samples with the preservative salt Provian NDV provided L. monocytogenes growth inhibition and low-level total viable counts (<2.8 log/g) dominated by Photobacterium and Carnobacterium during storage. Production of sodium-reduced CS salmon with inhibiting salts provides a simple method to achieve a healthier food product with increased food safety.

Analysing microbiome intervention design studies: Comparison of alternative multivariate statistical methods.

The diet plays a major role in shaping gut microbiome composition and function in both humans and animals, and dietary intervention trials are often used to investigate and understand these effects. A plethora of statistical methods for analysing the differential abundance of microbial taxa exists, and new methods are constantly being developed, but there is a lack of benchmarking studies and clear consensus on the best multivariate statistical practices. This makes it hard for a biologist to decide which method to use. We compared the outcomes of generic multivariate ANOVA (ASCA and FFMANOVA) against statistical methods commonly used for community analyses (PERMANOVA and SIMPER) and methods designed for analysis of count data from high-throughput sequencing experiments (ALDEx2, ANCOM and DESeq2). The comparison is based on both simulated data and five published dietary intervention trials representing different subjects and study designs. We found that the methods testing differences at the community level were in agreement regarding both effect size and statistical significance. However, the methods that provided ranking and identification of differentially abundant operational taxonomic units (OTUs) gave incongruent results, implying that the choice of method is likely to influence the biological interpretations. The generic multivariate ANOVA tools have the flexibility needed for analysing multifactorial experiments and provide outputs at both the community and OTU levels; good performance in the simulation studies suggests that these statistical tools are also suitable for microbiome data sets.

Does Responsiveness to Basic Tastes Influence Preadolescents' Food Liking? Investigating Taste Responsiveness Segment on Bitter-Sour-Sweet and Salty-Umami Model Food Samples.

The objective of this study was to investigate the relationships between taste responsiveness and food liking in preadolescents. Model food samples of grapefruit juice (GF) and vegetable broth (VB) modified with four additions of sucrose and sodium chloride, respectively, were employed. Intensity perception for sweetness, sourness, and bitterness were measured in GF while saltiness and umami were measured in VB. The children (N = 148) also completed food choice, familiarity, stated liking and neophobia questionnaires. The test was conducted at school, with instructions provided remotely via video call. Four segments were defined differing in basic taste responsiveness. Segments and sucrose concentrations significantly affected liking for GF, while no significant effect of segments and sodium chloride concentrations occurred on liking for VB. An increasing sucrose concentration was positively associated with liking for GF only in the segment with low responsiveness to bitter and sour tastes. No significant differences across segments were found for food choice, familiarity, stated liking, and neophobia. Conclusively, relationships between taste responsiveness and liking are product and basic taste-dependent in addition to being subject-dependent. Strategies to improve acceptance by using sucrose as a suppressor for warning sensations of bitterness and sourness can be more or less effective depending on individual responsiveness to the basic tastes.

High Oxygen Packaging of Atlantic Cod Fillets Inhibits Known Spoilage Organisms, but Sensory Quality Is Not Improved Due to the Growth of Carnobacterium/Carnobacteriaceae.

Improved quality control and prolonged shelf life are important actions in preventing food waste. To get an overview of the bacterial diversity of fillets from live stored mature Atlantic cod, bacterial isolates were identified before and after storage (air and vacuum) and freezing/thawing. Based on the load of dominating bacteria, the effect of different packaging methods and a short freezing/thawing process on prolonged shelf-life was evaluated (total viable counts, bacteriota, sensory attributes, and volatile components). Hand filleted (strict hygiene) cod fillets had a low initial bacterial load dominated by the spoilage organism Photobacterium, whereas industrially produced fillets had higher bacterial loads and diversity (Pseudomonas, Arthrobacter, Psychrobacter, Shewanella). The identified bacteria after storage in vacuum or air were similar to the initially identified bacteria. Bacteriota analysis showed that a short time freezing/thawing process reduced Photobacterium while modified atmosphere packaging (MAP; 60% CO2/40% O2 or 60% CO2/40% N2) inhibited the growth of important spoilage bacteria (Photobacterium,Shewanella, Pseudomonas) and allowed the growth of Carnobacterium/Carnobacteriaceae and Acinetobacter. Despite being dominated by Photobacterium, fresh fillets stored in MAP 60% CO2/40% N2 demonstrated better sensory quality after 13 days of storage than fillets stored in MAP 60% CO2/40% O2 (dominated by Carnobacterium/Carnobacteriaceae). Carnobacterium spp. or other members of Carnobacteriaceae may therefore be potential spoilage organisms in cod when other spoilage bacteria are reduced or inhibited.

