Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae) in the host Anaphes nitens (Hymenoptera: Mymaridae): first report of association with insects / Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae) no hospedeiro Anaphes nitens (Hymenoptera: Mymaridae): primeiro relato de associação com insetos

Endosymbiont bacteria can affect biological parameters and reduce the effectiveness of natural enemies in controlling the target insect. The objective of this work was to identify endosymbiont bacteria in Anaphes nitens (Girault, 1928) (Hymenoptera: Mymaridae), the main natural enemy used to manage Gonipterus platensis (Marelli, 1926) (Coleoptera: Curculionidae). Genomic DNA from six A. nitens populations was extracted and polymerase chain reactions (PCR) were performed with the primers to detect endosymbiont bacteria in this insect. The PCR products were amplified, sequenced, and compared with sequences deposited in the GenBank for the bacteria identification. All A. nitens populations had the bacterium Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae). This bacterium was originally described as free-living, and it is associated with and composes part of the A. nitens microbiota. This is the first report of Y. massiliensis in an insect host.

As bactérias endossimbiontes podem afetar os parâmetros biológicos e reduzirem a eficácia de inimigos naturais no controle do inseto alvo. O objetivo deste trabalho foi identificar bactérias endossimbiontes em Anaphes nitens (Girault, 1928) (Hymenoptera: Mymaridae), o principal inimigo natural usado no manejo de Gonipterus platensis (Marelli, 1926) (Coleoptera: Curculionidae). O DNA genômico de seis populações de A. nitens foi extraído e as reações em cadeia da polimerase (PCR) realizadas com os primers para detectar bactérias endossimbiontes neste inseto. Os produtos de PCR foram amplificados, sequenciados e comparados com as sequências depositadas no GenBank para identificação das bactérias. Todas as populações de A. nitens tinham a bactéria Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae). Esta bactéria foi originalmente descrita como de vida livre e está associada e compõe parte da microbiota de A. nitens. Este é o primeiro relato de Y. massiliensis em um hospedeiro.

Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae) in the host Anaphes nitens (Hymenoptera: Mymaridae): first report of association with insects / Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae) no hospedeiro Anaphes nitens (Hymenoptera: Mymaridae): primeiro relato de associação com insetos

Endosymbiont bacteria can affect biological parameters and reduce the effectiveness of natural enemies in controlling the target insect. The objective of this work was to identify endosymbiont bacteria in Anaphes nitens (Girault, 1928) (Hymenoptera: Mymaridae), the main natural enemy used to manage Gonipterus platensis (Marelli, 1926) (Coleoptera: Curculionidae). Genomic DNA from six A. nitens populations was extracted and polymerase chain reactions (PCR) were performed with the primers to detect endosymbiont bacteria in this insect. The PCR products were amplified, sequenced, and compared with sequences deposited in the GenBank for the bacteria identification. All A. nitens populations had the bacterium Yersinia massiliensis (Enterobacteriales:Enterobacteriaceae). This bacterium was originally described as free-living, and it is associated with and composes part of the A. nitens microbiota. This is the first report of Y. massiliensis in an insect host.

As bactérias endossimbiontes podem afetar os parâmetros biológicos e reduzirem a eficácia de inimigos naturais no controle do inseto alvo. O objetivo deste trabalho foi identificar bactérias endossimbiontes em Anaphes nitens (Girault, 1928) (Hymenoptera: Mymaridae), o principal inimigo natural usado no manejo de Gonipterus platensis (Marelli, 1926) (Coleoptera: Curculionidae). O DNA genômico de seis populações de A. nitens foi extraído e as reações em cadeia da polimerase (PCR) realizadas com os primers para detectar bactérias endossimbiontes neste inseto. Os produtos de PCR foram amplificados, sequenciados e comparados com as sequências depositadas no GenBank para identificação das bactérias. Todas as populações de A. nitens tinham a bactéria Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae). Esta bactéria foi originalmente descrita como de vida livre e está associada e compõe parte da microbiota de A. nitens. Este é o primeiro relato de Y. massiliensis em um hospedeiro.

Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae) in the host Anaphes nitens (Hymenoptera: Mymaridae): first report of association with insects

Abstract Endosymbiont bacteria can affect biological parameters and reduce the effectiveness of natural enemies in controlling the target insect. The objective of this work was to identify endosymbiont bacteria in Anaphes nitens (Girault, 1928) (Hymenoptera: Mymaridae), the main natural enemy used to manage Gonipterus platensis (Marelli, 1926) (Coleoptera: Curculionidae). Genomic DNA from six A. nitens populations was extracted and polymerase chain reactions (PCR) were performed with the primers to detect endosymbiont bacteria in this insect. The PCR products were amplified, sequenced, and compared with sequences deposited in the GenBank for the bacteria identification. All A. nitens populations had the bacterium Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae). This bacterium was originally described as free-living, and it is associated with and composes part of the A. nitens microbiota. This is the first report of Y. massiliensis in an insect host.

Resumo As bactérias endossimbiontes podem afetar os parâmetros biológicos e reduzirem a eficácia de inimigos naturais no controle do inseto alvo. O objetivo deste trabalho foi identificar bactérias endossimbiontes em Anaphes nitens (Girault, 1928) (Hymenoptera: Mymaridae), o principal inimigo natural usado no manejo de Gonipterus platensis (Marelli, 1926) (Coleoptera: Curculionidae). O DNA genômico de seis populações de A. nitens foi extraído e as reações em cadeia da polimerase (PCR) realizadas com os primers para detectar bactérias endossimbiontes neste inseto. Os produtos de PCR foram amplificados, sequenciados e comparados com as sequências depositadas no GenBank para identificação das bactérias. Todas as populações de A. nitens tinham a bactéria Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae). Esta bactéria foi originalmente descrita como de vida livre e está associada e compõe parte da microbiota de A. nitens. Este é o primeiro relato de Y. massiliensis em um hospedeiro.

Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae) in the host Anaphes nitens (Hymenoptera: Mymaridae): first report of association with insects.

Endosymbiont bacteria can affect biological parameters and reduce the effectiveness of natural enemies in controlling the target insect. The objective of this work was to identify endosymbiont bacteria in Anaphes nitens (Girault, 1928) (Hymenoptera: Mymaridae), the main natural enemy used to manage Gonipterus platensis (Marelli, 1926) (Coleoptera: Curculionidae). Genomic DNA from six A. nitens populations was extracted and polymerase chain reactions (PCR) were performed with the primers to detect endosymbiont bacteria in this insect. The PCR products were amplified, sequenced, and compared with sequences deposited in the GenBank for the bacteria identification. All A. nitens populations had the bacterium Yersinia massiliensis (Enterobacteriales: Enterobacteriaceae). This bacterium was originally described as free-living, and it is associated with and composes part of the A. nitens microbiota. This is the first report of Y. massiliensis in an insect host.

Administering 25-hydroxyvitamin D3 in vitamin D-deficient young type 1A diabetic patients reduces reactivity against islet autoantigens.

BACKGROUND & AIMS: We investigated whether improving 25-hydroxyvitamin D status in young type 1A diabetic patients reduces reactivity of peripheral blood mononuclear cells against islet autoantigens and associates with beta-cell functional changes. METHODS: Eight patients with 25-hydroxyvitamin D deficiency (<20 ng/ml), out of 15 consecutive young type 1A diabetic subjects received 25-hydroxyvitamin D3 to achieve and maintain levels above 50 ng/ml for up to one year. Peripheral blood mononuclear cell reactivity (Interferon-γ spots) against beta-cell autoantigens (glutamic acid decarboxylase 65-kD isoform, proinsulin and tyrosine phosphatase-like protein IA-2) and C-peptide during mixed meal were assessed before and after 25-hydroxyvitamin D3 replenishment. RESULTS: Target 25-hydroxyvitamin D blood levels were safely reached and maintained. Peripheral blood mononuclear cell reactivity against glutamic acid decarboxylase 65-kD isoform (3.8 ± 4.0 vs. 45 ± 16) and proinsulin (3.5 ± 3.2 vs. 75 ± 51) decreased significantly (p < 0.001 and p < 0.02) upon 25-hydroxyvitamin D3 replenishment, which was correlated with 25-hydroxyvitamin D concentrations. C-peptide values remained stable after one year of treatment. CONCLUSIONS: Safely restored and maintained 25-hydroxyvitamin D levels associated with reduced peripheral blood mononuclear cell reactivity against beta-cell autoantigens with no significant decrease of beta-cell function in this cohort of patients.

