Dry Fermented Sausages with Total Replacement of Fat by Extra Virgin Olive Oil Emulsion and Indigenous Lactic Acid Bacteria.

RESEARCH BACKGROUND: Formulations based on vegetable or fish oil and modifications in the production technology of dry fermented sausages have emerged in recent years aiming to achieve the desirable target of reducing the fat content of these meat products. However, previous efforts have confronted many difficulties, such as high mass loss and unacceptable appearance due to intensely wrinkled surfaces and case hardening. The objective of this study is to produce and evaluate dry fermented sausages by utilising a meat protein-olive oil emulsion as fat substitute and indigenous lactic acid bacteria (LAB) with probiotic properties isolated from traditional Greek meat products. EXPERIMENTAL APPROACH: A novel formulation with extra virgin olive oil and turkey protein was developed to totally replace the conventionally added pork fat. Probiotic and safety characteristics of autochthonous LAB isolates from spontaneously fermented sausages were evaluated and three LAB isolates were finally selected as starter cultures. Physicochemical, microbiological and sensory analyses were carried out in all treatments (control, Lactobacillus acidophilus, L. casei, L. sakei and Pediococcus pentosaceus) during fermentation. RESULTS AND CONCLUSIONS: Ready-to-eat sausages were found to be microbiologically stable. The olive oil-based formulation produced in this study generated a mosaic pattern visible in the sliced product simulating the fat in conventional fermented sausages and was regarded as an ideal fat substitute for the production of fermented sausages. An autochthonous isolate of Lactobacillus casei adapted the best to the final products as it was molecularly identified to be present in the highest counts among the LAB isolates used as starter cultures. NOVELTY AND SCIENTIFIC CONTRIBUTION: Α novel and high-quality dry fermented meat product was produced by replacing the added pork fat with a fat substitute based on a meat protein-olive oil emulsion. Autochthonous LAB with in vitro probiotic properties could have a potential use in large-scale novel dry fermented sausage production. Such isolates could be used as starters in an effort to standardise the production process and retain the typical organoleptic and sensory characteristics. Moreover, isolates like L. casei 62 that survived in high counts in the final products can increase the safety of fermented sausages by competing not only with pathogens but also with the indigenous microbiota and could have a potential functional value for the consumer.

Diversity of Campylobacter in retail meat and liver of lambs and goat kids.

The presence, genetic diversity, and antimicrobial susceptibility profile of Campylobacter spp. in retail lamb and goat kid carcasses were assessed. A total of 200 samples consisting of 100 meat and 100 liver surface swabs were collected from 47 lamb and 53 goat kid carcasses at 23 retail markets in Northern Greece, and 125 Campylobacter isolates were recovered from 32 meat surfaces (32%) and 44 liver surfaces (44%). Multiplex polymerase chain reaction and restriction fragment length polymorphism analysis specified Campylobacter coli as the most frequently detected species (59.2%) followed by C. jejuni (40.8%). Pulsed-field gel electrophoresis (PFGE) was applied in order to typify a subset of randomly selected isolates (n=80). SmaI-PFGE successfully clustered the 80 isolates in 38 SmaI-PFGE types, indicating high heterogeneity among the analyzed Campylobacter isolates, and provided data regarding the dissemination of Camplobacter among carcasses stored in the same retail market. Antimicrobial susceptibility profiles of Campylobacter isolates, assessed by the disk-diffusion method, indicated that 31 isolates (24.8%) were multidrug resistant, and the most common profile was the concurrent resistance to tetracycline and streptomycin. Overall, 56.8% of isolates (n=71, multidrug-resistant isolates included) exhibited resistance to at least one antimicrobial (tetracycline 34.4%, quinolones 27.2%, and streptomycin 20.8%). However, all isolates were susceptible to erythromycin and gentamicin. The findings of this study verify the contamination of retail lamb and goat kid carcasses with a heterogeneous population of thermotolerant campylobacters. These data underscore the fact that retail meat and liver of small ruminants could serve as vehicles for consumer contamination with Campylobacter and that further investigation is necessary in order to evaluate the risk imposed by such products within the epidemiology of human campylobacteriosis cases.

Characterization of Extended Spectrum Cephalosporin-Resistant Escherichia coli Strains Isolated from Raw Poultry Carcasses in Catering Services in Northern Greece.

Antimicrobial resistance is considered a topic of utmost interest under the concept of "One Health", having severe implications in both human and veterinary medicine. Among the antibiotic-resistant bacteria, gram-negative bacteria, especially those belonging to the order of Enterobacterales (such as Escherichia coli), hold a prominent position in terms of both virulence and possessing/disseminating antimicrobial resistance (AMR) traits. The aim of this study was to examine the presence of extended-spectrum ß-lactamase producing E. coli isolates in raw poultry carcasses collected from a university club. Five hundred raw poultry skin samples were collected from the Aristotle University of Thessaloniki (AUTh) club in Thessaloniki, Greece. A total of 64% of the samples were positive for the presence of extended-spectrum ß-lactamase (ESBL)-producing E. coli. The isolates were further examined for their susceptibility to selected antibiotics by the disc diffusion method and were characterized as true ESBL, as producing class C cephalosporinases (AmpC) or "of unknown etiology" by the combination disc test. The 86 of the 120 isolates (71.67%) were classified as true ESBL, 24 (20.00%) as AmpC, and 10 (8.33%) as "of unknown etiology". The isolates were screened for the occurrence of ß-lactamase genes (blaTEM, blaCTX-M, blaSHV, and blaOXA). Thirty-six isolates (32 ESBL- and 4 AmpC-phenotype) harbored both blaTEM and blaCTX-M genes, twenty-two isolates (among which 19 ESBL-phenotype and 2 AmpC-phenotype) harbored blaCTX-M only, whereas twenty-six (14 ESBL- and 12 AmpC-phenotype) isolates harbored blaTEM alone. No isolate harboring blaSHV or blaOXA was detected. The results demonstrate the existence of E. coli isolates producing extended-spectrum ß-lactamases in poultry carcasses from Greece, pausing a risk for antibiotic resistance transfer to humans.