Basic taste sensitivity, eating behaviour, food propensity and BMI of preadolescent children: How are they related?

Background: Taste sensitivity has been reported to influence children's eating behaviour and contribute to their food preferences and intake. This study aimed to investigate the associations between taste sensitivity, eating behaviour, food frequency and BMI (Body Mass Index) in preadolescents. Methods: Preadolescents' taste sensitivity was measured by detection threshold of sweetness (sucrose), sourness (citric acid), saltiness (sodium chloride), bitterness (caffeine, quinine), and umami (monosodium glutamate). In addition, the Child Eating Behaviour Questionnaire (CEBQ), the Food Propensity Questionnaire (FPQ) measuring food frequency, and the children's body weight and height were completed by the parents. A total of 69 child-parent dyads participated (preadolescents mean age =10.9 years). Results: Taste sensitivity to caffeine bitterness was significantly associated with eating behaviour in food responsiveness, emotional overeating, and desire to drink. The preadolescents who were less sensitive to caffeine bitterness had higher food responsiveness scores. Those who were less sensitive to caffeine bitterness and to sweetness had higher emotional overeating scores. In addition, preadolescents who were less sensitive to sourness and bitterness of both caffeine and quinine demonstrated to have higher scores in desire to drink. There was no association between taste sensitivity and FPQ, but significant differences were observed across preadolescents' BMI for FPQ of dairy food items, indicating higher consumption of low-fat milk in the overweight/obese compared to the underweight/normal-weight subjects. There was no significant difference in taste sensitivity according to BMI. Preadolescents' eating behaviour differed across BMI, demonstrating a positive association between BMI and food approach, and a negative association between BMI and food avoidance. Conclusions: This study contributes to the preliminary understanding of the relationships between taste sensitivity and eating behaviour in preadolescents. The results may be used to develop effective strategies to promote healthy eating practices by considering taste sensitivity in preadolescents.

Investigating the Relationships between Basic Tastes Sensitivities, Fattiness Sensitivity, and Food Liking in 11-Year-Old Children.

This study investigates the relationships between basic tastes and fattiness sensitivity and food liking in 11-year-old children. The basic taste sensitivity of 106 children was measured using different methods, namely detection (DT) and recognition (RT) thresholds, and taste responsiveness. Caffeine and quinine (bitter), sucrose (sweet), citric acid (sour), sodium chloride (salty), and monosodium glutamate (umami) were investigated for DT and RT at five concentrations in water solutions. In addition, taste responsiveness and liking were collected for the high-intensity concentrations. PROP (6-n-propylthiouracil) responsiveness was tested on paper strips. Fattiness sensitivity was measured by a paired comparison method using milk samples with varying fat content. Liking for 30 food items was recorded using a food-list questionnaire. The test was completed in a gamified "taste detective" approach. The results show that DT correlates with RT for all tastes while responsiveness to PROP correlates with overall taste responsiveness. Caffeine and quinine differ in bitterness responsiveness and liking. Girls have significantly lower DTs than boys for bitterness and sweetness. Food liking is driven by taste and fattiness properties, while fatty food liking is significantly influenced by fattiness sensitivity. These results contribute to a better holistic understanding of taste and fattiness sensitivity in connection to food liking in preadolescents.

Growth Behavior of Listeria monocytogenes in a Traditional Norwegian Fermented Fish Product (Rakfisk), and Its Inhibition through Bacteriophage Addition.

Listeria monocytogenes may persist in food production environments and cause listeriosis. In Norway, a product of concern is the traditional and popular fermented fish product "rakfisk", which is made from freshwater salmonid fish by mild-salting and brine maturation at low temperatures for several months. It is eaten without any heat treatment, and L. monocytogenes, therefore, poses a potential hazard. We investigated the effect of salt and temperature on the growth of L. monocytogenes in rakfisk during the 91 days of maturation. The amounts of organic acids produced during fermentation were too low to inhibit growth of L. monocytogenes. Temperature was clearly the most important parameter for controlling L. monocytogenes. At 7 °C, approximately 2 log growth was observed during the first 14 days of fermentation, and the level of L. monocytogenes thereafter remained constant. At 4 °C, only a little growth potential of the pathogen was recorded. We also investigated the effect of the anti-Listeria bacteriophage P100 on rakfisk with added L. monocytogenes. The phage was introduced to the L. monocytogenes-inoculated fish before fermentation, and an average of 0.9 log reduction was observed throughout the fermentation period. This is the first study of L. monocytogenes behavior in rakfisk and points to possible measures for increasing the product safety.

