Invited review: Plant polyphenols and rumen microbiota responsible for fatty acid biohydrogenation, fiber digestion, and methane emission: Experimental evidence and methodological approaches.

The interest of the scientific community in the effects of plant polyphenols on animal nutrition is increasing. These compounds, in fact, are ubiquitous in the plant kingdom, especially in some spontaneous plants exploited as feeding resources alternative to cultivated crops and in several agro-industry by-products. Polyphenols interact with rumen microbiota, affecting carbohydrate fermentation, protein degradation, and lipid metabolism. Some of these aspects have been largely reviewed, especially for tannins; however, less information is available about the direct effect of polyphenols on the composition of rumen microbiota. In the present paper, we review the most recent literature about the effect of plant polyphenols on rumen microbiota responsible for unsaturated fatty acid biohydrogenation, fiber digestion, and methane production, taking into consideration the advances in microbiota analysis achieved in the last 10 yr. Key aspects, such as sample collection, sample storage, DNA extraction, and the main phylogenetic markers used in the reconstruction of microbial community structure, are examined. Furthermore, a summary of the new high-throughput methods based on next generation sequencing is reviewed. Several effects can be associated with dietary polyphenols. Polyphenols are able to depress or modulate the biohydrogenation of unsaturated fatty acids by a perturbation of ruminal microbiota composition. In particular, condensed tannins have an inhibitory effect on biohydrogenation, whereas hydrolyzable tannins seem to have a modulatory effect on biohydrogenation. With regard to fiber digestion, data from literature are quite consistent about a general depressive effect of polyphenols on gram-positive fibrolytic bacteria and ciliate protozoa, resulting in a reduction of volatile fatty acid production (mostly acetate molar production). Methane production is also usually reduced when tannins are included in the diet of ruminants, probably as a consequence of the inhibition of fiber digestion. However, some evidence suggests that hydrolyzable tannins may reduce methane emission by directly interacting with rumen microbiota without affecting fiber digestion.

Milk fatty acid composition, rumen microbial population, and animal performances in response to diets rich in linoleic acid supplemented with chestnut or quebracho tannins in dairy ewes.

The aim of the study was to evaluate milk fatty acid (FA) profile, animal performance, and rumen microbial population in response to diets containing soybean oil supplemented or not with chestnut and quebracho tannins in dairy ewes. Eighteen Comisana ewes at 122±6 d in milking were allotted into 3 experimental groups. Diets were characterized by chopped grass hay administered ad libitum and by 800 g/head and day of 3 experimental concentrates containing 84.5 g of soybean oil/kg of dry matter (DM) and 52.8 g/kg of DM of bentonite (control diet), chestnut tannin extract (CHT diet), or quebracho tannin extract (QUE diet). The trial lasted 4 wk. Milk yield was recorded daily, and milk composition and blood parameters were analyzed weekly. At the end of the experiment, samples of rumen fluid were collected to analyze pH, volatile fatty acid profile, and the relative proportions of Butyrivibrio fibrisolvens and Butyrivibrio proteoclasticus in the rumen microbial population. Hepatic functionality, milk yield, and gross composition were not affected by tannin extracts, whereas milk FA composition was characterized by significant changes in the concentration of linoleic acid (CHT +2.77% and QUE +9.23%), vaccenic acid (CHT +7.07% and QUE +13.88%), rumenic acid (CHT -1.88% and QUE +24.24%), stearic acid (CHT + 8.71% and QUE -11.45%), and saturated fatty acids (CHT -0.47% and QUE -3.38%). These differences were probably due to the ability of condensed versus hydrolyzable tannins to interfere with rumen microbial metabolism, as indirectly confirmed by changes in the relative proportions of B. fibrisolvens and B. proteoclasticus populations and by changes in the molar proportions of volatile fatty acids. The effect of the CHT diet on the milk FA profile and microbial species considered in this trial was intermediate between that of QUE and the control diet, suggesting a differential effect of condensed and hydrolyzable tannins on rumen microbes. Compared with control animals, the presence of B. fibrisolvens increased about 3 times in ewes fed CHT and about 5 times in animals fed QUE. In contrast, the abundance of B. proteoclasticus decreased about 5- and 15-fold in rumen liquor of ewes fed CHT and QUE diets, respectively. The use of soybean oil and a practical dose of QUE or CHT extract in the diet of dairy ewes can be an efficient strategy to improve the nutritional quality of milk.

Rumen microbial community and milk quality in Holstein lactating cows fed olive oil pomace as part in a sustainable feeding strategy.

