Bovine lymph nodes as a source of Escherichia coli contamination of the meat.

Ground beef contamination with Escherichia coli is usually a result of carcass faecal contamination during the slaughter process. Carcasses are contaminated when they come into contact with soiled hides or intestinal leakage content during dressing and the evisceration processes. A more recent and compelling hypothesis is that, when lymph nodes are present in manufacturing beef trimmings, they can be a potential source of Enterobacteriaceae contamination of ground beef. The aim of this study was to investigate the occurrence of E. coli in lymph nodes from beef carcasses used for ground meat production, in six slaughter plants situated in central Italy A total of 597 subiliac (precrural) lymph nodes were obtained from 597 cattle carcasses and screened for E. coli by culture. Furthermore, E. coli isolates (one per positive carcass) were tested for stx1, stx2 eaeA and hlyA genes that are commonly used to identify and characterise shiga toxin-producing E. coli (STEC). In addition, the E. coli isolates were profiled for antimicrobial susceptibility. A proportion of 34.2% (204/597) carcasses were positive for E. coli. PCR revealed that 29% (59/204) of E. coli possessed stx1 or stx2 which corresponded to 9.9% of the cattle sampled. Moreover, a combination of stx1 or stx2 and eaeA was found in in 4 isolates (2% among E. coli positive samples and 1% among cattle sampled) and a combination of stx1 or stx2 and eaeA and hly in 1 isolate (0.5% and 0.2%). More than 95% of isolates were susceptible to gentamicin, ceftriaxone, cyprofloxacin and cefotaxime while high rates of resistance were recorded for cephalotin, ampicillin, tetracycline, tripe sulfa and streptomycin. The multivariate analysis identified "age" as the factor most closely related to E. coli positivity (either generic E. coli or STEC) in bovine lymph nodes. In conclusion, subiliac lymph nodes represent a source of E. coli for ground beef. These results are of major importance for risk assessment and improving good manufacturing practices during animal slaughter and ground meat production.

Characterization and Growth under Different Storage Temperatures of Ropy Slime-Producing Leuconostoc mesenteroides Isolated from Cooked Meat Products.

ABSTRACT: The presence of lactic acid bacteria can be detrimental when the abundant growth of slime-producing strains (Lactobacillus spp. and Leuconostoc spp.) causes spoilage of meat products. Two strains of lactic acid bacteria were isolated from vacuum-packed cooked hams that had been withdrawn from the market for the so-called ropy slime defect and identified as Leuconostoc mesenteroides. In an attempt to define the behavior of ropy slime-producing bacteria, two strains of L. mesenteroides were incubated in de Man Rogosa Sharpe broth at different storage temperatures and conditions of thermal abuse (4, 12, 20, 30, 37, and 44°C). Both strains showed a lack of growth at 44°C, a good level of development at 30 and 37°C, and evident growth ability at low temperatures, with a long stationary phase. In particular, the bacterial concentration at 4°C was >105 CFU mL-1 after more than 120 days of incubation. This study demonstrates that the refrigeration temperature for cooked meat products does not constitute a hurdle for ropy slime producers and their subsequent ability to spoil.

Cross-sectional study to identify risk factors associated with the occurrence of antimicrobial resistance genes in honey bees Apis mellifera) in Umbria, Central Italy.

The use antimicrobials for therapeutic and metaphylactic purpose in humans and agriculture exerts selective pressure on animal and environmental microbiota resulting in the survival and spread of antimicrobial resistance genes among bacteria and subsequent development of resistance in bacteria. Previous studies have shown that honey bees' microbiota (Apis mellifera) can accumulate antimicrobial resistance genes in their microbiome and act as collectors and disseminators of resistance genes. The aim of this study was to investigate to what extent honey bees act as reservoir of select antimicrobial resistance genes. This study was conducted on 35 groups of bees. Bees were collected from 35 sites in Umbria, Italy. PCR was used to screen pooled ground bees' specimens for genes that code for resistance against antimicrobials that are commonly used in humans and in veterinary medicine including aminoglycosides (aph), beta-lactams (blaZ), tetracycline (tetM) and sulphonamides (sul1 and sul2). Twenty-four samples out of 35 (68.57%) were positive for at least one antimicrobial resistance gene. Two samples were positive for the aph, 5.71%; eight for blaZ, 22.86%; three for tetM, 8.57%; ten for sul1, 28.57% and eighteen for sul2, 51.43%. Positivity to more than one antimicrobial resistance gene was observed in nine samples, 25.71%. The multivariate analysis identified "presence of farms nearby" as the factor most closely related to PCR positivity. Honey bees (Apis mellifera) from Umbria, Italy, carry antimicrobial resistance genes and can be used as indicators of the presence of resistance genes in the environment.

