Desenvolvimento pós-embrionário de Chrysomya putoria (Calliphoridae) em carne equina e sua importância para a entomologia e veterinária forenses / [Post embryonic development of Chrysomya putoria (Calliphoridae) using horse meat and its importance for forensic entomology and veterinary]

Devido à importância médica, veterinária, ambiental e forense do califorídeo Chrysomya putoria (Diptera), foi avaliado o desempenho apresentado por estágios imaturos criados em carne de cavalo. Os espécimes foram colocados em câmaras aclimatizadas reguladas a 30 ºC, 60 ± 10% UR e 14 horas de fotofase. Os estágios larval e pupal concluíram seu desenvolvimento em 4,16 e 4,12 dias, respectivamente. As larvas pós-alimentação e pupas com 24h de idade pesaram, em média, 58,06 e 40,10mg, respectivamente. As taxas de emergência registradas foram de 65% nas condições experimentais propostas. Como os compostos orgânicos voláteis são diferentes na decomposição de um animal para outro, este trabalho e outros realizados com a mesma dieta são os mais apropriados para estimativa de intervalo post mortem envolvendo cavalos.(AU)

Due to medical, veterinary, environmental and forensic importance of the calliphorid Chrysomya putoria (Diptera), the performance presented by immature stages, reared on horse meat, was evaluated. The specimens were placed in acclimatized chambers regulated at 30 ºC, 60 ± 10% RU and 14 hours of photo phase. The larval and pupal stages completed their development in 4, 16 and 4, 12 days, respectively. Post-feeding larvae and aged 24h pupae weight on average 58, 06 and 40, 10 mg, respectively. The emergence rates recorded were 65% under the experimental conditions proposed. As volatile organic compounds are different in the decomposition of one animal to another, this work and others carried out with the same diet are the most appropriate for estimating post-mortem interval involving horses.(AU)

Growth of Listeria monocytogenes in ready to eat salads at different storage temperatures and valuation of virulence genes expression.

BACKGROUND: Vegetables are major components of a healthy and balanced diet. However, 25% of foodborne diseases are linked to the consumption of vegetables. STUDY DESIGN: The aim of this work was to assess the microbiological risks associated with consumption of ready to eat salads (RTE). METHODS: Microbiological challenge tests were carried out for the evaluation of the L. monocytogenes growth potential in RTE salads stored at different temperatures. RESULTS: The results indicate that L.monocytogenes was able to grow (δ ≥ 0.5) in all storage conditions considered at the end of shelf life. In order to evaluate the virulence role of L. monocytogenes, the temperature-dependent transcription of major virulence genes was also investigated by RT-PCR. CONCLUSIONS: The microbiological challenge test allowed us to confirm, as also demonstrated by other authors, that RTE salads are able to support the growth of L. monocytogenes strains (d δ≥ 0.5) stored under different temperatures.

Microbiological Quality of the Water used in Agriculture in Sardinia.

BACKGROUND: The microbiological quality of the water used in irrigation is crucial for the safety of products, such as fruit and vegetables, especially when destined to be consumed raw. However, the microbiological quality of this water is not defined at a community regulatory level or at a national level. METHODS: With our present work, we wanted to investigate the microbiological quality of the water used for crop irrigation in various Sardinian provinces. Since in most fields the irrigation water is filtered to remove any impurities, the sample was processed twice - both before and after the filtering process. Furthermore, with the purpose of hypothesising the potential health risks attributable to the consumption of crops from the tested fields, samples of horticultural product were collect. Any eventual seasonal differences in the values of microbial concentration were assessed. Microorganism faecal contamination indicators (Escherichia coli, total coliform and faecal streptococci), but even the presence of the opportunistic pathogen such as Pseudomonas aeruginosa were researched in irrigation water. Total mesophilic counts (TMC) were assessed at 36°C and 22°C. On horticultural products we researched both the indicators of process parameters, such as Escherichia coli, Total mesophilic counts at 30°C, Enterobacteriaceae, Total Psychrophilic counts and Pseudomonas aeruginosa, and pathogens, such as Salmonella spp, Listeria monocytogenes and Yersinia enterocolitica. RESULTS: The number of target microorganisms, when present in irrigation water, was very limited: Escherichia coli, total coliform and faecal streptococci, were detected respectively in 48% and 67% of the water samples tested with average concentration values of 0.9, 1.2 and 1.4 log respectively. In fresh vegetable products, the total mesophilic counts (TMC) were found to have average values of 6.6x107 CFU/g. The average values of Enterobacteriaceae totalled 6.1x105 CFU/g; Escherichia coli was detected in only one sample (curly endive) with a value of 180 CFU/g. CONCLUSION: The data highlights the high quality of the water and how this contributed to achieving satisfactory quality on prime material. However the use of filters, to eliminate impurities, and reservoirs, may represent a crucial issue, if not managed correctly.

