An important signaling pathway involved in the regulation of auto

An important signaling pathway involved in the regulation of autophagy is the Ras/PKA pathway (Budovskaya et al., 2004). Inactivation of the Ras/PKA pathway, by overexpression of a dominant-negative allele of RAS2, known as RAS2ala22, resulted in increased induction of autophagy as compared with WT. However, additional inactivation of the genes encoding the PKA catalytic subunits, TPK1, TPK2 and TPK3, in the double Δipt1Δskn1 deletion mutant did not result in an enhanced autophagy phenotype (data not shown) as compared with the double Δipt1Δskn1 deletion mutant, indicating that Skn1, together with Ipt1, might act in the same pathway as Ras/PKA regarding induction/regulation

of autophagy. Moreover, PKA and Sch9 signaling pathways are known to regulate autophagy cooperatively in yeast (Yorimitsu et al., 2007). Long-chain bases including phytosphingosine are recognized as regulators of AGC-type protein

selleck compound kinase (where AGC stands for protein kinases A, G and C) Pkh1 and Pkh2, which are homologues of mammalian phosphoinositide-dependent protein kinase 1 (Sun et al., 2000). Based on in vitro data, Liu et al. (2005a, b) demonstrated that phytosphingosine stimulates Pkh1 to activate additional downstream kinases including Ypk1, Ypk2 and Sch9, and additionally, that phytosphingosine can directly activate Ypk1, Ypk2 and Sch9. In conclusion, it could be that the higher basal levels of phytosphingosine, which we observed in the double Δipt1Δskn1 mutant, affect Sch9 function directly or Urease indirectly,

and concomitantly, the authophagy response. Hence, future research will be directed towards determining whether Sch9 or other kinases are part of the link between sphingolipids and autophagy in yeast. In conclusion, all the data obtained in this study point to a negative regulation of autophagy by both Ipt1 and Skn1 in yeast, which could be mediated by sphingoid bases and might act in the same pathway as the Ras/PKA signaling pathway. Apparently, Ipt1 and Skn1 can functionally complement each other under nutrient limitation, not only regarding synthesis of the complex sphingolipid M(IP)2C upon nutrient limitation in half-strength PDB (Thevissen et al., 2005) but also regarding the negative regulation of autophagy under N starvation, as demonstrated in this study. This work was supported by a grant from FWO-Vlaanderen (research project G.0440.07) to B.P.A.C. Postdoctoral fellowships to A.M.A. (Research Council) and to K.T. (Industrial Research Found), both from K.U. Leuven, are gratefully acknowledged. F.M. and D.C.-G. are grateful to the FWF for SFB ‘Lipotox’ and NRN S-9304-B05. Lipidomics CORE at the Medical University of South Carolina is supported by NIH Grant No. C06 RR018823. D.J.K. is supported by National Institutes of Health Public Health Service grant GM53396.

This clinical audit and was conducted at a large teaching hospita

This clinical audit and was conducted at a large teaching hospital NHS Trust from February to March 2014. Adult inpatients receiving vancomycin during the study period were identified by a list that was generated daily by the microbiology deportment. Paediatric patients, patients receiving haemodialysis,

patients admitting to a ward that does not follow guidelines, patients with missing data on dosing were excluded from this audit. Patients’; medical charts were reviewed and information about patient demographics, the nature of infection and vancomycin dosing and monitoring were collected using a pre-designed data collection form, which was designed according STA-9090 in vitro to a literature review, expert opinions and a pilot study. Descriptive Epacadostat nmr statistics were used to describe the proportion of patients given the correct LD, correct MD, whose first PDL reached 10–20 mg/L. The time to maintaining within the therapeutic level was also calculated. This audit was conducted under the Trust’s research guidance and ethical approval was not required. Of the 104 eligible patients, 14 on dialysis, 8 from non-adherence wards and 5 had missing data were excluded. Of the 77 included patients, 55 (71.4%) were prescribed a LD according to guidelines; and 37 (67.3%) of the 54 patients were also prescribed a

MD according to guidelines. The overall adherence to the dosing guideline was 48.1%. Of the 37 patients whose LD and MD were prescribed correctly, 23 (62.2%) first PDL reached 10–20 mg/L. In contrast, of the 40 patients whose LD or MD was prescribed incorrectly, 21 (52.5%) patients’; first

