Publications by authors named "Giuseppe Merialdi"

42 Publications

Evidence of Common Isolates of in Bovines and Humans in Emilia Romagna Region (Northern Italy).

Front Microbiol 2021 11;12:673126. Epub 2021 Jun 11.

Experimental Zooprophylactic Institute in Lombardy and Emilia Romagna, Brescia, Italy.

(group B , GBS) is one of the most important agents of bovine mastitis and causes remarkable direct and indirect economic losses to the livestock sector. Moreover, this species can cause severe human diseases in susceptible individuals. To investigate the zoonotic potential of , 203 sympatric isolates from both humans and cattle, isolated in the same time frame (2018) and in the same geographic area (Emilia Romagna region, Northern Italy), were characterized by molecular capsular typing (MCT), pilus island typing (PI), and multi-locus sequence typing (MLST). In addition, antibiotic-resistant phenotypes were investigated. The distribution of the allelic profiles obtained by combining the three genotyping methods (MCT-PI-MLST) resulted in 64 possible genotypes, with greater genetic variability among the human compared to the bovine isolates. Although the combined methods had a high discriminatory power (>96,2%), five genotypes were observed in both species (20,9% of the total isolates). Furthermore, some of these strains shared the same antibiotic resistance profiles. The finding of human and bovine isolates with common genotypes and antibiotic resistance profiles supports the hypothesis of interspecies transmission of between bovines and humans.
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http://dx.doi.org/10.3389/fmicb.2021.673126DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8226232PMC
June 2021

A pathogenic HEXA missense variant in wild boars with Tay-Sachs disease.

Mol Genet Metab 2021 Jul 7;133(3):297-306. Epub 2021 May 7.

Department of Veterinary Science, University of Parma, Via Taglio, 8, 43100 Parma, Italy.

Gangliosidoses are inherited lysosomal storage disorders caused by reduced or absent activity of either a lysosomal enzyme involved in ganglioside catabolism, or an activator protein required for the proper activity of a ganglioside hydrolase, which results in the intra-lysosomal accumulation of undegraded metabolites. We hereby describe morphological, ultrastructural, biochemical and genetic features of GM2 gangliosidosis in three captive bred wild boar littermates. The piglets were kept in a partially-free range farm and presented progressive neurological signs, starting at 6 months of age. Animals were euthanized at approximately one year of age due to their poor conditions. Neuropathogens were excluded as a possible cause of the signs. Gross examination showed a reduction of cerebral and cerebellar consistency. Central (CNS) and peripheral (PNS) nervous system neurons were enlarged and foamy, with severe and diffuse cytoplasmic vacuolization. Transmission electron microscopy (TEM) of CNS neurons demonstrated numerous lysosomes, filled by parallel or concentric layers of membranous electron-dense material, defined as membranous cytoplasmic bodies (MCB). Biochemical composition of gangliosides analysis from CNS revealed accumulation of GM2 ganglioside; furthermore, Hex A enzyme activity was less than 1% compared to control animals. These data confirmed the diagnosis of GM2 gangliosidosis. Genetic analysis identified, at a homozygous level, the presence of a missense nucleotide variant c.1495C > T (p Arg499Cys) in the hexosaminidase subunit alpha gene (HEXA), located within the GH20 hexosaminidase superfamily domain of the encoded protein. This specific HEXA variant is known to be pathogenic and associated with Tay-Sachs disease in humans, but has never been identified in other animal species. This is the first report of a HEXA gene associated Tay-Sachs disease in wild boars and provides a comprehensive description of a novel spontaneous animal model for this lysosomal storage disease.
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http://dx.doi.org/10.1016/j.ymgme.2021.05.001DOI Listing
July 2021

Epidemiologic case investigation on the zoonotic transmission of Staphylococcus aureus infection from goat to veterinarians.

Zoonoses Public Health 2021 May 5. Epub 2021 May 5.

Department of Veterinary Medical Sciences, University of Bologna, Bologna, Italy.

Staphylococcus aureus infection led to a case of goat abortion, and four veterinarians contracted S. aureus infection from the goat during and after the abortion. Three veterinarians assisted a doe during the dystocic delivery of a dead foetus. Seventy-two hours after the dystocia, which ended with the goat's death, the veterinarians who assisted during the kidding and the veterinarian who performed the necropsy showed the presence of multiple, isolated, painful pustules 1-5 mm in diameter located along their forearms and knees. S. aureus was isolated from the pustules of the veterinarians, the placenta and uterus of the goat, the organs (brain, thymus gland, abomasum, liver and spleen) of the foetus, the scrotum and eye swabs of the buck, and mammary pustules of another goat from the same herd. Histological analysis revealed purulent metritis and inflammation of the placental cotyledons. Additional investigations eliminated the chances of other infections. S. aureus isolates recovered from the veterinarians, goats, foetus and buck were sensitive to the tested anti-microbials and did not encode staphylococcal enterotoxin genes (sea, ser, sep, see, seg and sei). The isolates were closely related, as indicated by the results of Fourier-transform infrared spectroscopy and comparative whole-genome sequencing analysis. The results of this study clearly support the hypothesis that an episode of professional zoonosis was caused by S. aureus infection during the abortion and also highlight the need for bacterial subtyping in epidemiological surveys.
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http://dx.doi.org/10.1111/zph.12836DOI Listing
May 2021

Antimicrobial Resistance Profile and ExPEC Virulence Potential in Commensal of Multiple Sources.

Antibiotics (Basel) 2021 Mar 26;10(4). Epub 2021 Mar 26.

Department of Veterinary Medical Sciences, Faculty of Veterinary Medicine, University of Bologna, Ozzano Emilia, 40064 Bologna, Italy.