Hygiene and health risks associated with the consumption of edible lamellibranch molluscs.

Edible lamellibranch molluscs are frequently the cause of very serious outbreaks of food poisoning. Microbiological suitability for human consumption, according to EC Reg. n. 1441/2007, is only based on two faecal bacteriological contamination parameters (Escherichia coli and Salmonella) and does not consider other microorganisms which are naturally present within the marine environment and are potentially pathogenic such as those belonging to the genus Vibrio. In this study, a total of 540 samples of edible lamellibranch molluscs (mussels, clams and oysters) bred in Sardinia were analyzed for E. coli and Salmonella, according to EC Reg n. 1441/2005, and for the presence of Vibrios of major public health interest. Environmental parameters (seawater temperature and salinity) were also recorded. The 40% (n=216) of samples were positive for Vibrio while the levels of E. coli were above the legal European limit only in 6.4% of samples. A correlation between the presence of micro-organisms of faecal origin and potentially harmful Vibrios to human health was not observed. Vibrio alginolyticus was the most common in 68% of isolated species, followed by Vibrio parahaemolyticus in 22.6%, Vibrio cholerae in 6% and Vibrio vulnificus in 1.1%. Virulence gene (trh sequence) was present in 1.6% of the V. parahaemolyticus-positive samples. The V. cholerae, V. vulnificus and V. alginolyticus strains detected were non toxigenic. The absence of genes encoding major virulence factors in the studied strains does not exclude the possibility of exposure to pathogenic strains. This study indicates the potential health risk associated with the presence of pathogenic Vibrios in edible lamellibranch molluscs bred in Sardinia, where shellfish production is very important to the local economy.

Evaluation of the level of domestic hygiene in household kitchens.

BACKGROUND: Each year in industrialized countries, 30-40% or more cases of food poisoning occur in the household. The aims of this study are to describe the aspects related to food safety in households by carrying out a microbiological characterization of the kitchens in residential dwellings and to increase consumers' awareness concerning the importance of good hygienic practices, which are required for preventing foodborne diseases at household level. METHODS: The collection of data involved taking 760 analytical samples of kitchen-counter tops and food from 80 kitchens of private dwellings; the manner in which food was treated was evaluated from the moment of purchase to its transformation and storage by means of a checklist; the questionnaire enabled us to determine the extent of consumers' knowledge concerning the food sector. RESULTS: Overall, the results showed a good level of hygiene concerning both food and kitchen-counter top food surfaces. The respondents of the questionnaire did not appear to be aware of various health risks especially concerning the management of the temperature and compartments of refrigerators as well as food storage times and the cooking or heating of food. CONCLUSIONS: The data indicate that final consumers should take more care when cleaning kitchen-counter tops and washing salad; special training programmes should be included in school curricular in order to increase citizens' awareness and knowledge concerning food risks within the household.

Amino acid deprivation regulates the stress-inducible gene p8 via the GCN2/ATF4 pathway.

In mammals, the GCN2/ATF4 pathway has been described as the main pathway involved in the regulation of gene expression upon amino acid limitation. This regulation is notably conferred by the presence of a cis-element called Amino Acid Response Element (AARE) in the promoter of specific genes. In vivo, the notion of amino acid limitation is not limited to nutritional context, indeed several pathological situations are associated with alteration of endogenous amino acid availability. This is notably true in the context of tumour in which the alteration of the microenvironment can lead to a perturbation in nutrient availability. P8 is a small weakly folded multifunctional protein that is overexpressed in several kinds of cancers and whose expression is induced by different stresses. In this study we have demonstrated that amino acid starvation was also able to induce p8 expression. Moreover, we brought the evidence, in vitro and in vivo, that the GCN2/ATF4 pathway is involved in this regulation through the presence of an AARE in p8 promoter.

Rheumatoid arthritis is the major risk factor for septic arthritis in rheumatological settings.

