Comparison of Selective Media for the Enumeration of Probiotic Enterococci from Animal Feed

The project »Methods for the Official Control of Probiotics Used as Feed Additives« has been undertaken to develop and validate methods for the selective enumeration and strain identification of six probiotic microorganism genera (enterococci, lactobacilli, bifidobacteria, pediococci, bacilli and yeast). A diversity of media has been used for the detection, isolation and enumeration of enterococci. Aiming at the selective enumeration of enterococci (mainly Enterococcus faecium) present in probiotic animal feeds, either as a single component or in combination with other microorganisms, an extensive screening of published methods for culturing and enumerating enterococci was carried out. A collection of enterococcal strains used as probiotics in animal feeds and of isolates as well as reference strains from culture collections was established. Moreover, selected strains of lactobacilli, pediococci and streptococci were included for reference purposes. Based on a multitude of publications, twelve commercially available media were selected for testing and then compared with regard to their usefulness and selectivity. Bile esculin azide (BEA) agar showed good selectivity and pronounced growth of most enterococcal strains. Good reproducibility and electivity (esculin hydrolysis) as well as no influence of the feed matrix on the colony counts and a simple preparation procedure formed the basis for the proposed enumeration protocol. This work formed the basis for the enumeration protocol that was adapted to ISO format and validated in a collaborative study involving twenty laboratories from twelve European countries

Mutter werden mit eingeschränkter Mobilität : das subjektive Erleben von Frauen mit mobilitätseinschränkenden Behinderungen in der Zeit von Schwangerschaft, Geburt und Wochenbett

Darstellung des Themas: Der Kinderwunsch bei Frauen mit mobilitätseinschränkenden Behinderungen nimmt immer mehr zu. Die Betreuung während der Schwangerschaft, der Geburt und dem Wochenbett verläuft allerdings oft nicht optimal. Für die Hebammen stellt die Betreuung von betroffenen Frauen eine besondere Herausforderung dar, da nur wenig Literatur und keine hebammenspezifischen Leitlinien zur Verfügung stehen. Ziel: Das Ziel dieser Arbeit ist es, zu ermitteln, wie Frauen mit mobilitätseinschränkenden Behinderungen die Schwangerschaft, die Geburt und das Wochenbett erleben und ob spezifische Wünsche an die Hebamme vorhanden sind. Daraus sollen Empfehlungen für die Hebammenarbeit abgeleitet werden. Methode: Anhand zuvor definierter Suchbegriffe wurde in vier Datenbanken eine systematische Literaturrecherche durchgeführt. Unter Berücksichtigung bestimmter Ein- und Ausschlusskriterien wurden vier qualitative Studien selektiert und ausgewertet. Relevante Ergebnisse: Frauen mit mobilitätseinschränkenden Behinderungen haben einen grossen Bedarf an Informationen und erleben Schwierigkeiten bei der Informationsbeschaffung. Den involvierten Fachpersonen mangelt es häufig an spezifischem Fachwissen und Erfahrung. Zudem treffen betroffene Frauen oft auf bauliche Hürden und nicht behindertengerecht ausgestattete Einrichtungen. Schlussfolgerung: Es braucht Leitlinien sowie eine individuelle und angepasste Betreuung in einem interprofessionellen Team, um eine optimale Versorgung gewährleisten zu können

Estimación de la edad relativa de crías de ballena franca austral (Eubalaena australis) a través de indicadores morfométricos y morfológicos