We recently described the genetic antimicrobial resistance and virulence profile of a collection of 279 commensal of food-producing animal (FPA), pet, wildlife and human origin. Phenotypic antimicrobial resistance (AMR) and the role of commensal as reservoir of extra-intestinal pathogenic (ExPEC) virulence-associated genes (VAGs) or as potential ExPEC pathogens were evaluated. The most common phenotypic resistance was to tetracycline (76/279, 27.24%), sulfamethoxazole/trimethoprim (73/279, 26.16%), streptomycin and sulfisoxazole (71/279, 25.45% both) among the overall collection. Poultry and rabbit were the sources mostly associated to AMR, with a significant resistance rate ( > 0.01) to quinolones, streptomycin, sulphonamides, tetracycline and, only for poultry, to ampicillin and chloramphenicol. Finally, rabbit was the source mostly associated to colistin resistance. Different pandemic (ST69/69*, ST95, ST131) and emerging (ST10/ST10*, ST23, ST58, ST117, ST405, ST648) ExPEC sequence types (STs) were identified among the collection, especially in poultry source. Both ST groups carried high number of ExPEC VAGs (pandemic ExPEC STs, mean = 8.92; emerging ExPEC STs, mean = 6.43) and showed phenotypic resistance to different antimicrobials (pandemic ExPEC STs, mean = 2.23; emerging ExPEC STs, mean = 2.43), suggesting their role as potential ExPEC pathogens. Variable phenotypic resistance and ExPEC VAG distribution was also observed in uncommon ExPEC lineages, suggesting commensal flora as a potential reservoir of virulence (mean = 3.80) and antimicrobial resistance (mean = 1.69) determinants.
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http://dx.doi.org/10.3390/antibiotics10040351DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8067153PMC
March 2021

Update on a model to describe Salmonella spp. population reduction in Italian salami during production and high-pressure processing.

Meat Sci 2021 Jul 18;177:108506. Epub 2021 Mar 18.

Department of Veterinary Medical Sciences, University of Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia (BO), Italy.

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http://dx.doi.org/10.1016/j.meatsci.2021.108506DOI Listing
July 2021

Distribution, virulence, genotypic characteristics and antibiotic resistance of Listeria monocytogenes isolated over one-year monitoring from two pig slaughterhouses and processing plants and their fresh hams.

Int J Food Microbiol 2021 Jan 9;336:108912. Epub 2020 Oct 9.

Department of Veterinary Medical Sciences, University of Bologna, Bologna, Italy. Electronic address:

Listeria monocytogenes contamination in raw pork and ready to eat foods is an important food safety concern, also for the increasing detection of antimicrobial-resistant isolates. Data on L. monocytogenes occurrence, persistence, distribution and genetic characterization in two different plants, namely in continuum from slaughtered pigs, environment and unfinished products (fresh hams) were observed by one-year monitoring and were integrated with their antimicrobial resistance patterns. A total of 98 samples out of the overall 1131 (8.7%) were positive for L. monocytogenes, respectively 2.6% and 13.2% in plants A and B: only three serotypes were identified, 1/2c (50%), 1/2b (36.7%) and 1/2a (13.27%), and strains were classified in 35 pulsotypes and 16 clusters by PFGE; a unique P-type was highlighted according to the detection of virulence genes. The contamination flow of L. monocytogenes has a low occurrence in slaughterhouse (Plant A = 1.1%, Plant B: 3.1%; p > 0.05) and increased throughout the processing chain with trimming area as the most contaminated (Plant A: 25%, Plant B: 57%; (p < 0.05)), both in the environment and in unfinished products (80% in hams before trimming in plant B). The dominant role of environmental contamination in post-slaughter processing is confirmed to be a significant cause of meat contamination by L. monocytogenes. Very high levels of resistance were observed for clindamycin (57%) and high resistance levels (>20-50%) to ciprofloxacin, oxacillin, levofloxacin and daptomycin, confirming the L. monocytogenes resistance trend to a wide range of antimicrobial agents. A total of 11 L. monocytogenes isolates were multidrug resistant and 7 out of them were isolated from slaughtered pigs. An interesting significant (p < 0.05) statistical correlation has been found between resistance to some antimicrobial agents and lineage/serotypes. Microbiological sampling of food and environments after sanitization are commonly used as verification procedure for the absence of L. monocytogenes in food plants and to give assurance of food safety, but strains characterization is necessary for industries to target specific control measures, like the enforcement of the hygiene program and of the control of operator activities, at least for permanent strains. The only presence of L. monocytogenes could not be considered as the conclusive assessment of a potential risk for public health, also in terms of emerging and emerged antimicrobial resistances.
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http://dx.doi.org/10.1016/j.ijfoodmicro.2020.108912DOI Listing
January 2021

Differences in larval survival and IgG response patterns in long-lasting infections by Trichinella spiralis, Trichinella britovi and Trichinella pseudospiralis in pigs.

Parasit Vectors 2020 Oct 16;13(1):520. Epub 2020 Oct 16.

Department of Infectious Diseases, Istituto Superiore di Sanità, viale Regina Elena 299, 00161, Rome, Italy.

Background: Domesticated and wild swine play an important role as reservoir hosts of Trichinella spp. and a source of infection for humans. Little is known about the survival of Trichinella larvae in muscles and the duration of anti-Trichinella antibodies in pigs with long-lasting infections.

Methods: Sixty pigs were divided into three groups of 20 animals and infected with 10,000 larvae of Trichinella spiralis, Trichinella britovi or Trichinella pseudospiralis. Four pigs from each group were sacrificed at 2, 6, 12, 18 and 24 months post-infection (p.i.) and the number of larvae per gram (LPG) of muscles was calculated. Serum samples were tested by ELISA and western blot using excretory/secretory (ES) and crude antigens.