Projective mapping based on choice or preference: An affective approach to projective mapping.

This work explores a new affective approach to projective mapping, based on consumers' choices or preferences. Two sessions, one week apart, were performed with the same consumers, using whole bread as a case study. Overall liking ratings (OL) were gathered in blind conditions and samples were also profiled by a trained panel using generic descriptive analysis. Three projective mapping tests were performed in different scenarios. Consumers' categorization and product descriptions were explored when consumers based their positioning on the products' similarities and differences (analytical approach, "classic napping") both in blind and informed conditions, and when consumers were focusing on their preference or choice (affective approach). The affective approach to projective mapping successfully revealed consumers' drivers of liking and choice from a holistic perspective, where consumers summarized their main drivers for categorizing products as they would do when choosing in real life situations, based on their preferences.

Influence of consumers' cognitive style on results from projective mapping.

Projective mapping (PM), one of the most holistic product profiling methods in approach, is increasingly being used to uncover consumers' perception of products and packages. Assessors rely on a process of synthesis for evaluating product information, which would determine the relative importance of the perceived characteristics they use for mapping them. Individual differences are expected, as participants are not instructed on the characteristics to consider for evaluating the degree of difference among samples, generating different perceptual spaces. Individual differences in cognitive style can affect synthesis processes and thus their perception of similarities and differences among samples. In this study, the influence of the cognitive style in the results of PM was explored. Two consumer studies were performed, one aimed at describing intrinsic sensory characteristics of chocolate flavoured milk and the other one looking into extrinsic (package only) of blueberry yogurts. Consumers completed the wholistic-analytic module of the extended Verbal Imagery Cognitive Styles Test & Extended Cognitive Style Analysis-Wholistic Analytic Test, to characterize their cognitive style. Differences between wholistic and analytic consumers in how they evaluated samples using projective mapping were found in both studies. Analytics separated the samples more in the PM perceptual space than wholistic consumers, showing more discriminating abilities. This may come from a deeper analysis of the samples, both from intrinsic and extrinsic point of views. From a sensory perspective (intrinsic), analytic consumers relied on more sensory characteristics, while wholistic mainly discriminated samples according to sweetness and bitterness/chocolate flavour. In the extrinsic study however, even if analytic consumers discriminated more between packs, they described the products using similar words in the descriptive step. One important recommendation coming from this study is the need to consider higher dimensions in the interpretation of projective mapping tasks, as the first dimensions could underestimate the complexity of the perceptual space; currently, most applications of PM consider two dimensions only, which may not uncover the perception of specific groups of consumers.

A novel role for pigment genes in the stress response in rainbow trout (Oncorhynchus mykiss).

In many vertebrate species visible melanin-based pigmentation patterns correlate with high stress- and disease-resistance, but proximate mechanisms for this trait association remain enigmatic. Here we show that a missense mutation in a classical pigmentation gene, melanocyte stimulating hormone receptor (MC1R), is strongly associated with distinct differences in steroidogenic melanocortin 2 receptor (MC2R) mRNA expression between high- (HR) and low-responsive (LR) rainbow trout (Oncorhynchus mykiss). We also show experimentally that cortisol implants increase the expression of agouti signaling protein (ASIP) mRNA in skin, likely explaining the association between HR-traits and reduced skin melanin patterning. Molecular dynamics simulations predict that melanocortin 2 receptor accessory protein (MRAP), needed for MC2R function, binds differently to the two MC1R variants. Considering that mRNA for MC2R and the MC1R variants are present in head kidney cells, we hypothesized that MC2R activity is modulated in part by different binding affinities of the MC1R variants for MRAP. Experiments in mammalian cells confirmed that trout MRAP interacts with the two trout MC1R variants and MC2R, but failed to detect regulation of MC2R signaling, possibly due to high constitutive MC1R activity.

Extrusion of barley and oat influence the fecal microbiota and SCFA profile of growing pigs.

