Gut microbiota and fecal volatilome profile inspection in metabolically healthy and unhealthy obesity phenotypes.

BACKGROUND: People with metabolically healthy (MHO) and metabolically unhealthy obesity (MUO) differ for the presence or absence of cardio-metabolic complications, respectively. OBJECTIVE: Based on these differences, we are interested in deepening whether these obesity phenotypes could be linked to changes in microbiota and metabolome profiles. In this respect, the overt role of microbiota taxa composition and relative metabolic profiles is not completely understood. At this aim, biochemical and nutritional parameters, fecal microbiota, metabolome and SCFA compositions were inspected in patients with MHO and MUO under a restrictive diet regimen with a daily intake ranging from 800 to 1200 kcal. METHODS: Blood, fecal samples and food questionnaires were collected from healthy controls (HC), and an obese cohort composed of both MHO and MUO patients. Most impacting biochemical/anthropometric variables from an a priori sample stratification were detected by applying a robust statistics approach useful in lowering the background noise. Bacterial taxa and volatile metabolites were assessed by qPCR and gas chromatography coupled with mass spectrometry, respectively. A targeted GC-MS analyses on SCFAs was also performed. RESULTS: Instructed to follow a controlled and restricted daily calorie intake, MHO and MUO patients showed differences in metabolic, gut microbial and volatilome signatures. Our data revealed higher quantities of specific pro-inflammatory taxa (i.e., Desulfovibrio and Prevotella genera) and lower quantities of Clostridium coccoides group in MUO subset. Higher abundances in alkane, ketone, aldehyde, and indole VOC classes together with a lower amount of butanoic acid marked the faecal MUO metabolome. CONCLUSIONS: Compared to MHO, MUO subset symptom picture is featured by specific differences in gut pro-inflammatory taxa and metabolites that could have a role in the progression to metabolically unhealthy status and developing of obesity-related cardiometabolic diseases. The approach is suitable to better explain the crosstalk existing among dysmetabolism-related inflammation, nutrient intake, lifestyle, and gut dysbiosis.

Sustainability evaluation of Sicily's lemon and orange production: an energy, economic and environmental analysis.

The island of Sicily has a long standing tradition in citrus growing. We evaluated the sustainability of orange and lemon orchards, under organic and conventional farming, using an energy, environmental and economic analysis of the whole production cycle by using a life cycle assessment approach. These orchard systems differ only in terms of a few of the inputs used and the duration of the various agricultural operations. The quantity of energy consumption in the production cycle was calculated by multiplying the quantity of inputs used by the energy conversion factors drawn from the literature. The production costs were calculated considering all internal costs, including equipment, materials, wages, and costs of working capital. The performance of the two systems (organic and conventional), was compared over a period of fifty years. The results, based on unit surface area (ha) production, prove the stronger sustainability of the organic over the conventional system, both in terms of energy consumption and environmental impact, especially for lemons. The sustainability of organic systems is mainly due to the use of environmentally friendly crop inputs (fertilizers, not use of synthetic products, etc.). In terms of production costs, the conventional management systems were more expensive, and both systems were heavily influenced by wages. In terms of kg of final product, the organic production system showed better environmental and energy performances.

Use of Limosilactobacillus reuteri DSM 17938 in paediatric gastrointestinal disorders: an updated review.

Strains of lactobacilli are the most widely used probiotics and can be found in a large variety of food products and food supplements throughout the world. In this study, the evidence on Limosilactobacillus reuteri DSM 17938 (LR DSM 17938) has been reviewed. This species secretes reuterin and other substances singularly or in microvesicles, inhibiting pathogen growth and interacting with the intestinal microbiota and mucosa, restoring homeostasis. The use of LR DSM 17938 has been exploited in several pathological conditions. Preclinical research has shown that this probiotic can ameliorate dysbiosis and, by interacting with intestinal mucosal cells, can raise the pain threshold and promote gastrointestinal motility. These aspects are amongst the significant components in functional gastrointestinal disorders, such as colic and regurgitation in infants, functional abdominal pain and functional constipation in children and adolescents. This strain can decrease the duration of acute diarrhoea and hospitalization for acute gastroenteritis but does not seem to prevent nosocomial diarrhoea and antibiotic-associated diarrhoea. Because of its ability to survive in the gastric environment, it has been tested in Helicobacter pylori infection, showing a significant decrease of antibiotic-associated side effects and a tendency to increase the eradication rate. Finally, all these studies have shown the excellent safety of LR DSM 17938 even at higher dosages. In conclusion data from various clinical trials here reviewed can guide the clinician to find the correct dose, frequency of administration, and therapy duration.