Results: Trichinella spiralis showed the highest infectivity and immunogenicity in pigs and larvae survived in pig muscles for up to 2 years p.i. In these pigs, the IgG level significantly increased at 30 days p.i. and reached a peak at about 60 days p.i., remaining stable until the end of the experiment. In T. britovi-infected pigs, LPG was about 70 times lower than for T. spiralis at 2 months p.i. and only very few infecting larvae were detected at 6 months p.i., whereas no larvae were detected at 12, 18 and 24 months p.i. At 6 months p.i., degenerated/calcified larvae and cysts were detected in the muscles by trichinoscopy and histology. The IgG pattern showed by T. britovi-infected pigs was similar to that of T. spiralis-infected pigs, although seroconversion occurred some days later. The larval burden of T. pseudospiralis was slightly greater than for T. britovi at 2 months p.i., but no larvae were detected at 6 and 12 months p.i. In T. pseudospiralis-infected pigs, seroconversion occurred slowly, as in T. britovi-infected pigs. The IgG level showed a significant drop at 6 months p.i. and declining to the cut-off value at 12 months p.i.

Conclusions: The longer survival of T. spiralis in pigs in comparison with the other two species highlights its exceptional dissemination potential. These results provide an explanation of the controversial data collected by parasitological and serological tools in the course of epidemiological investigations.
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http://dx.doi.org/10.1186/s13071-020-04394-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7566126PMC
October 2020

Modeling the behavior of Listeria innocua in Italian salami during the production and high-pressure validation of processes for exportation to the U.S.

Meat Sci 2021 Feb 17;172:108315. Epub 2020 Sep 17.

Department of Veterinary Medical Sciences, University of Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia (BO), Italy.

A model describing Listeria innocua evolution according to process parameters of 51 Italian salami processes and HPP in 31 companies was developed. A total of 51 challenge tests were performed. During processing a L. innocua reduction of 0.34-4.32 Log10 CFU/g was observed and HPP further reduced the count of 0.48-3.47 Log10 CFU/g; an overall reduction of 1.04-5.68 is reached. PH after acidification/drying process, a after seasoning, duration of the seasoning and caliber resulted associated (p < 0.05) with L. innocua decrease. HPP efficacy was associated (p < 0.05) with a and pH of the product: higher the pH and a after the acidification/drying and seasoning phases, higher resulted the L. innocua reduction after HPP. No significant association was observed between L.innocua and salt, nitrate and starter content and other characteristics of process. The model meets companies and Authorities needs and represents a useful tool to predict L. monocytogenes lethality, giving recommendations to food business operators interested in exportation to the U.S.
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http://dx.doi.org/10.1016/j.meatsci.2020.108315DOI Listing
February 2021

Effect of production process and high-pressure processing on viability of in traditional Italian dry-cured .

Ital J Food Saf 2020 Aug 19;9(2):9133. Epub 2020 Aug 19.

Istituto Zooprofilattico Sperimentale delle Lombardia e dell'Emilia Romagna, Sede Territoriale di Bologna.

In this study the effect of the application of High Pressure Treatment (HPP) combined with four different manufacturing processes on the inactivation of Listeria innocua, used as a surrogate for L. monocytogenes, in artificially contaminated samples was evaluated in order to verify the most suitable strategy to meet the Listeria inactivation requirements needed for the exportation of dry-cured meat in the U.S. Fresh anatomical cuts intended for production were supplied by four different delicatessen factories located in Northern Italy. Raw meat underwent experimental contamination with using a mixture of 5 strains. Surface contamination of the fresh anatomical cuts was carried out by immersion into inoculum containing spp The conditions of the HPP treatment were: pressure 593 MPa, time 290 seconds, water treatment temperature 14°C. was enumerated on surface and deep samples post contamination, resting, ripening and HPP treatment. The results of this study show how the reduction of the microbial load on during the production process did not vary among three companies (P>0.05) ranging from 3.73 to 4.30 log CFU/g, while it was significantly different (P<0.01) for the fourth company (0.92 log CFU/g). HPP treatment resulted in a significant (P<0.01) deep decrease of count with values ranging between 1.63-3.54 log CFU/g with no significant differences between companies. Regarding superficial contamination, HPP treatment resulted significant (P<0.01) only in Coppa produced by two companies. The results highlight that there were processes less effective to inhibit the pathogen; in particular for company D an increase of L. innocua count was shown during processing and HPP alone cannot be able to in reaching the Listeria inactivation requirements needed for exportation of dry-cured meat in the U.S. According to the data reported in this paper, HPP treatment increases the ability of the manufacturing process of in reducing count with the objective of a lethality treatment.
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http://dx.doi.org/10.4081/ijfs.2020.9133DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7459741PMC
August 2020

Effect of production process and high-pressure processing on viability of in traditional Italian dry-cured .

Ital J Food Saf 2020 Aug 19;9(2):8445. Epub 2020 Aug 19.

Istituto Zooprofilattico Sperimentale delle Lombardia e dell'Emilia Romagna, Sede Territoriale di Bologna.

The aim of the study was to investigate the combined effect of the manufacturing process followed by HPP treatment on the inactivation of spp. in artificially contaminated samples, in order to verify the ability of the combined processes to achieve the objective of a 5-log reduction of spp. needed for exportation to the U.S. Fresh anatomical cuts intended for production were supplied by four different delicatessen factories located in Northern Italy. Raw meat underwent experimental contamination with spp. using a mixture of 3 strains. Surface contamination of the fresh anatomical cuts was carried out by immersion into inoculum containing spp The conditions of the HPP treatment were: pressure 593 MPa, time 290 seconds, water treatment temperature 14°C. Surface and deep samples were performed post contamination (T0), end of the cold phase (T1), end of process (Tend), and after HPP treatment (postHPP) and spp. Enumerated. The results of this study show a significant reduction of spp. all through the production process (P<0.01) for all companies, followed by an additional reduction of bacterial counts due to HPP treatment (P<0.01), both in superficial and deep contaminations (P<0.01). The superficial overall reduction resulted of 1.58 to 5.04 log CFU/g during the production process. HPP treatment resulted in a significant (P<0.01) superficial and deep decrease in spp. enumeration varying from 0.61 to 4.01 log and from 1.49 to 4.13 log. According to the data presented in this study, only the combined approach of manufacturing process followed by HPP treatment always led to a 5-log reduction of Salmonella spp. required by USDA/FSIS guidelines.
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http://dx.doi.org/10.4081/ijfs.2020.8445DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7459750PMC
August 2020

Who killed my dog? Use of forensic genetics to investigate an enigmatic case.