Characterization of the Mediterranean Italian buffaloes melatonin receptor 1A (MTNR1A) gene and its association with reproductive seasonality.

The aim of this study was to examine the polymorphism in MTNR1A gene and its relation to reproductive seasonality in Mediterranean Italian buffaloes reared in Sardinia. The mating period and calving of 100 multiparous buffalo-cows were recorded for three years (2005-2008). Genomic DNA was subjected to PCR for the amplification of the exon II, then 40 amplicons were sequenced. The obtained sequence was deposited in GeneBank database (accession number GU817415). PCR products were checked for the presence of HpaI restriction sites and assigned to genotypes "C/C", "C/T" or "T/T". Allelic frequency of C and T alleles was 0.44 and 0.56 and genotypic frequency was 26% for genotype C/C, 40% for C/T and 34% for T/T. In the three observed years the animals with C/C genotype showed the highest number of mating in the semester between August and January and their calving mainly occurred from August to September. On the other hand animals with T/T genotype showed mating mostly in the semester between February and July and calving occurred largely from March to May in all the three years. Heterozygous, in all the three years, showed about the same number of animals mated within each six-month period. The results of the present study provide for the first time a partial sequence as well as one polymorphic site of the MTNR1A receptor gene from buffaloes. Moreover our data showed an association between Single Nucleotide Polymorphism and seasonal reproductive activity in these animals.

Stakeholder appraisal of policy options for responding to obesity in Italy.

This article deals with the phenomenon of obesity in contemporary Italian society. It is based on the fieldwork carried out during the year 2005 within the European Union-funded project 'PorGrow: policy options for responding to obesity' (see: http://www.sussex.ac.uk/spru/PorGrow). The most recent statistical data on the spread of overweight and obesity in Italian society reveal that the phenomenon has reached a dimension that, albeit not as serious as in other Western countries, constitutes a serious threat to public health and to the national budget. The panel of stakeholders interviewed for this research showed awareness of this issue and generally agreed on the necessity to adopt a very multifaceted portfolio of policy measures to address the problem. Participants frequently regarded educational policies as the highest priority, followed by informational measures and infrastructural actions. Fiscal policy options were widely considered ineffective and unacceptable, and little enthusiasm was shown for technological innovations. In sum, while interviewees saw a real need for improved food habits on the part of Italian citizens, nonetheless in a country affected by many criticalities, the weakness of political will and the pressure of fast food culture remain severe obstacles in the fight against overweight and obesity.

Mercury- and copper-induced lysosomal membrane destabilisation depends on [Ca2+]i dependent phospholipase A2 activation.