The use of alternative feed ingredients from the Agro-industry could be an efficient tool to improve the sustainability of dairy cow production. Since the richness in polyphenols, olive oil pomace (OOP), produced during olive oil milling, seems a promising by-product to ameliorate milk's nutritional value. The aim of this study was to test the use of OOP produced by means of a new technology (biphasic with stone deprivation) in dairy cow feeding strategy to evaluate the effect on animal performances, rumen microbiota, biohydrogenation processes and milk quality by a multidisciplinary approach. Forty multiparous Italian-Friesian dairy cows, at middle lactation, were randomly allotted into two homogenous groups and fed respectively a commercial diet (CON) and the experimental diet (OOPD) obtained by adding OOP to CON as partial replacement of maize silage. The two diets were formulated to be isoproteic and isoenergetic. The same diets were tested also in an in vitro trial aimed to evaluate their rumen degradability (% DEG). The dietary supplementation with OOP did not affect DM intake, rumen % DEG and milk production. The milk's nutritional quality was improved by increasing several important functional fatty acids (FAs; i.e., linoleic acid, conjugated linoleic acid, oleic acid, vaccenic acid). This finding was related to a decrease in rumen liquor biohydrogenation rate of unsaturated FAs. The stochiometric relation between volatile FA production in the rumen and methanogenesis suggested that OOP lowers the methane potential production (CON = 0.050 mol/L vs OOPD = 0.024 mol/L, SEM = 0.005, P = 0.0011). Rumen microbiota and fungi community did not be strongly altered by OOP dietary inclusion because few bacteria were affected at the genus level only. Particularly, Acetobacter, Prevotellaceae_UCG-004, Prevotellaceae_UCG-001, Eubacterium coprostanoligenes, Lachnospira, Acetitomaulatum, Lachnospiraceae_NK3A20 group were more abundant with OOPD condition (P < 0.05). Data reported in this study confirm that the use of OOP in dairy cow feeding can be an interesting strategy to improve milk nutritional quality increasing functional FA content without compromising the rumen degradability of the diet or causing strong perturbation of rumen ecosystem and maintaining animal performances.

A novel approach combining the Calgary Biofilm Device and Phenotype MicroArray for the characterization of the chemical sensitivity of bacterial biofilms.

A rapid method for screening the metabolic susceptibility of biofilms to toxic compounds was developed by combining the Calgary Biofilm Device (MBEC device) and Phenotype MicroArray (PM) technology. The method was developed using Pseudomonas alcaliphila 34, a Cr(VI)-hyper-resistant bacterium, as the test organism. P. alcaliphila produced a robust biofilm after incubation for 16 h, reaching the maximum value after incubation for 24 h (9.4 × 10(6) ± 3.3 × 10(6) CFU peg(-1)). In order to detect the metabolic activity of cells in the biofilm, dye E (5×) and menadione sodium bisulphate (100 µM) were selected for redox detection chemistry, because they produced a high colorimetric yield in response to bacterial metabolism (340.4 ± 6.9 Omnilog Arbitrary Units). This combined approach, which avoids the limitations of traditional plate counts, was validated by testing the susceptibility of P. alcaliphila biofilm to 22 toxic compounds. For each compound the concentration level that significantly lowered the metabolic activity of the biofilm was identified. Chemical sensitivity analysis of the planktonic culture was also performed, allowing comparison of the metabolic susceptibility patterns of biofilm and planktonic cultures.

Exploring the relationship between bacterial genera and lipid metabolism in bovine rumen.