Int J Legal Med 2021 Mar 11;135(2):387-392. Epub 2020 Aug 11.

Department of Veterinary Medical Sciences, University of Bologna, Ozzano dell'Emilia, Italy.

Genetic testing of animal biological material has become a valuable tool in forensic investigations, and it is successfully used to identify unknown crime perpetrators, to unmask food frauds, or to clarify cases of animal attacks on humans or other animals. When DNA profiling is not possible due to inadequate amounts of nuclear DNA, mitochondrial DNA (mtDNA) testing is the only viable alternative, as in the case of shed hair samples. In this case, a dog was allegedly killed by wild animals while being hosted in a boarding house. Extraneous hair fragments recovered from the dog's mouth and paws were subjected to genetic analysis: the cytochrome b gene located on mtDNA was amplified and sequenced in order to determine the species responsible for the killing. The mtDNA analysis provided evidence that the dog was killed by other dogs, thus unmasking a false wild animal attack and putting the case in an entirely different perspective.
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http://dx.doi.org/10.1007/s00414-020-02388-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7870635PMC
March 2021

Factors affecting the microbiological load of Italian hunted wild boar meat (Sus scrofa).

Meat Sci 2020 Feb 17;160:107967. Epub 2019 Oct 17.

Local Health Authority - Veterinary Area, Bologna, Italy. Electronic address:

This study investigates the microbiological conditions before maturation of wild boar meat (Sus scrofa) processed in approved game handling establishments in Italy. Fillets and legquarters of 37 carcasses were tested to assess Aerobic Colony Count (ACC), Enterobacteriaceae Count (EC) and Salmonella presence. Salmonella was never found and mean values of ACC and EC were 4.67 ± 1.78 SD and 2.60 ± 1.58 SD log CFU/cm, respectively. Both ACC and EC increased with time between evisceration and skinning, were significantly higher in fillets and when meat was processed by untrained operators. ACC also increased with boars' weight and when carcasses were cleaned with running potable water. Based on limits set by EU Regulation No 1441/2007 for pork meat, most legquarters resulted satisfactory or acceptable (59% for ACC and 70% for EC), while most fillets were unsatisfactory (76% ACC, 78% EC). Results show that the wild game meat supply chain can be a safe process when handling practices reported in European and National regulations are met.
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http://dx.doi.org/10.1016/j.meatsci.2019.107967DOI Listing
February 2020

A Probable Fatal Case of Oleander (Nerium oleander) Poisoning on a Cattle Farm: A New Method of Detection and Quantification of the Oleandrin Toxin in Rumen.

Toxins (Basel) 2019 07 25;11(8). Epub 2019 Jul 25.

Pavia Poison Control Centre-National Toxicology Information Centre, 27100 Pavia, Italy.

Oleander (Nerium oleander) is an ornamental plant common in tropical and sub-tropical regions that is becoming increasingly widespread, even in temperate regions. Oleander poisoning may occur in animals and humans. The main active components contained in the plant are cardiac glycosides belonging to the class of cardenolides that are toxic to many species, from human to insects. This work describes a case of oleander poisoning that occurred on a small cattle farm and resulted in the fatality of all six resident animals. Furthermore, the investigation of the poisonous agent is described, with particular focus on the characterization of the oleandrin toxin that was recovered from the forage and rumen contents. The innovation of this study is the first description of the detection and quantification of the oleandrin toxin by liquid chromatography-high resolution mass spectrometry (LC-HRMS) in rumen.
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http://dx.doi.org/10.3390/toxins11080442DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6723884PMC
July 2019

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) spa type t127, Sequence Type (ST)1, quickly spreads and persists among young pigs.

Pathog Dis 2019 04;77(3)

Istituto Zooprofilattico Sperimentale del Lazio e della Toscana "M. Aleandri", Department of General Diagnostics, National Reference Laboratory for Antimicrobial Resistance, Via Appia Nuova, 1411, 00178, Rome, Italy.

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) clones other than Clonal Complex (CC)398, as CC1, have been isolated in pigs in some countries, and appeared to be prevalent in Italy. The aim of this study was to evaluate the capability of Sequence Type (ST)1, CC1, LA-MRSA clone to colonize and to be transmitted among piglets. Eighteen caesarean-derived/colostrum-deprived piglets of 35 days of age were assigned randomly to three groups: four seeder piglets were contaminated with a spa type t127, ST1, SCCmec V, MRSA (Group A), 10 MRSA-negative piglets were exposed to Group A after 2 days post-contamination, dpc (Group B) and 4 piglets were used as control group (Group C). Piglets were evaluated until 44 dpc (Group A) or at 42 days post-exposure, dpe (Group B) and then euthanized and necropsied. All nasal and skin cultures of Group A resulted MRSA-positive throughout the experiment starting from two dpc, while Group C tested always MRSA-negative. At first sampling, all Group B piglets became positive and remained positive throughout the experiment. This is the first colonization/transmission study with a CC1 LA-MRSA in pigs. The results add further knowledge on the ability of CC1 LA-MRSA to colonize pigs, and on colonization/transmission patterns, both suggesting good host adaptation.
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http://dx.doi.org/10.1093/femspd/ftz033DOI Listing
April 2019

Reduction of Salmonella spp. populations in Italian salami during production process and high pressure processing treatment: Validation of processes to export to the U.S.

Meat Sci 2019 Nov 16;157:107869. Epub 2019 Jun 16.

Department of Veterinary Medical Sciences, University of Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia (BO), Italy.