Fil: Domig, Natalia L. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Carrera de Ciencias Biológicas; Argentina.Las técnicas o métodos conocidos para estimar la edad en ballenas vivas se basan principalmente en la evaluación de proporciones corporales, en particular el largo y el ancho. En el presente trabajo se evaluaron potenciales indicadores morfológicos y morfométricos para caracterizar y estimar la edad relativa de las crías de ballena franca austral (Eubalaena australis) en Península Valdés, Argentina. Las cuatro categorías etarias se determinaron a partir de la proporción del largo relativo de la cría respecto al largo de su madre (LR) las cuales se asociaron a un rango de edad estimada en días (EED). Se analizaron indicadores cualitativos como la definición del patrón de callosidades, su coloración, la ocurrencia de ciámidos, la apariencia y coloración del lomo. Asimismo, se evaluaron variables cuantitativas en relación al área que ocupan los ciámidos en la cabeza, el área con ecdisis en el lomo, y las proporciones entre anchos corporales y el ancho de la aleta caudal. Los resultados mostraron que en las categorías etarias 1 y 2 las callosidades son grisáceas, con escasa cantidad de ciámidos, en tanto que en las categorías 3 y 4, son más bien blancas por la presencia de ciámidos. La distribución de estos organismos varía con la edad del ballenato, y mientras que en la categoría 1 se presentan en abundancia sobre la zona de los labios (5% del total del área de la cabeza), en las categorías 2 y 3 se encuentran ciámidos en las mejillas (4.9% y 5.2%; particularmente naranjas, C. erraticus) y en el rostro (7.8% y 10.6%), que disminuyen levemente en la cuarta categoría (4.1% en mejilla y 10.2% en rostro). Por otro lado, el proceso de ecdisis es característico de las dos primeras categorías (13.2% y 9.2% del total del área del lomo, respectivamente), que presentan una espalda de aspecto áspero y coloración oscura o gris opaco, mientras que en las categorías 3 y 4 está prácticamente ausente (0.9% y 0%) y el aspecto de su piel es más bien variado o suave con coloración negra. Por último, se observó que tanto el ancho de la mitad del cuerpo (A50%) como el ancho de la cabeza (A25%) respecto al ancho de la aleta caudal (Aac) presentan una proporción que aumenta con el crecimiento, siendo de 1.7 en la categoría 1, y de 2 en la categoría 4. La mayoría de las características estudiadas, al utilizarlas de manera integrada mostraron ser buenas indicadores, para diferenciar las categorías etarias. La determinación y la caracterización de categorías etarias resulta compleja en grandes cetáceos, por lo que su uso como herramienta para identificar individuos a campo o a través de fotografías es relevante en diversas investigaciones, como 6 el estudio de la estructura y dinámica poblacional. El presente estudio contribuye al conocimiento sobre la ontogenia de las crías de ballena franca austral, aportando indicadores que pueden utilizarse para describir las distintas etapas del desarrollo durante sus primeros meses de vida.Fil: Domig, Natalia L. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Carrera de Ciencias Biológicas; Argentina

Susceptibility of Bifidobacteria of Animal Origin to Selected Antimicrobial Agents

Strains of the genus Bifidobacterium are frequently used as probiotics, for which the absence of acquired antimicrobial resistance has become an important safety criterion. This clarifies the need for antibiotic susceptibility data for bifidobacteria. Based on a recently published standard for antimicrobial susceptibility testing of bifidobacteria with broth microdilution method, the range of susceptibility to selected antibiotics in 117 animal bifidobacterial strains was examined. Narrow unimodal MIC distributions either situated at the low-end (chloramphenicol, linezolid, and quinupristin/dalfopristin) or high-end (kanamycin, neomycin) concentration range could be detected. In contrast, the MIC distribution of trimethoprim was multimodal. Data derived from this study can be used as a basis for reviewing or verifying present microbiological breakpoints suggested by regulatory agencies to assess the safety of these micro-organisms intended for the use in probiotics

Species identification of enterococci by biochemical test and molecular-genetic methods

The aim of this study was comparison different methods of species identification of enterococci. One hundred and fifty three suspected colonies were isolated from milk and dairy products (cheeses from cow´s, ewe´s and goat´s milk). On the bases of their growth on BEA agar, microscopic characteristic, results of Gram staining, catalase test and PYRAtest was thirty four isolates assigned to the genus Enterococcus. These isolates were identified by commercial biochemical test EN-COCCUS. 52.9% of them were included in species E. faecalis, 29.4% in E. faecium, 14.7% in E. durans and 2.9% in E. group III. This group includes 3 species: E. durans, E. hirae, E. faecalis asaccharolytic var. Then 16S rRNA sequencing nucleotide of all isolates was realized. Results of sequencing were compared with NCBI database. Only 14.7% of isolates were in 100% accordance. One from them was species E. durans and others were designated as E. faecium. For 20.6% of detected isolates was in accordance with more reference strains. Other isolates were identical with reference strain on 99%. For verification of all results species-specific PCR was used and 52.9% isolates were identified as species E. faecalis, 32.4% as E. faecium and 14.7% as E. durans. Strains belonging to the species E. faecalis were identified the most reliable by all used methods

Uporaba lokalnih sojeva bakterije Lactococcus lactis kao starter kultura u proizvodnji tradicionalnih crnogorskih sireva