PDL reached 10–20 mg/L. Sixteen (20.8%) of the 77 patients were excluded from the calculation for time to reach maintaining therapeutic 4��8C range due to missing data on dates and times for LD or only one PDL reading available. Of the 61 patients who had more than one PDL measure, 29 (47.5%) were given the correct LD and MD, and 14 (48.3%) of them maintained at the therapeutic level, and it took an average of 4.8 days to reach. Of the 32 patients given the incorrect LD or MD, only 10 maintained at the therapeutic level, it took 6.7 days to reach the level. Almost half of patients were prescribed vancomycin following the dosing guideline, and adherence to the guidelines increased the likelihood that therapeutic level was obtained and the therapeutic level was reached quicker. Further research is needed to explore reasons for non-adherence to vancomycin dosing guideline and evaluate the clinical outcomes related to appropriate dosing. R. Sivam, I. Sanghera, F. Turnbull Northwick Park Hospital, Harrow, UK The Nursing and Midwifery Council (NMC) ‘Standards for Medicines Management’ recommends that nurses should carry several checks before administering an injectable medicine. Only 51% (n = 90) of drug charts were documented with two signatures.

Clinical diagnosis is often difficult, as infectious exanthematou

Clinical diagnosis is often difficult, as infectious exanthematous diseases such as measles, rubella, Palbociclib research buy human parvovirus B19, dengue, human herpes virus (HHV)-6, roseola infantum, and scarlet fever have overlapping clinical symptoms. In Brazil, from 1994 to 1998, 327 patients presenting with pathologies characterized by variable combinations of exanthema, cough, conjunctivitis, coryza, and fever were studied. A laboratory-confirmed diagnosis was achieved in 71.3% of cases: 33% were diagnosed with dengue fever, 20% with rubella, 9.2% with human parvovirus B19, 6.7% with measles,

and 2.1% with HHV-6.[4] These results underline the important proportion of cosmopolitan febrile exanthemas. In France, Hochedez and colleagues screened 62 returning travelers presenting with fever and exanthema for exotic (if returning from endemic areas) and cosmopolitan infections. They found a specific etiology in over 90% of the patients. The three main diagnoses were chikungunya, dengue, and African tick bite fever, followed by infectious mononucleosis, human immunodeficiency virus-1 primary infection, cytomegalovirus primary infection, MDV3100 nmr measles, rubella, chicken pox, streptococcal infections, primary toxoplasmosis, acute schistosomiasis, and adverse drug reactions.[1]

Travelers presenting with febrile exanthema should therefore be screened not only for arboviral infections according to the area visited but also for more common infections. The diagnosis of dengue fever is based on the detection of NS1 Ag, antibodies (IgM and IgG) or reverse transcription (RT)-PCR (virus isolation is used less often). For early diagnosis (onset < 5 days), detection of NS1 Ag may

be used, but its moderate sensitivity requires the presence of both NS1 Ag and IgM for a definite diagnosis.[5] IgM are positive 4 to 5 days after disease onset and remain so for up to 3 to 6 months. IgG appear approximately 7–10 days after onset and are detectable thereafter for life. RT-PCR detection of viral RNA is a very reliable technique for patients presenting within 5 to 7 days of the onset of symptoms, but this method is more expensive, nonstandardized, and only a few centers in France use it C-X-C chemokine receptor type 7 (CXCR-7) routinely.[6] Consequently, serological tests are commonly used to establish or confirm a diagnosis of dengue. Currently available commercial rapid tests offer good sensitivity, but they lack specificity, which may lead to false-positive results as in our index case. Overall, possible explanations for false-positive results include cross-reactive flavivirus-specific antibodies, nonspecific binding of antibodies secreted in the course of various infections such as mononucleosis or hepatitis, and rheumatoid factor.[7] Cross-reactivity with measles antibodies is not commonly assumed by biologists and, to our knowledge, has only been reported once in Belgium.

Pujol for advice This work was supported in part by the project

Pujol for advice. This work was supported in part by the project with reference AGL2011-30461-C02-02 by the Ministerio de Ciencia e Innovación (Spain). Please note: Wiley-Blackwell is not responsible for the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article. “
“The Gram-negative anaerobe Dichelobacter nodosus is the causative agent of footrot in sheep. Different strains of D. nodosus cause disease of differing severities, ranging from benign to virulent.

Virulent strains have greater twitching motility and secrete proteases that are more thermostable than those secreted by benign strains. We have identified polynucleotide phosphorylase (PNPase) as a putative virulence regulator and have proposed that PNPase expression is modulated by the adjacent integration of genetic elements. In this study, GSK126 manufacturer we compared PNPase activity in three virulent and four benign strains of D. nodosus and found that PNPase activity is lower in virulent learn more strains. We disrupted the pnpA gene in three benign D. nodosus strains and two virulent strains and showed that deletion of the S1 domain of PNPase reduced catalytic activity. In all but one case,