This study involved ten enterprises producing Italian salami, 20 different samples of fermented sausages underwent challenge tests to assess and record the following parameters: time, temperature, pH, a, and Salmonella counts. A linear regression model was used to describe the Salmonella spp. decay: at the end of the process the result of total Salmonella reduction was 0.97-5.84 Log CFU/g and it was significantly associated with pH at the end of acidification/drying process, a at the end of seasoning period, the duration of seasoning, and the caliber of salami respectively. High Pressure Processing (HPP) further reduced the Salmonella level by 2.41-5.84 Log CFU/g with an efficacy that resulted inversely associated with a of salami at the end of seasoning; the objective of 5-Log reduction was always reached in all the cases tested by the production process plus HPP. This model could be a useful tool for enterprises and Authorities to evaluate the efficacy of the processes to reduce Salmonella load for exportation to the U.S.
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http://dx.doi.org/10.1016/j.meatsci.2019.06.005DOI Listing
November 2019

Antimicrobial resistance, biofilm synthesis and virulence genes in Salmonella isolated from pigs bred on intensive farms.

Ital J Food Saf 2018 07 3;7(2):7223. Epub 2018 Jul 3.

Department of Veterinary Sciences, University of Parma.

is the second cause of food-borne infection in humans in the USA and Europe. Pigs represent the second most important reservoir for the pathogen and the consumption of pork meat is a major risk factor for human salmonellosis. Here, we evaluated the virulence patterns of eleven isolated from pigs (carcasses and faces) bred in intensive farms in the north of Italy. The two serotypes identified were . Typhimurium and its monophasic variant 1,4,5,12:i:-. None of the isolates was an ESBL producer, as confirmed also by PCR. However, the presence of a multi-drug resistant pattern was evident, with all the isolates being resistant to at least to five antimicrobial agents belonging to various classes. Moreover, six out of eleven isolates showed important resistance profiles, such as resistance against colistin and ciprofloxacin, with nine to twelve recorded resistances. The isolates were negative for the biofilm synthesis test, while four different virulotypes were characterized. All the isolates showed the presence of A, A, , A, C. One sample also harbored R and C genes. One strain was positive for all the virulence genes tested and was resistant to 12 antimicrobial agents. The present study contributes new data to the surveillance program for antibiotic resistance. Furthermore, the presence of eleven highly virulent isolates poses concern for human health in relation to their diffusion in the environment.
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http://dx.doi.org/10.4081/ijfs.2018.7223DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6036996PMC
July 2018

Multilocus microsatellite typing (MLMT) reveals host-related population structure in Leishmania infantum from northeastern Italy.

PLoS Negl Trop Dis 2018 07 5;12(7):e0006595. Epub 2018 Jul 5.

Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia-Romagna, Brescia, Italy.

Background: Visceral leishmaniasis (VL) caused by Leishmania infantum is an ongoing health problem in southern Europe, where dogs are considered the main reservoirs of the disease. Current data point to a northward spread of VL and canine leishmaniasis (CanL) in Italy, with new foci in northern regions previously regarded as non-endemic.

Methodology/principal Findings: Multilocus microsatellite typing (MLMT) was performed to investigate genetic diversity and population structure of L. infantum on 55 samples from infected humans, dogs and sand flies of the E-R region between 2013 and 2017. E-R samples were compared with 10 L. infantum samples from VL cases in other Italian regions (extra E-R) and with 52 strains within the L. donovani complex. Data displayed significant microsatellite polymorphisms with low allelic heterozygosity. Forty-one unique and eight repeated MLMT profiles were recognized among the L. infantum samples from E-R, and ten unique MLMT profiles were assigned to the extra E-R samples. Bayesian analysis assigned E-R samples to two distinct populations, with further sub-structuring within each of them; all CanL samples belonged to one population, genetically related to Mediterranean MON-1 strains, while all but one VL cases as well as the isolate from the sand fly Phlebotomus perfiliewi fell under the second population. Conversely, VL samples from other Italian regions proved to be genetically similar to strains circulating in dogs.

Conclusions/significance: A peculiar epidemiological situation was observed in northeastern Italy, with the co-circulation of two distinct populations of L. infantum; one population mainly detected in dogs and the other population detected in humans and in a sand fly. While the classical cycle of CanL in Italy fits well into the data obtained for the first population, the population found in infected humans exhibits a different cycle, probably not involving a canine reservoir. This study can contribute to a better understanding of the population structure of L. infantum circulating in northeastern Italy, thus providing useful epidemiologic information for public health authorities.
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http://dx.doi.org/10.1371/journal.pntd.0006595DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6057669PMC
July 2018

Case report of a pustular dermatitis outbreak in sheep: Clinical and food safety considerations.

Ital J Food Saf 2018 Mar 11;7(1):6980. Epub 2018 Apr 11.

Department of Veterinary Medical Sciences, University of Bologna.

The objective of this report is to describe an outbreak of pustular dermatitis in a flock of about 200 sheep, its clinical evolution and food safety implications. The onset of the symptoms was sudden and the lesions spread very quickly from ewe to ewe, so that in about 3 days almost all of the lactating sheep were stricken. Pustules from 5 different animals, six milk samples, two cheese samples, teat cup samples from the milking machine and farmer's hands were analysed. A pure culture of , producing staphylococcal enterotoxin (SE) C, was isolated from pustules. Milk and cheese showed a contamination by coagulase positive staphylococci <15 and 30 colony forming units respectively and the absence of SE. Farmer's hands and teat cups samples resulted negative for coagulase positive staphylococci. Therapy with daily topical medicaments was prescribed and a prophylactic intervention was suggested by the administration of an autovaccine. The low level of milk and cheese contamination and the absence of SE in cheese supported the decision to not advise the farmer to recall cheese produced with milk from affected animals.
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http://dx.doi.org/10.4081/ijfs.2018.6980DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5913706PMC
March 2018

Shiga Toxin-Producing in Slaughtered Pigs and Pork Products.

Ital J Food Saf 2017 Apr 9;6(2):6584. Epub 2017 May 9.

Institute for Experimental Veterinary Medicine of Lombardy and Emilia Romagna, Bologna, Italy.

