RICERCHE EFFETTUATE
IGIENE DEGLI ALIMENTI AD USO UMANO
Accurso° D, Menotta° S, Fedrizzi° G
Studio sull'origine della contaminazione di Sulfadiazina nel miele
XIV Congresso Nazionale SIDiLV : 24 - 26 Ottobre 2012, Sorrento (NA) : volume degli atti / [s.l. :
Societa' Italiana Diagnostica di Laboratorio Veterinaria ( SIDiLV ), 2012]. - p 123-124. - 2 bib ref [Nr.
Estr. 5104]
Congresso Nazionale Societa' Italiana Diagnostica di Laboratorio Veterinaria (SIDiLV) (14. :
Sorrento (NA) : 24 - 26 Ottobre 2012)
The bees's diseases induce beekeeper to uncontrolled use of antibiotics. Occasionally the not
justified antibiotics presence in honey occur. Fraudulent use of veterinary medicinal products is the
most common practice but the origin of contamination is hard to find. This is the case where the
presence in honey of Sulfadiazine (SDZ) is not proven by the beekeeper's use. The investigation
extends outside the beekeeping. The first hypothesis consider the contiguity rela-tions with
neighboring farms or livestock but finally the more probable hypothesis is the robbers bees's activity
on nearby hives.
Arrigoni° N, Weiss C
Latte crudo in Emilia Romagna : risultati del progetto Sibilla
Convegno Microbiologia Predittiva: possibili utilizzi nell'attività di controllo ufficiale degli operatori
del settore alimentare / [s.l. : s.n., 2012]. - 19 p.
http://www.ausl.re.it/Home/DocumentViewer.aspx?ID=1759&TIPODOC=COMUNICAZIONE [Nr.
Estr. 5224]
Convegno Microbiologia Predittiva: possibili utilizzi nell'attività di controllo ufficiale degli operatori del
settore alimentare : Reggio Emilia : 30 Novembre 2012)
Bertasi° B, D'Amico° S, Tilola° M, Ferrari° M, Pant eghini° C, D'Incau° M, Finazzi° G,
Daminelli° P, Bonomini A, Pedroni P, Losio° MN
Caratterizzazione molecolare di ceppi del genere salmonella in un focolaio di tossinfezione
alimentare
XIV Congresso Nazionale SIDiLV : 24 - 26 Ottobre 2012, Sorrento (NA) : volume degli atti / [s.l. :
Societa' Italiana Diagnostica di Laboratorio Veterinaria ( SIDiLV ), 2012]. - p 148-149. - 5 bib ref [Nr.
Estr. 5103]
Congresso Nazionale Societa' Italiana Diagnostica di Laboratorio Veterinaria (SIDiLV) (14. :
Sorrento (NA) : 24 - 26 Ottobre 2012)
The transmission of Salmonella to humans mainly occurs through the ingestion of animal origin
contaminated food. Salmonella infections are, in most industrialized countries, a major cause of
foodborne humans illness. First aim of this work was to assess the existence of a correlation
between the Salmonella strains isolated from patients and those isolated from food, through two
molecular techniques (PFGE and Ribotyping). Moreover, we have compared the discriminative
power and the consequent usefulness in cases food-borne of these two molecular techniques.
Bertasi° B, Galuppini° E, Consoli° M, Fusini° F, Pa voni° E, Saetti° F, Rubini° S,
Daminelli° P, Finazzi° G, Losio° MN
Monitoraggio di Vibrio spp. in molluschi eduli lamellibranchi e acqua di mare
XIV Congresso Nazionale SIDiLV : 24 - 26 Ottobre 2012, Sorrento (NA) : volume degli atti / [s.l. :
Societa' Italiana Diagnostica di Laboratorio Veterinaria ( SIDiLV ), 2012]. - p 150-151. - 4 bib ref [Nr.
Estr. 5102]
Congresso Nazionale Societa' Italiana Diagnostica di Laboratorio Veterinaria (SIDiLV) (14. :
Sorrento (NA) : 24 - 26 Ottobre 2012)
The aim of this work was to confirm, by Polymerase chain reaction (PCR), the samples of seawater
and clams positive with cultural methods, in order to evaluate the presence of Vibrio cholera, Vibrio
parahaemolyticus and Vibrio vulnificus in cosatal areas adjacent to the Po outfall.
Bertocchi° L, Scalvenzi° A, Santini S, Fusi° F
Aflatossine dal mais al latte? La siccità alimenta i timori
Inf Zootec. - Vol. 59 no 15 ( 2012). - p 6-18 [Nr. Estr. 5063]
Biancardi° A, Aimo° C, Cremaschini° L, Piro° R
I rischi emergenti : presenza di ocratossina A in alimenti "non convenzionali"
4° Congresso nazionale "Le micotossine nella filier a agro-alimentare" : 11-13 giugno 2012 Roma :
riassunti / a cura di Carlo Brera ... [et al.]. - Roma : Istituto Superiore di Sanità, c2012. - (ISTISAN
Congressi ; 12/C3) p 47 [Nr. Estr. 5020]
Congresso nazionale "Le micotossine nella filiera agro-alimentare" (4. : Roma : 11-13 giugno
2012)
L'Ocratossina A (OTA) è una micotossina nefrotossica prodotta da specie tossinogene dei generi
Aspergillus e Penicillium. Sebbene cereali e derivati siano considerati i principali substrati di
contaminazione da OTA, anche il caffè, l'uva e i prodotti derivati (vino, aceto) costituiscono una
secondaria ma rilevante fonte di esposizione (circa 25%). È stato condotto uno studio di
monitoraggio per valutare la presenza di OTA in caffè, vino e aceto. Per i campioni di vino è stato
usato un metodo IAC-HPLC normato [OIV- Ed. 2009 (MA¬E-AS315-10-0CHRAT)]. Il metodo, previo
opportuno aggiustamento del pH nella fase iniziale di diluizione, è stato anche applicato ai campioni
di aceto. Dal medesimo è stata poi messa a punto una variante adatta per la matrice caffè. La
concentrazione minima rilevabile del metodo è 0,02 pg/kg per vino e aceto, mentre è 1 pg/kg per il
caffè. La normativa vigente (Regolamento CE 1881/2006) stabilisce tenori massimi di OTA pari a 2
pg/kg per il vino, 5 µg/kg per il caffè torrefatto e 10 pg/kg per il caffè solubile. Per la matrice aceto
non è previsto alcun limite di legge. Per quanto riguarda i campioni di caffè esaminati (n=104), il
95% è negativo ovvero <1 pg/kg. Il restante 5% è risultato >1 pg/kg; i valori riscontrati vanno da un
minimo di 2,45 pg/kg ad un massimo di 6,35 pg/kg (un solo valore è risultato superiore al limite di 5
pg/kg). La situazione è dunque del tutto rassicurante circa il rischio di esposizione ad OTA
attraverso il caffè; ciò è essenzialmente da attribuire alle drastiche condizioni di tostatura che
comportano una significativa riduzione del tenore di OTA eventualmente presente nelle materie
prime. Per quanto riguarda i campioni di vino esaminati (n=122), 1'84% è <0,02 µg/kg, mentre nel
restante 16% è stata riscontrata la presenza di OTA tra 0,02 pg/kg e 1,5 pg/kg con un massimo di
incidenza dell'8% nell'intervallo 0,021+0,10 µg/kg. I vini rossi sono risultati più contaminati dei
bianchi, e comunque tutti al di sotto del limite di tolleranza. Il panorama complessivo pare quindi
rassicurante; nessun campione è risultato positivo al di sopra del limite normato (il valore più alto
riscontrato è 1,49 p.g/kg). Al contrario, solo il 10% dei campioni di aceto (non balsamico, balsamico
e balsamico tradizionale per un totale di n=100) è risultato negativo ovvero <0,02 µg/kg. Nel 90% dei
campioni è stata riscontrata la presenza di OTA tra 0,02 pg/kg e 2 p.g/kg con un massimo di
incidenza del 40% nell'intervallo 0,11+0,50 pg/kg. Questa diffusa presenza di OTA, che potrebbe
essere ascrivibile ad un eventuale uso di materie prime di scarsa qualità ovvero già contaminate, è
stata riscontrata più nell'aceto balsamico che nel comune aceto di vino non balsamico; ciò potrebbe
essere spiegato sulla base di un possibile effetto di concentrazione di OTA durante la fase di cottura
del mosto e di maturazione del prodotto.
Biancardi° A, Aimo° C, Piazza° P, Piro° R
Tecniche di screening e conferma: il caso aflatossina M1
4° Congresso nazionale "Le micotossine nella filier a agro-alimentare" : 11-13 giugno 2012 Roma :
riassunti / a cura di Carlo Brera ... [et al.]. - Roma : Istituto Superiore di Sanità, c2012. - (ISTISAN
Congressi ; 12/C3) p 20 [Nr. Estr. 5019]
Congresso nazionale "Le micotossine nella filiera agro-alimentare" (4. : Roma : 11-13 giugno
2012)
Il Regolamento CE n. 1881/2006 per il controllo della presenza di residui di aflatossina Ml nel latte
richiede metodologie analitiche sensibili, precise ed accurate. Le più diffuse sono basate su metodi
normati: ISO 14675-2003 (metodo ELISA solitamente usato per analisi di screening) e ISO
14501-2007 (metodo IAC-HPLC impiegato per conferme e considerato tecnica di elezione). Per
avere un dato oggettivo di affidabilità, sono stati confrontati i dati di controllo di qualità prodotti dal
laboratorio nell'arco di 4 anni di attività. È stato sempre utilizzato lo stesso tipo di kit ELISA, avente
unico fornitore e caratterizzato da prestazioni analitiche costantemente garantite e certificate. Per
quanto riguarda la tecnica ELISA, sono stati raccolti 338 dati di accuratezza ottenuti nel periodo
2007-2010 utilizzando 7 diverse matrici di riferimento nell'intervallo da 11 a 69 ng/kg. I valori di
z-score medio, ottenuti per ogni livello, vanno da un minimo di -0,22 ad un massimo di 0,08. Per
quanto riguarda la tecnica IAC-HPLC, sono stati raccolti 151 dati di accuratezza ottenuti nello stesso
periodo con 7 diverse matrici di riferimento nell'intervallo da 32 a 69 ng/kg. I valori di z-score medio,
ottenuti per ogni livello, vanno da un minimo di -0,43 ad un massimo di 0,16. Dal confronto ANOVA
delle due rette di correlazione tra le medie delle concentrazioni trovate vs le concentrazioni teoriche
attese (r2=0,9968 per ELISA; r2=0,9784 per HPLC) non si evidenzia una differenza significativa tra i
2 metodi. Quindi si può anzitutto affermare che il metodo ELISA ha prestazioni analoghe al metodo
di riferimento IAC-HPLC. Inoltre, da un'attenta valutazione dei risultati in termini di accuratezza
(z-score; rette di correlazione) e precisione (CV%) si può aggiungere che il metodo ELISA è migliore
del metodo IAC-HPLC. Questa apparentemente anomala conclusione è dovuta al fatto che nel
metodo ELISA la manipolazione del campione è minima (il campione è centrifugato ed
eventualmente diluito prima del test). Ciò comporta importanti e innumerevoli vantaggi quali velocità
di analisi e costi ridotti, ma anche un'elevata produttività e soprattutto una minore probabilità di
introdurre errori casuali e sistematici. Al contrario, il metodo HPLC comporta varie fasi, tra cui una
purificazione e concentrazione mediante IAC; aumenta quindi la probabilità di introdurre errori,
influendo significativamente sulle caratteristiche di accuratezza e precisione. Inoltre il metodo è più
dispendioso sia in termini di tempo che di costi e non si presta all'analisi di un numero elevato di
campioni per seduta analitica. Tali risultanze possono essere particolarmente utili per quei laboratori
che, per scelta strategica o istituzionale o per mancanza di idonea strumentazione cromatografica e
quindi di risorse, si limitano ad uno screening ELISA. Opportunamente condotta l'analisi mediante
ELISA è in grado di dare risultati quantitativi precisi ed accurati a qualsiasi livello di contaminazione,
a patto di utilizzare un kit ELISA idoneo e di operare secondo ISO 17025. Queste conclusioni sono
state applicate ad uno studio di monitoraggio su latte formulato per la prima infanzia usando
unicamente la tecnica ELISA. I campioni (n=114) sono risultati negativi (<5 ng/kg) e ne è stata
dimostrata la reale negatività mediante prove mirate ELISA, senza far ricorso al metodo HPLC.
Bolzoni° G, Marcolini° A, Delle_ Donne° G, Varisco ° G
Le innovazioni in Lombardia : sicurezza alimentare, art 68, quote
Inf Zootec. - Vol. 59 no 19 ( 2012). - p 40-46 [Nr. Estr. 5141]
Bonilauri° P
Prodotti trasformati a base di carne di pecora Cornella
Convegno Microbiologia Predittiva: possibili utilizzi nell'attività di controllo ufficiale degli operatori
del settore alimentare / [s.l. : s.n., 2012]. - 26 p
http://www.ausl.re.it/Home/DocumentViewer.aspx?ID=1763&TIPODOC=COMUNICAZIONE [Nr.
Estr. 5227]
Convegno Microbiologia Predittiva: possibili utilizzi nell'attività di controllo ufficiale degli operatori del
settore alimentare : Reggio Emilia : 30 Novembre 2012)
Boschetti L, Rubini° S, Losio° MN, Pavoni° E, Berta si° B, Berardelli C
Caratterizzazione della qualità microbiologica nei molluschi bivalvi vivi sottoposti a ciclo
depurativo, prima della commercializzazione, nei centri depurazione molluschi
Convegno Microbiologia Predittiva: possibili utilizzi nell'attività di controllo ufficiale degli operatori
del settore alimentare / [s.l. : s.n., 2012]. - 21 p.
http://www.ausl.re.it/Home/DocumentViewer.aspx?ID=1764&TIPODOC=COMUNICAZIONE [Nr.
Estr. 5223]
Convegno Microbiologia Predittiva: possibili utilizzi nell'attività di controllo ufficiale degli operatori del
settore alimentare : Reggio Emilia : 30 Novembre 2012)
Bronzo V, Locatelli C, Cremonesi P, Castiglioni B, Varisco° G, Bertocchi° L, Moroni
P
MRSA detection in bulk milk from Italian dairy farms
XXVII World Buiatrics Congress : 3 - 8 June, 2012 Lisbon, Portugal : abstract book / [s.l. : s. n.,
2012]. - p 130 (P:416) [Nr. Estr. 5022]
World Buiatrics Congress (27th : Lisbon, Portugal : 3 - 8 June, 2012)
Objectives: Methicillin-Resistant variant of Staph. aureus (MRSA) is an important issue in human
infection and has been recently becoming a veterinary matter since it has been isolated in pets and
livestock as well. Poultry and swine holdings seem to be important MRSA reservoir but also lactating
cows can be positive. As a mastitis pathogen it is an issue in the regards of antimicrobial treatment
and zoonotic potential. Materials and Methods: In order to investigate MRSA pressure and
prevalence in an area characterized by a high livestock density, bulk milk samples from 270 Staph.
aureus positive dairy farms were collected in Brescia area. Ali samples underwent search for MRSA
both by molecular and cultural methods. An aliquot from each sample was incubated overnight
before DNA extraction. Alter a milk sample pre-treatment, in order to remove protein and fat content,
the DNA extraction was done following a protocol describe in literature. PCRs amplifying Staph.
aureus 23S specific region and mecA gene were performed in two independent reactions. One
hundred microliters of milk were plated onto Baird Parker agar, in order to confirm Staph. aureus
positivity, and onto MRSA Chromogenic Agar (Pronadisa) after incubation firstly in Mueller Hinton
broth added with 6,5 % NaCI and then in Tryptic Soy broth added with cefoxitin and aztreonam. Both
cultural media were incubated for 48 hours at 37°C. Suspected colonies were isolated and
subcultured onto Mueller Hinton agar with 6 g/I of oxacillin. Isolates able to grow underwent DNA
extraction and a duplex-PCR targeted to nuc gene and mecA gene in order to confirm the results
obtained on bulk milk samples. Results: To our knowledge this is the first survey in Italy searching
for MRSA in bulk milk both by PCR and classic bacteriological methods. Up to now 119 samples
have been processed and 5 MRSA positive farms have been recognized both by PCR and culturing.
