Studio di un nuovo algoritmo diagnostico per l’identificazione dei micobatteri non tubercolari (NTM)

NTM (Non tuberculous mycobacteria) count more than 160 species, widely dispersed in the environment as saprophites, but they can cause severe infections in human. The identification of NTM to the species level is extremely important to determine whether it is or not a NTM infection and the proper treatment. Therefore, many techniques have been developed for the accurate and rapid identification of NTM, among which PCR-RFLPs (PRAs) are easy, rapid and inexpensive tools for identifying mycobacterial species. The aim of this work was to realise a new PCR-RFLP technique with a high discriminatory power, cost effective, easy to be performed and able to overcome the limits of those used nowadays, using a new target, called dnaK, described as a powerful potential target for the identification of NTM. 30 mycobacterial species were used to develop a novel algorithm targeting locus dnaK and using restriction enzymes Fau I and BstMW I, which was successfully blind tested on 45 clinical micobacterial isolates that had been already identified to the species level with other techniques. Moreover, dnaK-PRA was tested on 40 clinical samples whose cultures were positive for mycobacteria and correctly detect and identified mycobacterial species in up to 82% of them. The results obtained showed that dnak-PRA is a reliable tool for the identification of mycobacterial isolates to the species level and can be helpful to detect and identify mycobacteria directly from clinical samples

Tuberculosis in Sardinia: An investigation into the relationship between natives and immigrants

AbstractObjective/background: Tuberculosis (TB) has had a recrudescence in the last few decades in Italy as a result of many factors, among which migration from countries where TB is endemic is one of them. In Sardinia, a major island of Italy, there was no knowledge of the mechanisms of transmission of TB in the immigrant subpopulation and the impact it may have on the native subpopulation and on the community as a whole. Therefore, a molecular epidemiological study was carried out to get a clearer picture of the number and genetic features of Mycobacterium tuberculosis strains isolated from immigrants and from natives in Sardinia. Methods: Two groups of clinical isolates of M. tuberculosis, one collected from immigrants and the other one from Sardinians, were analyzed in this study. The genotyping was executed through the variable number tandem repeat-mycobacterial interspersed repetitive units technique and a first-line antimycobacterial drug-susceptibility test was also carried out. Results: Thirty-six clinical isolates from immigrants and 25 from Sardinians were analyzed. Variable number tandem repeat-mycobacterial interspersed repetitive units technique showed that all of them belonged to different strains and there was a quite high allelic diversity among them. Moreover, data collected allowed the finding of, with a good approximation, the phylogenetic relations among the strains isolated and the best-known phylogenetic groups. Conclusion: The study pointed out that since every strain is different, there was no TB transmission in any of the subpopulations and between immigrants and natives. This showed that the presence of immigrants was not a risk factor for contracting TB in the community

Epidemic of tuberculosis in a high school in Northern Sardinia

Abstract The aim of this study was to investigate the Mycobacterium tuberculosis transmission among high school student and teacher populations in a high school in Northern Sardinia. Tuberculin skin-test screening, chest-X-rays, QuantiFERON-TB Gold, microbiological examination, spoligotyping and variable numbers of tandem repeats (VNTR) analysis of M. tuberculosis isolates were performed. This study indicates the effectiveness of the epidemiological investigation