During the years 2015-2016, 83 faecal samples were collected at slaughter from pigs reared in farms located in Central-Northern Italy. During the years 2014-2016 a total of 562 pork products [465 not-readyto-eat (NRTE) and 97 ready-to-eat (RTE) products] were collected from retail outlets, large retailers and processing plants. The samples were analysed according to ISO TS 13136:2012. Out of 83 swine faecal samples, 77 (92.8%) resulted stx-positive by real time polymerase chain reaction (PCR), 5 and 1 Shiga toxin-producing (STEC) strains were isolated. Among the 465 NRTE samples, 65 (14.0%) resulted stx-positive by real time PCR and 7 STEC strains were isolated. The gene was detected more frequently than the gene both in faecal samples (90.4 vs 8.4%) and in NRTE pork products (13.3 vs 1.3%). All the RTE samples included in the analysis resulted negative. Among the samples resulted positive for and genes, serogroup-associated genes were detected at high frequency: O26 resulted the most frequent in faecal samples (81.3%) and O145 in pork products (88.1%). The O157 serogroup resulted positive in 83.3 and 78.1% of pork products and faecal samples, respectively. Despite the frequent detection by real time PCR of genes indicating the possible presence of STEC strains belonging to the six serogroups, the bacteriological step did not confirm the isolation of any such strains.
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http://dx.doi.org/10.4081/ijfs.2017.6584DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5505102PMC
April 2017

Distinct Leishmania infantum Strains Circulate in Humans and Dogs in the Emilia-Romagna Region, Northeastern Italy.

Vector Borne Zoonotic Dis 2017 06 16;17(6):409-415. Epub 2017 Mar 16.

1 Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia-Romagna "Bruno Ubertini", Brescia, Italy .

Human leishmaniasis is an emerging problem in Italy and is on the increase in the Emilia-Romagna region, northeastern part of the country. Nevertheless, studies dealing with the molecular characterization of Leishmania spp. circulating in these areas are limited. In the present work, we explored the genetic polymorphism of Leishmania isolates from 28 cases of canine leishmaniasis and three cases of human visceral leishmaniasis (VL), which occurred in 2013-2014 in the Emilia-Romagna region. The characterization was carried out in comparison with nine human isolates of Leishmania from other VL endemic Italian regions and two reference strains. Nucleic acid from 31 Leishmania-positive phlebotomine sandfly pools, sampled in 2012-2013 in the Emilia-Romagna region, were also evaluated. DNA amplification and sequencing of the ribosomal internal transcribed spacer-1 and of a repetitive nuclear region on chromosome 31 were carried out for genotyping. Two size polymorphic targets were also analyzed by PCR, the cpb E/F-gene and the k26-gene. Altogether, the analysis showed the circulation of different Leishmania infantum genotypes in the Emilia-Romagna region: two genotypes found in dogs from public kennels were similar to VL isolates from other Italian regions, whereas a third genotype was detected in VL cases of the Emilia-Romagna region and in all but one of the sandfly pools. The combined molecular tools applied in this study can constitute a helpful support for parasite tracking (e.g., in outbreak investigations) and for a better understanding of the epidemiological evolution of leishmaniasis in northeastern Italy.
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http://dx.doi.org/10.1089/vbz.2016.2052DOI Listing
June 2017

Study on Potential Growth and Toxin Production in Parma Ham.

Ital J Food Saf 2016 Apr 19;5(2):5564. Epub 2016 Apr 19.

Department of Public Health, Parma Local Health Unit , Parma, Italy.

The objective of this study was to investigate growth and toxin production in the industrially manufactured Italian Parma ham. The study focuses on the Parma ham production phase identified as maximum risk to proliferation, . the transition from cold phase (salting and resting) to a phase carried out at temperature between 15 and 23°C (drying). A preliminary test was carried out in order to verify the capability of 6 strains (1 type A, 4 type B, and 1 type E strains) to grow in conditions of temperature, pH and NaCl concentration comparable to those of the beginning stage of ham drying. Five strains grew at 20°C and pH 6, four strains produced toxin when inoculated at a concentration equal to 10 cfu/mL at NaCl concentration of 4%, while when the inoculum concentration was 10 cfu/mL, NaCl concentration of 3% resulted the toxin-genesis limiting factor. An experimental contamination with a mixture of the 5 strains selected by the preliminary test was performed on 9 thighs inoculated at the end of the resting phase. The study was designed to evaluate the potential growth and toxin production in extremely favourable conditions for the bacterium. Type B proteolytic toxin was produced after 14 days of incubation at 20°C in 2 thighs characterised by high weight, low number of days of resting and anomalous physiochemical characteristics [one for very low NaCl concentration (1.59%), the other for elevated pH (6.27) and both for high water activity values (>0.970)]. The results of this research confirm that the cold resting step is a critical phase in the production process of Parma ham for the investigated hazard. Based on the present study, the long resting phase adopted in the manufacturing of Parma ham is proven effective to prevent the growth of , an event which could not otherwise be excluded if the hams were processed under less stringent technological conditions.
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http://dx.doi.org/10.4081/ijfs.2016.5564DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5076734PMC
April 2016

Detection of Food Hazards in Foods: Comparison of Real Time Polymerase Chain Reaction and Cultural Methods.

Ital J Food Saf 2016 Jan 28;5(1):5641. Epub 2016 Jan 28.

Institute for Experimental Veterinary Medicine of Lombardy and Emilia Romagna , Brescia.