Beside them, other 33 farms resulted mecA gene-positive through total milk PCR but no MRSA was
isolated. In those cases only mecA gene but not nuc gene has been detected by PCR on
isolates.Conclusions: That justifies total milk PCR results and identifies the presence of
coagulase-negative Staphylococci (CNS) harboring the same MRSA resistance determinant. A
provisional MRSA prevalence of 4,2 % has so been found, that is a relevant one in consideration of
the threat to public and animai health.
Capuano F, Picotto P, Delibato E, Di_Pasquale S, Lena R, Losio° MN, Durante G,
Mancini L, Caponigro V, Dionisi AM, De_Giusti M, Luzzi I, De_Medici D, Borrello S
Allerta europee e prodotti di origine vegetale: il caso salmonella Napoli nella Piana del Sele
Infezioni trasmesse da alimenti e acqua : diagnostica ed epidemiologia : VIII workshop nazionale
Enter-net Italia, Sistema di sorveglianza delle infezioni enteriche : 31 maggio-1 giugno 2012, Vietri
sul Mare, Salerno / a cura di Caterina Graziani, Francesca Manciini e Ida Luzzi. - Roma : Istituto
Superiore di Sanità, 2012. - (ISTISAN congressi ; 12/C2) p 36 [Nr. Estr. 4995]
Workshop nazionale Enter-net Italia (8 : Vietri sul Mare, Salerno : 31 maggio-1 giugno 2012)
L'aumento del consumo di vegetali freschi ha recentemente causato diversi focolai di tossinfezione
dovuti alla contaminazione di questi prodotti da parte di microrganismi patogeni da ricondurre
principalmente alle pratiche di coltivazione, manipolazione e lavorazione. Le fonti d'acqua
contaminate, impiegate per irrigare le colture e lavare i raccolti prima del conferimento alla
trasformazione, rappresentano uno dei principali fattori di rischio implicati in un grande numero di
questi focolai. Inoltre, in questi ultimi anni il sistema europeo di allerta RASFF, ha più volte
richiamato l'attenzione dei Paesi membri riguardo al pericolo Salmonella nei prodotti vegetali, e
diverse sono state le allerta comunitarie che hanno interessato vegetali coltivati in Italia. Nei periodi
2004-2005 e 2008-2009, tali allerta hanno riguardato principalmente la rucola prodotta in Campania
e alcuni vegetali contaminati da S. Napoli, un sierotipo responsabile del 2-3% di casi di infezione
nell'uomo e raramente isolato da animali da reddito. Per rispondere alle allerta comunitarie, il
Ministero della Salute ha costituito un gruppo di studio multidisciplinare per affrontare le
problematiche relative alla contaminazione microbiologica dei prodotti vegetali. In particolare,
l'attività del gruppo si è focalizzata sulla valutazione del rischio associato al consumo di vegetali
freschi e sulla definizione di strategie di prevenzione e protezione da rischi correlati a pericoli
microbiologici veicolati dai vegetali di I e IV gamma. Per definire il rischio correlato alla presenza di
S.' Napoli nei vegetali coltivati nella Piana del Sele è stato effettuato un monitoraggio su campioni di
acqua irrigua e campioni vegetali di I gamma che sono stati analizzati sia con metodiche colturali sia
di real-time PCR. I ceppi isolati sono stati sottoposti a tipizzazione fenotipica e genotipica attraverso
elettroforesi in campo pulsato (PFGE).
In 2004-2005 and 2008-09 the European Rapid Alert System Food and Feeds (RASFF) notified
different alerts involving rocket salads produced in Campania, a south-central Italian region. Most of
the alerts reported vegetable contamination by Salmonella Napoli, a serovar which in Italy accounts
for 3% of human infections, frequently isolated from surface water but rarely isolated from foodstuff
of animal origin.
In order to manage these repeated alerts the Italian Ministry of Health organized
a multidisciplinary taskforce aiming at defining strategies to reduce or eliminate the risk of
contamination of fresh vegetables. In particular the studies were focused on: determining the
sources of contamination by S. Napoli in Campania region; validating a rapid method (Real Time
PCR) to detect microbial contamination on vegetables; defining the prevalence of microbial
contamination in Italian vegetables.
Cirillo G, Mangiavillano A, Biserni R, Luzzi I, Manuppella A, Molina M, Scuota S,
Staffolani M, Stenico A, Bandettini G, Pongolini° S
Salmonella e ambiente: dati 2009-2011
Infezioni trasmesse da alimenti e acqua : diagnostica ed epidemiologia : VIII workshop nazionale
Enter-net Italia, Sistema di sorveglianza delle infezioni enteriche : 31 maggio-1 giugno 2012, Vietri
sul Mare, Salerno / a cura di Caterina Graziani, Francesca Manciini e Ida Luzzi. - Roma : Istituto
Superiore di Sanità, 2012. - (ISTISAN congressi ; 12/C2) p 13 [Nr. Estr. 4993]
Workshop nazionale Enter-net Italia (8 : Vietri sul Mare, Salerno : 31 maggio-1 giugno 2012)
La ricerca del genere Salmonella su matrici ambientali, attraverso un monitoraggio periodico, viene
effettuata nei Laboratori di Microbiologia delle Agenzie Regionali di Prevenzione Ambientale
(ARPA). Nelle matrici ambientali si assiste (a differenza delle matrici di derivazione clinica e
veterinaria) a una marcata eterogeneità di specie. In Emilia-Romagna la sezione ARPA di Forlì
ospita dal 1997 (e prima ancora come progetto Salmnet) il Centro di Riferimento Enter-net. In questi
anni il Centro è riuscito ad avere come clienti la quasi totalità dei Laboratori di Microbiologia della
Regione, questo rende molto attendibili e vicini alla realtà i dati che annualmente pubblichiamo in un
Report. Dal 2011 poi il Centro è divenuto partner dell'IZS di Parma in un programma che prevede
che i ceppi umani siano sottoposti sierotipizzazione e successiva elettroforesi in campo pulsato
(PFGE) al fine di definirne il genotipo. Questo migliora la tracciabilità rispetto alla sola
sierotipizzazione permettendo una più accurata identificazione dei focolai di MTA. Si è costituita poi
una minirete dedicata solo ai dati di origine ambientale cui partecipano Piemonte, Liguria, Prov.
Autonoma di Bolzano, Emilia-Romagna; Molise, Lazio, Veneto. Nel triennio 2009/2011, i Laboratori
sopra indicati, hanno isolato e tipizzato 4.640 sierotipi di cui 2.687 ambientali, 1.870 umani. Dei
2.687 ceppi ambientali, circa il 75% proviene da acque superficiali, il 15% da acque di scarico, 1'8%
da fanghi e il 2% da altre fonti (indoor, ecc). Si può notare l'alta variabilità di specie che si verifica
negli isolamenti da matrici ambientali con 168 sierotipi diversi anche se i Gruppi B/C/D/E/F sono i
maggiormente rappresentati. I primi dieci sierotipi (circa il 55% contro 1'84% dei top-ten umani)
isolati da matrici ambientali sono S. Typhimurium (22,4%), Veneziana (20,3%), Derby (12,0%), Var.
Monofasica (10,1%), Infantis (8,1%), Arizona (6,5%), Rissen (6,1%), London (5,7%), Enteritidis
(4,4%), Hadar (3,4%). S. Veneziana, dopo S. Typhimurium, è ancora la specie di maggior
isolamento nell'ambiente, anche se, per ora, non trova validi riscontri nell'uomo e negli alimenti.
Interessante il costante isolamento anche da matrici ambientali della Var., Monofasica. Esiste,
invece, una convergenza nei primi dieci sierotipi di 5/10 fra umani e ambientali. S. Enteritidis, isolata
dal 12,3% delle infezioni umane, rappresenta solo il 4,5% degli isolamenti ambientali. La variabilità
di sierotipi ambientali testimonia la presenza di questo microorganismo in serbatoi naturali non
costituiti soltanto da insediamenti agroalimentari, ma anche da specie selvatiche. Se da una parte è
noto che solo alcuni sierotipi parassitano uomo e animali diventando agenti di MTA non possiamo
escludere che anche altri sierotipi possano causare MTA come accaduto in questi tre anni in Europa
con le piccole epidemie, documentate dalla rete Enter-net, sostenute da S. Goldcoast e S.
Stratchona.
Consoli° M, Galuppini° E, Losio° MN, Di_Bartolo I, Ruggeri FM, Varisco° G
Human norovirus infection : an Italian case
Norovirus and other Caliciviruses on the Rise International Conference : March 20-22, 2012,
Luebeck, Germany / [s.l. : s.n.], 2012. - p 60 - 3 bib ref [Nr. Estr. 4970]
Rise International Conference : Luebeck, Germany : March 20-22, 2012)
Noroviruses (NoVs) are small, non-enveloped, icosahedral viruses members of Caliciviridae family
and are a major cause of outbreaks of acute gastroenteritis world-wide.1 NoVs have a single-strand,
positive sense RNA genome of 7.5-7.7 kb in length with three open-reading¬frames (ORFs), that
encode for structural and non-structural proteins. NoVs are genetically different and have been
classified finto five genogroups based in homology sequence of capsid gene; NoVs GGI, GGII and
GGIV have the most significance to human health.2 These viruses are commonly identified in
different foodmatrices and mussels; red fruits and vegetables are often contaminated. Recently, the
central role of water as an important route of human NoVs transmission was demonstrated. NoVs
are the third most frequent pathogenes for waterborne disease. Several large foodborne and
waterborne outbreaks due to NoVs have been described, but there are only a few reports in which
both epidemiological and environmental data have been confirmed by molecular data on the
waterborne NoVs.3 The present work describes the epidemic case occurred in the Alps village of
Malonno in Valle Trompia (Lombardy) on September 2011, when a group of 30 children between 13
and 18 years old, showed the typical symptoms of gastroenteritis during a summer camp. Three
persons were also hospitalised. At the Food Safety Department of Istituto Zooprofilattico in Brescia,
one faecal sample and one rectal swab from a female patient, and three water samples, collected
from a drinking tube near the river of the camp, were analysed. The faecal sample and the rectal
swab were clarified in sterile distilled water (1 g in 10 ml), while water samples were filtered in acid
conditions, also with AICI3(6H20) 0.5 mM addition . After the RNA extraction with a commercial kit,
the retrotranscription of the nucleic acid was performed and the cDNA was amplified with two
Real-Time PCRs, specific for GGI e GGII NoVs genogroups. The faecal sample, the rectal swab,
and one of the three water samples were positive for NoV GGI. These results were also confirmed
performing a "booster PCR" giving 326 bp amplicons. Sequencing analysis of the amplified DNA,
executed in Brescia and in the Istituto Superiore di Sanità in Rome, confirmed the presence of NoV
GGI, strain GI.4, in all the three samples. The comparison of the GI.4 NoV sequences, showed
100% homology. These data support the hypothesis that water could had a role in the transmission
of Noroviruses to the children attending the summer camp. Water from the collecting site, probably,
derived from the river and contamination could arose by pastures located upstrea m..
Daminelli° P
Il progetto Ars alimentaria
Convegno Microbiologia Predittiva: possibili utilizzi nell'attività di controllo ufficiale degli operatori
del settore alimentare / [s.l. : s.n., 2012]. - 22 p
http://www.ausl.re.it/Home/DocumentViewer.aspx?ID=1768&TIPODOC=COMUNICAZIONE [Nr.
Estr. 5228]
Convegno Microbiologia Predittiva: possibili utilizzi nell'attività di controllo ufficiale degli operatori del
settore alimentare : Reggio Emilia : 30 Novembre 2012)
Daminelli° P, Cosciani_Cunico° E, Fierro A, Finazzi ° G, Bertasi° B, Dalzini° E,
Varisco° G
Sviluppo e validazione di un modello predittivo per stimare la crescita di Bacillus cereus
durante il raffreddamento post-pastorizzazione del mascarpone
XIV Congresso Nazionale SIDiLV : 24 - 26 Ottobre 2012, Sorrento (NA) : volume degli atti / [s.l. :
Societa' Italiana Diagnostica di Laboratorio Veterinaria ( SIDiLV ), 2012]. - p 237-239. - 8 bib ref [Nr.
Estr. 5098]
Congresso Nazionale Societa' Italiana Diagnostica di Laboratorio Veterinaria (SIDiLV) (14. :
Sorrento (NA) : 24 - 26 Ottobre 2012)
B. cereus is a spore forming gram-positive bacterium (1). There are numerous research reports on
the prevalence of B. cereus in dairy products (2). Mascarpone is an unripened, soft spread cheese
obtained from thermal-acidic coagulation of milk cream (3). The presence of spore-forming bacteria
able to survive the heat treatments applied to milk cream during the manufacturing represents the
most important problem (1). The objective of this work was to development and validation of a
mode) to predict growth of B. cereus in mascarpone cheese during the post-pasteurization cooling
process.
Daminelli° P, Losio° MN, Gianfranceschi M, Decastel li L, Comin DP, Fischetti R,
Valiani A, Fadda A, Goffredo E, Nava D, Cardamone C, Prencipe VA, Varisco° G
La rete degli II.ZZ.SS al servizio della sicurezza alimentare dei prodotti tradizionali italiani
XIV Congresso Nazionale SIDiLV : 24 - 26 Ottobre 2012, Sorrento (NA) : volume degli atti / [s.l. :
Societa' Italiana Diagnostica di Laboratorio Veterinaria ( SIDiLV ), 2012]. - p 67-68. - 7 bib ref [Nr.
Estr. 5108]
Congresso Nazionale Societa' Italiana Diagnostica di Laboratorio Veterinaria (SIDiLV) (14. :
Sorrento (NA) : 24 - 26 Ottobre 2012)
At Community level, actually there are over 240 Italian products, of which 151 registered PDO and
90 branded PGI. The enhancement of the traditional agro-food must necessarily respond to the
need to ensure food safety in relation to the protection of human health, in order to ensure the food
bu-siness operators the ability to export products made in Italy absolving the requirements imposed
in particular also from Third Countries. The predictive microbiology models are important tools for
managing food safety and provide a scientific basis for addressing some key aspects of HACCP
plans. The combination of science and technology is essential for the effective application of
predictive microbiology. The creation of a network between Istituti Zooprofilattici Sperimentali (II. ZZ.
SS.) and the National Institute of Health (ISS) for the sharing of knowledge of the microbiological
and physico-chemical characteristics of traditional Italian products and the potential applications of
the tools predictive microbiology, represents a fundamental tool to ensure a uniform approach to the
safety problems of food from animal origin, proposing experimental protocols and scientifically viable
internationally.