High occurrence of Helicobacter pylori in raw goat, sheep and cow milk inferred by glmM gene: a risk of food-borne infection?

Helicobacter pylori is an organism widespread in humans and sometimes responsible for serious illnesses, such as gastric and duodenal ulcers, MALToma and even gastric cancer. It has been hypothesized that the infection route by H. pylori involves multiple pathways including food-borne transmission, as the microorganism has been detected from foods such as sheep and cow milk. This work reports the results of a survey conducted in order to investigate the presence of H. pylori in raw goat, sheep and cow milk produced in Southern Italy, employing a Nested Polymerase Chain Reaction (Nested-PCR) assay for the detection of the phosphoglucosamine mutase gene (glmM), as screening method followed by conventional bacteriological isolation. Out of the 400 raw milk samples examined, 139 (34.7%) resulted positive for the presence of glmM gene, but no strains were isolated. In this work H. pylori DNA has been firstly detected from 41 (25.6%) raw goat milk samples. The results deserve further investigations on the contamination source/s of the milk samples and on the major impact that it may have on consumers.

Feasibility of biodegradable based packaging used for red meat storage during shelf-life: A pilot study.

This study was designated to ascertain the effectiveness of polylactic acid (PLA) based packaging solution to store red fresh meat during its refrigerated shelf-life. Recently the attention in the packaging industry regarding the use of bioplastics has been shifting from compostable/biodegradable materials toward biobased materials. Steaks obtained from semimembranous muscle of Piemontese beef were packaged in PLA trays closed with a lid made of PLA film and for comparison purposed in a conventional reference package consisting of a amorphous polyethylene terephthalate/polyethylene (APET/PET) trays and wrapped in plastic film of polyvinyl chloride (PVC). The packaging under modified atmosphere MAP was carried out by using a gas mixture of 66% O2, 25% CO2 and 9%N2. By using PLA packaging combination it was possible to maintain an optimum red colour together with a reduced content of volatile compounds associated to off-flavours of meat samples particularly related to the oxidation phenomena.

Escherichia coli O26 in minced beef: prevalence, characterization and antimicrobial resistance pattern.

Verocytotoxin-producing Escherichia coli (VTEC) non-O157 serogroups are among the most important emerging food-borne pathogen groups. In particular, the O26 serogroup is able to cause a large spectrum of illnesses in humans which have a significant public health impact as they may range from haemorrhagic colitis (HC) to haemolytic uremic syndrome (HUS) and thrombotic thrombocytopenic purpura (TTP). It is known that VTEC organisms are associated with animal reservoirs, i.e. ruminants, and foods of animal origin, especially undercooked meat and raw milk, are often involved in outbreaks. In this study, 250 minced beef samples collected at retail outlets in southern Italy were tested for the presence of E. coli O26 and the isolates were characterized and studied for their antimicrobial resistance properties. Three minced beef samples (1.2%) tested positive for E. coli O26; one isolate per positive sample was characterized. One isolate harboured the genes encoding for virulence factors intimin (eaeA) and enterohaemolysin (hlyA), while none presented verocytotoxin-encoding genes (stx1 and stx2) and all were negative at the verotoxicity assay. All the isolates showed resistance properties to at least four antimicrobial agents tested and two were multi-drug resistant (MDR). Although no verocytotoxin-encoding genes were found in the isolates, the presence of potentially pathogenic E. coli O26 strains in minced beef points to the need for proper hygiene during meat production to reduce the risk of food-borne illnesses and transmission of MDR organisms via foods to humans. This paper is the first report on the presence and characterization of E. coli O26 in minced beef marketed in Italy.