deletion of the PNPase S1 domain had no effect on the thermostability of extracellular proteases. However, this deletion resulted in an increase in twitching motility in benign, but not in virulent strains. Reconstruction of the pnpA gene in two mutant benign strains reduced twitching motility to the parental level. These results support the hypothesis that PNPase is a virulence repressor in benign strains of D. nodosus. Footrot is a mixed bacterial infection Parvulin of the hooves of sheep, goats and deer that leads to lameness. The Gram-negative anaerobic bacterium Dichelobacter nodosus is the principal causative agent (Beveridge, 1941). Different strains of D. nodosus cause disease of differing severities, ranging from benign to virulent. The extracellular proteases secreted by virulent

strains are more thermostable than proteases secreted by benign strains (Depiazzi & Richards, 1979). Virulent strains also have greater twitching motility, generated by polar type IV fimbriae, than benign strains (Depiazzi & Richards, 1985), and twitching motility is essential for virulence (Kennan et al., 2001; Han et al., 2008). Comparative analysis of DNA from virulent and benign strains has led to the identification of a series of genetic elements that integrate into the D. nodosus chromosome. These include the intA (Katz et al., 1991, 1992, 1994; Cheetham et al., 1995; Billington et al., 1996), intB (Bloomfield et al., 1997), intC (Bloomfield et al., 1997) and intD elements (Tanjung et al., 2009), each of which contains an integrase gene. A fifth integrated element, the virulence-related locus, vrl (Katz et al., 1991; Haring et al., 1995; Billington et al., 1999), lacks an integrase gene.

According to Buchner (1960), many hatching larvae and nymphs (eg

According to Buchner (1960), many hatching larvae and nymphs (e.g. Regorafenib weevils, stink bugs) own biting mouthparts with which they feed parts of the eggshell during its burst and are thus infected with the bacteria. Larval infection of P. riparius with endosymbionts most probably takes place in the same manner. However, where exactly the endosymbiotic bacteria of P. riparius are located inside the beetles was not resolved in this work and requires further FISH investigations with the novel oligonucleotide probes developed in this study. We thank J. Piel (Kekulé Institute, University of Bonn) for the supply with the ketosynthase-specific primer

pair KS1F/KS1R, H. Rödel (Institute of Animal Physiology, University of Bayreuth) for the kind introduction in sigmaplot 9.0, R. Grotjahn (Institute of Electron Microscopy, University of Bayreuth) for numerous electron-microscopical exposures, E. Helldörfer (Institute of Animal Ecology II, University of Bayreuth) for the creation of scientific figures of Paederus beetles’ anatomy, W. Nowak and I. Nowak for

the provision of several P. riparius specimens and Harold L. Drake for provision of a LINUX-based network for arb. Financial support by the Deutsche Forschungsgemeinschaft (DFG) is gratefully acknowledged (GRAKO 678). “
“Bacteria secrete small signal molecules into the environment that induce self and neighbour gene expression. This phenomenon, termed quorum sensing, allows cooperative Apoptosis inhibitor behaviours that increase the fitness of the group. The best-studied signal molecules are the N-acylhomoserine lactones (AHLs), Montelukast Sodium secreted by a growing number of bacterial species including important pathogen species such as Pseudomonas aeruginosa. These molecules have recently been proposed to have properties other than those of signalling, functioning as iron quelants or antibiotics. As the presence of an acylase capable of inactivating long-chain AHLs in Anabaena sp. PCC7120 could constitute a defence mechanism against these molecules,

in this work we analyse the effects of different AHLs varying in length and substitutions on the growth and nitrogen metabolism of the cyanobacterium Anabaena sp. PCC7120. All the AHLs tested strongly inhibited nitrogen fixation. The inhibition seems to take place at post-transcriptional level, as no effect on heterocyst differentiation or on the expression of nitrogenase was observed. Moreover, N-(3-oxodecanoyl)-l-homoserine lactone (OC10-HSL) showed a specific cytotoxic effect on this cyanobacterium in the presence of a combined nitrogen source, but the mechanism involved seems to be different from that described so far for tetramic acid derivatives of oxo-substituted AHLs. These results suggest a variety of new unexpected activities for AHLs, at least on cyanobacterial populations. The term ‘quorum sensing’ (QS) (Fuqua et al.