Septic arthritis (SA) is a clinical emergency with considerable morbidity and mortality that can lead to rapid joint destruction and irreversible functional loss. The reported incidence varies from 2-5 cases/100,000 person-years in the general population to 70 cases/100,000 person-years among patients with rheumatoid arthritis. In fact, individuals with rheumatoid arthritis are at particular risk for developing SA. This may be due to several reasons: joint disease predisposes to bacterial joint colonization and RA itself and its treatment with corticosteroids, disease-modifying antirheumatic drugs (DMARDs) and biological therapies may decrease the immune function required for protection from pathogens. Steroids and DMARDs seem to affect the leukocyte synovial count; indeed, RA patients with SA have a leukocyte count in synovial fluid (SF) lower than patients with SA without underlying rheumatic diseases. The diagnosis of SA in RA patients can be difficult because the development of a hot painful joint is often confused with a relapse of the underlying joint disease leading to delay in diagnosis. For this reason the microscopic analysis and culture of synovial fluid are crucial to exclude septic arthritis.

[Septic arthritis: what is the role for the rheumatologist?]. / Artrite settica: il ruolo del reumatologo.

Septic arthritis (SA) is a clinical emergency with considerable morbidity and mortality that can lead to rapid joint destruction and irreversible loss of function. The reported incidence varies from 2-5 cases per 100.000 individuals per year in the general populations to 70 cases per 100.000 individuals annually among patients with rheumatoid arthritis (RA). Predisposing factors are immunosuppressive and corticosteroids therapy and RA "itself". The expected decrease in incidence of SA was not seen over the last 20 years period but we can, on the contrary, expect an increase in the frequency of its appearance because of the population ageing, the increasingly prosthetic joint replacement, the ability of the bacteria to evade clearance by the host immune response and the rapidly growing number of patients with RA, ankylosing spondylitis and psoriatic arthritis treated with tumour necrosis factor alpha (TNF alpha) antagonists. Up to now there have been conflicting reports regarding joint infections in patients under anti-TNF therapy but according to data from Deutsch as well as the British register there might be an increase in the incidence of joint infections in anti-TNF treated patients. Microscopic analysis and culture of synovial fluid are fundamental diagnostic tools in the evaluation of possible joint sepsis. Sonographic guidance of arthrocentesis led to successful aspiration of difficult-to-access joints as shoulder and hip. There is controversy over which mode of drainage of septic synovial fluid should be employed but needle aspiration appear to be preferable to surgical treatment as an initial mode of treatment of SA. Rheumatologists should have a central role in the diagnosis and management of SA.

[Septic arthritis: a 12 years retrospective study in a rheumatological university clinic]. / Artriti settiche: studio retrospettivo di 12 anni in un unico centro reumatologico.

BACKGROUND: Septic arthritis is a disabling and potentially life-threatening condition that requires prompt diagnosis and treatment. The most important risk factors are joint prosthesis, pre-existing joint disease and immunosuppressive drugs. The aim of our study therefore was to revaluate all septic arthritis cases discharged from our Rheumatologic Unit in the last 12 years, to assess the risk factors, the clinical and laboratory characteristics, the causative microorganisms and its possible increase in frequency. METHODS: The medical records of 42 consecutive patients with septic arthritis discharged from our Rheumatology Unit between January 1995 and December 2006 were reviewed. The patients ranged in age from 23 to 90 and there isn't gender predominance. Septic arthritis was diagnosed based on the finding of purulent material in the joint space and/or the isolation of a bacterial pathogen from joint fluid. Demographic data, risk factors, co-morbidity, clinical manifestations, time interval between symptoms onset and diagnosis, treatment and laboratory data including serum white blood cell count, erythrocyte sedimentation rate (ESR), C reactive protein (CRP), synovial white blood cells and culture results were analysed. We considered these parameters in the whole population and in two different age groups (< or =60, >60) and tried to determine if there was a change of microorganisms involved in septic arthritis during the years. RESULTS: Of 42 patients, 47% were aged 60 and younger. Only 10 patients were admitted to our unit before 2001. A predisposing factor was recorded in 90,5% of cases: 15 patients had rheumatoid arthritis, 8 were diabetic, 6 had seronegative arthritis, 4 had a connective tissue disease, 8 patients had a prosthetic infection and 3 were subjected recently to arthrocentesis. We found that patients aged 60 and younger were more frequently affected by joint disease and had a synovial white blood cell count lower than patients older than 60. Staphylococcus aureus caused septic arthritis in 70% of cases before 2001, and only in 35,8 % after 2001. Also, after 2001, some infections were caused by more unusual pathogens, prevalently in patients treated with TNFa inhibitors. Instead Streptococcus infections were found only in patients aged 70 and older. CONCLUSION: The incidence of bacterial arthritis has increased in the last six years and there was a modification of microorganisms involved, possibly related to a greater therapeutic aggressiveness. The increased frequency of joint disease and the use of immunosuppressive drugs in patients under the age of 60 could be responsible for a lower synovial white blood cell count in these patients.