Effect of floor type on carcass and meat quality of pen raised growing rabbits

[EN] The aim of the experiment was to compare the carcass and meat quality traits of growing rabbits housed on different floor types. At the age of 35 d, rabbits (n=126) were randomly sorted into 3 groups and housed in pens with different floor types: plastic-mesh, deep-litter straw or wire-mesh. Slaughter weight, carcass and its parts’ weight, meat (Longissimus thoracis et lumborum [LL] muscle and hind leg) pH and colour, oxidative status and fatty acid profile were measured and correlations calculated. The deep-litter straw rabbits showed the lowest pHu and b* values of LL muscle and oxidation of the both muscles. The fatty acid profile of LL muscle of deep-litter straw rabbits showed a higher percentage of monounsaturated fatty acids and long chain n-3 polyunsaturated (PUFA) fatty acids, whereas the content of  18:2n-6 and total PUFA was lower. We concluded that housing the growing rabbits on wire- or plastic-mesh floors showed no substantial differences, while housing rabbits on deep-litter negatively affected certain qualitative traits.This research was supported by the GOP-1.3.1-11/B-2011-0045 projectDal Bosco, A.; Szendrö, Z.; Matics, Z.; Castellini, C.; Ruggeri, S.; Szendrö, K.; Martino, M.... (2015). Effect of floor type on carcass and meat quality of pen raised growing rabbits. World Rabbit Science. 23(1):19-26. https://doi.org/10.4995/wrs.2015.3270SWORD1926231Dalle Zotte, A., Cullere, M., Sartori, A., Szendrő, Z., Kovàcs, M., Giaccone, V., & Dal Bosco, A. (2014). Dietary Spirulina (Arthrospira platensis) and Thyme (Thymus vulgaris) supplementation to growing rabbits: Effects on raw and cooked meat quality, nutrient true retention and oxidative stability. Meat Science, 98(2), 94-103. doi:10.1016/j.meatsci.2014.05.005Enser M., 1999. Nutritional effects on meat flavour and stability. In: Richardson, R.I., Mead, G.C. (Eds.). Poultry Meat Sci. Poultry Science Symposium Series. Vol. 25. CABI Publishing, Oxon, 197-215.Folch J., Lees M., Sloane-Stanley H. 1961. A simple method for the isolation and purification of total lipids from animal tissues. J. Biol. Chem., 226: 497-502.Jekkel G., Milisits G. 2009. Comparison of the behaviour of growing rabbits reared on wire net or combined floor at different stocking densities. Ital. J. Anim. Sci., 3: 202- 204.Jekkel G., Milisits G., Nagy I., Biró-Németh E. 2008. Analysis of the behaviour of growing rabbits housed in deep litter at different stages of rearing. In: Proceeding 9th World Rabbit Congress, June 10-13, 2008. Verona, Italy, pp. 1189-1193.Korkeala, H., Mäki-Petäys, O., Alanko, T., & Sorvettula, O. (1986). Determination of pH in meat. Meat Science, 18(2), 121-132. doi:10.1016/0309-1740(86)90088-4Matics Zs., Szendrő Zs., Radnai I., Biró-Németh E., Gyovai M. 2003. Examination of free choice of rabbits among different cage-floors. Agric. Consp. Scient., 68: 265-269.Szendrő, Z., & McNitt, J. I. (2012). Housing of rabbit does: Group and individual systems: A review. Livestock Science, 150(1-3), 1-10. doi:10.1016/j.livsci.2012.09.017Trocino A., Xiccato G., Queaque P.I., Sartori A. 2004. Group housing of growing rabbits: Effect of stocking density and cage floor on performance, welfare, and meat quality. In: Proceedings of the 8th World Rabbit Congress, September 7-, 2004. Puebla City, Mexico, pp. 1277-1281.Trocino A., Xiccato G., Majolini D., Fragkiadakis M. 2008. Effect of cage floor and stocking density on growth performance and welfare of group-housed rabbits. In: Proc. of the 9th World Rabbit Congress, June 10-13, 2008. Verona, Italy, pp. 1251-1255

Safety evaluation of buffered vinegar as a food additive

The EFSA Panel on Food Additives and Flavourings (FAF) provides a scientific opinion on the safety of buffered vinegar as a new food additive. Buffered vinegar is a liquid or dried product prepared by adding sodium/potassium hydroxides (E 524 to E 525) and sodium/potassium carbonates (E 500 to E 501) to vinegar, compliant with European Standard EN 13188:2000 and exclusively obtained from an agricultural source origin (except wood/cellulose). The primary constituents of buffered vinegar are acetic acid and its salts. No biological or toxicological data obtained with the proposed food additive were submitted by the applicant as part of the dossier as, following oral ingestion, buffered vinegar dissociates into the acetic anion and acetate a natural constituent of the diet, and of the human body for which extensive data on their biological effects exist and for which EFSA in 2013 has previously concluded that the establishment of an acceptable daily intake (ADI) is not considered necessary. At the proposed maximum/typical use levels, the mean exposure to buffered vinegar from its use as a food additive expressed as acetic acid equivalents ranged from 8.9 mg/kg body weight (bw) per day in infants to 280.3 mg/kg bw per day in children. The 95th percentile of exposure to buffered vinegar ranged from 27.9 mg/kg bw per day in infants to 1,078 mg/kg bw per day in toddlers. The Panel concluded that there is no safety concern for the use of buffered vinegar as a food additive at the proposed maximum/typical use levels. The Panel could not conclude on the safety for the proposed uses at quantum satis as Group I food additive since the resulting exposure could not be estimated