De_Cicco° C, Ricchi° M, Garbarino° C, Cammi° G, Arr igoni° N
Ricerca di Mycobacterium avium subsp. paratuberculosis vitale mediante cattura
peptido-magnetica e trattamento con Propidium monoazide in campioni di latte
XIV Congresso Nazionale SIDiLV : 24 - 26 Ottobre 2012, Sorrento (NA) : volume degli atti / [s.l. :
Societa' Italiana Diagnostica di Laboratorio Veterinaria ( SIDiLV ), 2012]. - p 75-77. - 7 bib ref [Nr.
Estr. 5107]
Congresso Nazionale Societa' Italiana Diagnostica di Laboratorio Veterinaria (SIDiLV) (14. :
Sorrento (NA) : 24 - 26 Ottobre 2012)
The detection of viable cells of Mycobacterium avium subsp. paratuberculosis (Map) in milk and its
derivatives is currently achieved by cultural methods, which are time consuming and show a lack of
sensitivity. In this study we have developed a new technique to detect viable Map ín milk based on
the peptide magnetic separation (PMS) followed by qPCR after sample treatment with Propidium
monoazide (PMA). This innovative and promising method allowed us to obtain a detection of viable
Map by amplification of specific sequence element (target F57).
De_Medici D, De_Giusti M, Picotto P, Capuano F, Losio° MN, Delibato E,
Di_Pasquale S, Lena R, Durante G, Marcheggiani S, Dionisi AM, Pavoni° E,
Marinelli L, Cottarelli A, Mancini L, Luzzi I, Mendito A, Borrello S
Salmonella and vegetables: the Italian management and multidisciplinary approach
Eur J Public Health. - Vol. 22 suppl 2 ( 2012). - p 78 [Nr. Estr. 5122]
European Public Health Conference (5th : Malta : 8-10 November 2012)
Background In 2004-2005 and 2008-09 the European Rapid Alert System for Food and Feed
(RASFF) notified different alerts involving rocket salads produced in Campania, a south-central
Italian region. Most of the alerts reported vegetable contamination by Salmonella Napoli, a serovar
which in Italy accounts for 3% of human salmonellosis. In order to manage these repeated alerts the
Italian Ministry of Health organized a multidisciplinary taskforce aiming at defining strategies to
reduce or eliminate the risk of contamination of fresh vegetables. Methods In 2009-2011 a
microbiological survey was performed on vegetables from 21 producers located in "Piana del Sele",
Campania. Water irrigation samples and vegetables samples (rocket salads) were analyzed by Real
Time PCR for detection of Salmonella spp. PCR positive samples were tested for Salmonella by
means of the standard microbiological ISO method (ISO 6579:2002/Cor.1:2004). Molecular
subtyping by PFGE was performed to evaluate the genetic relationship among Salmonella isolates.
In addition vegetable samples from the Italian market were analyzed for the presence of Salmonella
using the Real Time PCR and the positive results were confirmed by means of the ISO method.
Results Fifty-eight out of 234 water samples were found positive for Salmonella spp and S. Napoli
was isolated from 25% of positive samples. PFGE revealed a dose genetic correlation of S. Napoli
isolates. All vegetable samples analyzed were negative for Salmonella. Concerning leafy vegetables
sampled from the market and analysed by means of Real Time PCR, seventeen samples out of
2002 were found positive for Salmonella with four samples confirmed by the ISO method.
Conclusion This study demonstrates that a multidisciplinary approach is needed to guarantee an
adeguate control of raw materials and water for irrigation aiming at preventing/eliminating/ reducing
biological hazards. This is of particular relevance for ready to eat vegetables where initial
contamination is very difficult to remove by means of common washing procedures. In addition the
results of the study stress the need to apply very strict Good Agriculture Practices for vegetables
intended to be eaten raw.
De_Medici D, Fiore A, Vilmercati A, Fenicia L, Losio° MN, Anniballi F
Assessment of activity of probiotic bacteriocins against bont-producing Clostridia and
Cronobacter Spp
International scientific conference Probiotics and Prebiotics : conference proceedings : 12th - 14th
June 2012 Kosice, Slovakia / [s.l. : s.n., 2012]. - p 69-70 [Nr. Estr. 5008]
International scientific conference Probiotics and Prebiotics : Kosice, Slovakia : 12th - 14th June
2012)
Bacterial antagonism of probiotic bacteria is exerted by competition for nutrient and for attachment
sites to the epithelium of the colon, and also by production of antimicrobial substances such as
organic acids, hydrogen peroxide (H 202) and proteinaceus toxins like bacteriocins, that inhibit
growth of similar bacterial strains and/or kill pathogens microbes. Bacteriocin are ribosomally
synthesized and are carried out by plasmid or transposons. A lot of research work has proved the
antibacterial activity of bacteriocins against food borne pathogens, like Listeria spp, Bacillus spp,
Staphylococcus aureus, Clostridium perfrigens, but very few studies have been performed against
Botulinum neurotoxin (BoNT) producing Clostridia and Gram negatives bacteria. like Cronobacter
spp. In this study, the antimicrobial potential of bacteriocins produced by phenotypically and
genotypically well characterized strains of Lactococcus lactis, Streptococcus thermophilus and
Bacillus coagulans were evaluated, by in vitro assay. Particularly, the bacteriocins activity were
assessed to inactivate (BoNT) producing Clostridia (Clostridium botulinum and neurotoxigen
Clostridium butyricum) and Cronobacter spp strains, all potential agents of intestinal colonization.
Methods - Twenty-four probiotic strains, from both food supplement and ATCC collection, were
tested for bacteriocin production. Particularly, antimicrobial activity of 6 Lactococcus lactis, 9
Streptococcus thermophilus and 9 Bacillus coagulans, bacteriocin — producing, were assessed by
spot on the lawn method and zones of inhibition surrounded colonies were visualized. To eliminate
antimicrobial effect due to hydrogen peroxide and organic acids production, we used specific device.
Thirty-five strains of BoNT-producing Clostridia (isolated in Italy by National Reference Centerfor
Botulism) and 10 strains of Cronobacter spp (from both ATCC collection and proficiency test
performed at the Microbiological Foodborne Hazard Unit) were used as potential bacteriocin —
sensitive organisms. Each probiotic species used in this study were further investigated by
Real-Time PCR reaction to confirm the presence of the specific gene encoded bacteriocin.
Particularly, we used 3 primer pair for nisin, thermophilin and coagulin,respectively. Real-Time PCR
reaction were performed using SYBR Green scheme.
Results - Results of this study show that: all 24 probiotic strains are provided of gene encoded for
specific bacteriocin, all 35 BoNT-producing Clostridia strains are sensitive to nisin and coagulin;
about 86% of strains are sensitive to thermophilin. Distinct inhibition zones were obtained only for
one of Cronobacter spp strains tested. Well standardized Real-Time PCR reaction is a useful tool to
investigate on probiotic activities.
Discussion - The information in BACTIBASE reported that nisin, coagulin and thermophilin are active
against Gram positive bacteria but, generally, not against Gram-negative. Results of our study show,
for the first time, the sensibility of one strain of Cronobacter spp to all bacteriocin tested, without
necessity of any treatment. We demonstrated that the use of probiotic can reduce the risk of
intestinal colonization of BONt producing Clostridia and Cronobacter spp. The manipulation of the
flora, by use of food containing probiotícs may allow promotion of beneficiai flora, especially in
formula-fed infants that are more at risk of contracting infections.
Delibato E, Sinibaldi_Vallebona P, Anniballi F, Filetici E, Luzzi I, Losio° MN,
De_Medici D
Yersinia enterocolitica patogena nei vegetali: valutazione della sopravvivenza e della
persistenza al processi di sanificazione mediante real-time PCR
Infezioni trasmesse da alimenti e acqua : diagnostica ed epidemiologia : VIII workshop nazionale
Enter-net Italia, Sistema di sorveglianza delle infezioni enteriche : 31 maggio-1 giugno 2012, Vietri
sul Mare, Salerno / a cura di Caterina Graziani, Francesca Manciini e Ida Luzzi. - Roma : Istituto
Superiore di Sanità, 2012. - (ISTISAN congressi ; 12/C2) p 44 [Nr. Estr. 4996]
Workshop nazionale Enter-net Italia (8 : Vietri sul Mare, Salerno : 31 maggio-1 giugno 2012)
Negli ultimi anni Yersinia enterocolitica ha acquistato sempre maggiore importanza quale agente di
tossinfezioni alimentari. Infatti si è registrato un aumento nell'incidenza di tale patogeno in diversi
Paesi europei, probabilmente, non solo in relazione alla crescente diffusione dell'allevamento
intensivo ed ai sistemi di produzione ad esso correlati, ma anche dovuta al consumo di alimenti di
origine vegetale sottovalutati fino ad oggi, come probabili veicoli. Il metodo colturale di riferimento,
per la ricerca di tale patogeno, richiede più di 7 giorni, ed è quindi necessario sviluppare nuovi
metodi sensibili e rapidi per la sua determinazione. A tale scopo, nel presente lavoro, durante una
prima fase, è stato ottimizzato e validato, in accordo alla ISO 16140, un metodo di real-time PCR
per la determinazione della Yersinia enterocolitica patogena nei vegetali. Dai risultati della
validazione le performance analitiche del metodo hanno evidenziato il 100% di selettività, specificità
relativa, sensibilità relativa e accuratezza relativa. Quindi, tale metodica è stata utilizzata per
valutare la sopravvivenza, alla temperatura di refrigerazione, di Yersinia enterocolitica nei vegetali
durante la shelf-lifè del prodotto, e per verificare l'efficacia dei processi di sanificazione in ambito
domestico e durante la fase di lavorazione della IV gamma. Dall'osservazione delle curve di
amplificazione, ottenute utilizzando la real-time PCR, attestanti la positività di tutte le aliquote
contaminate e conservate alla temperatura di refrigerazione, si evince che il patogeno sopravvive e
cresce fino al 6° giorno dopo la contaminazione. I risultati ottenuti durante la fase di valutazione
della resistenza ai differenti processi di sanificazione, mostrano in tutti i campioni trattati, la presenza
di cellule vitali e resistenti ai vari processi di sanificazione, con una riduzione parziale della carica
del patogeno nelle concentrazioni più basse quando trattati con 220 ppm di ipoclorito di sodio per 15
minuti. Dai risultati ottenuti, durante la prima fase, si evidenzia che l'utilizzo della real¬time PCR
riveste un ruolo fondamentale nel controllo microbiologico dei prodotti vegetali di I e IV gamma, in
quanto è in grado di fornire i risultati nell'arco delle 24 ore, consentendo così, sia nell'ambito del
controllo ufficiale che nell'autocontrollo, di prendere decisioni tempestive riguardanti il rilascio di lotti
di prodotto o la possibilità del loro ritiro dal commercio.
Delle_ Donne° G, Bolzoni° G, Varisco° G
Nicchie : latte di bufala in Lombardia
Inf Zootec. - Vol. 59 no 9 ( 2012). - p 63-67 [Nr. Estr. 5143]
Delle_ Donne° G, Marcolini° A, Zanardi° G, Bolzoni° G
Rapporto tra acidi grassi saturi e insaturi nel latte bovino : studio dell'Istituto zooprofilattico
di Brescia
Inf Zootec. - Vol. 59 no 19 ( 2012). - p 91-97 [Nr. Estr. 5142]
Dottori° M,
Progetto regionale Sibilla: formaggio Parmigiano Reggiano
Convegno Microbiologia Predittiva: possibili utilizzi nell'attività di controllo ufficiale degli operatori
del settore alimentare / [s.l. : s.n., 2012]. - 12 p
http://www.ausl.re.it/Home/DocumentViewer.aspx?ID=1770&TIPODOC=COMUNICAZIONE [Nr.
Estr. 5230]
Convegno Microbiologia Predittiva: possibili utilizzi nell'attività di controllo ufficiale degli operatori del
settore alimentare : Reggio Emilia : 30 Novembre 2012)
Dusi° G, Arioli F, Ghidelli V, Casati A, Hathaway T , Pompa G, Bertocchi° L
Investigation on the origin of prednisolone residue in cow urine
Residues of veterinary drugs in food : proceeding of the Euroresidue VII conference : Egmond aan
Zee, the Netherlands, 14-16 May, 2012 / [s.p. : s.n., 2012]. - p 297-302. - 3 bib ref [Nr. Estr. 5000]
Euroresidue : conference on residues of veterinary drugs in food (7th : Egmond aan Zee
(Netherlands) : 14-16 May, 2012)
Prednisolone is a steroid belonging to the corticosteroid group. The results obtained in the
application of the 2008 and 2009 Italian Residue Control Plans show a frequent detection of
prednisolone traces in cow urine. Since most of positive samples were detected at the
slaughterhouse, researchers hypothesized that, together with an increase of cortisol and cortisone
concentrations, traces of prednisolone could be produced endogenously during the stressful
situations due to the handling before slaughter. In a preliminary study three dairy cows were treated
i.m. with a synthetic analogue of adrenocorticotropic hormone (ACTH) to simulate stress. The results
published (Pompa et al., 2011) suggested that prednisolone could be produced endogenously. In
the present trial, 52 lactating cows housed in 7 different Lombardy farms were studied. Urine
samples were collected at the farm (after urethral catheterisation and immediately after the slaughter
(from urinary bladder). The urine samples were analyzed using an LC-MS/MS confirmatory method
validated according to the Commission Decision 2002/657/ EC criteria. The results demonstrated
that prednisolone can be endogenously produced by dairy cows and furthermore its endogenous
presence in bovine urine seems to be strongly related to a state of stress in the animals (at farm and
slaughterhouse).
Dusi° G, Vago G, Ghidelli° V, Pellegrino RM, Galari ni R
Analytical investigations about the presence of prednisolone in cow urine
J Mass Spectrom. - Vol. 47 ( 2012). - p 1221-1231. - 31 bib ref [Nr. Estr. 5050]
Since 2008, the analyses carried out in the Lombardia region as part of National Residue Control
Plans have evidenced unexpected frequent detection of the corticosteroid prednisolone (PRED) in
cow urine samples taken to the slaughterhouse. Considering the scarce plausibility of these high
frequent findings, analytical investigations were started to ascertain the real presence of this
corticosteroid. The applied confirmatory method involved liquid-chromatography low-resolution
tandem mass spectrometry (triple quadrupole) as instrumental technique, and it was validated in
compliance with the requirements of the Commission Decision 2002/657/EC. However, recently
some criticism regarding Commission Decision 2002/657/EC identification criteria has been pointed
out, experimentally demonstrating false positive results (wrong identification) although these criteria
have been strictly observed. Therefore, considering the serious implications (i.e. the possibility that
PRED could be considered endogenous in particular animal conditions), studies were carried out to
investigate the reliability of PRED identification through the change of the chromatographic
conditions (mobile phases, gradient and analytical column) of the confirmatory procedure routinely
applied. Further confirmation came from the application of high-resolution mass spectrometry
technique (MS2 and MS3 experiments) to analyze incurred cow urines samples. All the obtained
results confirmed definitively the real presence of this corticosteroid excluding false-positive findings
in routine analysis. In addition, other experiments demonstrated that high-resolution mass
spectrometers (Time of Flight and Orbitrap technologies) could be successfully applied to routine
determination of steroid residues in biological fluids at very low concentrations (< 1 µg L-1).