The aim of this study is to characterise and examine the biochemical properties of 40 Lactococcus lactis strains isolated from indigenous Montenegrin dairy products in order to explore their potential to be used as starter cultures for producing typical Montenegrin cheese, such as ‘bijeli sir’, ‘masni sir’ and ‘njeguški sir’. Their safety regarding the production of biogenic amines, the presence of antimicrobial resistance and the antibacterial activity against relevant pathogens and spoilage microorganisms has also been tested. Based on the characterisation, all strains belong to L. lactis ssp. lactis. Out of these 40 strains, 23 displayed rapid acidification ability and proteolysis. However, none of the strains exhibited the ability of lipid degradation. Most of the strains were not associated with any health risk investigated. Summing up, a large percentage (27.5 %) of the tested strains showed good properties. These strains should be further examined for their possible application as specific starter cultures in the production of indigenous cheese in MontenegroSvrha je ovoga rada bila okarakterizirati i ispitati biokemijska svojstva 40 sojeva bakterije Lactococcus lactis, izoliranih iz autohtonih crnogorskih mliječnih proizvoda, te istražiti mogućnost njihove uporabe kao starter kultura u proizvodnji tipičnih crnogorskih sireva, kao što su bijeli sir, masni sir i njeguški sir. Također je ispitana sigurnost primjene tih sojeva, tj. sposobnost proizvodnje biogenih amina, antimikrobna rezistencija i sposobnost suzbijanja rasta važnih patogenih mikroorganizama i uzročnika kvarenja hrane. Karakterizacijom je utvrđeno da svih 40 sojeva pripadaju podvrsti L. lactis ssp. lactis. Njih 23 imala su sposobnost brzog zakiseljavanja i proteolize. Međutim, niti jedan soj nije pokazao sposobnost razgradnje lipida. Većina sojeva nije predstavljala nikakav rizik za zdravlje. Velik postotak sojeva (27,5 %) pokazao je dobra proizvodna svojstva, pa treba dodatno ispitati mogućnost njihove uporabe kao specifičnih starter kultura u proizvodnji autohtonih crnogorskih sireva

Suitability of Different PCR-DGGE Primer Sets for the Monitoring of Lactic Acid Bacteria in Wine

Lactic acid bacteria (LAB) play a dual role in winemaking as they are the main effectors of malolactic fermentation, but some members can also cause wine spoilage. PCR-DGGE has proved to be a quick tool to study the LAB community and their fluctuation in wine. For detecting wine-associated LAB by PCR-DGGE, the primer sets WLAB1/WLAB2GC, WBAC1/WBAC2GC, Lac1/Lac1o/Lac2GC, 341fGC/518r and rpoB1/rpoB1o/rpoB2GC were tested and evaluated in this study. The primer systems were assessed by the separation of LAB reference strains on DGGE gels and by attributing the resulting amplicons to defined species. Subsequently, the detection of LAB in wine samples and enrichments thereof was compared. While the primer systems WBAC1/WBAC2GC and 341fGC/518r were not appropriate, the Lac1/Lac1o/Lac2GC primer set performed well. However, multiple bands complicated the evaluation. The rpoB1/rpoB1o/rpoB2GC set seemed to be promising for the detection of LAB in wine, although further improvements in terms of the detection limit need to be done. Due to the pronounced sensitivity and the sufficient discrimination of LAB at species level, the WLAB1/WLAB2GC primer system was found to be most suitable for studying the occurrence of LAB in wine

Ropiness in Bread—A Re-Emerging Spoilage Phenomenon

As bread is a very important staple food, its spoilage threatens global food security. Ropy bread spoilage manifests in sticky and stringy degradation of the crumb, slime formation, discoloration, and an odor reminiscent of rotting fruit. Increasing consumer demand for preservative-free products and global warming may increase the occurrence of ropy spoilage. Bacillus amyloliquefaciens, B. subtilis, B. licheniformis, the B. cereus group, B. pumilus, B. sonorensis, Cytobacillus firmus, Niallia circulans, Paenibacillus polymyxa, and Priestia megaterium were reported to cause ropiness in bread. Process hygiene does not prevent ropy spoilage, as contamination of flour with these Bacillus species is unavoidable due to their occurrence as a part of the endophytic commensal microbiota of wheat and the formation of heat-stable endospores that are not inactivated during processing, baking, or storage. To date, the underlying mechanisms behind ropy bread spoilage remain unclear, high-throughput screening tools to identify rope-forming bacteria are missing, and only a limited number of strategies to reduce rope spoilage were described. This review provides a current overview on (i) routes of entry of Bacillus endospores into bread, (ii) bacterial species implicated in rope spoilage, (iii) factors influencing rope development, and (iv) methods used to assess bacterial rope-forming potential. Finally, we pinpoint key gaps in knowledge and related challenges, as well as future research questions