Heavy metals are environmental pollutants able to produce different cellular effects, such as an alteration of Ca2+ homeostasis and lysosomal membrane destabilisation. The latter is one of the most used stress indices in biomonitoring programs. Recently, it has been demonstrated that cytosolic calcium increase can modulate lysosomal membrane destabilisation via activation of Ca(2+)-dependent phospholipase A2 (cPLA2). The aim of this work was to investigate the possible involvement of Ca(2+)-activated PLA2 in lysosomal membrane destabilisation induced by heavy metals in mussel haemolymph cells. We have studied the effects of Hg2+ and Cu2+ on free cytosolic calcium using Fura2/AM-loaded cells and lysosomal membrane destabilisation using neutral red (NR) staining. Hg2+ induced a [Ca2+]i rise from 100 to 780 nM in 30 min, and a lysosome destaining of 70% after 60 min that indicates destabilisation of lysosomal membranes. Both effects were reduced in a Ca(2+)-free medium, suggesting a cause-effect relationship. Exposure to Cu2+ produced the same effects, but with an intensity of about 50% respect to Hg2+. Metal-induced lysosomal destabilisation was also reduced in cells pre-exposed to a specific Ca(2+)-dependent cPLA2 inhibitor (AACOCF3). Conversely, haemocyte pretreatment with a Ca(2+)-independent PLA2 inhibitor (bromoenol-lactone (BEL)) did not prevent the destabilizing effect of heavy metals on lysosomes. Exposure to heavy metals also produced an increase in lysosomal volume of 1.8-2-folds, that was prevented by pre-incubation with AACOCF3 but not with BEL. These data indicate an involvement of cPLA2 in lysosomal membrane destabilisation induced by heavy metals.

Free radical-dependent Ca2+ signaling: role of Ca2+-induced Ca2+ release.

Previously we have shown that Fe3+/ascorbate-induced Ca2+ release from scallop sarcoplasmic reticulum (SR) is due to Ca2+-channel gating by free radicals. This study is aimed at demonstrating that Ca2+-induced Ca2+ release (CICR) plays a role in this kind of Ca2+ release. Scallop SR vesicles were incubated with fluo-3 and exposed to Fe3+/ascorbate. Fluorimetric recordings showed massive Ca2+ release, with maximum rate and 50% release occurring at 30 min after exposure. Conversely, the use of the probe for reactive oxygen species dihydrorhodamine or the assay of malondialdehyde allowed oxyradical production to be traced for approximately 5 min only. Hence, although Ca2+ release started just after exposure to Fe3+/ascorbate, most release occurred after free radical exhaustion. Ruthenium red addition after Fe3+/ascorbate slowed down the Ca2+ release, whereas cyclic adenosine 5'-diphosphoribose addition accelerated it, indicating that the free radical-induced Ca2+ release from SR vesicles triggers a mechanism of CICR that dramatically increases the initial effect.

Interference of heavy metal cations with fluorescent Ca2+ probes does not affect Ca2+ measurements in living cells.

In studies about the effects of heavy metals on intracellular Ca2+, the use of fluorescent probes is debated, as metal cations are known to affect the probe signal. In this study, spectrofluorimetric experiments in free solution, using Fluo-3 and Fura-2, showed that Zn2+ and Cd2+ enhanced the probe signal, Cu2+ quenched it, and Hg2+ had no effect. Addition of GSH prevented most of these effects, suggesting the occurrence of a similar protective role in living cells. Digital imaging of living mussel haemocytes loaded with Fura-2/AM or Fluo-3/AM showed that Hg2+, Cu2+ and Cd2+ induced a rise in probe fluorescence, whereas up to 200 microM Zn2+ had no effect. In particular, Cd2+ produced the strongest probe signal rise in free solution, but the lowest fluorescence increase in cells. Probe calibration yielded [Ca2+]i values characteristic of resting levels in control and Zn2+-exposed cells, and, as expected, indicated Ca2+ homeostasis impairment in cells exposed to Cd2+, Cu2+ and Hg2+. Our results show that Ca2+ probe responses to heavy metals in living cells are completely different from those obtained in free solution, indicating that fluorescent probes can be a suitable tool to record the effects of heavy metals on [Ca2+]i.

Learning from citizens: a Venetian experience.