Ferretti° E, Gasparini° M, Angelone° B, Castrezzati ° G, Pellegrini° E, Sarasini° S
Monitoring pesticide residues in different vegetal matrices by LC-MS/MS and GC-MS/MS
9th European Pesticide Residues Workshop : June 25th-28th, 2012 Vienna : book of abstracts / [s.l.
: s.n., 2012]. - p 254 [Nr. Estr. 5159]
European Pesticide Residues Workshop (9th : Vienna : June 25th-28th, 2012)
During 2011 the Italian Ministry of Health has agreed with the Institute Zooprofilattico of Lombardy
and Emilia Romagna, the activation of monitoring for the determination of pesticide residues in plant
foods. Mon than 500 samples were analyzed and different types of matrices have been studied
(fruit, vegetables cereals, baby food vegetables). The samples were both from domestic and foreign,
they were collected frati the wholesale fruit and vegetable producers of organic food, organic food
stores and markets. The analysis was performed by QuEChERS with determination by LC-MS/MS
and GC-MS/MS; the whole method, including 162 molecules, was validated and accredited
according to ISO 17025. Pesticide residues were detected in 37% of samples; 0.8% of them
exceeded LMR considered the uncertainty. Residues over 0.01 mg/kg weren’t found in organic
samples and baby foods. The frequency of molecules presence was: 13% one pesticides, 11% two,
6% three, 5°h four, 1% five and 1% over five. The m atrices more contaminated were: peaches
apples, oranges, lemons as fruit and salad, peppers, tomatoes, celery as vegetables. 66 different
pesticides were detected, the ten most frequently detected analytes were: chlorpyrifos-ethyl imazalil,
etofenprox, boscalid, imidacloprid, pyraclostrobin, azoxystrobin, tebuconazole, thiabendazole
difenoconazole.
Finazzi° G, Losio° MN, Daminelli° P, Bertasi° B. Va risco° G
Sicurezza alimentare : le precauzioni di filiera
Inf Zootec. - Vol. 59 no 18 ( 2012). - p 54-67 [Nr. Estr. 5056]
The work describes the main biological agents associated to episode of food outbreak in Europe.
The work also describes the role of IZSLER in the different control programs established in northern
Italy.
Fiorentini° L, Lilliu° E, Tosi° G, Taddei° R, Lonta ni B, Gaspari P, Massi° P
Prevalenza dei portatori asintomatici di Listeria monocytogenes nei polli regolarmente
macellati, valutazione dell’antibioticoresistenza dei ceppi isolati: risultati preliminari
LI Convegno annuale Societa' Italiana di Patologia Aviare (SIPA) : 11-12 Ottobre 2012
Salsomaggiore Terme (PR) : atti / [s.l. : s.n., 2012]. - p 284-292. - 12 bib ref [Nr. Estr. 5242]
Convegno annuale Societa' Italiana Patologia Aviare (SIPA) (51 : Salsomaggiore Terme (PR) :
11-12 Ottobre 2012)
In order to study the prevalence of the enteric infection of Listeria spp. in broiler chickens a study
was carried out in a slaughterhouse during 2011. The study was performed on 50 ß ocks. The
overall prevalence of the enteric Listeria spp. Infection was 82% of the tested ß ocks. The
prevalence of the Listeria monocytogenes infection was 6% of the tested ß ocks. In the second part
of the study the in-vitro susceptibility of the isolated strains to 15 antibiotics was carried out. All
isolated strains of L. innocua, L. welshimeri and L. monocytogenes were fully susceptible to
amoxicillin, doxycycline, tetracycline, neomycin and colistin and to lincomycin/spectinomycin and
trimethoprim/sulphonamides combinations. 3 of 28 L.innocua isolated strains and 1 of 3
L.monocytogenes isolated strains were resistant to oxacillin. The antimicrobial susceptibility of the L.
grayi isolated strains and of the L. ivanovii isolated strains showed a considerable variability and a
resistance to the most tested antibiotics.
Galarini R, Dusi° G, Caroccia A, Fioroni L, Cristof ani E, Saluti G
Development and validation of a multiresidue method for the determination of macrocyclic
lactones in food and feed
Residues of veterinary drugs in food : proceeding of the Euroresidue VII conference : Egmond aan
Zee, the Netherlands, 14-16 May, 2012 / [s.l. : s.n., 2012]. - p 849-854. - 8 bib ref [Nr. Estr. 4999]
Euroresidue : conference on residues of veterinary drugs in food (7th : Egmond aan Zee
(Netherlands) : 14-16 May, 2012)
A confirmatory high performance liquid chromatography method with fluorescence detection
(HPLC-FLD) was developed and validated for the símultaneous determination of the following
anti-parasitic veterinary drugs in foodstuffs (liver, muscle, and milk) and feed: eprinomectin,
moxidectin, emamectin, abamectin, doramectin and ivermectin. Samples were extracted, purified
and analysed by HPLC-FLD previous derivatization with N-methylimidazole, trifluoracetic anhydride
and acetic acid to a stable fluorescent derivative. Found recoveries were in the range of 73-98%
(liver), 82-93% (muscle), 77-84% (milk) and 58-73% (feed). In all matrices the CV (coefficient of
variation evaluated in intra-laboratory reproducibility conditions) were lower than or equal to 15%.
Only for emamectin in liver worst performances were observed. Due to high acceptability regarding
the required criteria and applicability both in food and in feed, this multi-analyte method has been
successfully implemented in residue monitoring Italy.
Gambarin P, Magnabosco C, Losio° MN, Pavoni° E, Gat tuso A, Arcangeli G,
Favretti M
Listeria monocytogenes in Ready-to-Eat seafood and potential hazards for the consumers
Int J Microbiol. - Vol. 2012, Article ID 497635 ( 2012). - 10 p.
http://www.hindawi.com/journals/ijmb/2012/497635/ (ultimo accesso 02/07/2012) [Nr. Estr. 5013]
The risk of exposure to Listeria monocytogenes (L, monocytogenes) when consuming Ready-to-Eat
(RTE) seafood was assessed in the Veneto Regina (Italy). Thirty-eight samples were analyzed, each
sample consisted of three subunits belonging to the same batches. The first of the three units was
examined immediately, the second was stored at +4`C (for all of its shelf-fife) and the third at +10'C
(for the latter third of its shelf-life) before the analysis. Chemical-physical and microbiological
parameters were tested simultaneously. Culture results showed the presence of viable L.
monocytogenes in 9 (23,68%) of the 38 samples analysed, 3 (33,33%) of which with a coneentration
>100 cfu/g. PCR tests yielded 12 L. monocytogenes positive samples. Semipreserves with aw
(water activity) and pH values that favour L. monocytogenes growth were the only ones to result
positive to microbiological and PCR tests. Temperature proved to be an important factor as it limits
the growth of L, monocytogenes, including products with potentially high competitive microbial
charges. Four different serotypes were recovered and ribotyping has helped to highlight the genomic
variability of L. monocytogenes strains in food. This supports the hypothesis that L. monocytogenes
continues to evolve genetically to the detriment of phenotypic conservation.
Giacometti F, Serraino A, Bonilauri° P, Ostanello F , Daminelli° P, Finazzi° G, Losio°
MN, Marchetti G, Liuzzo G, Zanoni RG, Rosmini R
Quantitative risk assessment of verocytotoxin-producing Escherichia coli O157 and
Campylobacter jejuni related to consumption of raw milk in a Province in Northern Italy
J Food Prot. - Vol. 75 no 11 ( 2012). - p 2031-2038. - 43 bib ref [Nr. Estr. 5120]
A quantitative risk assessment was developed to describe the risk of campylobacteriosis and
hemolytic uremic syndrome (HUS) linked to consumption of raw milk sold in vending machines in
Northern Italy. Exposure assessment considered the microbiological status of dairy farms, expected
milk contamination, storage conditions from bulk tank to home storage, microbial growth during
storage, destruction experiments, consumption frequency of raw milk, age of consumers, serving
size, and consumption preference. The differential risk between milk handled under regulation
conditions (4°C throughout all phases) and the wors t field handling conditions was considered. The
probability of Campylobacter jejuni infection was modeled with a single-hit dose-response
beta-Poisson model, whereas for HUS an exponential dose-response model was chosen and two
probabilities were used to model the higher susceptibility of children younger than 5 years old. For
every 10,000 to 20,000 consumers each year, the models predicted for the best and worst storage
conditions, respectively, 2.12 and 1.14 campylobacteriosis cases and 0.02 and 0.09 HUS cases in
the 0- to 5-year age group and 0.1 and 0.5 HUS cases in the >5-year age group. The expected
pediatric HUS cases do not differ considerably from those reported in Italy by the Minister of Health.
The model developed may be a useful tool for extending the assessment of the risk of
campylobacteriosis and HUS due to raw milk consumption at the national level in Italy. Considering
the epidemiological implications of this study, the risk of illness linked to raw milk consumption
should not be ignored and could be reduced by the use of simple measures. Boiling milk before
consumption and strict control of temperatures by farmers during raw milk distribution have
significant effects on campylobacteriosis and HUS and are essential measures for risk management.
Giacometti F, Serraino A, Finazzi° G, Daminelli° P, Losio° MN, Arrigoni° N, Piva S,
Florio D, Riu R, Zanoni RG
Sale of raw milk in Northern Italy: food safety implications and comparison of different
analytical methodologies for detection of foodborne pathogens
Foodborne Patog Dis. - Vol. 9 n 4 ( 2012). - p 293-297. - 25 bib ref [Nr. Estr. 4983]
The safety of raw milk sold in Northern Italy was investigated in relation to hygiene quality
parameters and presence of Salmonella spp., Listeria monocytogenes, thermotolerant
Campylobacter, and Verocytotoxin producing Escherichia coli O157:H7. The performance of
different analytical methods used—official culture method (ISO), modified Bacteriological Analytical
Manual cultural method (mBAM), and polymerase chain reaction (PCR)— was evaluated. The
presence of Mycobacterium avium subsp. paratuberculosis (Map) was investigated only by PCR. All
samples met regulations for alkaline phosphatase and inhibitory substance, while 18% and 44.8% of
samples collected from vending machines had, respectively, somatic cell count (SCC) > 300,000/mL
and total bacterial count (TBC) > 50,000CFU/mL. The correlation between hygienic quality
parameters in samples collected from bulk tank and vending machines showed a significant
increase of TBC in vending machines meaning that raw milk was mishandled during distribution and
sale. All pathogens investigated were detected in raw milk sold at vending machines; a total of five
samples (5%) had at least one pathogen, of which two were detected by PCR and three by mBAM.
None of the samples was positive by cultural ISO methods. Even if the comparison of analytical
methods showed that none performs significantly better than the others, testing a higher volume of
milk (25 versus 210 mL) affects significantly the detection rate of pathogens. Three samples (3%)
were positive for Map, suggesting that raw milk is a significant source of Map exposure for
consumers. The observed TBC increase and the detection of several pathogenic bacteria pose
questions on the safety of raw milk; the use of ISO seems inefficient in detecting a low
contamination level of pathogens in milk and consequently not appropriate as official method for
testing. In order to ensure consumer’s safety, a new approach for the raw milk chain is required.
Giacometti F, Serraino A, Finazzi° G, Daminelli° P, Losio° MN, Bonilauri° P,
Arrigoni° N, Garigliani A, Mattioli R, Alonso S, Pi va S, Florio D, Riu R, Zanoni RG
Foodborne pathogens in in-line milk filters and associated on-farm risk factors in dairy farms
authorized to produce and sell raw milk in Northern Italy
J Food Prot. - Vol. 75 no 7 ( 2012). - p 1263-1269. - 68 bib ref [Nr. Estr. 5021]
All dairy farms authorized to produce and sell raw milk in a province of Northern Italy were
investigated to determine the presence of Campylobacter spp., verocytotoxin-producing Escherichia
coli (VTEC), Listeria monocytogenes, and Salmonella spp. in in-line milk filters and to assess their
association with suspected risk factors on farms. A logistic regression mode) was used to analyze
data collected describing the characteristics and management practices of 27 farms and the
microbiological status of 378 in-line mille filters by both culture-based and molecular methods.
Thermotolerant Canzpylobacter. VTEC, and L. monocytogenes were detected in 24 (6.45%), 32
(8.4%), and 2 (0.5%) samples, respectively. No Salmonella spp. were detected. For risk analysis,
data of L. monocytogenes and Salmonella spp. were not included in the model because of the low
prevalence or absence of these organisms. The invariate analysis disclosed that the presence of
VTEC and/or Campylobacter spp. in milk filters was associated with lack of cleanliness of bedding,
water trough, and feed trough; nonevaluation of water hardness; lack of cleanliness of milk tank; and
nonapplication of forestripping. After multivariate analysis, an association was observed with
inadequate cleanliness of bedding and milk tank and the nonapplication of forestripping. PCR
analysis of milk filters was a rapid and sensitive method for the microbiological evaluation of herd
contamination status and shoutd be included among the registration reguirements for the
authorization to produce and sell raw milk. Specific control actions must be incorporated into the
farmer's daily practices to ensure the low-risk production of raw milk.
Giacometti F, Serraino A, Finazzi° G, Daminelli° P, Losio° MN, Tamba° M,
Garigliani A, Mattioli R, Riu R, Zanoni RG
Field handling conditions of raw milk sold in vending machines: experimental evaluation of
the behaviour of Listeria monocytogenes, Escherichia coli O157:H7, Salmonella Typhimurium
and Campylobacter jejuni
Ital J Anim Sci. - Vol. 11 e24 ( 2012). - p 132-136. - 31 bib ref [Nr. Estr. 4964]
The direct sale by farmers of raw milk for human consumption has been allowed in Italy since 2004.
The aim of this study was to evaluate the behaviour of selected foodborne pathogens in raw milk
sold in vending machines, in field handling conditions, and during shelf-life from production to
consumption. Temperature of storage of raw milk in 33 farms authorized to produce and sell raw
milk were investigated from farm to vending machine delivery, together with consumer habits in one
province of the Emilia-Romagna region of northern Italy. Failure to maintain appropriate low
temperatures during shelf-life was recorded and 43% of consumers did not boil milk before
consumption. Listeria monocytogenes, Escherichia coli O157:H7, Salmonella Typhimurium and
Campylobacter jejuni strains were inoculated into raw milk samples, and the best (4°C as
established by law) and worst temperature storage conditions detected (variable temperature) were
simulated. Boiling tests were performed for each pathogen considered at high and low levels of
contamination. Results showed an increase in L. monocytogenes in milk stored at 4°C and at
variable temperatures recorded in shelf-life monitoring, an increase in E. coli O157:H7 and S.
Typhimurium at variable temperatures but not at 4°C , and a decrease in C. jejuni in all storage
conditions. Boiling milk is effective in making it safe for consumers. This study provides evidence
that appropriate handling of raw milk, maintaining low temperatures, together with consumer
education concerning boiling raw milk before consumption are key factors in preventing foodborne
infections linked to raw milk consumption, and helps assess the risk of foodborne infection linked to
raw milk consumption.
Guberti E, Navacchia P, Veronesi M, Magnavita P, Bardasi° L
Caratterizzazione della qualità microbiologica delle paste ripiene con prodotti a base di
carne: tortellini prodotti a Bologna
Convegno Microbiologia Predittiva: possibili utilizzi nell'attività di controllo ufficiale degli operatori
del settore alimentare / [s.l. : s.n., 2012]. - 27 p
http://www.ausl.re.it/Home/DocumentViewer.aspx?ID=1774&TIPODOC=COMUNICAZIONE [Nr.