The effect of extrusion of barley and oat on the fecal microbiota and the formation of SCFA was evaluated using growing pigs as model system. The pigs were fed a diet containing either whole grain barley (BU), oat groat (OU), or their respective extruded samples (BE and OE). 454 pyrosequencing showed that the fecal microbiota of growing pigs was affected by both extrusion and grain type. Extruded grain resulted in lower bacterial diversity and enrichment in operational taxonomic units (OTUs) affiliated with members of the Streptococcus, Blautia and Bulleidia genera, while untreated grain showed enrichment in OTUs affiliated with members of the Bifidobacterium and Lactobacillus genera, and the butyrate-producing bacteria Butyricicoccus, Roseburia, Coprococcus and Pseudobutyrivibrio. Untreated grain resulted in a significant increase of n-butyric, i-valeric and n-valeric acid, which correlated with an increase of Bifidobacterium and Lactobacillus. This is the first study showing that cereal extrusion affects the microbiota composition and diversity towards a state generally thought to be less beneficial for health, as well as less amounts of beneficial butyric acid.

Unveiling an abundant core microbiota in the human adult colon by a phylogroup-independent searching approach.

The potential presence of widespread and stable bacterial core phylogroups in the human colon has promoted considerable attention. Despite major efforts, no such phylogroups have yet been identified. Therefore, using a novel phylogroup- and tree-independent approach, we present a reanalysis of 1,114,722 V2 region and 71,550 near full-length 16S rRNA sequences from a total of 210 human beings, with widespread geographic origin, ethnic background and diet, in addition to a wide range of other mammals. We found two highly prevalent core phylogroups (cores 1 and 2), belonging to the clostridial family Lachnospiraceae. These core phylogroups showed a log-normal distribution among human individuals, while non-core phylogroups showed more skewed distributions towards individuals with low levels compared with the log-normal distribution. Molecular clock analyses suggest that core 2 co-evolved with the radiation of vertebrates, while core 1 co-evolved with the mammals. Taken together, the stability, prevalence and potential functionality support the fact that the identified core phylogroups are pivotal in maintaining gut homeostasis and health.

Transcript profiling of candidate genes in testis of pigs exhibiting large differences in androstenone levels.

BACKGROUND: Boar taint is an unpleasant odor and flavor of the meat and occurs in a high proportion of uncastrated male pigs. Androstenone, a steroid produced in testis and acting as a sex pheromone regulating reproductive function in female pigs, is one of the main compounds responsible for boar taint. The primary goal of the present investigation was to determine the differential gene expression of selected candidate genes related to levels of androstenone in pigs. RESULTS: Altogether 2560 boars from the Norwegian Landrace and Duroc populations were included in this study. Testicle samples from the 192 boars with most extreme high or low levels of androstenone in fat were used for RNA extraction, and 15 candidate genes were selected and analyzed by real-competitive PCR analysis. The genes Cytochrome P450 c17 (CYP17A1), Steroidogenic acute regulatory protein (STAR), Aldo-keto reductase family 1 member C4 (AKR1C4), Short-chain dehydrogenase/reductase family member 4 (DHRS4), Ferritin light polypeptide (FTL), Sulfotransferase family 2A, dehydroepiandrosterone-preferring member 1 (SULT2A1), Cytochrome P450 subfamily XIA polypeptide 1 (CYP11A1), Cytochrome b5 (CYB5A), and 17-beta-Hydroxysteroid dehydrogenase IV (HSD17B4) were all found to be significantly (P < 0.05) up-regulated in high androstenone boars in both Duroc and Landrace. Furthermore, Cytochrome P450 c19A2 (CYP19A2) was down-regulated and progesterone receptor membrane component 1 (PGRMC1) was up-regulated in high-androstenone Duroc boars only, while CYP21 was significantly down-regulated (2.5) in high-androstenone Landrace only. The genes Nuclear Receptor co-activator 4 (NCOA4), Sphingomyrlin phosphodiesterase 1 (SMPD1) and 3beta-hydroxysteroid dehydrogenase (HSD3B) were not significantly differentially expressed in any breeds. Additionally, association studies were performed for the genes with one or more detected SNPs. Association between SNP and androstenone level was observed in CYB5A only, suggesting cis-regulation of the differential transcription in this gene. CONCLUSION: A large pig material of highly extreme androstenone levels is investigated. The current study contributes to the knowledge about which genes that is differentially expressed regard to the levels of androstenone in pigs. Results in this paper suggest that several genes are important in the regulation of androstenone level in boars and warrant further evaluation of the above mentioned candidate genes, including analyses in different breeds, identification of causal mutations and possible gene interactions.

Changes in lipid metabolism associated gene transcripts during porcine adipogenesis.