Methicillin-resistant Staphylococcus aureus (MRSA) in foods of animal origin product in Italy.

Methicillin-resistant Staphylococcus aureus (MRSA) strains are a global health concern. The present study regarded 160 S. aureus strains that had been isolated from 1634 foodstuff samples of animal origin in a previous survey conducted in Italy during 2003-2005. The strains were characterized by detecting the mecA gene, the production of type A to D staphylococcal enterotoxins (SEs), and studying their resistance properties against several antibiotics; their ecological origin was determined by biotyping. Of the 160 analyzed S. aureus strains six (3.75%) were mecA positive and derived from six different samples; four isolates were from bovine milk and two from dairy products (pecorino cheese and mozzarella cheese). Two strains isolated from milk belonged to the non-host-specific biovar while the others to the ovine biovar. The strain isolated from mozzarella cheese belonged to the non-host-specific biovar and the strain isolated from pecorino cheese to the ovine biovar. All the MRSA strains isolated were enterotoxigenic; two strains synthesized SEA/SED two SED and one SEC. All the strains showed resistance to at least one of the antibiotics tested but none was resistant to glycopeptides.

Occurrence, characterization and antimicrobial resistance of enterotoxigenic Staphylococcus aureus isolated from meat and dairy products.

Staphylococcus aureus is considered to be one of the leading causes of food-borne illnesses. Milk, dairy products and meats are often contaminated with enterotoxigenic strains of this bacterium. Foodstuff contamination may occur directly from infected food-producing animals or may result from poor hygiene during production processes, or the retail and storage of foods, since humans may carry the microorganism. The number of S. aureus strains that exhibits antimicrobial-resistance properties has increased, together with the potential risk of transmitting the same properties to the human microflora via foods or inducing infections hard to be treated. This paper reports the results of a 3-year survey (2003-2005) on the occurrence of S. aureus in meat and dairy products. Of 1634 samples examined, 209 (12.8%) were contaminated with S. aureus. A total of 125 enterotoxigenic S. aureus strains were biotyped and their antimicrobial resistance pattern tested. Most of the isolated strains produced SED (33.6%), followed by SEA (18.4%), SEC (15.2%), SEB (6.4%) and belonged mainly to the Human ecovar (50.4%), followed by Ovine (23.2%), Non-Host-Specific (17.6%), Bovine (7.2%) and Poultry-like (1.6%) ecovars. Finally, the 68.8% analysed strains showed antimicrobial resistance properties at least at one of antibiotics tested. Human biotype showed antimicrobial resistance at more than one antibiotic than the other biotypes (p<0.05). The results provided evidence that the presence of enterotoxigenic and antimicrobial resistant strains of S. aureus has become remarkably widespread in foods. This calls for better control of sources of food contamination and of the spread of antimicrobial-resistance organisms.

Composition and seasonal variation of soluble cuticular waxes in Actinidia deliciosa leaves.

This research examines the annual evolution and composition of soluble cuticular waxes of Actinidia deliciosa Chev. cv Hayward leaves. Soluble cuticular waxes of foliar blade were extracted in chloroform and analysed by GC-MS. The seasonal weighted mean of the wax coverage was about 24 microg cm(-2). The alkyl alkanoates were the main class of components (10 microg cm(-2)) followed by hydrocarbons (6 microg cm(-2)), terpenes (3 microg cm(-2)), alkanols (1 microg cm(-2)), ketones (1 microg cm(-2)), alkanoic acids (1 microg cm(-2)), alkanals (0.7 microg cm(-2)), and sterols (0.6 microg cm(-2)). The concentration of the soluble cuticular components reached a peak (43 microg cm(-2)) on the 83rd day after bud break. Different causes were proposed to explicate the seasonal evolution of the leaf waxes: biosynthesis of the waxes prevalently during rapid leaf growth; natural wax erosion and evaporation; progressive reduction in the extractability of the intracuticular free compounds due to the slow polymerization of the cutin matrix.