Estr. 5233]
Convegno Microbiologia Predittiva: possibili utilizzi nell'attività di controllo ufficiale degli operatori del
settore alimentare : Reggio Emilia : 30 Novembre 2012)
Lichtenberg_Kraag B, Piro° R, Brinkmann B, Piana L,
Kast C
Beaune P, Kilchenmann V,
Determination of physicochemical characteristics of honey comparing reference methods
and fourier transforminfrared spectroscopy performed in four different laboratories
II International Symposium on Bee Products : annual meeting of the International Honey
Commission : September 9-12, 2012 Braganca, Portugal : book of abstracts / [s.l. : s.n., 2012]. - p
34 [Nr. Estr. 5121]
International Symposium on Bee Products : Annual meeting of the International Honey Commission
(2 : Braganca, Portugal : September 9-12, 2012)
Fourier Transform Infrared Spectroscopy (FT-IR) determines several physicochemical measurands
simultaneously and hence is faster and less laborious than the current reference methods. FT-IR is
the ordinary analytical method used for analysis of milk and other food products. In several
laboratories, FT-IR has recently been established as routine method for honey analysis. With this
study we aimed to evaluate if FT-IR analysis leads to comparable results even if instrument types
and sample preparation (diluted and undiluted samples) are different. Hence we were interested to
investigate if FT-IR can be considered equivalent to the established methods. Therefore, the
measurands fructose, glucose, pH, electrical conductivity and free acidity were determined by
reference and FT-IR analysis in four different laboratories using Foss or Bruker instrumentation and
the measurands water and sucrose in two and three laboratories, respectively. So far, we analyzed
several honey samples from German ring triais, as well as six different European unifloral honeys.
The methods were analyzed and compared using statistical z-score evaluation and other common
statistical test (eg. Cochran). The reference methods of all laboratories showed good accuracy for all
the honey types. Average values of the reference methods were subsequently used for the
evaluation of the quality of the FT-IR results. The accuracy of these results were related to the
honey type and differed among the measurands. Most measurands were comparable for FT-IR and
reference methods when acacia, rape, lavender, tilia and honeydew honeys were analyzed,
however the some measurands determined by FT-IR deviated a lot when chestnut honey was
examined. In general, fructose, glucose, pH and free acidity gave mostly comparable values with
both methods, while the values of the electrical conductivity diverged more from the reference
method. Different results obtained from FT-IR and reference methods may be related to the honey
types used for FT-IR calibration in the various laboratories, rather than the different FT-IR
instrumentation or sample preparation. This first interlaboratory testing shows, that FT-IR is a
promising approach even for a product with a high variability of components like honey. Knowing the
outliers in the ring trials, each laboratory can improve or correct its specific calibrations. Improved
correlation should be verified in an additional forthcoming ring trial.
Lontani B, Dell’orfano G, Montanari E, Ronconi A, Libretto M, Ponzoni N, Cecchini
A, Fiorentini° L
Caratterizzazione del formaggio di Fossa DOP e non DOP (risultati preliminari)
Convegno Microbiologia Predittiva: possibili utilizzi nell'attività di controllo ufficiale degli operatori
del settore alimentare / [s.l. : s.n., 2012]. - 42 p
http://www.ausl.re.it/Home/DocumentViewer.aspx?ID=1769&TIPODOC=COMUNICAZIONE [Nr.
Estr. 5229]
Convegno Microbiologia Predittiva: possibili utilizzi nell'attività di controllo ufficiale degli operatori del
settore alimentare : Reggio Emilia : 30 Novembre 2012)
Losio° MN, Pavoni° E, Consoli° M, Saetti° F, Suffre dini
Battistini R, Rossini I, Arcangeli G, Croci L
E, Serracca L,
Norovirus nei prodotti della pesca: nove anni di monitoraggio in Italia
XIV Congresso Nazionale SIDiLV : 24 - 26 Ottobre 2012, Sorrento (NA) : volume degli atti / [s.l. :
Societa' Italiana Diagnostica di Laboratorio Veterinaria ( SIDiLV ), 2012]. - p 344-346. - 7 bib ref [Nr.
Estr. 5095]
Congresso Nazionale Societa' Italiana Diagnostica di Laboratorio Veterinaria (SIDiLV) (14. :
Sorrento (NA) : 24 - 26 Ottobre 2012)
Noroviruses (NoVs) are increasingly important as aetiological agents of gastrointestinal infections.
They belong to the family Caliciviridae, and are composed by small round structured viruses with a
positive-sense, single-stranded RNA genome. Due to their high levet of genetic diversity, they are
divided into five genogroups (G). Consumption of bivalve molluscs and ready-to-eat fishery products
is the most common way of NoVs foodborne infections, and the rise of outbreaks of viral
gastroenteritis represents an important health problem which is also responsible for economic
losses.
Losio° MN, Pavoni° E, Cosciani_Cunico° E, Daminelli ° P, Bertasi° B
The role of lactic acid bacteria against Listeria monocytogenes, Salmonella typhimurium, E.
coli O157:H7 and Staphylococcus aureus, during the production of bagòss cheese
International scientific conference Probiotics and Prebiotics : conference proceedings : 12th - 14th
June 2012 Kosice, Slovakia / [s.l. : s.n., 2012]. - p 85 [Nr. Estr. 5009]
International scientific conference Probiotics and Prebiotics : Kosice, Slovakia : 12th - 14th June
2012)
Bagòss cheese is made with raw milk in the district of Brescia (North of Italy). It is a hard cheese
characterized by a long maturation period, from 12 to 36 months. The milk and the transformation
technology were subjected to studies to understand the evolution of various pathogenic
microorganisms, potentially present during the curing. Bagòss was also artificially contaminated and
the behavior of pathogens was described.
Methods One sample of milk, one of curd and a slice of cheese were collected once a month for 12
months. These samples were analyzed to determine the pH value and water activity (aw). In
addition, the concentration of indigenous lactic acid bacteria (LAB) and enterobacteria, indicative of
fecal contamination or mastitis, were determined. During the experimental contamination, milk was
artificially contaminated with L. monocytogenes, Salmonella Typhimurium, E. coli 0157: 117, and S.
aureus. The concentration of bacteria, was very high (107 cfu/ml) in order to evaluate the evolution
of pathogens during the 12 months of seasoning. Samples were collected and monitored to evaluate
the behavior of indigenous populations and artificially added pathogens, depending on pH and free
water (aw). However, the synergy between the pH lowering, due to the metabolic activity of LAB,
and the significant decrease of aw, after two months, were factors that significantly affected the
viability of pathogens.
Results For not contaminated cheese, Staphylococcus aureus was not isolated already after one
month, even as its toxin. After six months, the presence of Escherichia coli and Enterobacteriaceae
was not detected. The technology of production, enhanced the activity of some indigenous LAB;
thermophilic bacteria, characterized by an intense metabolic production lowering the milk pH, and
prodUcing organic acids, were selected. The concentration of LAB also maintained constant and
high (107 cfu/g) throughout all the period of ripening. For artificially contaminated cheese, data
showed that the first two months were significant for the pathogens decrease (L. monocytogenes
from 107 cfu/g to 104 cfu/g; S. typhimurium from 107 cfu/g to 102 cfu/g; E. coli 0157:H7 from 107
cfu/g to 102 cfu/g; S. aureus from 107 cfu/g to 102 cfu/g).
Discussion The temperature alone did not influence the behavior of pathogens. It was
demonstrated that the milk cooking (> 48°C) and the stop period of the curd under the whey, did not
significantly decrease the concentration of pathogens. It was necessary to consider also the synergy
between the pH lowering, the aw reduction, the bio-competition, and the production of bacteriocins
by different strains of LAB. The international protocols for the export of many food products require
process parameters related to the milk pasteurization or to the cooking, that actually would not
permit to export raw milk products. However, this work, showed that Bagòss processing and
seasoning generated a decrease of at least 100,000 cfu/g of pathogens, confirming that it is
considered to be safe even if produced with raw milk.
Luini° M, Pozzi° F, Battisti° A, Benedetti° V, Fabb i° M, Franco° A, Vezzoli° F
Methicillin-resistant staphylococcus aureus (MRSA) in dairy cattle of Northern Italy
XXVII World Buiatrics Congress : 3 - 8 June, 2012 Lisbon, Portugal : abstract book / [s.l. : s. n.,
2012]. - p 143 (P:461) [Nr. Estr. 5235]
World Buiatrics Congress (27th : Lisbon, Portugal : 3 - 8 June, 2012)
Objectives: Recent studies show the isolation from cases of bovine mastitis of methicillin resistant
Staphylococcus aureus (MRSA ) belonging to the classical genotype Livestock Associated (ST 398).
The aim of our work was to investigate the presence of MRSA in milk samples from cases of bovine
mastitis and bulk milk samples of dairy farms in Northern Italy. Materials and Methods: We
considered (i) 699 strains of S. aureus isolated from bovine milk samples during the routine
diagnosis of bovine mastitis between November 2008 and September 2011 and (ii) 155 strains
isolated from bulk milk samples of the 351 dairy farms of the Lodi province. Other 7 isolated from
bulk milk of dairies of other provinces of Lombardy were also considered. Up to four strains of S.
aureus for each farm were tested by disk diffusion method. Oxacillin resistant or intermediate strains
were tested with a specific multiplex PCR for nuc and mecA genes to confirm the specie and
methicillin resistance. Thirteen MRSA strains isolated from mastitic milk and 21 from bulk milk were
genotyped with MLST (Multi Locus Sequence Typing). Results: Our analysis showed 73 strains of
MRSA out of 699 S. aureus isolates from mastitic milk belonging to 25 dairy farms. Twenty-one out
of 155 strains isolates from bulk milk of farms in the province of Lodi were MRSA belonging to 19
dairy farms. In addition MRSA were dimonstrated in 3 samples of bulk milk from other provinces of
North. In total we showed MRSA in 44 different cattle farms. MLST analysis of 13 MRSA strains
isolated from mastitis showed that CC 398 (ST 398) was spreading in 6 of 9 problem herds, while
CC 1, CC 97 and CC 152 in one herd each. Of the 21 MRSA strains isolated from bulk milk, 12
belonged to the CC 97 group, 5 to CC 398, 2 to CC 1, 1 to CC 22 and one to ST 152 (singleton CC
). Conclusions: Our investigations show that MRSA are involved in a large number of S. aureus
mastitis and that are also frequently isolated from bulk milk samples taken routinely. In contrast to
previous reports ST 398 genotype was not the more frequent. Apparently, MRSA ST 398 occurred
more frequently in diagnostic samples of mastitis, while in samples of bulk milk MRSA CC 97
resulted more prevalent. Although the two types of studies are not easily comparable, further insight
in molecular epidemiology of MRSA in the territories of Northern Italy would be useful, along with
further efforts in the control of mastitis in the herds affected by MRSA.
5.
Maccabiani° G, Zanardini° N, D'Amico° S, Galuppini° E, Pavoni° E, Finazzi° G,
Giuradei° F, Losio° N, Varisco° G
Utilizzo di Fagi come agenti bio-decontaminanti in prodotti caseari
XIV Congresso Nazionale SIDiLV : 24 - 26 Ottobre 2012, Sorrento (NA) : volume degli atti / [s.l. :
Societa' Italiana Diagnostica di Laboratorio Veterinaria ( SIDiLV ), 2012]. - p 80-82. - 2 bib ref [Nr.
Estr. 5111]
Congresso Nazionale Societa' Italiana Diagnostica di Laboratorio Veterinaria (SIDiLV) (14. :
Sorrento (NA) : 24 - 26 Ottobre 2012)
In recent years decontamination processes and "functional packaging" have been developed to
assure healthy and quality food products. Substances considered "biodecontaminants" are the
bacteriocins, molecules produced by lactic bacteria strains able to inhibit the growth of pathogen
bacteria. Moreover, bacteriophages are considered potentially biodecontaminants, thanks to their
ability to selectively kill bacteria strains. In this work, the behavior of the phage IZS100 against
Listeria monocytogenes have been evaluated in dairy products.
Martini G, Dosa G, Giani G, Massirio I, Carobbi D, Giacometti B, Spighi P, Usberti
R, Milandri C, Frasnelli° M
Selvaggina cacciata di ungulati : parametri per la valutazione dell'igienicità delle carni : dati
preliminari
Convegno Microbiologia Predittiva: possibili utilizzi nell'attività di controllo ufficiale degli operatori
del settore alimentare / [s.l. : s.n., 2012]. http://www.ausl.re.it/Home/DocumentViewer.aspx?ID=1773&TIPODOC=COMUNICAZIONE [Nr.
Estr. 5160]
Convegno Microbiologia Predittiva: possibili utilizzi nell'attività di controllo ufficiale degli operatori del
settore alimentare : Reggio Emilia : 30 Novembre 2012)
Meletti° F, Merigo° D, Kaware Y, Peroni° S, Andreol i° M, Pavoni° E, Losio° MN,
Maccabiani° G, Mioni RM, Bertasi° B
La ristorazione collettiva: studio delle problematiche sanitarie associate alle allergie
XIV Congresso Nazionale SIDiLV : 24 - 26 Ottobre 2012, Sorrento (NA) : volume degli atti / [s.l. :
Societa' Italiana Diagnostica di Laboratorio Veterinaria ( SIDiLV ), 2012]. - p 390-391. - 3 bib ref [Nr.
Estr. 5093]
Congresso Nazionale Societa' Italiana Diagnostica di Laboratorio Veterinaria (SIDiLV) (14. :
Sorrento (NA) : 24 - 26 Ottobre 2012)
The change in lifestyle and eating behaviours means that an increasing number of people have a
significant number of meals outside home and consume different and ethnic products. Food safety in
the mass catering is becoming very important to protect the health of consumers and to support this
important economic field. Also food allergies are an important health problem in mass catering of
industrialized countries. In this study the results of the monitoring plan of allergens in food from
mass catering are presented. Analyses are carried out with ELISA tests and Real-Time PCR.
Merla C, Andreoli° G, Vicari° N, Dalla_Valle C, Cav anna C, Manfredini° A, Pajoro°
M, Guglielminetti ML, Biancardi° A, Fabbi° M
Identificazione morfologica e molecolare della flora fungina isolata da salami tradizionali
della Provincia di Pavia
XIV Congresso Nazionale SIDiLV : 24 - 26 Ottobre 2012, Sorrento (NA) : volume degli atti / [s.l. :
Societa' Italiana Diagnostica di Laboratorio Veterinaria ( SIDiLV ), 2012]. - p 392-394. - 11 bib ref
[Nr. Estr. 5092]
Congresso Nazionale Societa' Italiana Diagnostica di Laboratorio Veterinaria (SIDiLV) (14. :
Sorrento (NA) : 24 - 26 Ottobre 2012)
Moulds play an important role in the ripening of salami, even if the presence of species or strains
which can produce mycotoxins, like ochratoxin A, could represent a health risk for the consumers.
The aim of this study was to evaluate the biodiversity of moulds isolated from traditional fermented
salami produced in Pavia province (Italy) during 2011. Our data show that mycoflora on salami is
composed mostly by Penicillium (95/142), in particular by Penicillium species with roughwalled
conidiophores (43/95 strains). An Aspergillus strain ochratoxin A - producing was also isolated.