Antimycotic activity of Myrtus communis L. towards Candida spp. from isolates.

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Interferon-γ release assay in people infected with immunodeficiency virus

The present study aimed to use QuantiFERON TB Gold in tube (Cellestis Limited, Carnegie, Victoria, Australia) as a tool for the screening for tubercular infection in HIV-positive patients. Seventy-three HIV-positive subjects were tested. For each individual, QuantiFERON TB in tube was performed. The immunoassay was negative in 53 subjects, positive in eight and indeterminate in 12. The data obtained indicate that factors such as the CD4 cell count and their percentage, as well as the stage of the disease, could affect the performance of the interferon-γ release assay in populations at risk such as HIV-positive subjects

Usefulness of the QuantiFERON-TB-Gold <i>in tube</i> in a population at risk of bovine tubercular infection

The effectiveness of the QuantiFERON-TB-Gold in tube was compared with PPD to detect tuberculosis infection in the staff of cattle-farms, as a consequence of an outbreak of bovine tuberculosis in livestock.The data revealed the advantage of the immunological test for the specific detection of subjects infected by Mycobacterium tuberculosis complex

Usefulness of the QuantiFERON-TB-Gold in tube in a population at risk of bovine tubercular infection

The effectiveness of the QuantiFERON-TB-Gold in tube was compared with PPD to detect tuberculosis infection in the staff of cattle-farms, as a consequence of an outbreak of bovine tuberculosis in livestock.The data revealed the advantage of the immunological test for the specific detection of subjects infected by Mycobacterium tuberculosis complex

Follow‐up of the re‐evaluation of glycerol esters of wood rosins (E 445) as a food additive

Abstract Glycerol esters of wood rosin (GEWR) (E 445) were re‐evaluated in 2018. On the toxicity database and given the absence of reproductive and developmental toxicity data, the acceptable daily intake (ADI) of 12.5 mg/kg body weight (bw) per day for GEWR (E 445) established by the Scientific Committee on Food (SCF) in 1994 was considered temporary. The conclusions of the assessment were restricted to GEWR derived from Pinus palustris and Pinus elliottii and with a chemical composition in compliance with GEWR used in the toxicological testing. Following a European Commission call for data to submit data to fill the data gaps, the present follow‐up opinion assesses data provided by interested business operators (IBOs). Considering the technical data submitted by IBOs, the EFSA Panel on Food Additives and Flavourings (FAF Panel) recommended some modifications of the existing EU specifications for E 445, mainly a revision of the definition of the food additive and lowering the limits for toxic elements. Considering the available toxicological database evaluated during the re‐evaluation of E 445 by the ANS Panel in 2018, and the toxicological studies submitted by the IBOs, the Panel established an ADI of 10 mg/kg bw per day based on the no observed adverse effect level (NOAEL) of 976 mg/kg bw per day from the newly available dietary reproduction/developmental toxicity screening study in rats and applying an uncertainty factor of 100. Since GEWR from P. palustris and P. elliottii were tested in the toxicity studies considered to establish the ADI and in the absence of detailed information on the chemical composition (major constituents) in GEWR generated from other Pinus species, thus not allowing read across, the ADI is restricted to the GEWR (E 445) manufactured from P. palustris and P. elliottii. The Panel concluded that there was no safety concern for the use of GEWR (E 445), at either the maximum permitted levels or at the reported uses and use levels