Foodstuffs should not contain microorganisms or their toxins or metabolites in quantities suggesting an unacceptable risk for human health. The detection of food hazards in foods is performed by several tests that produce results dependent on the analytical method used: an analytical reference method, defined as standard, is associated with each microbiological criterion laid down in Regulation 2073/2005/EC, but, analytical methods other than the reference ones, in particular more rapid methods, could be used. Combined screening methods performed by real time-polymerase chain reaction (RT-PCR) are currently validated as alternative methods according to the ISO 16140:2003 and certified by the n. However, the positive results obtained with these alternative methods, the investigated molecular relations that resulted positive have to be confirmed with cultural methods using the same enrichment media in which the molecular screening was performed. Since it is necessary to assess if these testing schemes provide equivalent guarantees of food safety, the aim of this retrospective study is to analyse the data collected, from 2012 to 2014 by Emilia Romagna Region in the field of (Food Regional Plan) during official controls monitoring food samples of animal and other than animal origin. Records performed by combined methods of molecular screening of spp., and thermophilic and cultural confirmation results were gathered together and the results were compared in order to assess the sensitivity of the methods. A total of 10,604 food samples were considered in this study: the comparison of the data revealed that the RT-PCR method detected , and thermophilic in 2.18, 3.85 and 3.73% of the samples, respectively, whereas by using cultural method these pathogens were isolated in 0.43, 1.57 and 1.57% of samples, respectively. In spite of the use of the same enrichment broth, the RT-PCR method disclosed a percentage of positive samples that was negative to cultural examination ranging between 20 and 43%, with a PCR/culture ratio between 2.37 to 5.00. In conclusion, the results of this study pose a doubt about the sensitivity of the official cultural methods regarding the isolation of the three investigated foodborne pathogens. Moreover this study may be a useful tool for veterinary authorities to assess appropriate sampling plans to control the risk relating to the consumption of contaminated foods.
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http://dx.doi.org/10.4081/ijfs.2016.5641DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5076710PMC
January 2016

Aflatoxin B1 Risk Management in Parmigiano Reggiano Dairy Cow Feed.

Ital J Food Saf 2016 Jan 18;5(1):5291. Epub 2016 Jan 18.

Department of Veterinary Sciences, Alma Mater Studiorum-University of Bologna , Ozzano Emilia (BO).

This study investigated aflatoxin B1 (AFB1) contamination in dairy cow feed and the risk management of AFB1 content in concentrates undertaken by feed industries in the area. Data on aflatoxin contamination risk management applied in 29 feed industries were collected and the AFB1 content of 70 feed samples was analysed. Data were collected within the framework of a quality control programme promoted by the Consortium in 2013 and 2014. Audit results showed that the control procedures to prevent AFB1 contamination mainly focused on maize and its by-products. AFB1 concentration resulted lower than 5 ppb [legal European Union (EU) limit] in all samples; in one out of 70 samples, AFB1 content was 3.8 ppb and in all the other samples it was lower than 3 ppb. Results showed that AFB1 risk management applied by Italian feed industries effectively monitors AFB1 levels in feed below the EU legal limit.
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http://dx.doi.org/10.4081/ijfs.2016.5291DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5076703PMC
January 2016

Prevalence of Salmonella strains in wild animals from a highly populated area of north-eastern Italy.

Ann Ist Super Sanita 2016 Apr-Jun;52(2):277-80

Dipartimento di Scienze Mediche, Sezione di Medicina di Sanità Pubblica, Università degli Studi di Ferrara, Ferrara, Italy.

Introduction: Salmonella is a ubiquitous pathogen that can infect host species, like wild birds, rodents, and/or arthropods, which may transmit infection to domestic animals and human population.

Aim: In order to assess the related risk, a cross-sectional study was performed on 1114 carcasses of wild animals from a north-eastern area of the Emilia-Romagna Region, Italy.

Materials And Methods: During post mortem examination, intestine samples were cultured. A statistical analysis demonstrated that there is no correlation between the presence of sub-clinically infected animals and greater human population density. In contrast, a significant correlation between the number of carcasses positive for Salmonella spp. and greater spatial density of pig, poultry, and cattle farms was observed (p < 0.01).

Results: The results of the present study show that wild animals with omnivorous feeding habits are particularly exposed to Salmonella colonization and, consequently, to spreading the organism. Regarding drug resistance, this study confirms the resistance to antimicrobials is increasing in commensal and environmental isolates.
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http://dx.doi.org/10.4415/ANN_16_02_21DOI Listing
April 2017

Shelf life of donkey milk subjected to different treatment and storage conditions.

J Dairy Sci 2016 Jun 16;99(6):4291-4299. Epub 2016 Mar 16.

University of Bologna, Department of Veterinary Medical Sciences, Via Tolara di Sopra 50-40064 Ozzano Emilia, Bologna, Italy.

The aim of this study was to investigate the effects of different treatment conditions on microbiological indicators of donkey milk hygiene and their evolution during shelf life at 4 and 12°C from 3 to 30d, simulating a farm-scale pasteurization and packing system. Four treatment conditions were tested: no treatment (raw milk), pasteurization (65°C × 30 min), high-pressure processing (HPP), and pasteurization plus HPP. The microbiological quality of the raw donkey milk investigated was not optimal; our results highlight the importance of raw milk management with the need for animal hygiene management and good dairy farming practices on donkey farms to improve handling procedures. The raw milk treated with HPP alone showed visible alterations with flocks, making the milk unfit for sale. The microbiological risk posed by consumption of raw donkey milk was significantly reduced by heat treatment but farm-scale packing systems cannot guarantee an extended shelf life. In contrast, the pasteurization plus HPP treatment was the most effective method to maintain microbiological milk quality. Microflora growth had little effect on pH in donkey milk: pH values were significantly different only between raw milk and pasteurized and pasteurized plus HPP milk stored at 12°C for 3d. Alkaline phosphatase activity and furosine could be used as indicators of proper pasteurization and thermal processing in donkey milk. Moreover, the presence and growth of Bacillus cereus in the case of thermal abuse hamper the wide-scale marketing of donkey milk due to the potential consequences for sensitive consumers and therefore further tests with time/temperature/high-pressure protocols associated with B. cereus are needed. Finally, our study shows that an HPP treatment of pasteurized milk after packing extends the shelf life of donkey milk and assures its microbial criteria up to 30d if properly stored at 4°C until opening; therefore, combined heat treatment and storage strategies are recommended to enhance the shelf life of donkey milk.
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http://dx.doi.org/10.3168/jds.2015-10741DOI Listing
June 2016

Human campylobacteriosis related to the consumption of raw milk sold by vending machines in Italy: Quantitative risk assessment based on official controls over four years.