Milandri C, Usberti R, Mambelli M, Fiorentini° L
Ciccioli di suino parametri per la valutazione della qualita’ igienico-sanitaria delle produzioni
Convegno Microbiologia Predittiva: possibili utilizzi nell'attività di controllo ufficiale degli operatori
del settore alimentare / [s.l. : s.n., 2012]. - 25 p
http://www.ausl.re.it/Home/DocumentViewer.aspx?ID=1771&TIPODOC=COMUNICAZIONE [Nr.
Estr. 5231]
Convegno Microbiologia Predittiva: possibili utilizzi nell'attività di controllo ufficiale degli operatori del
settore alimentare : Reggio Emilia : 30 Novembre 2012)
Nogarol C, Gallina S, Musicanti P, Bianchi DM, Buonincontro G, Fragassi S,
Corvonato M, Zuccon F, Ramon E, Bertasi° B, Losio° MN, Decastelli L
PCR END-POINT per la determinazione di Y. enterocolitica presunta patogena in matrici
vegetali crude: validazione secondo ISO 16140:2003
XIV Congresso Nazionale SIDiLV : 24 - 26 Ottobre 2012, Sorrento (NA) : volume degli atti / [s.l. :
Societa' Italiana Diagnostica di Laboratorio Veterinaria ( SIDiLV ), 2012]. - p 415-417. - 7 bib ref [Nr.
Estr. 5090]
Congresso Nazionale Societa' Italiana Diagnostica di Laboratorio Veterinaria (SIDiLV) (14. :
Sorrento (NA) : 24 - 26 Ottobre 2012)
New trends on food consuming report highly diffusion of vegetables ready-to-eat (RTE) products.
Yenterocolitica is one of the zoonotic agents that may colonize this products: species of this genus
are classified in serotypes and biotypes due to phenotypic and biochemical features. Virulence of
pathogenic strains is related to the presence of gene sequences coding for invasive proteins. Thís
study shows the validation process, accordíng to IS016140:2003, of a duplex PCR protocol that is
able to differen-tiate non pathogen biotype (1A) from pathogen biotypes.
Oliverio° E, Finazzi° G, Bertasi° B, Ruggiero° V, M onastero° P, Cosciani_Cunico°
E, Dalzini° E, Losio° MN, Daminelli° P
Dinamica di comportamento di Listeria monocytogenes durante il processo produttivo e la
shelf-life del formaggio erborinato porzionato e confezionato in vaschetta
XXII Convegno Nazionale Associazione Italiana Veterinari Igienisti (AIVI) "Autorità competente e
operatore del settore alimentare: armonizzazione degli strumenti di gestione del rischio" : abstracts /
[s.l. : s.n., 2012]. - p 43 [Nr. Estr. 5074]
Convegno Nazionale Associazione Italiana Veterinari Igienisti (AIVI) (22. : Torino : 19 - 21
Settembre 2012)
Questo lavoro ha lo scopo di valutare il comportamento di Listeria monocytogenes .durante la
produzione e la shelf-life del formaggio erborinato porzionato e confezionato in vaschetta. E' stato
eseguito un challenge test utilizzando una miscela con tre ceppi di Listeria monocytogenes. Sono
state prodotte quattro forme dí formaggio erborinato: due forme di controllo e due forme
contaminate. La contaminazione è avvenuta dopo il trattamento di pastorizzazione. Per valutare il
comportamento di L.monocytogenes durante la shelf-life, le forme sono state porzionate in fette di
circa 1oog e confezionate ín vaschette plastiche termosaldate. Le confezioni sono state conservate
per 66 giorni a temperatura variabile: 8°C per 7 gi orni e 12°C per altri 59. Per la prova sperimentale
è stato utilizzato un metodo di campionamento distruttivo. Durante la produzione e la stagionatura le
analisi sono state effettuate prelevando 2 campioni di latte, cagliata e formaggio ad intervalli
prestabiliti. Durante la shelf-life le analisi sono state eseguite su 3 vaschette ad intervalli prestabiliti.
La pasta e la crosta di ciascun campione sono state analizzate separatamente. Sui campioni
contaminati è stato eseguita la numerazione di Listeria monocytogenes; sui campioni di controllo
sono state invece eseguite la ricerca di Listeria spp e L. monocytogenes, numerazione dei
lattobacilli mesofili, misurazione del pH e dell'A„. Al termine della lavorazione, nella pasta del
formaggio, L. monocytogenes ha subito una riduzione di carica del 96% rispetto alla concentrazione
iniziale. La prova sperimentale durante la shelf-life è stata condotta simulando le condizioni di
conservazione più sfavorevoli per la sicurezza del prodotto, nelle quali si è osservato che la pasta
del formaggio rappresenta un substrato idoneo alla crescita del patogeno. Questo studio costituisce
la base per un lavoro di caratterizzazione microbiologica e chimico fisica durante il processo dí
trasformazione di questa tipologia di formaggio, per individuare le fasi critiche della contaminazione
e ridurre la prevalenza di Listeria monocytogenes.
This study was aimed at evaluating the behaviour of Listeria monocytogenes during production and
shelf-life of blue-veined cheese after cutting and packaging. The experimental trial was a challenge
test using a mixture of three strains of Listeria monocytogenes to contaminate the milk before the
production (after pasteurization).Four cheeses have been manufactured: two negative control
cheeses and two contaminated cheeses.Cheeses have been portioned into slices of about 100 g
and packed.The packs have been stored for 66 days: 7 days at 8°C and 59 days at 12°C. It has
been used a destructive sampling.During production and maturation, 2 samples of milk, curd and
cheese have been analysed at setted intervals. During shelf-life 3 samples of cheese have been
analysed at setted intervals.The dough and the rind of each sample have been analysed
separately.Contaminated cheese has been analysed enumerating Listeria monocytogenes; control
cheese has been analysed enumerating Listeria spp., Listeria monocytogenes, Lactic Acid Bacteria
and measuring of pH and Aw.Results showed that, at the end of production and maturation, Listeria
monocytogenes has been reduced charge of 96%, compared to the concentration at the
beginning, in the cheese dough.The experimental test has been carried out, during shelf-life,
simulating the most adverse storage conditions, in which it has been observed that the cheese
dough is a good substrate for the growth of Listeria monocytogenes. This study provides the basis
for a characterization of this type of cheese, to identify the critical point of contamination and to
reduce the prevalence of Listeria monocytogenes.
Olivo F, Abete MC, Stella P, Ferro GL, Dusi° G, Gil i M
Development of a confirmatory method for 16 (beta)-lactam antibiotic residues in milk by
LC-MS/MS
Residues of veterinary drugs in food : proceeding of the Euroresidue VII conference : Egmond aan
Zee, the Netherlands, 14-16 May, 2012 / [s.p. : s.n., 2012]. - p 315-321. - 3 bib ref [Nr. Estr. 5001]
Euroresidue : conference on residues of veterinary drugs in food (7th : Egmond aan Zee
(Netherlands) : 14-16 May, 2012)
A rapid and very effective analytical procedure for the simultaneous quantitative determination of 16
Plactam antibiotic residues (ampicillin, amoxicillin, penicillin G, nafcillin, oxacillin, cloxacillin,
dicioxacillin, penicillin V, cefadroxil, cefoperazone, cefazolin, ceftiofur, cefalexin, cefapirin,
cefquinome and cefuroxime) in milk was developed. During method development a range of
LC-MS/MS conditions were tested. Matrix effect was also evaluated and its correlation with
ionization conditions was tested. The final procedure involved defatted sample extraction with
acetonitrile, centrifugation and supernatant evaporation to dryness, residue dissolution with water.
The analytes were detected using a LC-MS/MS (QQQ) equipped with a H-ESI, operating in positive
and negative multiple reaction monitoring (MRM) mode. The separation of analytes was performed
on a Phenomenex Synergy polar column with gradient elution with aqueous formic acid/ammonium
formiate and methanol. As internal standards ampicillin-d5, amoxicillin-d4, penicillin V-d5,
cephadroxii-d4, and cefuroxime-d3 were used. The validation according to Commission Decision
2002/657/EC criteria for confirmatory method is in progress.
Pavoni° E, D'Amico° S, Galuppini° E, Giuradei° F, B ozzo F, Nolli° G, Losio° MN
Analysis of functional activity by lactic acid bacteria isolated from Silter cheese
International scientific conference Probiotics and Prebiotics : conference proceedings : 12th - 14th
June 2012 Kosice, Slovakia / [s.l. : s.n., 2012]. - p 90 [Nr. Estr. 5007]
International scientific conference Probiotics and Prebiotics : Kosice, Slovakia : 12th - 14th June
2012)
The porpoise of this study was to indentify new methods to increase the levels of food safety,
ensuring a control over the concentration of pathogens in food, and integrating with self-control
systems currently implemented by food makers (HACCP). The set up of possible techniques for bio
decontamination by lactic acid bacteria producing bacteriocins have been selected. Biocompetitive
and lytic properties of acid lactic bacteria (LAB) were evaluated against 8 pathogens: Listeria
monocytogenes, Salmonella enterica-enterica, Salmonella Derby, Salmonella Thyphimurium,
Salmonella Napoli, Salmonella Enteritidis; E. coli 0157:H7, and Staphylococus aureus. The LAB
were obtained from the natural flora of a typical Lombard (North of Italy) hard cheese called Silter
camuno-Sebino, which has valuable organoleptic and sensory characteristics, due to the microbial
flora present in raw milk, produced by cows (Brown Alpine breed) grazing at high altitudes. Several
samples of cheese, at different aging stages, were homogenized and seeded on appropriate media
to obtain the isolation of LAB, and subsequent classification by molecular characterization.
Methods To test which of these LAB had probiotic activity, two important characteristics were
considered: the production of bacteriocins, molecules that inhibit the growth of certain pathogens,
and the adhesion of lactic acid bacteria to intestinal cells. All isolates were also subjected to
molecular characterization by ARDRA (Amplified Ribosomal DNA Restriction Analysis) method and
RiboPrinting (Qualicon Du-Pont). The production of bacteriocins by each LAB strain against
pathogens above mentioned was assessed as mentioned by Blom et al., 1997. For the celi adhesion
to the substrate, Caco-2 cells (Human adenocarcinoma), that can reproduce in vitro the
physiological conditions of the host, were employed. The method provided: 1) preparation of ten fold
dilutions of the LAB in saline buffer, previously grown in MRS broth 2) seeding of dilutions onto
Caco-2 cell line, 3) trypsinization, ten fold dilution in saline buffer, and seeding of cells in MRSA for
inclusion, 4) LAB colony counts before and alter the contact with Caco-2.
Results From different batches of cheese 1,156 strains were isolated. Each of these had a
concentration of, at least, 107 CFU/g. Based on the characterization by ARDRA, 426 strains
belonging to the group of lactic flora were identified. The two most prevalent were Enterococcus
faecalis (16%), and Enterococcus faecium durans (14%). Minor species, composed 40% of the
analyzed strains. Regarding the production of bacteriocins, 274 LAB strains (64%) produced a halo
of inhibition against at least one pathogen, due to the bacteriocins. For the cell adhesion to
thesubstrate, 211 strains (77%) on the 274 LAB producing bacteriocins, showed adhesion to
Caco-2, competing with pathogens for membrane receptors.
Discussion Data obtained showed that over 50% of LAB produced bacteriocins, and adhered to
enterocytes in competition to pathogens for cellular receptor proteins. It could therefore concluded
that the biocompetition and the bacteriocins production, are natural tools against food pathogens,
especially Listeria monocytogenes.
Pavoni° E, Di_Pasquale S, De_Medici D, Losio° MN, S uffredini E, Bertasi° B, Croci
L
Norovirus GI and GII prevalence in bivalve molluscs and vegetables in Italy
Norovirus and other Caliciviruses on the Rise International Conference : March 20-22, 2012,
Luebeck, Germany / [s.l. : s.n.], 2012. - p 57 [Nr. Estr. 4969]
Rise International Conference : Luebeck, Germany : March 20-22, 2012)
In recent years, Noroviruses (NoV) are considered the major cause of non bacterial gastroenteritis in
industrialized countries, although there is still reason to believe that their number is underestimated.
According to the nucLeotide sequence of the NoV capsid region, they are divided into 5 genogroups
(GI, GII, GIII, GIV and GV), which in turn contain several clusters or genotypes. The majority of
strains infecting humans belongs to genogroup I, with 14 genotypes, and the genogroup II, with 17
genotypes (1). Transmission may occur by "person to person" contact or by consumption of
contaminated water or food. In this context, bivalve molluscs and raw or ready to eat (RTE)
vegetables play a special role. It is important to note that Italy is the third European producer of
molluscs, after Spain and France and every year 180,000 tons of molluscs are harvested in Italy (2).
Despite the epidemiological evidences, the current European legislation (EC Reg. 1441/2007)
included no microbiological criteria for the control of viral contamination in foodstuffs. The availability
of molecular methods based both on conventional PCR (i.e. RT-booster-PCR), or on Real-time PCR,
permitted to assess the prevalence of NoV GI and GII in these types of foods. We report data
obtained from surveys carried out on bivalve molluscs collected on import sites and on markets.
Moreover, two surveys were carried on to verify NoV contamination in vegetables. Of these, the first
was conducted in Lombardia region (Northern Italy) on samples from the market, while the second
was done in Lazio region (Central Italy) on samples from two companies, differing for their microbial
stabilization technology (cryogenic removal and disinfection with halogens respectively). The total
number of analysed bivalve mollusc samples was 872. The 6.4% of samples collected on markets
was positive, while 29.3% of samples collected on import sites were contaminated by NoV.
Vegetables samples from Lombardia region were 297, and 6 samples (2.02%) were positive for
NoV: in three samples was confirmed the presence of NoV G11/4. In the monitoring carried out in
Lazio region 124 samples were analysed (46 raw materiale and 78 packaged RTE products
analysed on their last day of sheif life). All samples were negative for Norovirus. These data
underline the need to integrate the European Community legislation on microbiological criteria for
possible virus contamination in foodstuffs. Furthermore, it is important that the food business
operators, both in case of import or in case of primary production, take into account the possibility of
a viral contamination, and improve their HACCP programs in order to ensure that food safety is not
compromised.
Pistone D, Marone P, Pajoro° M, Fabbi° M, Vicari° N , Daffara S, Dalla_Valle C,
Gabba S, Sassera D, Verri A, Montagna M, Epis S, Monti C, Strada EG, Grazioli V,
Arrigoni° N, Giacosa A, Bandi C
Mycobacterium avium paratuberculosis in Italy: commensal or emerging human pathogen?
Dig Liver Dis. - Vol. 44 ( 2012). - p 461-465. - 45 bib ref [Nr. Estr. 5033]
Background: Specific bacterial infections or alterations of the gut microbiota likely trigger
immunopathological phenomena associated with Crohn’s disease and ulcerative colitis.
Mycobacterium avium subspecies paratuberculosis is a candidate etiological agent of Crohn’s
disease. Definitive causal connection between Mycobacterium avium subspecies paratuberculosis
infection and Crohn’s disease has not been demonstrated. Aims: To determine the circulation of
Mycobacterium avium subspecies paratuberculosis in Crohn’s disease patients and water supplies
in an Italian region where this bacterium is endemic in cattle farms. Methods: Mycobacterium avium
subspecies paratuberculosis screening was performed on biopsies from human patients, and from
water samples, using two different PCR procedures. Results: In hospitals where multiple specimens
were obtained from different sites in the intestine, the prevalence of Mycobacterium avium
subspecies paratuberculosis infection was 82.1% and 40% respectively in Crohn’s disease and
ulcerative colitis patients; in another hospital, where single specimens were obtained from patients,
the bacterium was not detected. Control subjects also harboured Mycobacterium avium subspecies
paratuberculosis, but at a lower prevalence. Tap water samples collected in the study area
contained Mycobacterium avium subspecies paratuberculosis DNA. Discussion: The results of
screenings for Mycobacterium avium subspecies paratuberculosis in humans are deeply influenced
by both the number and location of the collected biopsies. There is a wide circulation of the
organism in the study area, considering the prevalence in humans and its presence in drinking
water.