Coagulase-positive Staphylococci and Staphylococcus aureus in food products marketed in Italy.

Staphylococcus aureus is a very common organism capable of producing several enterotoxins (SEs) that cause intoxication symptoms of varying intensity in humans when ingested through contaminated food. This paper reports the results of an investigation on the presence of Coagulase-Positive Staphylococci (CPS) and S. aureus in several food products marketed in Italy and on food contact surface swabs sampled from the food industry. A total of 11,384 samples were examined and 1971 of them (17.3%) were found to contain CPS. The assays performed on 541 CPS strains led to the identification of 537 S. aureus strains on which characterization of type A, B, C and D staphylococcal enterotoxins (SEA, SEB, SEC and SED) was performed. A total of 298 S. aureus strains (55.5%) produced one or more SEs: 33.9% of the strains produced SEC, 26.5% SEA, 20.5% SEA+SED, 13.4% SED, 2.7% SEB, 1.7% SEA+SEB, 0.7% SEC+SED and 0.3% produced SEA+SEC and SEB+SEC. The investigation highlighted that these organisms are very common and constitute a potential risk for consumers' health.

Multiplex-touchdown PCR assay for the detection and genotyping of Helicobacter pylori from artificially contaminated sheep milk.

Helicobacter pylori (Hp) is an organism commonly present worldwide in the human population, sometimes causing serious illnesses such as duodenal and gastric ulcers, adenocarcinoma of the stomach, and low-grade B-cell mucosa-associated lymphoid tissue lymphoma of the stomach. This article describes a multiplex-touchdown PCR method for the identification and genotyping (vacA-s1/m1, sl/m2, and s2/m2-and cagA genes) of Hp directly from sheep milk artificially contaminated with Hp strains from human gastric biopsies and with Hp ATCC 43504. The strains from humans carried sl/m2 cagA+ and s2/m2 cagA allelic combinations, while the ATCC strains carried an sl/ml cagA+ allelic combination. The technique showed a sensitivity of 15 CFU/ml for species identification and of 1,500 CFU/ml for the detection of genes encoding for VacA and CagA. It has proven to be specific and rapid, and the authors suggest that it be used as a rapid screening method to ensure that sheep milk is uncontaminated with this organism.

Market survey of Vibrio spp. and other microrganisms in Italian shellfish.

A survey was conducted of Vibrio spp., Escherichia coli, fecal coliforms, and Salmonella in 644 molluscan shellfish samples marketed in the Apulia region of southern Italy. Vibrios were found in 278 samples (43%), and levels of E. coli and fecal coliforms were above the Italian legal limit in 27 and 34 samples (4 and 5%), respectively. Salmonella was not detected in any of the samples. Because the majority of the vibrio isolates were found in samples that were compliant with Italian regulations, there appears to be no relationship between the presence of microorganisms of fecal origin and the presence of vibrios potentially harmful to human health.

Distribution of Trichinella spiralis larvae in muscles from a naturally infected horse.

Epidemiological investigations conducted during 10 trichinellosis outbreaks between 1975 and 1994 showed that horse-meat was the probable source of infection. Though hundreds of thousands of horses have been examined at abattoirs in America and Europe to detect Trichinella infection by artificial digestion or trichinelloscopy, an infected horse has never been detected during routine analysis, which consists of examining 1 g of tissue muscle from the diaphragm. In November 1996, a naturally infected horse imported from Romania was detected in Southern Italy. The parasite was identified as Trichinella spiralis by random amplified polymorphic DNA analysis. Artificial digestion of tissue samples from 60 different muscles from 13 different sites of the infected horse carcass showed that M. levator Labii maxillaris, M. hyoideus transversus, and M. buccinator were the 3 most infected muscles. Muscles from the tongue, the masseter, and the diaphragm, which have normally been considered the muscles of choice for diagnosis, were the 4th, 6th and 13th most infected muscles, respectively. When comparing body sites, muscle tissues from the head showed the highest level of infection, followed by muscles from the neck. This finding may explain the negative results that have been obtained in the past during routine examination of the diaphragm of horses.