The rumen is characterised by a complex microbial ecosystem, which is particularly active in lipid metabolism. Several studies demonstrated a role of diet and breed on bacterial community profile, with the effect on metabolic pathways. Despite the knowledge achieved on metabolism and the bacterial profile, little is known about the relationship between individual bacteria and metabolic pathways. Therefore, a multivariate approach was used to search for possible relationships between bacteria and products of several pathways. The correlation between rumen bacterial community composition and rumen lipid metabolism was assessed in 40 beef steers (20 Maremmana and 20 Aubrac) reared with the same system and fed the same diet. A canonical discriminant analysis combined with a canonical correlation analysis (CCA) was performed to explore this correlation. The variables showing a Pearson correlation higher than 0.6 as absolute value and significant were retained for CCA considering the relationship of bacterial composition with several metabolic pathways. The results indicated that some bacterial genera could have significant impacts on the presence of several fatty acids. However, the relationship between genera and fatty acid changes according to the breed, demonstrating that the metabolic pathways change according to the host genetic background, related to breed evolution, although there is also an intra-breed genetic background which should not be ignored. In Maremmana, Succiniclasticum and Rikenellaceae_RC9_gut_group showed a high positive correlation with dimethylacetals (DMAs) DMAC13:0, DMAC14:0, DMAC14:0iso, DMAC15:0, DMAC15:0iso, and DMAC18:0. Prevotellaceae_UCG-003 correlates with C18:3c9c12c15 and C18:1t11, while Fibrobacter and Succiniclasticum correlate with C18:2c9t11 and Lachnospiraceae_NK3A20_group correlates with C18:1c12. Prevotellaceae_UCG-003, Ruminococcaceae_UCG-010, and Oribacterium showed a positive correlation with C13:0iso, and C17:0. Conversely, in Aubrac, Treponema_2 and Rikenellaceae_RC9_gut_group correlated with DMAC14:0iso, DMAC16:0iso, DMAC17:0iso, while Ruminococcaceae_UCG-010, Christensenellaceae_R-7_group and Ruminococcaceae_NK4A214_group correlated with DMAC18:1t11, DMAC14:0, DMAC18:1c12. Acetitomaculum correlated with C18:2c9c12, C18:1c12, C18:1c13, C18:1t12 and Lachnospiraceae_NK3A20_group with C18:1t6-8 and C18:1t9. Saccharofermentas, Ruminococcaceae_UCG-010 and Rikenellaceae_RC9_gut_group correlated with C18:2c9t11 while, Prevotellaceae_UCG-001 and Ruminococcus_1 correlated with C14:0iso, C15:0, C15:0iso, C17:0. Saccharofermentans, Rikenellaceae_RC9_gut_group, Ruminococcaceae_NK4A214_group, and Ruminococcaceae_UCG-010 correlated with C13:1c12 and C16:0iso. These results lead to hypothesise a possible association between several metabolic pathways and one or a few bacterial genera. If these associations are confirmed by further investigations that verify the causality of a bacterial genus with a particular metabolic process, it will be possible to deepen the knowledge on the activity of the rumen population in lipid metabolism. This approach appears to be a promising tool for uncovering the correlation between bacterial genera and products of rumen lipid metabolism.

A histidine-kinase cheA gene of Pseudomonas pseudoalcaligens KF707 not only has a key role in chemotaxis but also affects biofilm formation and cell metabolism.

A histidine-kinase cheA gene in Pseudomonas pseudoalcaligenes KF707 plays a central role in the regulation of metabolic responses as well as in chemotaxis. Non-chemotactic mutants harboring insertions into the cheA gene were screened for their ability to form biofilms in the Calgary biofilm device. Notably, ≥95% decrease in the number of cells attached to the polystyrene surface was observed in cheA mutants compared to the KF707 wild-type biofilm phenotype. The ability to form mature biofilms was restored to wild-type levels, providing functional copies of the KF707 cheA gene to the mutants. In addition, phenotype micro-arrays and proteomic analyses revealed that several basic metabolic activities and a few periplasmic binding proteins of cheA mutant cells differed compared to those of wild-type cells. These results are interpreted as evidence of a strong integration between chemotactic and metabolic pathways in the process of biofilm development by P. pseudoalcaligenes KF707.

From hazardous asbestos containing wastes (ACW) to new secondary raw material through a new sustainable inertization process: A multimethodological mineralogical study.

Nowadays, asbestos-containing wastes (ACW) still represent an important environmental problem and a severe health hazard due to the well known pulmonary diseases derived from asbestos fibers inhalation. Except for a very few cases, ACW are currently confined in controlled landfills, giving rise to increasingly high amounts of still hazardous wastes. A promising alternative to landfill confinement is represented by ACW inertization, but the high cost of the inertization processes so far proposed by the scientific community have hampered the creation of actually operative plants. In this paper, we explore the possibility to use an innovative process that ensures the obtainment of asbestos-free inert material in an exceptionally short processing time, thus greatly reducing cost-related problems. The efficacy of the inertization process has been verified through accurate mineralogical investigations on both chrysotile and crocidolite de-activated fibers, through X-ray diffraction, scanning and transmission electron microscopy. Overall mineralogical, microstructural and granulometric characteristics of the inert bulk material suggest that it could be successfully re-used as a secondary raw material in ceramic industries. This innovative inertization procedure could therefore provide an effective and economically sustainable solution for ACW management.

Garnet, the archetypal cubic mineral, grows tetragonal.