Rossi S, Bertocchi° L, Compiani R
Ormoni : boldenone e prednisolone, il punto sulla ricerca
Inf Zootec. - Vol. 59 no 2 ( 2012). - p 40-45 [Nr. Estr. 4954]
Rubini° S, Bergamini M, Stefanati A, Lupi S, Costan tini E, Gregorio P
Definizione del rischio di parassitosi da nematodi legate al consumo umano di pesce crudo
Prevenzione e sanità pubblica al servizio del paese, l'igenista verso le nuove esigenze di salute : 45°
Congresso Nazionale SITI : 3 - 6 Ottobre 2012 Cagliari : atti / [s.l. : s.n.], 2012. - p 719-720 [Nr. Estr.
5207]
Congresso Nazionale Società Italiana di Igiene (SITI) (45. : Cagliari : 3 - 6 Ottobre 2012)
Rubini° S, Boschetti L, Berardelli C, Bianchi° L, M elloni° R, Santi° A, Galletti° G,
Tamba° M
Confronto tra due metodi normati per la numerazione di Escherichia coli in molluschi bivalvi
Atti del I° Convegno Nazionale Società Italiana di Ricerca Applicata alla Molluschicoltura "Ricerca e
produzione a confronto" : 10 novembre, Teramo / [s.l. : s.n., 2012]. - p 75-76. - 4 bib ref [Nr. Estr.
5194]
Convegno Nazionale Società Italiana di Ricerca Applicata alla Molluschicoltura (1. : Teramo : 10
novembre 2012)
I criteri da applicare nella classificazione delle aree di allevamento e/o raccolta dei molluschi bivalvi
sono ampiamente legiferati dalla Unione Europea. I Regolamenti europei 853/2004, 854/2004 e
s.m.i. suddividono le aree di allevamento e/o raccolta in 3 classi, A, B e C. Mentre i molluschi
provenienti da zone di classe A possono essere destinati al consumo umano diretto, quelli di zona di
classe B devono subire una fase di decontaminazione in un Centro di depurazione (CDM). I bivalvi
raccolti in zona C devono invece essere stabulati in zone A per lungo tempo o subire un trattamento
termico. Il Reg. 2073/2005 indica i metodi di analisi da applicare nella ricerca e nella determinazione
dei parametri microbiologici. Per la numerazione di Escherichia coli viene indicato come metodo
analitico da utilizzare la norma ISO 16649-3:2005 (Microbiology of food and animai feeding stuffs Horizontal method for the enumeration of (3-glucuronidase-positive Escherichia coli Part 3: Most
probable number technique using 5- bromo-4-chloro-3-indolyl-beta-D-glucuronidase). Questo
metodo richiede due diversi terreni di coltura: un terreno di arricchimento liquido, MMGM
(Mineral-Modified Glutamate Medium) e un terreno di conferma in piastra, TBX (Tryptone Bile
Glucuronide Medium). II risultato si ottiene dopo 48 ore dall'inizio delle analisi. II metodo ISO
16649-2:2001 (Microbiology of food and animai feeding stuffs - Horizontal method for the
enumeration of 3-glucuronidase-positive Escherichia coli Part 2: Colony-count technique at 44'C
using 5- bromo-4-chloro-3-indolyl-beta-D-glucuronidase) è una conta in piastra che richiede un unico
terreno di coltura, il TBX e riduce i tempi di analisi a 24 ore. Scopo di questo lavoro è quello di
mettere a confronto i due metodi normati per la determinazione di E. coli 3-glucuronidasi positivi nei
molluschi bivalvi.
The classification criteria of the areas where shellfish are harvested, are laid down by the UE
Regulations 853/2004, 854/2004, 2073/2005. According to European Legislation, the enumeration of
Escherichia coli in bivalve molluscs must be performed with the ISO method 16449-3:2005 (MPN).
The aim of this work is to compare MPN and pour plate on TBX tests (both ISO methods) to
ascertain whether the latter could substitute the former. Because of the very short shelf-life of
bivalve molluscs the analysis to determine their fitness to human consumption should be as fast as
possible. The MPN method requires two media and a two-day analysis, whereas the TBX method
requires one medium and one-day analysis only. We tested 427 samples of molluscs with both
methods. The results were compared with McNemar's test and by calculating relative Sensitivity and
Specificity. The two methods show significant differences in sensitivity. Although TBX could be
usefully used as screening for MPN, applying a cutoff of 7 colonies (in 5 plates).
Rubini° S, Casaro S, Bianchi° L, Melloni° R, Govoni G, Galletti° G
Comparison of two official methods for the enumeration of Escherichia coli in Manila clams
(Tapes philippinarum)
International Symposium on rapid methods chemical, microbiological and sensory analysis of foods :
7 - 8 May 2012 Copenhagen, Denmark / [Oslo : Nordic Committee on Food Analysis (NMKL), 2012].
- p 43 [Nr. Estr. 5070]
International Symposium on rapid methods chemical, microbiological and sensory analysis of foods
: Copenhagen, Denmark : 7 - 8 May 2012)
The classification criteria of the areas where shellfish are harvested, are laid down by the UE
Regulations 853/2004, 854/2004, 2073/2005. According to European Legislation, the enumeration of
Escherichia coli in bivalve molluscs must be performed with the ISO method 16449-3:2005 (MPN).
The aim of this work is to compare MPN and pour plate on TBX tests (both ISO methods) to
ascertain whether the latter could substitute the former. Because of the very short shelf-life of
bivalve molluscs the analysis to determine their fitness to human consumption should be as fast as
possible. The MPN method requires two media and a two-day analysis, whereas the TBX method
requires one medium and one-day analysis only. We tested 56 samples of Manila clams with both
methods. The results were compared with McNemar's test and by calculating relative Sensitivity and
Specificity. The two methods do not show any significant differences.
Ruggiero° V, Finazzi° G, Bertasi° B, Stroppa A, Noc etti M, Cosciani_Cunico° E,
Oliverio° E, Dalzini° E, Monastero° P, Losio° MN, D aminelli° P
Valutazione della dinamica di comportamento di microrganismi tossigeni durante
l'affioramento del latte per la produzione di Grana Padano DOP e Parmigiano Reggiano DOP
XXII Convegno Nazionale Associazione Italiana Veterinari Igienisti (AIVI) "Autorità competente e
operatore del settore alimentare: armonizzazione degli strumenti di gestione del rischio" : abstracts /
[s.l. : s.n., 2012]. - p 26 [Nr. Estr. 5073]
Convegno Nazionale Associazione Italiana Veterinari Igienisti (AIVI) (22. : Torino : 19 - 21
Settembre 2012)
La sperimentazione è stata realizzata allo scopo di valutare l'andamento di microrganismi patogeni
per l'uomo e tossigeni (B.cereus, C.perfringens, S.aureus) durante il processo di affioramento
naturale della panna da latte crudo utilizzato per la produzione di Grana Padano DOP e Parmigiano
Reggiano DOP. E'stato utilizzato latte vaccino crudo proveniente da una stalla in provincia di
Brescia, che è stato in parte pastorizzato e contaminato allo stesso modo del latte crudo, per
verificare se la presenza di flora indigena nel latte potesse influire sul comportamento dei
microrganismi in fase di affioramento della panna. Il latte è stato suddiviso in io vasche da io litri
ciascuna, contaminate con io° ml di sospensione di Staphylococcus aureus, Bacillus cereus
produttore di tossine diarroiche, Bacillus cereus produttore di tossine emetiche e Clostridium
perfringens, ciascuno alla concentrazione iniziale di io4 UFC/ml. Il latte è stato lasciare affiorare per
circa 12 ore a temperature di 18 e 27°C (i vasca di controllo e 4 vasche contaminate, una per ogni
patogeno, per ciascuna temperatura) e sono stati effettuati prelievi sia del latte che della panna, al
tempo o e a intervalli di tempo di 4,7, 9,1112 ore. Ad ogni prelievo è stata effettuata la numerazione
sia dei diversi microrganismi inoculati, utilizzando metodiche ISO ( UNI EN ISO 7932:2oo5 per
Bacillus cereus, UNI EN ISO 7937:2005 per Clostridium perfringens e UNI EN ISO 6888:1999 per
Staphylococcus aureus) sia delle flore lattiche indigene utilizzando un metodo interno. E' stata
ricercata la presenza di enterotossine e tossina emetica su latte e panna a fine affioramento, con
esito negativo. Sul latte crudo di controllo è stata effettuata inoltre la misurazione del pH tramite
strumento Hanna Instrument HI 223, risultato essere 6.62. Dalle analisi effettuate è emerso che a
fine affioramento la popolazione di batteri lattici è notevolmente maggiore nel latte crudo, rispetto al
latte pastorizzato; tale differenza fa si che la concentrazione del Bacillus cereus aumenti di 4 cicli
logaritmici nel latte pastorizzato mentre nel latte crudo non aumenta neppure di un logaritmo. La
presenza di flore lattiche indigene nel latte crudo svolge dunque un ruolo determinante nel
contenimento di specie come B.cereus, capace invece di sviluppare e crescere nel latte
pastorizzato, in assenza di antagonismo batterico. Tale fenomeno si riconferma per S.aureus, che
mantiene la sua concentrazione iniziale per l'intero periodo di affioramento grazie allo sviluppo di
flore lattiche. Cl.perfringens è presente ad una concentrazione numerabile solo al To. Nei campioni
di controllo di latte e panna pastorizzata è stata rilevata la sola presenza di B.cereus, sia a 18 che
27°C. Nei campioni di controllo di latte e panna cr udi, è stata riscontrata la presenza di stafilococchi
coagulasi positivi e B.cereus, ma non di C.perfringens.
The aim of this study was to evaluate the behaviour of Bacillus cereus, Clostridium perfringens and
Staphylococcus aureus and the presence of Lactic Acid Bacteria during milk outcrop. In detail this
experimental trial shows the distribution of the pathogens and LAB in cream and skimmed milk. The
study was done using raw and pasteurized milk contaminated with pathogens. During outcrop (12
hours) at 18°C and 27°C, different samples were col lected and analyzed The main result obteneid
was the different behaviour of Bacillus cereus in raw milk and pasteurized milk: in pasteurized milk
the lack of LAB allows a 2 log cfu/ml increase of Bacillus cereus. The use of raw milk, with presence
of LAB, is confirmed as a safety factor in the phase of autcrop of milk during the production of
Parmigiano Reggiano DOP and Grana Padano DOP.
Rugna° G, Carra° E, Gelmini° L, Bassi° S
Riscontro di Listeria monocytogenes in latte crudo per il consumo umano: indagine
epidemiologica con il supporto dell'analisi genomica degli isolati mediante PFGE
Infezioni trasmesse da alimenti e acqua : diagnostica ed epidemiologia : VIII workshop nazionale
Enter-net Italia, Sistema di sorveglianza delle infezioni enteriche : 31 maggio-1 giugno 2012,Vietri
sul Mare, Salerno / a cura di Caterina Graziani, Francesca Manciini e Ida Luzzi. - Roma : Istituto
Superiore di Sanità, 2012. - (ISTISAN congressi ; 12/C2) p 67 [Nr. Estr. 4998]
Workshop nazionale Enter-net Italia (8 : Vietri sul Mare, Salerno : 31 maggio-1 giugno 2012)
Introduzione. È noto che nella maggior parte dei casi la contaminazione del latte crudo con Listeria
monocytogenes (L. monocytogenes) è di origine ambientale. Esistono tuttavia casi in cui la
contaminazione è di tipo primario conseguente, in genere, a mastiti sostenute da L. monocytogenes
con eliminazione diretta del patogeno attraverso il latte. Dal punto di vista epidemiologico stabilire
l'origine della contaminazione è molto importante perché consente di intervenire in maniera mirata
per prevenirla o interromperla. Viene descritto l'approfondimento epidemiologico fatto a seguito di
una positività per L. monocytogenes in un campione di latte crudo prelevato da un distributore
automatico della provincia di Modena nell'ambito di una indagine riguardante la qualità
igienico-sanitaria di questo alimento. Metodi. Dopo il riscontro della presenza di L. monocytogenes
in un campione di latte crudo destinato al consumo umano, al fine di escludere una contaminazione
verificatasi in fase di distribuzione, è stato esaminato il latte di massa prelevato dalla cisterna
aziendale con il quale veniva alimentato il distributore automatico. Successivamente sono stati
esaminati campioni individuali di latte delle 107 vacche in lattazione presenti in azienda. Tutti i
campioni sono stati sottoposti alla ricerca microbiologica di L. monocytogenes mediante ISO
11290-1:1996/Amd 1:2004. I ceppi di L. monocytogenes isolati dai campioni in esame sono stati
genotipizzati mediante PFGE. Risultati. Il latte di massa è risultato positivo per L. monocytogenes.
Un numero significativo di colonie di L. monocytogenes isolate dal latte di massa è stato sottoposto
ad indagine biomolecolare per valutare la presenza o meno di diversi profili genomici. Dal campione
individuale di latte di una bovina è stata isolata L. monocytogenes con valori di carica pari a 2.000
UFC/ml. La vacca è stata riformata. L'esame anatomo-isto-patologico ha rilevato la presenza di una
mastite interstiziale di tipo cronico in due quarti e l'esame batteriologico ha portato all'isolamento di
L. monocytogenes dai linfonodi sopramammari. Il mancato isolamento dalla mammella è ascrivibile
al fatto che la bovina era stata trattata con antibiotici per via endocanalicolare poco prima della
macellazione. La caratterizzazione molecolare di tutti i ceppi testati ha evidenziato la presenza di un
unico profilo genetico. Conclusioni. La presenza di un unico genotipo tra i ceppi di L.
monocytogenes isolati dalle diverse matrici ne conferma la probabile origine clonale supportando
l'ipotesi della presenza di un'unica fonte di contaminazione identificabile nella bovina con mastite. I
successivi controlli sul latte di massa eseguiti dopo eliminazione della bovina positiva hanno dato
esito favorevole confermando di fatto tale ipotesi.
Sabatucci° G, Zanardini° N, Berta° V, D'Ippolito° N , Coffinardi° F, Titola° M, Fusini°
F, D'Amico° S, Bertasi° B
Messa a punto di metodiche di biologia molecolare per la ricerca di cianobatteri
XIV Congresso Nazionale SIDiLV : 24 - 26 Ottobre 2012, Sorrento (NA) : volume degli atti / [s.l. :
Societa' Italiana Diagnostica di Laboratorio Veterinaria ( SIDiLV ), 2012]. - p 480-481. - 2 bib ref [Nr.
