Safety, beneficial and technological properties of enterococci for use in functional food applications - a review.

Enterococci are ubiquitous lactic acid bacteria (LAB) that predominantly reside in the gastrointestinal tract of humans and animals but are also widespread in food and the environment due to their robust nature. Enterococci have the paradoxical position of providing several benefits of technological interest in food fermentations but are also considered as opportunistic pathogens capable of causing infection in immunocompromised patients. Several species of the genus have been correlated with disease development in humans such as bacteremia, urinary tract infections, and endocarditis. The pathogenesis of enterococci has been attributed to the increasing incidence of antibiotic resistance and the possession of virulence determinants. On the contrary, enterococci have led to improvements in the aroma, texture, and flavor of fermented dairy products, while their beneficial use as probiotic and protective cultures has also been documented. Furthermore, they have emerged as important candidates for the generation of bioactive peptides, particularly from milk, which provide new opportunities for the development of functional foods and nutraceuticals for human nutrition and health. The detection of pathogenic traits among some species is compromising their use in food applications and subsequently, the genus neither has Generally Regarded as Safe (GRAS) status nor has it been included in the Qualified Presumption of Safety (QPS) list. Nevertheless, the use of certain enterococcal strains in food has been permitted on the basis of a case-by-case assessment. Promisingly, enterococcal virulence factors appear strain specific and food isolates harbor fewer determinants than clinical isolates, while they also remain largely susceptible to clinically relevant antibiotics and thus, have a lower potential for pathogenicity. Ideally, strains considered for use in foods should not possess any virulence determinants and should be susceptible to clinically relevant antibiotics. Implementation of an appropriate risk/benefit analysis, establishment of a strain's innocuity, and consideration for relevant guidelines, legislation, and regulatory aspects surrounding functional food development, may help industry, health-staff and consumers accept enterococci, like other LAB, as important candidates for useful and beneficial applications in food biotechnology.

A 3-dimensional model for teaching local flaps using porcine skin.

The European Working Time Directive and streamlined training has led to reduced training time. Surgery, as an experience-dependent craft specialty is affected more than other medical specialties. Trainees want to maximize all training opportunities in the clinical setting, and having predeveloped basic skills acquired on a simulated model can facilitate this.Here we describe the use of a novel model to design and raise local flaps in the face and scalp regions. The model consists of mannequin heads draped with porcine skin which is skewered with pins at strategic points to give a 3-dimensional model which closely resembles a cadaveric head.The advantages of this model are that it is life size and incorporates all the relevant anatomical features, which can be drawn on if required.This model was used on a recent course, Intermediate Skills in Plastic Surgery: Flaps Around the Face, at the Royal College of Surgeons England. The trainees found that practicing on the porcine skin gave them an opportunity to master the basics of flap design and implementation.In summary, this innovative 3-dimensional training model has received high levels of satisfaction and is currently as close as we can get to cadaveric dissection without the constraints and cost of using human tissue.

Three new species of the barracudina genus Lestidium (Aulopiformes: Paralepididae) from the Indo-West Pacific.

Three new species of the genus Lestidium with complete lateral line are described from the Indo-west Pacific Ocean. Lestidium longilucifer sp. nov., from Western Australia and Taiwan, belongs to the Lestidium atlanticum species complex and can be separated from other congeners by having 41-43 prehaemal vertebrae, 85-88 total vertebrae and 126-146 total lateral-line scales; and body proportions. Lestidium australis sp. nov. from eastern Australia and Lestidium rofeni sp. nov. from Taiwan and the Philippines together with Lestidium prolixum form the L. prolixum species complex. These three species can be separated from each other by a combination of different fin positions, vertebral formula, number of lateral-line scales and pigmentation.

Review of the Stemonosudis rothschildi species complex, with descriptions of two new species from the Indo-west Pacific Ocean (Aulopiformes: Paralepididae).

Three barracudina species are recognized in the Stemonosudis rothschildi species complex, which includes Stemonosudis rothschildi Richards, 1967, and two new species described herein. Stemonosudis multifasciatus sp. nov. is described based on five specimens collected off northwestern Australia and Myanmar, Eastern Indian Ocean. It is characterized by having 16 brownish blotches on dorsum (10 before dorsal-fin origin); 49-51 caudal vertebrae; 93‒95 total vertebrae; dorsal-fin origin relatively forward in position, distance between origins of pelvic and dorsal fins 52.3‒63.0% of distance between origins of pelvic and anal fins; and combination of body proportions. Stemonosudis retrodorsalis sp. nov. is described based on 15 specimens collected from off the Philippines, Indonesia and northwestern Australia. It is characterized by having dorsal-fin origin at about vertical through anal-fin origin, insertion of anal fin relatively forward, preanal length 71.5‒79.5% SL; 6 blotches on dorsum before DFO and 4 on abdominal ridge before VFO and unique combination of body proportions. A redescription of S. rothschildi, based on specimens collected from off Dongsha (Pratas) Islands, Australia and West Indies, is also included.