Prev Vet Med 2015 Sep 24;121(1-2):151-8. Epub 2015 Jun 24.

University of Bologna, Department of Veterinary Medical Sciences, Via Tolara di Sopra, 50, Bologna, 40064 Ozzano Emilia, Italy. Electronic address:

A quantitative risk assessment (RA) model was developed to describe the risk of campylobacteriosis linked to consumption of raw milk sold in vending machines in Italy. Exposure assessment was based on the official microbiological records of raw milk samples from vending machines monitored by the regional Veterinary Authorities from 2008 to 2011, microbial growth during storage, destruction experiments, consumption frequency of raw milk, serving size, consumption preference and age of consumers. The differential risk considered milk handled under regulation conditions (4°C throughout all phases) and the worst time-temperature field handling conditions detected. Two separate RA models were developed, one for the consumption of boiled milk and the other for the consumption of raw milk, and two different dose-response (D-R) relationships were considered. The RA model predicted no human campylobacteriosis cases per year either in the best (4°C) storage conditions or in the case of thermal abuse in case of boiling raw milk, whereas in case of raw milk consumption the annual estimated campylobacteriosis cases depend on the dose-response relationships used in the model (D-R I or D-R II), the milk time-temperature storage conditions, consumer behaviour and age of consumers, namely young (with two cut-off values of ≤5 or ≤6 years old for the sensitive population) versus adult consumers. The annual estimated cases for young consumers using D-R II for the sensitive population (≤5 years old) ranged between 1013.7/100,000 population and 8110.3/100,000 population and for adult consumers using D-R I between 79.4/100,000 population and 333.1/100,000 population. Quantification of the risks associated with raw milk consumption is necessary from a public health perspective and the proposed RA model represents a useful and flexible tool to perform future RAs based on local consumer habits to support decision-making on safety policies. Further educational programmes for raw milk consumers or potential raw milk consumers are required to encourage consumers to boil milk to reduce the associated risk of illness.
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http://dx.doi.org/10.1016/j.prevetmed.2015.06.009DOI Listing
September 2015

Reduction of Contamination in Vacuum-Packaged Dry-Cured Italian Pork Products After High Hydrostatic Pressure Treatment.

Ital J Food Saf 2015 May 17;4(2):4515. Epub 2015 Jun 17.

Institute for Experimental Veterinary Medicine of Lombardy and Emilia Romagna , Brescia, Italy.

The present work aims to present the results of the application of a treatment with high hydrostatic pressure (HHP) on Italian fermented and dry-cured pork products. The products used in this study were portioned cured ham, portioned bacon and salami, vacuumpackaged and produced by a single processing company. Two studies were conducted on a single batch of the three products by means of an artificial contamination with as a surrogate of . In the first trial a superficial contamination was obtained by immersion for 3 min in the culture broth with a concentration of approximately 9 log cfu/mL. At the end of the inoculum step, the pieces were dred at room temperature and vacuum packaged. In the second trial 50 kg of minced pork meat were contaminated before production of salami. In both cases the inoculum contained 5 strains of . Subsequently, in both trials, 10 samples were randomly divided into two groups of 5 pieces each: i) TH group, samples treated with HHP; ii) group C, control samples, not subjected to any treatment. All samples were stored at refrigeration temperature at the end of HHP treatments (if applied), and analyzed for the determination of the surface (1st trial) and deep (2nd trial) quantitative contamination of . pH and a were also determined on 3 pieces of each products belonging to group C. The difference between the medians of the log cfu/cm2 or g established between controls and treated were compared using the non-parametric test (Kruskal-Wallis test) with P<0.01. In all products and in both trials the level of contamination detected in treatment groups was always significantly lower than in controls (P<0.01). In particular, in vacuum-packaged ham, bacon and salami viability logarithmic viability reductions equal to -2.29, -2.54 and -2.51 were observed, respectively. This study aimed to evaluate a not-thermal treatment on Italian cured or fermented pork products. The results of this study need to be confirmed in different products and in a greater number of lots, but they appear promising, also because of the considerable literature available for different categories of products (cheese, vegetables and fruit).
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http://dx.doi.org/10.4081/ijfs.2015.4515DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5076645PMC
May 2015

Gossypol Content of Cotton Free Commercial Feed for Dairy Cows.

Ital J Food Saf 2015 May 3;4(2):5174. Epub 2015 Jun 3.

Department of Veterinary Medical Sciences, Alma Mater Studiorum - University of Bologna, Ozzano dell'Emilia (BO) ; Italy.

Gossypol is a yellow pigment occurring in all parts of cotton plants, with the highest levels found in seeds, and it exhibits a variety of toxic effects. Few data are available on the content of gossypol in the commercial complementary feed and in feed raw materials. The present study was focused on the investigation of the presence of free gossypol in commercial complementary feed not containing cotton. A total of 50 samples of commercial complementary feed for dairy cows were performed in 29 feed mills both using and not using cotton as feed material. The free gossypol contamination resulted under the detection limit of the technique (4 mg/kg) in 12 out of 50 samples analysed and ranged from 4 to 20 mg/kg in 28 samples. In 10 samples the level of free gossypol ranged from 20 to 29.5 mg/kg. Average contamination of samples was 12.2±9.2 SD mg/kg. No significant difference (P=0.571) was shown in free gossypol concentration between feed produced in plants and in plants where cotton is used as feed material. Free gossypol content detected in the present study allows considering complementary feed for dairy cows not at risk. On the other hand, the detection of free gossypol in complementary feed, probably attributable to cross contamination of feed materials upstream of the feed mill, should be further investigated.
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http://dx.doi.org/10.4081/ijfs.2015.5174DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5076657PMC
May 2015
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