The influence of mannan oligosaccharides, acidifiers and their combination on caecal microflora of Japanese quail (Coturnix japonica).

The aim of this study was to investigate the effects of the dietary supplementation of mannan oligosaccharides (MOS) extracted from yeast Saccharomyces cerevisiae, acidifiers-calcium formate (CF), calcium propionate (CP)- and their combination on the caecal microflora of Japanese quail (Coturnix japonica). Four hundred and fifty 1-day old quail where divided in six groups with three replicates each. One group that served as control received the basal diet. The five experimental diets consisted of the basal diet to which either 1 g MOS/kg, or 6 g CF/kg, or 6 g CP/kg, or 1 g MOS plus 6 g CF/kg or 1 g MOS plus 6 g CP/kg were added. The body weight was examined at weekly intervals and mortality was recorded daily. At days 21 and 42 of age, the total count of aerobic bacteria, lactic acid bacteria, enterobacteriaceae and coliforms in the caecal content of one bird of each replicate was determined. Also, at day 42 of age, two birds of each replicate were slaughtered and their carcass weight was determined. The results showed that MOS significantly (P ≤ 0.050) increased the total aerobic plate and lactic acid bacteria counts on day 21. Furthermore, CP significantly (P ≤ 0.050) decreased the total aerobic plate and lactic acid bacteria counts compared to controls on day 21. Significant interaction between MOS and acidifiers was noticed on total aerobic plate count on day 21. No significant (P > 0.050) difference was found in the caecal microflora on day 42. Finally, no significant (P > 0.050) difference was noticed on mortality, body and carcass weight.

Campylobacter in small ruminants at slaughter: prevalence, pulsotypes and antibiotic resistance.

The present study aimed to address the prevalence, pulsotypes, and antimicrobial susceptibility patterns of Campylobacter species present in sheep and goat carcasses at slaughter. In total, 851 samples were collected (343 meat surfaces, 282 ileum contents, 226 liver surfaces) and 835 Campylobacter isolates were detected in 274 out of 343 carcasses (116 kids, 110 lambs, 63 goats and 54 sheep). The contamination rates per carcass category were 78.4% for kids, 94.5% for lambs, 63.5% for goats, and 72.2% for sheep. On average, 30% of the intestinal content samples and more than 70% of carcass and liver surfaces yielded the presence of campylobacters. Multiplex-PCR and RFLP analysis identified Campylobacter coli as the most prevalent species (76.2%) followed by Campylobacter jejuni (21.4%), albeit 2.4% of selected colonies yielded the concurrent presence of both these species. Macrorestriction profiling by pulsed-field gel electrophoresis (PFGE) was applied in order to characterise a subset of isolates. SmaI-PFGE successfully clustered 222 isolates in 82 SmaI-PFGE types indicating high heterogeneity among the campylobacter isolates (67 types among 174C. coli isolates and 15 types among 48C. jejuni isolates). No carcass-type (lamb, kid, sheep, and goat) specific PFGE clusters were recognised since there was a general overlapping of PFGE patterns regarding ovine and caprine isolates. Multiple pulsotypes were simultaneously present on single carcasses in the majority of tested animals. PFGE provided data regarding the potential routes of meat and liver contamination such as spillage of faecal material and cross-contamination during slaughter. Antimicrobial susceptibility patterns of Campylobacter isolates (n=240), determined by disk diffusion method, revealed resistance to tetracycline (47.9%) followed by streptomycin (22.9%) and ciprofloxacin along with nalidixic acid (18.3%). Isolates exhibited low resistance to erythromycin (2.5%) and were susceptible to gentamicin. The findings of the present study confirm the contamination of sheep and goats at slaughter with thermophilic campylobacters and underline their potential input in the epidemiology of human campylobacteriosis.

Effect of chitosan treatments on quality parameters of fresh refrigerated swordfish (Xiphias gladius) steaks stored in air and under vacuum conditions.

The present study examined the effect of chitosan (1.0% w/v) in combination with packaging on the shelf-life of fresh swordfish steaks. Treatments included the following: A (untreated, control samples stored in air), A-CH (treated with chitosan 1.0% w/v, stored in air), VP (untreated, stored under vacuum packaging) and VP-CH (treated with chitosan, stored under vacuum packaging). VP-CH significantly affected Total Viable Counts (TVC), Pseudomonas spp., H(2)S-producing bacteria (including Shewanella putrefaciens), lactic acid bacteria and Enterobacteriaceae. Production of TMA-N and TVB-N for A-CH, VP and VP-CH swordfish samples was significantly lower than for control (A) samples at day 8 of storage. Histamine formation for all treatments was low. A-CH and VP-CH resulted in significantly lower levels of putrescine, cadaverine and tyramine (day 8) as compared to A and VP swordfish samples. Results of this study indicate that the shelf-life of swordfish steaks can be extended using, either aerobic or vacuum packaging and in combination with chitosan, by approximately 4 (A-CH), 8 (VP) and 12 (VP-CH) days. Swordfish steaks treated with chitosan and stored under VP were sensorially acceptable up to 17days. The presence of chitosan (A-CH and VP-CH) did not negatively influence the taste of cooked swordfish.