Study on the Growth and Enterotoxin Production by Staphylococcus aureus in Canned Meat before Retorting.

Possible contamination by Staphylococcus aureus of the production environment and of the meat of a canned meat production factory was analysed. A total of 108 samples were taken from nine critical control points, 13 of them were positive for S. aureus. None of the isolates produced enterotoxins. To determine how much time can elapse between can seaming and sterilisation in the autoclave without any risk of enterotoxin production by S. aureus, the growth and enterotoxin production of three enterotoxin A producing strains of S. aureus (one ATCC strain and two field strains) in canned meat before sterilisation was investigated at three different temperatures (37, 20 and 10 °C). Two types of meat were used, one with and one without sodium nitrite. In the canned products, the spiked bacteria spread throughout the meat and reached high levels. Enterotoxin production was shown to start 10 hours after incubation at 37 °C and after 48 h after incubation at 20 °C; the production of enterotoxin was always detected in the transition between the exponential and the stationary growth phase. At 10 °C, the enterotoxin was never detected. The statistical analysis of the data showed that the difference between the two different types of meat was not statistically significant (p value > 0.05). Since it is well known that following heat treatment, staphylococcal enterotoxins, although still active (in in vivo assays), can be undetectable (loss of serological recognition) depending on the food matrix and pH, it is quite difficult to foresee the impact of heat treatment on enterotoxin activity. Therefore, although the bacteria are eliminated, the toxins may remain and cause food poisoning. The significance of the results of this study towards implementing good manufacturing practices and hazard analysis critical control points in a canned meat factory are discussed with reference to the management of pre-retorting steps after seaming.

Short communication: Characterization of enterotoxin-producing Staphylococcus aureus isolated from mastitic cows.

Staphylococcus aureus is not only a common cause of bovine mastitis, but also an agent of food poisoning in humans. In an attempt to determine whether staphylococci causing bovine mastitis could also cause food poisoning, 60 isolates of presumed S. aureus were isolated in the period between March and August 2017 from 3,384 routine, composite, quarter milk samples of individual cows raised on 12 dairy farms in central Italy. Seventeen out of 60 isolates were confirmed as S. aureus after coagulase, thermonuclease, and biochemical tests. These isolates were analyzed by PCR for the presence of the nuc, sea, seb, sec, sed, and see genes. The positive isolates were nuc, 100% (17); sea, 35.29% (6); seb, 5.88% (1); sec, 5.88% (1); sed, 29.41% (5); and see, 47.06% (8). The isolates were also tested with 2 enzyme immunoassay diagnostic kits, one for the screening detection of the production of staphylococcal enterotoxins (SEA, SEB, SEC, SED, SEE) and one for the detection of specific enterotoxin produced by each isolate. Seven out of 17 (41.18%) were enterotoxin producers: 7 produced SEA (41.18%), 1 SEB (5.88%), 1 SEC (5.88%), 5 SED (29.41%), and 6 SEE (35.29%). To further characterize the isolates, they were analyzed by the Kirby Bauer test for susceptibility to 13 antimicrobials (ampicillin, ciprofloxacin, kanamycin, tetracycline, gentamicin, methicillin, nalidixic acid, erythromycin, amoxicillin/clavulanic acid, streptomycin, vancomycin, neomycin, and enrofloxacin), and we detected resistance to ampicillin (52.94%), nalidixic acid (70.59%), erythromycin (5.88%), and amoxicillin/clavulanic acid (17.65%). The isolates were sensitive to the main classes of antimicrobials used for the treatment of bovine subclinical mastitis. The presence of enterotoxin-producing isolates of S. aureus in bovine milk means that a temperature abuse or a breakdown in the thermal treatment of the milk could present a food safety risk, particularly if all enterotoxigenic isolates could potentially produce SEA in milk.