In the context of different projects, some participatory methods were used, together with more traditional ones, in a research process aimed at eliciting local knowledge about risks derived from industrial activities. The policy implications of this approach are towards promoting citizen awareness and encouraging their involvement in actions addressed to reducing community vulnerability to risks. The paper illustrates the objectives and methodology of a project on public risk perception and comments on some of its findings. Drawing also from other research experiences, it outlines a set of 'principles' which can guide the interpretation of citizens' concerns and aspirations.

Role of metallothionein against oxidative stress in the mussel Mytilus galloprovincialis.

Metallothionein (MT) is a sulfhydryl-rich protein involved mainly in heavy metal homeostasis and detoxification. In this study, the use of the mussel as an experimental model allowed us to test MT antioxidant properties at the molecular, cellular, and organism level. MT induction was achieved by mussel exposure to Cd (200 microg/l) in aquaria for 7 days followed by detoxification in the sea for 28 days. Cd-preexposed and nonexposed mussels were then treated with Fe (300-600 microg/l) in aquaria for 3 days. Biochemical assays on digestive gland tissue showed that treatment with Fe led to a significant increase in oxyradical production and malondialdehyde level only in mussels not preexposed to Cd. The Cd-dependent resistance to oxidative stress was ascribed to MT induction, as Cd produced no significant variation of reduced glutathione and major antioxidant enzymes. Digital imaging of isolated digestive gland cells showed lower oxyradical rise and higher viability in cells from Cd-preexposed mussels after treatments with 0.5-5 mM H2O2. Analyses on whole organisms showed that anoxic survival was lowered in mussels that had been treated with Fe, but such an effect was less pronounced in Cd-preexposed mussels compared with nonpreexposed ones. In conclusion, data suggest an antioxidant role for MT, which seems to occur through oxyradical scavenging and is able to protect both isolated cells and the entire organism from oxidative stress.

Occurrence of Na(+)-Ca2+ exchange in the ciliate Euplotes crassus and its role in Ca2+ homeostasis.

Ciliates possess diverse Ca2+ homeostasis systems, but little is known about the occurrence of a Na(+)-Ca2+ exchanger. We studied Na(+)-Ca2+ exchange in the ciliate Euplotes crassus by digital imaging. Cells were loaded with fura-2/AM or SBF1/AM for fluorescence measurements of cytosolic Ca2+ and Na+ respectively. Ouabain pre-treatment and Na+o substitution in fura-2/AM-loaded cells elicited a bepridil-sensitive [Ca2+]i rise followed by partial recovery, indicating the occurrence of Na(+)-Ca2+ exchanger working in reverse mode. In experiments on prolonged effects, ouabain, Na+o substitution, and bepridil all caused Ca2+o-dependent [Ca2+]i increase, showing a role for Na(+)-Ca2+ exchange in Ca2+ homeostasis. In addition, by comparing the effect of orthovanadate (affecting not only Ca2+ ATPase, but also Na(+)-K+ ATPase and, hence, Na(+)-Ca2+ exchange) to that of bepridil on [Ca2+]i, it was shown that Na(+)-Ca2+ exchange contributes to Ca2+ homeostasis. In electrophysiological experiments, no membrane potential variation was observed after bepridil treatment suggesting compensatory mechanisms for ion effects on cell membrane voltage, which also agrees with membrane potential stability after ouabain treatment. In conclusion, data indicate the presence of a Na(+)-Ca2+ exchanger in the plasma membrane of E. crassus, which is essential for Ca2+ homeostasis, but could also promote Ca2+ entry under specific conditions.

Evidence of Toscana virus infections without central nervous system involvement: a serological study.