Estr. 5087]
Congresso Nazionale Societa' Italiana Diagnostica di Laboratorio Veterinaria (SIDiLV) (14. :
Sorrento (NA) : 24 - 26 Ottobre 2012)
L'eutrofizzazione dei bacini idrici, risultato dell'implemento esogeno e endogeno di nutrienti nelle
acque, può causare imponenti fioriture di Cianobatteri. Un aumento massivo di Cianobatteri induce
potenziali effetti negativi sul-la qualità ambientale delle acque e può provocare distorsioni ecologiche
pesanti come la moria dei pesce. L'accumulo di microrganismi produttori dí tossine in acque
potabili ed eventualmente nei tessuti dei pesci, rendono le fioriture dei Cianobatteri potenzialmente
pericolose per l'uomo; inoltre, nel corso degli ultimi anni, si è verificato un implemento del rischio di
esposi-zione a causa dei cambiamenti climatici. Tra le varie specie di Cianobatteri, Microcystis è la
più studiata, in quanto comunemente presente negli ecosistemi acquatici di tutto il mondo e
maggiormente rappresentativa dei Cianobatteri che producono Microcistine, tossine che minacciano
la salute. dell'uomo e degli animali (epatotossine e promotori tumorali). Quantificare, nei campioni
ambientali, i cianobatteri potenzialmente tossici è molto importante per monitorare le fioriture alghe
nocive ed effettuare, nei limiti del possibile, interventi di prevenzione. Il diretto conteggio
microscopico viene normalmente utilizzato per l'enumerazione dei cianobatteri. Tuttavia, a volte,
risulta difficile l'identificazione e la conta di queste specie. Nonostante la presenza dei microrganismi
non sia indicativa della produzione di tossine, recentemente sono state sviluppate tecniche
molecolari, per stimare l'abbondanza di cellule che possiedano geni specifici per la produzione di
tossine. Tali tecniche sono comunque importanti per la valutazione preliminare dei rischi associati
alle fioriture. Il presente lavoro è stato intrapreso con l'obiettivo della messa a punto di metodiche
biomolecolari quantitative, per la verifica della presenza di Cianobatteri potenzialmente tossigeni, in
campioni di acqua, con la possibilità di applicare la metodica anche al controllo di matrici più
complesse (prodotti ittici).
The aim of this work will be to set up molecular biology methods to detect cyanobacteria in fresh
water The importance of cyanobacteria identification is related to their toxins productions ability, at
different hazard level for human and animals. Molecular biology techniques to detect toxin producers
microorganisms could be advantageous compared to morphological identification or chemical
methods, respectively first poorly discriminating about species level and more expensive. So it is
possible to detect potentially dangerous cyanobacteria in water, before the consequently bloom.
Saletti C, Mazzetti C, Bardasi° L, Cosciani_Cunico° E
Verifica della sicurezza alimentare dei prodotti di IV Gamma
Convegno Microbiologia Predittiva: possibili utilizzi nell'attività di controllo ufficiale degli operatori
del settore alimentare / [s.l. : s.n., 2012]. - 20 p
http://www.ausl.re.it/Home/DocumentViewer.aspx?ID=1761&TIPODOC=COMUNICAZIONE [Nr.
Estr. 5225]
Convegno Microbiologia Predittiva: possibili utilizzi nell'attività di controllo ufficiale degli operatori del
settore alimentare : Reggio Emilia : 30 Novembre 2012)
Scordella G, Bresolin R, Baldi D, Rubini° S
Risk assessment on Listeria monocitogens policy management
AQUA 2012 Global Aquaculture securing our future : Prague, Czech Republic Sep 1-5 2012 :
abstracts / [s.l. : s.n, 2012]. - p 1006 [Nr. Estr. 5071]
AQUA 2012 : Prague, Czech Republic : Sep 1-5, 2012)
Listeria monocytogenes is widely distributed in the general environment including fresh water,
coastal water and live fish from these areas and has been isolated from both domestic and imported,
fresh, frozen, and processed seafood products, including crustaceans, molluscs, and finfish.
Although L. monocytogenes has been recovered from fresh and smoked mussels, the incidence in
other molluscs is low. Human listeriosis has been associated with consumption of food including
seafood. The organism has been recovered from ready-to-eat products such as cooked crab meat,
shrimp and finfish, and from hot¬smoked and cold-smoked mussels, salmon, and other finfish.
Surveillance information on the presence of L. monocytogenes in seafood products is required as a
background for steps that are being taken to inform risk managers of risks associated with particular
food products. Risk assessment uses quantitative scientific and epidemiological information in a
structured format to determine the risks associated with particular hazards. The questions being
considered include those on level of consumption, epidemiology, dose response, and the virulente,
biology, and ecology of the organism. A change in regulatory policy on L. monocytogenes should be
made within the framework of a risk analysis scheme. Risk management decisions about L.
monocytogenes policy should be made on the basis of a detailed review of the risk assessment. The
European Regulation 2073/2005 and next amendment differs from regulations operating in other
foreign countries (such as Japan and USA). Considering the different food habits on the different
populations, a wider agreement in laws should be considered.
Scordella G, Gustinelli A, Caffara M, Prearo M, Rubini° S; Bresolin R, Baldi D,
Fioravanti ML
Occurrence of zoonotic parasites in wild freshwater fish from Italy
AQUA 2012 Global Aquaculture securing our future : Prague, Czech Republic Sep 1-5 2012 :
abstracts / [s.l. : s.n, 2012]. - p 1005 [Nr. Estr. 5072]
AQUA 2012 : Prague, Czech Republic : Sep 1-5, 2012)
Foodborne parasitic infection affect more than 40 million people throughout the world, and
particularly prevalent in South-East Asia, Western Pacific Regions and East Europe. In particular,
liver fluke infection caused by Opisthorchiidae is a major public health problem in many parts of the
Far East, Southeast Asia, and eastern Europe. The parasites' mode of transmission is closely linked
to human behavioral patterns, and specifically to methods of food production and preparation in the
endemic countries. The recent changes in eating habits registered in Italy, with an increasing
consumption of raw,cold smoked or marinated freshwater fish, has resulted in re-emerging and
emerging fish-borne parasitic zoonosis. A recrudescence of human Diphyllobothriasis by the
pseudophyllidean cestode Diphyllobothriuni latum has been observed in Como Lake district, with
some dozen cases diagnosed from 2001 up to now, and some outbreaks of human Opistorchiasis
by the digenean Opisthorchis felineus have been described in central Italy alter consumption of
marinated lacustrine fish, with over 50 cases reported from 2003 to 2010. In order to understand the
epidemiology of these parasites in Italy, a parasitological survey in fish populations from several
lakes has been carried out in 2009-2011. The results of this survey showed a wide diffusion
ofD.latum plerocercoid larvae in European perch (Perca fluviatilis) from Como and Iseo Lakes. All
the perch from Maggiore and Garda Lakes were negative. Concerning O. felineus metacercariae,
almost all the tench (Tinca tinca) coming from Vico, Bracciano and Bolsena Lakes in centrai Italy
resulted positive, while the 92 tench from Trasimeno Lake were negative. The consumption of raw
perch ("perch carpaccio") and marinated tench in the areas taken into consideration has been
identified as the main food source of viable infective stages of D. latum and O. felineus respectively.
The analysis of risk factors influencing the epidemiology of these zoonotic helminthes indicate the
Jack of efficient urban wastewater depuration plants around Como and Iseo Lakes shores as the
main aspect influencing the maintenance of D. latum cycle in these aquatic environments.
Concerning O. felineus, the presence of parasitised stray cats, main reservoirs of the infection, and
of Bythinia spp., suitable first intermediate host, seem to be the key factor in the subsistence of this
zoonotic helminth described in Italy since XIX century.
Serraino A, Finazzi° G, Marchetti G, Daminelli° P, Riu R, Giacometti F, Losio° MN,
Rosmini R
Behaviour of Salmonella Typhimurium during production and storage of artisan water buffalo
mozzarella cheese
Ital J Anim Sci. - Vol. 11 e53 ( 2012). - p 285-289. - 37 bib ref [Nr. Estr. 5052]
Water buffalo mozzarella cheese (WBMC) is a fresh pasta filata cheese produced from whole chilled
buffalo milk. Although pasteurization of milk and the use of defined starter cultures are
recommended, traditional technology involving the use of unpasteurized milk and natural whey
cultures is still employed for WBMC production in Italy. The aim of this study were to assess the
behaviour of Salmonella Typhimurium during the production of artisan water buffalo mozzarella
cheese and during its shelf life under different temperature conditions. Raw milk was inoculated with
S. Typhimurium and the evolution of S. Typhimurium count during production and shelf life was
monitored. In artisan WBMC production technology S. Typhimurium multiplied in the curd during
ripening, but its growth rate expressed in log CFU/g/h was lower than the growth rate reported by
theoretical predictions. Stretching proved to be a process with good repeatability and able to reduce
S. Typhimurium contamination by 5.5 Log CFU/g. The intrinsic characteristics of traditional WBMC
proved to be unable to obstacolate the growth of S. Typhimurium during storage in the case of
thermal abuse. Control of raw milk contamination and a proper refrigeration temperature are key
factors in reducing the risk for consumers
.
Taddei° R, Tosi° G, Boniotti° MB, Casadio° M, Fiore ntini° L, Fabbi° M, Massi° P
Caratterizzazione molecolare di ceppi del virus della bronchite infettiva aviare isolati in Italia
tra il 1963 ed il 1989
LI Convegno annuale Societa' Italiana di Patologia Aviare (SIPA) : 11-12 Ottobre 2012
Salsomaggiore Terme (PR) : atti / [s.l. : s.n., 2012]. - p 332-341. - 19 bib ref [Nr. Estr. 5241]
Convegno annuale Societa' Italiana Patologia Aviare (SIPA) (51 : Salsomaggiore Terme (PR) :
11-12 Ottobre 2012)
A retrospective study was carried out on 123 Infectious Bronchitis Virus strains isolated in Italy
between 1963 and 1989. A portion of the S1 gene was amplified ed and sequenced in order to
explore the genetic variability of the IBV strains circulating in those years. Genotype Massachussets
was the prevalent genotype in the mid .60s, since then 624/I became the prevalent genotype until
the end of the .80s. D1466 was Þ rst detected in 1979, D274 in 1981 and 793B in 1989. During the
.60s and .70s the presence of some variant strains was also highlighted.
Zanardi° G, Bolzoni° G, Marcolini° A, Giuseppe° B,
Bertocchi° L
Fatty acids profile by Fourier Transform Infrared spectroscopy (FTIR) of fat: validation as
screening test in dairy herds of Lombardy Region, Italy
XXVII World Buiatrics Congress : 3 - 8 June, 2012 Lisbon, Portugal : abstract book / [s.l. : s. n.,
2012]. - p 119-120 (P:381) [Nr. Estr. 5011]
World Buiatrics Congress (27th : Lisbon, Portugal : 3 - 8 June, 2012)
Objectives: Aim of this work has been to determine the precision and the accuracy of FTIR method
in profiling the fatty acids in bovine bulk tank milk samples, analyzed within the payment quality
system in Lombardy region, Italy. Distribution of fatty acids profiles was outlined from about 5,000
dairy herds over 36 months. Materials and Methods: Equipment FTIR spectrometry
MilkoscanFT6000® (Foss, DK) with management software Foss Integrator®. Calibration curves
developed by producer for satured (SAT), unsatured (UNSAT), monounsat¬ured (MONO),
polyunsatured (POLY) fatty acids. Gas chromatography (GC) - HP 6890 GC System (Agilent
Technologies Inc.) as reference method measurements of milk fatty acid composition. Samples
Refrigerated bulk milk samples processed by milk laboratory in Brescia within the payment quality
system were selected to carry out the validation, Bovine, goat and buffalo milk as internal reference
material was used to determine the intra-laborato-ry repeatability and reproducibility. Repeated
measurements of these reference samples have been gathered over a 30-45 days period. Accuracy
has been estimated on bovine milk samples collected ad hoc and analyzed by FTIR spectrometry as
raw milk and by GC as frozen milk. Results: On each parameter the intra-laboratory values of
Repeatability [r], Reproducibility [R] and Standard Error of Prediction with slope and intercept
correction [SEPcorr] were estimated. Fatty acids profile screening was performed on more than
5,000 dairy herds over a 36 months period for a total of about 320,000 milk samples. Analysis of
data was carried out on a sub-population of 3,093 dairy herds, eligible for at least one sampling per
month. The results calculated as g/100g were the following: SAT: r = 0.054, R = 0.076, SEPcorr =
0.040; UNSAT: r = 0.048, R = 0.065, SEPcorr = 0.021; MONO: r = 0,042, R = 0.065, SEPcorr =
0.029; POLY: r =0.014 , R = 0.014, SEPcorr = 0.023. The relative distribution in percentage of fatty
acids shows the following results (mean ± 1SD): SAT = 0.692% ± 0.024%; UNSAT = 0.313% ±
0.023%; MONO = 0.276% ± 0.021%; POLY = 0.037% ± 0,004%. Conclusions: The estimated
precisíon and accuracy agree with previous studies and instrumental performances specification.
FTIR spectrometry is a useful screening tool to determine the mean fatty acids profile in dairy herds
submitted to the payment quality system. In the next future it will be of interest to evaluate the
relationship between the feeling of cattle and fatty acids composition in the milk and in the dairy
products.
Zanardi° G, Finazzi° G, Daminelli° P, Bonometti° G,
Losio° N
Programma di controllo ispirato al principi dell'HACCP nella produzione primaria: risultati
preliminari
XIV Congresso Nazionale SIDiLV : 24 - 26 Ottobre 2012, Sorrento (NA) : volume degli atti / [s.l. :
Societa' Italiana Diagnostica di Laboratorio Veterinaria ( SIDiLV ), 2012]. - p 529-530. - 3 bib ref [Nr.
Estr. 5086]
Congresso Nazionale Societa' Italiana Diagnostica di Laboratorio Veterinaria (SIDiLV) (14. :
Sorrento (NA) : 24 - 26 Ottobre 2012)
Il regolamento (CE) n. 852/2004 non obbliga il produttore primario o operatore del settore alimentare
a realizzare e ad applicare un sistema HACCP, tuttavia lo responsabilizza nell'esercitare le buone
pratiche di allevamento in materia d'igiene, di registrazione attraverso l'attuazione di una corretta
prassi igienica. L'inerente realizzazione di manuali va integrata con un adeguato programma di
autocontrollo della produzione primaria negli allevamenti di bovine da latte per documentare
l'applicazione di valide procedure, finalizzate a controllare i rischi per garantire la sicurezza
alimentare del consumatore. In quest'ambito, l'IZSLER e il Centro Miglioramento Qualità Latte
(CMQL) di Brescia hanno realizzato e collaudato nel 2011 un modello di sorveglianza degli
allevamenti da latte ispirato al principio dell'HACCP, imperniato sul monitoraggio sanitario,
attraverso l'uso di indicatori microbiologici e chimici (1, 2). L'obiettivo è di creare conoscenza ed
evidenza scientifica di dinamiche sanitarie nella produzione primaria, funzionalí a un sistema di
auto-controllo che consenta di identificare i pericoli, valutare e controllare i rischi, attraverso idonei
protocolli di assistenza aziendali. Il lavoro presenta i risultati preliminari relativi al primo anno di
attuazione del sistema, progettato su base triennale.
This work describes the preliminary results of a control programme based on HACCP principles and
carried out in 2011 on 250 daity farms in the province of Brescia. The aim of this surveillance is to
assess the risk of pathogens like Listeria monocytogenes, Escherichia coli VTEC, Streptococcus
agalactiae, Staphilococcus aureus and dioxins, PCB dioxin relevant to food safety during the milk
production. Polimerase Chain Reaction, microbiological and chemical analysis were carried out on
bulk milk samples; a questionnaire concerning the presence of microbiological risk factors was
realized.