Noise assessment in slaughterhouses by means of a smartphone app.

Regulation (EC) No. 1099/2009 on the protection of animals at the time of killing establishes an obligation to spare animals any avoidable suffering or stress prior to their slaughter. Although it has been pointed out that stressors also include noise, which can cause suffering and affect the quality of the meat, current legislation does not set a limit for environmental noise in slaughterhouses. This study was conducted in three slaughterhouses in central Italy to assess the environmental acoustic level using a smartphone app. The selected, medium-sized slaughterhouses for pigs and cattle were subjected to measurements using a sound-level meter (Noise Meter for iOS) during working hours at the unloading area and lairage, along the chute to the restraining pen, at the time of stunning and at the slaughter hall. For the bovine lines the average values expressed in dB ranged from 76.33 (SD 2.08) to 93.00 (SD 2.14) for abattoir 1, from 75.00 (SD 1.87) to 92.33 (SD 4.89) for abattoir 2 and from 75.67 (SD 7.09) to 88.83 (SD 4.79) for abattoir 3. For the pig lines the average values expressed in dB ranged from 77.50 (SD 3.11) to 100.33 (SD 1.53) for abattoir 1, from 83.00 (SD 2.00) to 99.75 (SD 2.63) for abattoir 2 and from 71.20 (SD 6.49) to 99.50 (SD 1.31) for abattoir 3. Data show that the pig slaughter line was always noisier than the cattle line and the slaughter hall always showed the highest values (i.e. 100 dB), when compared to the unloading area (i.e. 79 dB).

Analysis of the causes of the seizure and destruction of carcasses and organs in a slaughterhouse in central Italy in the 2010-2016 period.

Meat safety and hygiene are fundamental issues for producers and endusers. To achieve these goals, the inspection of carcasses and organs at slaughter is critical. The results of post-mortem inspection are the basis for planning preventive measures against consumer risks and for limiting economic losses. In this retrospective study, the causes of the seizure and destruction of carcasses and organs were analysed at a slaughterhouse in central Italy from 2010 to 2016. 436,646 slaughtered animals were taken into consideration, for a total of 61,799 seizures (73.29% pigs, 23.87% cattle, 2.77% sheep and 0.07% horses). The organs or groups of organs that most frequently showed lesions in pigs were liver (72.38%), heart (10.77%) and pluck (10.20%); in cattle, lungs (64.86%), liver (31.20%) and stomachs (11.63%); in sheep, liver (77.15%), pluck (18.70%) and lung (3.80%); in horses, liver (75.56%), kidney (68.89%) and lung (31.11%). Among the diagnoses, parasitic diseases of the liver (ascariasis and distomatosis) were especially frequent in all species, followed by pericarditis and polyserositis in pigs and diseases affecting the respiratory system in cattle. The data obtained show that postmortem inspection is of fundamental importance for limiting risks for the consumer and ensuring meat safety. It is also evident, even more than ten years after the coming into force of the so-called hygiene package regulations, that the slaughterhouse can still act as an epidemiological observatory to provide the data needed for the development of plans for the control and eradication of the most frequent diseases in the area.

Prevalence and characterisation of shigatoxigenic Escherichia coli isolated from beef cattle fed with prebiotics.