Garnet is the archetypal cubic mineral, occurring in a wide variety of rock types in Earth's crust and upper mantle. Owing to its prevalence, durability and compositional diversity, garnet is used to investigate a broad range of geological processes. Although birefringence is a characteristic feature of rare Ca-Fe3+ garnet and Ca-rich hydrous garnet, the optical anisotropy that has occasionally been documented in common (that is, anhydrous Ca-Fe2+-Mg-Mn) garnet is generally attributed to internal strain of the cubic structure. Here we show that common garnet with a non-cubic (tetragonal) crystal structure is much more widespread than previously thought, occurring in low-temperature, high-pressure metamorphosed basalts (blueschists) from subduction zones and in low-grade metamorphosed mudstones (phyllites and schists) from orogenic belts. Indeed, a non-cubic symmetry appears to be typical of common garnet that forms at low temperatures (<450 °C), where it has a characteristic Fe-Ca-rich composition with very low Mg contents. We propose that, in most cases, garnet does not initially grow cubic. Our discovery indicates that the crystal chemistry and thermodynamic properties of garnet at low-temperature need to be re-assessed, with potential consequences for the application of garnet as an investigative tool in a broad range of geological environments.

Selectivity in the heavy metal removal by exopolysaccharide-producing cyanobacteria.

AIMS: The aim of this study was to assess the selective removal of Cu(II), Cr(III) and Ni(II) by strains of exopolysaccharide (EPS)-producing cyanobacteria, and to investigate the interaction of sorption in solutions with multiple-metals. METHODS AND RESULTS: Nine EPS-producing cyanobacteria were tested for their ability to remove Cr, Cu and Ni in both single- and multiple-metal solutions. In the single-metal solutions, some of the strains showed very high values of metal uptake, however, only two of them showed the capability to selectively remove one or two of the metals present in the multiple-metal solutions. In multi-metal systems, the binding process was either noninteractive, synergistic or competitive between metal ions for different strains of cyanobacteria. CONCLUSIONS: Cyanothece 16Som 2 showed significantly greater sorptive capacity for Cu (1.5-20x) and Cr (2-50x) than all other strains tested. The Nostoc PCC 7936 strain showed high specific and almost exclusive selectivity towards Cu, which suggests its use aimed at recovering this metal from multiple-metal solutions. SIGNIFICANCE AND IMPACT OF THE STUDY: To find out microbial sorbents with good metal selectivity may be very useful for building up processes aimed to recover valuable metals from industrial wastewaters.

Anterolateral thigh cutaneous flap vs. radial forearm free-flap in oral and oropharyngeal reconstruction: an analysis of 48 flaps.

Microvascular free-flaps have been in clinical use for nearly 3 decades becoming the gold standard in oral and oropharyngeal reconstruction. In this study, a comparison has been made between the radial forearm fasciocutaneous flap with the thinned anterolateral thigh cutaneous flap (tALT), showing the advantage of the anterolateral thigh cutaneous flap in oral and oropharyngeal reconstruction. Between January 2003 and January 2007, 48 reconstructions were performed in patients submitted to surgery for oral and oropharyngeal carcinoma using, in 17 cases, a radial forearm fasciocutaneous flap and, in 31, an anterolateral thigh cutaneous flap. In patients treated with the radial forearm fasciocutaneous flap, results showed 94.1% flap survival; in cases treated with the anterolateral thigh cutaneous flap, 93.5% flap survival (p < 0.9). Functional results, at receiving site, were comparable in both groups. Functional results, at donor site, were less successful in the radial forearm fasciocutaneous flap group, with permanent forearm movement impairment in 35.3% of cases; in the anterolateral thigh cutaneous flap group, only transitory gait impairment occurred in 12.9% of patients. In conclusion, in our experience, the thinned anterolateral thigh cutaneous flap is comparable to radial forearm fasciocutaneous flap in terms of functional results at receiving site, but, having no limitation in availability of donor tissue, it allows a more extended resection of the tumour. Moreover, the donor site can be closed primarily with only an inconspicuous curvilinear scar left over the thigh and with significantly reduced functional impairment.

Intraparotid facial nerve schwannoma: two case reports and a review of the literature.