Evidence of reduced poly-B-hydroxybutyrate biosynthesis in free-living nitrogen-fixing bacteria, Azotobacter chroococcum, following acquired resistance to the fungicide captan.

Some biological activities of Azotobacter chroococcum, strain Azcap 1, (spontaneous mutant, captan resistant up to 300 micrograms/ml) were assayed on RM medium with and without the presence of the fungicide. Comparisons were also carried out with Az. chroococcum sensitive strains Azwt, Azcan 10 and 14. The hydrolysis of captan, incorporated in agar plates of RM at 100 micrograms/ml, was rapid, since on 4-day plates, no effect was found on the strain Azwt, while on freshly prepared ones its growth was completely blocked. As for Azcap 1, grown on RM only, the behaviour was similar to that of sensitive strains, whereas when grown on captan the results of experiments showed: (i) a lag of approximately 12 h to reach the maximum nitrogen-fixing activity; (ii) delay of 12-24 h in the full consumption of glucose present in the medium, although the invertase activity did not present differences; (iii) high ATP culture content during the 50 h of the experiment; (iv) approximately 6-10-fold lower production of PHB (poly-B-hydroxybutyrate); (v) lack of typical encystment phase, for the tested 96 h and reduced viability in developing colonies on agar RM medium. In contrast, when captan was added to cultural medium at sublethal concentration, 50 micrograms/ml for sensitive strain Azwt and 200 micrograms/ml for Azcap 1, the amount of glutathione produced (to remove the fungicide toxicity) was several times higher for the former.

Enterotoxins of Staphylococcus aureus induce a cytopathic effect in cell lines.

We describe the use of cell cultures for the detection of Staphylococcus aureus enterotoxins. Madin Darby Bovine Kidney (MDBK), Bovine Embryo lung cells (PEB) and Dog Carcinoma Cell line (A-72) cell monolayers were tested. MDBK, A-72 and PEB cell lines proved to be susceptible to the cytotoxic effect induced by staphylococcal enterotoxins. A cytopathic effect was observed in some cases after 2 hours of incubation. The PEB cell line was the most susceptible and may be used as a useful and cheap method for screening enterotoxigenic S. aureus strains.

Survival of Helicobacter pylori in artificially contaminated ultrahigh temperature and pasteurized milk.

Helicobacter pylori (H. pylori) is a very important bacterial pathogen of humans which may cause gastrointestinal illnesses ranging from gastric and duodenal ulcers to neoplastic diseases such as MALToma and gastric cancer. Transmission via contaminated food is still uncertain but several authors believe this can realistically occur and milk may act as a vehicle of infection. This paper reports the results of H. pylori survival trials in pasteurized and ultrahigh temperature (UHT) milks artificially contaminated and aerobically stored at 4 degrees C. The results obtained showed that the four strains used in this study (H. pylori nat 18-19-20 and H. pylori ATCC 43504), had a progressive reduction in bacterial load with a median survival of 9 days in pasteurized milk and 12 days in UHT milk, with approximate average of initial inoculum of 10(5) and 10(6)cfu/ml, respectively. These findings are very important to clarify the route of transmission of H. pylori to humans via food and for implementation of a correct risk analysis for food safety purposes.

Infantile condylomata of the oral cavity.

A child had condylomata acuminata localized to the oral cavity. Main points of interest were this exclusive localization, the extremely high number of papillary lesions, not reported in the literature until now, and the excellent response to interferon and local applications of podophyllin. Histologic, ultrastructural, and in situ molecular hybridization techniques were performed to make a correct diagnosis. Transmission of the etiologic agent and therapeutic approaches are discussed.