Ten Holstein Friesian calves were divided into two groups of five: one group was given prebiotics in their food, while the other group served as the control group. Every two weeks from birth up to 18 months, samples of feces were taken from the rectal ampulla to determine the concentration of E. coli. At each sampling session, three aliquots per sample were collected. The arithmetic mean was calculated and all values (converted into logs) were analysed with GraphPad InStat for analysis of variance, followed by the Tukey-Kramer test. A total of 69 E. coli strains were detected, 29 (42.03%) from treated animals and 40 (57.97%) from the control group. The isolates were analysed by PCR for the presence of the stx-1, stx-2, hly and eae genes and by the Kirby Bauer test for susceptibility to the most commonly used antimicrobials in cattle breeding. Hierarchical clustering of the isolates was done using Ward's method. Thirty samples were positive for the stx-1 gene, 18 for stx- 2, 12 for both stx-1 and stx-2, 8 for hly, and 10 for eae. 4.3% were resistant to sulfamides, 8.6% to tetracycline, 1.4% to gentamicin, 94.6% to cephalothin, 2.8% to chloramphenicol, 13% to ampicillin, 13% to amoxicillin/clavulanic acid, 7.2% to sulphonamides, 4.3% to ceftriaxone, 5.7% to nalidixic acid, 34.7% to ticarcillin, 88.5% to erythromycin, and 5.7% to streptomycin. The isolates from the samples taken from day 210 to day 300 were grouped into a single cluster. Bacteriological examinations showed a reduction in the concentration of E. coli in the feces of the treated animals compared to the control group. The presence of strains with shigatoxigenic Escherichia coli virulence profiles and the reduction of these in the treated animal group demonstrated that diet can play an important role in reducing E. coli prevalence in cattle.

Animal Welfare: Data from an Online Consultation.

This paper analyses data obtained from an online survey related to animal welfare and religious slaughter topics. The questionnaire was conducted with the purpose of examining the purchase behaviour of a group of consumers (with different religious orientation) and their views on animal protection and ritual slaughter. The main results of the consultation were two. The first evidenced the respondents' great interest about the question on animal welfare, which is in accordance with the growing interest of European citizens concerning this issue. The second was the demand for a more transparent labelling of animal products, which would also reflect animal welfare and the slaughter method used. These results are in contrast with marketing analysis, which finds that consumers want to only receive positive information. Paradoxically, the more information is transmitted to reassure consumers, the higher is the risk to alarm them.

Seroprevalence and risk factors for Toxoplasma gondii in sheep in Grosseto district, Tuscany, Italy.

BACKGROUND: Serum samples from 630 milk sheep, in 33 dairy flocks representative of the southern area of the Tuscany region, were tested for the presence of antibodies to Toxoplasma gondii using an indirect immunofluorescence antibody test (IFAT). Questionnaires exploring the management system were completed by the veterinarian in charge of the flocks. RESULTS: At least one seropositive animal was found in 32 of the 33 flocks tested (97.0%; 95% CI: 84.2%, 99.9%). In the positive flocks, median seroprevalence was 29.4% (interquartile range: 15.9%-46.1%). Overall animal-level seroprevalence, adjusted for sampling weights and test sensitivity and specificity, was 33.3% (95% CI: 24.8%, 42.7%). In a multivariable negative binomial regression model the number of seropositive animals in a flock decreased with increasing flock size (for >400 vs. <300 animals: count ratio (CR) = 0.62; 95% CI: 0.41, 0.95; P = 0.028) and was greater on farms where stray cats had access to animals' water (CR = 1.54; 95% CI: 1.05, 2.26; P = 0.027). CONCLUSIONS: Small flock size and access of cats to water are potential risk factors for Toxoplasma infection in sheep in the Grosseto district in Tuscany, Italy. Sheep could be an important source of T. gondii infection in humans, since we estimate that between 25% and 43% of sheep in the district were seropositive. Toxoplasmosis is also likely to be an important cause of abortion in sheep in the district. Control and prophylactic measures must be adopted to improve the rearing system and the implementation of health promoting programmes in a joint effort between sheep farmers, farmers' associations and veterinarians to inform about the means of transmission of the infection and for a better understanding of the disease.

Toxoplasma in animals, food, and humans: an old parasite of new concern.

All hosts, including humans, can be infected by any one of the three forms of the parasite Toxoplasma gondii that correspond to three morphological stages: tachyzoite, bradyzoite, and sporozoite form. Felids are definitive hosts for T. gondii, which is an intracellular pathogen that infects a wide range of warm-blooded intermediate hosts. Toxoplasmosis is a disease where the interest of the diverse medical and veterinary specialties converge. Awareness needs to be increased that toxoplasmosis can induce clinical disease not only in immunocompromised patients or through congenital infections, but also in healthy patients. This is a review article that aims at illustrating why toxoplasmosis should be regarded a veterinary public health issue and how veterinary practitioners can contribute in controlling the infection.