Use of quantitative ultrasound of the hand phalanges in the diagnosis of two different osteoporotic syndromes: Cushing's syndrome and postmenopausal osteoporosis.

The aim of this study was to assess the ability of the quantitative ultrasound of the hand phalanges to detect different types of osteoporosis resulting from different pathogenetic mechanisms. For this purpose, postmenopausal and glucocorticoid-induced osteoporosis was studied. Thirteen female patients with Cushing's syndrome (CS) resulting from pituitary-dependent bilateral adrenal hyperplasia (10 patients) and from adrenal adenoma (3 patients), and 32 postmenopausal osteoporotic (OP) women, were examined. The two groups of patients were comparable for body mass index (BMI), but CS patients were significantly younger than OP ones (CS 44.5+/-11.6; OP: 73.9+/-3.6). All the patients had femoral neck bone mineral density (BMD) T-score less than -2.0. Cushing patients had a femoral neck BMD similar to that of OP patients (CS: 603+/-66 mg/cm2; OP: 628+/-69 mg/cm2; p=0.19). In contrast, amplitude-dependent speed of sound (AD-SoS) was significantly higher in CS patients than in OP patients (CS: 1997+/-91 m/s; OP: 1707+/-114 m/s; p<0.0001). By adjusting DXA and ultrasound parameters according to age, femoral neck BMD was significantly lower in CS patients and AD-SoS remained significantly higher than in OP patients. These findings indicate that these two different kinds of osteoporosis can be distinguished by ultrasonography and that ultrasound parameters alone cannot be used for evaluating skeletal status in CS patients.

Inhibition of CHOP translation by a peptide encoded by an open reading frame localized in the chop 5'UTR.

Chop is a ubiquitously expressed mammalian gene encoding a small nuclear protein related to the CCAAT/enhancer-binding protein (C/EBP) family of transcription factors. CHOP protein plays an important role in various cellular processes such as growth, differentiation and programmed cell death. CHOP expression is strongly increased in response to a large variety of stresses including perturbation of the endoplasmic reticulum function, DNA damage and nutrient deprivation. Multiple mechanisms including transcriptional and post-transcriptional controls are involved in the regulation of CHOP expression. We show here that the 5'UTR of the Chop transcript plays an important role in controlling the synthesis of CHOP protein. In particular, the 5'UTR contains a conserved uORF which encodes a 31 amino acid peptide that inhibits the expression of the downstream ORF. Mutational analysis of the 5' leader region and peptide coding sequences suggests that the peptide itself inhibits expression of the downstream ORF. Such results suggest a role for uORF in limiting ribosomal access to downstream initiation sites. With respect to the importance of CHOP protein in the regulation of cellular functions, the mechanisms that regulate its basal level are of considerable interest.

Amino acids control mammalian gene transcription: activating transcription factor 2 is essential for the amino acid responsiveness of the CHOP promoter.

In mammals, plasma concentration of amino acids is affected by nutritional or pathological conditions. It has been well established that nutrients, and particularly amino acids, are involved in the control of gene expression. Here we examined the molecular mechanisms involved in the regulation of CHOP (a CCAAT/enhancer-binding protein [C/EBP]-related gene) expression upon amino acid limitation. We have previously shown that regulation of CHOP mRNA expression by amino acid concentration has both transcriptional and posttranscriptional components. We report the analysis of cis- and trans-acting elements involved in the transcriptional activation of the human CHOP gene by leucine starvation. Using a transient expression assay, we show that a cis-positive element is essential for amino acid regulation of the CHOP promoter. This sequence is the first described that can regulate a basal promoter in response to starvation for several individual amino acids and therefore can be called an amino acid response element (AARE). In addition, we show that the CHOP AARE is related to C/EBP and ATF/CRE binding sites and binds in vitro the activating transcription factor 2 (ATF-2) in starved and unstarved conditions. Using ATF-2-deficient mouse embryonic fibroblasts and an ATF-2-dominant negative mutant, we demonstrate that expression of this transcription factor is essential for the transcriptional activation of CHOP by leucine starvation. Altogether, these results suggest that ATF-2 may be a member of a cascade of molecular events by which the cellular concentration of amino acids can regulate mammalian gene expression.