In central Italy, acute lymphocytic meningitis and meningoencephalitis due to a Phlebotomus transmitted virus (Phlebovirus Toscana, TOSv) occurring throughout the summer are frequently observed. Several serum specimens of patients hospitalized with a clinical picture of viral meningitis/meningoencephalitis showed anti TOS-IgG reactivity suggestive of a previous infection occurring at an unknown time in the past. This observation led us to design a serological investigation of 83 household contacts of 46 summertime CNS infection patients (index cases) with the purpose of evaluating the percentage of both IgG and IgM in seropositive healthy individuals, living, like the index cases, in areas at high risk of phlebotomine sandfly bites. The serological study was carried out using an ELISA method: 22% of the sera showed a reactivity for anti-TOS IgG antibodies and 6% resulted IgG/IgM positive; none of them reported having had any symptoms of CNS involvement; the distribution of seropositive cases was similar in contacts of both TOS-confirmed and TOS-negative cases. Our results indicate: (1) that phlebovirus Toscana is frequently implicated in cases with CNS disease as well as in infections occurring without neurological involvement; (2) in our country the ecological requirements encompass the conditions in and around the human settlements for phlebotomine sandflies to become peridomestic thus amplifying the risk of TOSv infections, which are in fact widespread and frequent in Siena and its surroundings.

The Seveso Directive: an Italian pilot study in enabling communication.

In 1982 the Council of the European Communities issued a Directive on the Major Accident Hazards of Certain Industrial Activities, commonly known as the Seveso Directive. Besides other requirements, the Directive specifies that members of the public liable to be affected by a major accident from certain industrial activities must be informed of safety measures and how they should behave in the event of an accident. The Directive was subsequently amended in an extensive way. Upon the transposition of the Seveso Directive and its amendments into national legislations, the 12 European Member States are faced with the task of "appropriately and effectively" informing the public about major accident hazards. Delays are observed throughout the community, both in transposing the Directives and implementing public information. A pilot study on "Risk Information Needs" was commissioned and carried out at two Seveso sites in Italy in order to begin to resolve some of the problems involved in the provision of public information and to suggest appropriate guidelines. One hundred subjects answered an experimental protocol designed to assess their "information needs." This article presents the theoretical background, the pragmatic aims, and the design of the study. It also summarizes its most relevant findings and suggests some recommendations for the provision of information to the public.

Effective Communication between the Scientific Community and the Media.

There are many accounts of hazard warnings which, although received by the target population, have not been appropriately understood and/or acted upon. In all such cases, what needs to be explored is a social relational failure rather than a technological one. Although the relationship between the mass media and the general public has been thoroughly explored, that between the media and the scientific community has been generally neglected. Scientists who communicate warning information to the media must recognise the background, commitments, values, needs and expectations of those they communicate with. Their efforts should be directed at conveying information in a way which is useful to the target population and, at the same time, which appeals to the media. Various "communication strategies" for the achievement of this goal are outlined.

[A case of erythema nodosum in tularemia]. / Descrizione di un caso di eritema nodoso in corso di tularemia.

Since 1980 some epidemics of tularemia occurred, involving large areas of Tuscany (Italy). Among the different clinical features described in the classic form of tularemia, erythema nodosum never has been reported. One case of erythema nodosum, the first reported in course of tularemia, is described. The correct diagnosis is based on careful question about exposure to mammals and ticks. The importance of early treatment is stressed.

[Comparative study and time changes in the resistance to gentamycin, tobramycin, sisomicin and amikacin of Gram-negative nosocomial bacterial strains isolated from the urinary tract, in relation to the annual use of the aminoglycosides tested]. / Studio comparativo e variazioni nel tempo delle resistenze a gentamicina, tobramicina, sisomicina ed amikacina di ceppi batterici nosocomiali Gram-negativi isolati dalle vie urinarie, in relazione ai consumi ospedalieri annuali degli aminoglucosidi esaminati.

Changes in susceptibility to four aminoglycosides (gentamycin, amikacin, tobramycin, sisomicin) of bacterial strains, isolated from in-patients with urinary tract infections at the "Santa Maria della Scala" Hospital of Siena, in the period January-April in three consecutive years (1980, 1981, 1982) were studied. The change susceptibility patterns were related to the use of the four antimicrobial drugs in the same periods. Emergence of antibiotic-resistant strains was seen after a wider use, whereas an increasing number of susceptible strains was observed after reducing use of antimicrobial drugs.