Development of a rapid, one-step screening method for the isolation of presumptive proteolytic enterococci.

Enterococci show higher proteolytic activities than other lactic acid bacteria and thus have received considerable attention in scientific literature in recent years. Proteolytic enzymes of enterococci have warranted the use of some species as starter, adjuncts or protective cultures and as probiotics, while in some strains they have also been linked with virulence. Consequently, the isolation and identification of proteolytic enterococci is becoming of increasing interest and importance. However, current screening methods for proteolytic enterococci can be time consuming, requiring a two-step procedure which may take up to 96h. This study describes a method, utilising Kanamycin Skim Milk Aesculin Azide (KSMEA) agar, for the isolation of proteolytic enterococci in one-step, thereby significantly reducing screening time. KSMEA combines the selective properties of Kanamycin Aesculin Azide Agar (KAA) with skim milk powder for the detection of proteolytic enterococci. Enterococci produced colonies with a black halo on KSMEA which were accompanied by a zone of clearing in the media when enterococci were proteolytic. KSMEA medium retained the selectivity of KAA, while proteolytic enterococci were easily distinguished from non-proteolytic enterococci when two known strains were propagated on KSMEA. KSMEA also proved effective at isolating and detecting enterococci in raw milk, faeces and soil. Isolates recovered from the screen were confirmed as enterococci using genus-specific primers. Proteolytic enterococci were present in the raw milk sample only and were easily distinguishable from non-proteolytic enterococci and other microorganisms. Therefore, KSMEA provides a rapid, one-step screening method for the isolation of presumptive proteolytic enterococci.

Therapeutic applications of the larvae for wound debridement.

It has been known for centuries that application of larvae is useful to heal certain wounds by facilitating debridement of necrotic tissue. Their therapeutic use was popularised in the beginning of the 19th century, but waned in the 1940s with the advent of antiseptic wound management and antibiotics. In more recent years, larvae are once again in vogue for management of difficult wounds. The mechanism of wound debridement by larvae includes the complete wound by continuous larval motion, secretion of proteolytic enzymes and antibacterial substances, effects on epidermal growth factor and interleukin-6 (IL-6) and ingestion and digestion of bacteria and necrotic tissue. In our study, wound debridement was achieved satisfactorily in 29 of 34 patients (85%) with chronic wounds. In the remaining five patients, failures occurred due to inadequate sealing in two patients (6%), death of larvae in two patients (6%) and treatment intolerance in one patient (3%). Larval therapy should be considered as a therapeutic option in the management of certain difficult wounds.

Cytochrome P450 inhibition using recombinant proteins and mass spectrometry/multiple reaction monitoring technology in a cassette incubation.

Detailed cytochrome P450 (P450) inhibition profiles are now required for the registration of novel molecular entities. This method uses combined substrates (phenacetin, diclofenac, S-mephenytoin, bufuralol, and midazolam) with combined recombinant P450 enzymes (CYP1A2, 2C9, 2C19, 2D6, and 3A4) in an attempt to limit interactions with other more minor P450s and associated reductases. Kinetic analysis of single substrate with single P450 (sP450) yielded apparent Km values of 25, 2, 20, 9, and 3 microM, for CYP1A2, 2C9, 2C19, 2D6, and 3A4, respectively. Combined substrates with combined P450s (cP450) yielded apparent Km values of 65, 4, 19, 7, and 2 microM. Selectivity of the substrates for each P450 isoform was checked. Phenacetin proved to be the least selective substrate. However, the ratio of the various P450s was modified in the final assay such that metabolism of phenacetin by other enzymes was approximately 20% of the metabolism by CYP1A2. IC50 determinations with alpha-naphthoflavone (0.04 microM), sulfaphenazole (0.26 microM), tranylcypromine (9 microM), quinidine (0.02 microM), and ketoconazole (0.01 microM) were similar for sP450 and cP450 enzymes. The assay was further evaluated with 11 literature compounds and 52 in-house new chemical entities, and the data compared with radiometric/fluorescent values. The overall protein level of the assay was reduced from the original starting point, as this led to some artificially high IC50 measurements when compared with existing lower protein assays (radiometric/fluorometric). This method offers high throughput P450 inhibition profiling with potential advantages over current radiometric or fluorometric methods.