SUMMARY: Schwannomas are rare benign tumours that arise from Schwann cells. The most known and studied is the intracranial vestibular schwannoma, even if it is not the most frequent. More often schwannomas arise from peripheral sensitive nerves, and the vagous is most involved among the cranial nerves. Intraparotid schwannomas account for just 10% of all facial involvement, so they are an extremely rare localisation. At present, there are less than 100 cases described in the literature. We performed a retrospective analysis of parotidectomy in two Italian hospitals and present two cases of intraparotid schwannoma and a review of the literature. In the first case, we performed a parotidectomy with a stripping of tumour from the nerve. In the other case, a hypoglossal-facial neurorrhaphy was performed. Follow-up was 24 months in the first (House-Brackmann II degree in temporal-ocular and III in facial-cervical branches) and 30 months in the second case (House-Brackmann III degree in both temporal-ocular and facial-cervical branches). Preoperative diagnosis of facial nerve schwannoma is a challenge; however, it is extremely important since post-operative palsy is common and often higher grade. Unfortunately, schwannoma has similar radiologic finding as more common pleomorphic adenoma and often FNAC is not helpful. Due to its rarity and benign nature, there is debate in the literature on the need for surgical removal. Wait-and-see is a valid option, but may could give problems in secondary surgery. Stripping or near-total removal can be useful in cases of limited involvement of the nerve. Neurorrhaphy can provide good functional results when facial sacrifice is needed.

Species attribution and strain typing of Oenococcus oeni (formerly Leuconostoc oenos) with restriction endonuclease fingerprints.

In several wines, malolactic fermentation is required to improve the organoleptic characters and to stabilize the final product. In order to establish a controlled malolactic fermentation in wine, easy identification and sensitive typing of strains of Oenococcus oeni (new name of the malolactic bacterium Leuconostoc oenos) used as starter cultures are necessary. To accomplish these tasks, several strains of Oenococcus oeni isolated from wines of the Chianti region (Italy), along with reference strains and strains of L. mesenteroides subsp. mesenteroides, L. carnosum, L. fallax, L. pseudomesenteroides, L. lactis and Weisella paramesenteroides, were studied with RFLP of ribosomal genes and ultrasensitive total DNA restriction pattern analysis performed on polyacrylamide gel. With each of four restriction endonucleases used, identical restriction profiles of ribosomal genes were obtained for all strains of O. oeni. These ribopatterns, being strongly dissimilar to profiles of the other lactic acid bacteria tested, appear to be well suited for the attribution of wine lactic acid bacteria to the species O. oeni. Cluster analysis performed on two total DNA restriction profile data sets showed that the species O. oeni possesses a good degree of genomic homogeneity. Very sensitive typing of strains of O. oeni was obtained with total DNA restriction profiles. The potential of an integrated approach using restriction profiles for species assignment and typing of selected malolactic bacteria is demonstrated.

Genotypic diversity and typing of cyanobacterial strains of the genus Arthrospira by very sensitive total DNA restriction profile analysis.

Arthrospira maxima and A. platensis are two species of cyanobacteria cultivated and sold as health food, animal feed and source of food additives and fine chemicals. The genotypic diversity of several strains attributed to these two species on the basis of morphological criteria was investigated using very sensitive total DNA restriction profile analysis. The restriction profiles were obtained after sodium dodecyl sulphate polyacrylamide gel electrophoresis and silver staining. The unweighted pair-group method using arithmetic averages applied to the matrix of Dice similarity coefficient values clustered the electropherograms of the strains in two well-separated genotypic groups. These clusters corresponded to those obtained with morphological criteria. The molecular approach used was also able to type the examined strains.

A single-centre, before-after study of the short- and long-term efficacy of Narivent(®) in the treatment of nasal congestion.

OBJECTIVE: Nasal congestion is a common symptom in allergic and nonallergic rhinitis, rhinosinusitis and nasal polyposis. The present study evaluated the clinical effectiveness of Narivent(®), an osmotically-acting medical device with anti oedematous and anti-inflammatory effects, in nasal congestion. METHODS: A single-centre, prospective study with a pre- post design and consecutive patient enrolment was conducted in an Italian otolaryngology department. Patients with persistent nasal congestion were allocated to treatment groups as follows: group 1 (n = 36) treated for 7 days; group 2 (n = 56) treated for 30 days. In each group, patients received two puffs of Narivent(®) into each nostril twice daily. Symptom severity was assessed subjectively on a 0-10 visual analogue scale, and objectively by the presence/absence of signs and symptoms. Differences in subjective and objective severity measures before and after treatment were compared using Wilcoxon's signed rank test. RESULTS: All symptoms and objective scores improved after treatment with Narivent(®) for 7 or 30 days and no adverse effects were reported by the patients in either group. CONCLUSION: Narivent(®) appears to be efficacious in treating nasal congestion over a 7- or a 30-day period.