Pigs are recognised as suitable biomedical models to study obesity and obesity-related diseases; however, little is known about adipose tissue development and adipogenesis in pigs. In this study, the temporal expression of key genes involved in lipid metabolism was investigated during porcine adipogenesis and the metabolic fate of exogenously administered palmitic acid (16:0) was examined in differentiating preadipocytes. The expression of genes encoding elongases and desaturases increased simultaneously with those involved in fatty acid and triacylglycerol synthesis during porcine adipogenesis, and a high biosynthesis of monounsaturated fatty acids was measured prior to storage in differentiating preadipocytes. Although the total fatty acid oxidation in differentiating preadipocytes was low, differentiating cells showed increased expression of hormone-sensitive lipase and mitochondrial and peroxisomal genes. These data provide new insight into the temporal expression of genes involved in lipid metabolism during porcine adipogenesis and suggest a possible role of elongation and desaturation events prior to lipid accumulation in porcine adipocytes.

MALDI-TOF mass spectrometry for quantitative gene expression analysis of acid responses in Staphylococcus aureus.

Microorganisms are constantly exposed to new and altered growth conditions, and respond by changing gene expression patterns. Several methods for studying gene expression exist. During the last decade, the analysis of microarrays has been one of the most common approaches applied for large scale gene expression studies. A relatively new method for gene expression analysis is MassARRAY, which combines real competitive-PCR and MALDI-TOF (matrix-assisted laser desorption/ionization time-of-flight) mass spectrometry. In contrast to microarray methods, MassARRAY technology is suitable for analysing a larger number of samples, though for a smaller set of genes. In this study we compare the results from MassARRAY with microarrays on gene expression responses of Staphylococcus aureus exposed to acid stress at pH 4.5. RNA isolated from the same stress experiments was analysed using both the MassARRAY and the microarray methods. The MassARRAY and microarray methods showed good correlation. Both MassARRAY and microarray estimated somewhat lower fold changes compared with quantitative real-time PCR (qRT-PCR). The results confirmed the up-regulation of the urease genes in acidic environments, and also indicated the importance of metal ion regulation. This study shows that the MassARRAY technology is suitable for gene expression analysis in prokaryotes, and has advantages when a set of genes is being analysed for an organism exposed to many different environmental conditions.

Gene expression profiles in liver of pigs with extreme high and low levels of androstenone.

BACKGROUND: Boar taint is the unpleasant odour and flavour of the meat of uncastrated male pigs that is primarily caused by high levels of androstenone and skatole in adipose tissue. Androstenone is a steroid and its levels are mainly genetically determined. Studies on androstenone metabolism have, however, focused on a limited number of genes. Identification of additional genes influencing levels of androstenone may facilitate implementation of marker assisted breeding practices. In this study, microarrays were used to identify differentially expressed genes and pathways related to androstenone metabolism in the liver from boars with extreme levels of androstenone in adipose tissue. RESULTS: Liver tissue samples from 58 boars of the two breeds Duroc and Norwegian Landrace, 29 with extreme high and 29 with extreme low levels of androstenone, were selected from more than 2500 individuals. The samples were hybridised to porcine cDNA microarrays and the 1% most significant differentially expressed genes were considered significant. Among the differentially expressed genes were metabolic phase I related genes belonging to the cytochrome P450 family and the flavin-containing monooxygenase FMO1. Additionally, phase II conjugation genes including UDP-glucuronosyltransferases UGT1A5, UGT2A1 and UGT2B15, sulfotransferase STE, N-acetyltransferase NAT12 and glutathione S-transferase were identified. Phase I and phase II metabolic reactions increase the water solubility of steroids and play a key role in their elimination. Differential expression was also found for genes encoding 17beta-hydroxysteroid dehydrogenases (HSD17B2, HSD17B4, HSD17B11 and HSD17B13) and plasma proteins alpha-1-acid glycoprotein (AGP) and orosomucoid (ORM1). 17beta-hydroxysteroid dehydrogenases and plasma proteins regulate the availability of steroids by controlling the amount of active steroids accessible to receptors and available for metabolism. Differences in the expression of FMO1, NAT12, HSD17B2 and HSD17B13 were verified by quantitative real competitive PCR. CONCLUSION: A number of genes and pathways related to metabolism of androstenone in liver were identified, including new candidate genes involved in phase I oxidation metabolism, phase II conjugation metabolism, and regulation of steroid availability. The study is a first step towards a deeper understanding of enzymes and regulators involved in pathways of androstenone metabolism and may ultimately lead to the discovery of markers to reduce boar taint.