Revision of species delineation in the genus Ectothiorhodospira.

When the type strains and other strains of the six currently defined species of the genus Ectothiorhodospira were examined by DNA-DNA reassociation and RFLP of 16S/23S rDNA (ribotype), only four genospecies could be found. The possibility of defining taxonomically meaningful species corresponding to these four genospecies was investigated by combining DNA relatedness and ribotype data with other genotypic and phenotypic characters already described in the literature, an approach known as polyphasic taxonomy. Following this comparison, the type strain and another strain of Ectothiorhodospira vacuolata were found to be very similar to the type strain of Ectothiorhodospira shaposhnikovii and have been transferred to this latter species. Also, the type strain of Ectothiorhodospira marismortui and another previously unidentified strain were found to be very similar to the type strain of Ectothiorhodospira mobilis and have been transferred to this latter species. Due to the limited degree of reciprocal DNA relatedness, strains belonging either to Ectothiorhodospira marina or to Ectothiorhodospira haloalkaliphila are still considered as belonging to separate species, even though they show a remarkable phenotypic similarity. This revision has led to the delineation of only four species in the genus Ectothiorhodospira, namely E. mobilis, E. shaposhnikovii, E. marina and E. haloalkaliphila. E. vacuolata is recognized as a junior synonym of E. shaposhnikovii and E. marismortui as a junior synonym of E. mobilis.

Suppression of acute and relapsing experimental allergic encephalomyelitis with mitoxantrone.

The effect of treatment with the antineoplastic, immunomodulatory agent mitoxantrone on the course of acute and relapsing experimental allergic encephalomyelitis (EAE) in the mouse has been studied. Untreated mice immunized to produce acute EAE had an 81% incidence of clinical disease and 100% incidence of pathologic disease. Mice treated with mitoxantrone at a dose of 0.5 mg/kg daily for the 10 days following immunization did not develop any clinical signs and had minimal pathologic signs of disease. A dose of 0.25 mg/kg gave an intermediate response. Untreated mice immunized for relapsing EAE had a 100% incidence of disease with an average onset of disease on Day 148. Mice treated with mitoxantrone at a dose of 0.05 mg/kg three times weekly for 12 weeks following immunization had a 67% incidence of clinical disease with a significant delay in the average onset date to Day 279. These results indicate that mitoxantrone was highly effective in suppressing development of acute EAE. Mitoxantrone delayed the onset of relapsing EAE in mice, but did not fully inhibit the eventual expression of the disease. These studies suggest that the use of cytotoxic therapies in the treatment of autoimmune diseases may require periodic cycles of therapy to block disease expression.

Saccharomyces cerevisiae biodiversity in spontaneous commercial fermentations of grape musts with 'adequate' and 'inadequate' assimilable-nitrogen content.

AIM: To evaluate whether intraspecific diversity of Saccharomyces cerevisiae in wine fermentations is affected by initial assimilable-nitrogen content. METHODS AND RESULTS: Saccharomyces cerevisiae isolates from two spontaneous commercial wine fermentations started with adequate and inadequate nitrogen amounts were characterized by mitochondrial DNA restriction analysis. Several strains occurred in each fermentation, two strains, but not the same ones, being predominant at frequencies of about 30%. No significant differences were detected by comparing the biodiversity indices of the two fermentations. Cluster analysis demonstrated that the strain distribution was independent of nitrogen content, the two pairs of closely related dominant strains grouping into clusters at low similarity. CONCLUSIONS: The genetic variability of S. cerevisiae in wine fermentations seemed not to depend on the nitrogen availability in must. SIGNIFICANCE AND IMPACT OF THE STUDY: Nitrogen content did not affect the genetic diversity but may have induced a 'selection effect' on S. cerevisiae strains dominating wine fermentations, with possible consequences on wine properties.

Environmental household exposures to asbestos and occurrence of pleural mesothelioma.

We reviewed the certificates of 39,650 deaths which occurred in the period 1975-1988 in Leghorn and of 45,900 in La Spezia (Italy) in the period 1958-1988. In total 262 cases have been recorded as pleural mesothelioma. The main occupational exposures occurred in the shipbuilding industry. Regarding non-occupational exposures to asbestos, 13 cases of mesothelioma were found in women who had washed the work clothes of their relatives at home; we also found other domestic uses of asbestos which were rarely or never discussed previously in the literature: six cases might be explained by the installation of fireproof or non-conductive materials in the domestic environment. These exposures probably are more frequent than realized until now.