This finding was not surprising, as higher expression levels of m

This finding was not surprising, as higher expression levels of mexB and mexD are to be expected in mexR or nfxB mutants (Dumas et al., 2006). The nature of the specific beneficial adaptive mechanisms in the tolerant bacterial population is under investigation in our lab. To gain insight into the effect of inactivation of genes involved in the GO system on selleck chemicals the global gene expression, we studied the transcriptional changes produced by inactivation of the two genes. The inactivation of both mutY and mutM caused significant changes (more than twofold and P-value < 0.05 compared with PAO1) in six genes (Table 4). Remarkable was the up-regulation of pfpI whose product has been shown to have a protective role against

DNA damage caused by the oxidative stress (Rodriguez-Rojas & Blazquez, 2009). This can be considered a compensatory mechanism for the protection of the DNA in PAOMY-Mgm with impaired repair of the DNA oxidative damage. Interestingly, the most significantly down regulated gene was PA5148 involved in iron selleck trafficking. The modified expression of pfpI and PA5148 expression in PAOMY-Mgm compared with PAO1 was confirmed using RT-PCR, which showed up-regulation of pfpI (9 ± 2.3-fold) and down-regulation of PA5148 (4.04 ± 2.7-fold). Complementation of PAOMY-Mgm,

which showed high expression levels of pfpI and low expression levels of PA5148 compared with PAO1 with wild-type mutM or mutY, reduced the level of pfpI up-regulation to 6 ± 2.4-fold and 4.6 ± 2.4-fold, respectively and the level of PA5148 down-regulation to 1.6 ± 0.09-fold and 2.1 ± 0.25-fold, respectively. Pseudomonas aeruginosa, which colonizes and persists within the highly ROS-rich CF airways has to protect itself against the mutagenic effect of ROS and it uses the GO system, consisting of MutT, MutY and MutM to prevent or eliminate the oxidized form of guanine, which is a mutagenic lesion. Homologue proteins are present in other microorganisms as well as in eukaryotic cells. Inactivation

of each of the three genes encoding for the respective proteins led to various degree of increase in the spontaneous MF with mutants in mutY and mutM exhibiting a moderate and weak mutator phenotype (increase in MF < 20 times the MF of PAO1) (Mandsberg et al., Dipeptidyl peptidase 2009; Morero & Argarana, 2009; Sanders et al., 2009). In the present study, we show for the first time that the mutY and mutM double mutant (PAOMY-Mgm) showed a strong mutator phenotype providing evidence for the cooperation of MutM and MutY to prevent mutagenesis in P. aeruginosa, in a similar manner as in E. coli (Michaels et al., 1992; Tajiri et al., 1995). It has been shown that hypermutability plays an important role in the adaptive evolution of P. aeruginosa in the CF lung (Mena et al., 2008), and it has been demonstrated that mutator populations are amplified by hitchhiking with adaptive mutations. The selective pressure exerted by antibiotics plays an important role in the adaptive process of P.

The ghrelin-mimetic drug growth-hormone releasing peptide 6 (GHRP

The ghrelin-mimetic drug growth-hormone releasing peptide 6 (GHRP-6) has been shown to inhibit light-induced cFos expression in the SCN and attenuate a light induced phase shift (Yi et al., 2006; Yi et al., 2008), suggesting that ghrelin can act as a non-photic stimulus to alter the timing of light-signaled behaviour. Therefore, it is not surprising that the absence of ghrelin could alter the timing of activity, especially in LL, where photic Zeitgebers are also absent. In this situation, the absence of ghrelin activity at the GHRS receptor did not have a significant effect on comsummatory behaviour, as the two groups ate the same amount of food and there were no differences

in body weight. One question that must be addressed is the surprising lack of food anticipatory activity in WT mice housed in LL. Indeed, food anticipatory activity has been previously demonstrated in rats housed Cabozantinib clinical trial in LL (Bolles & Stokes, 1965; Edmonds & Adler, 1977a,b; Lamont et al., 2005). In Lamont et al. (2005), no attempt

was made to quantify the amount of anticipatory activity, but certainly overall activity levels were very low after an extended period in LL, as can be seen in the actograms presented in that article. Species differences may selleck compound account for the lack of food anticipatory activity observed in the present study in WT mice. In one study using spiny mice, Acomys cahirinus, wheel-running activity was reduced dramatically in LL compared to LD and only two of the 11 mice studied actually showed entrainment to a restricted feeding schedule under LL, although all 11 had shown significant food anticipatory activity on an LD schedule prior to exposure to

LL (Chabot et al., 2012). In the current experiment, Celecoxib 30 days in LL reduced daily activity levels in WT mice to fewer than 200 wheel revolutions per day, as compared to 600 in KO mice. With such a low level of activity in WT mice, it may simply be difficult to detect food anticipatory activity in these animals. Sampling of brain and peripheral tissues for clock gene protein and RNA at different time points during the temporal feeding period would have demonstrated whether central and peripheral circadian oscillators were entrained to the time of food availability, although the large number of animals required for this type of study was prohibitive. Alternately, a circadian-controlled measurement that is suppressed by light to a lesser degree, such as body temperature, may have been useful in detecting food anticipation in these mice. Regrettably, these data were not collected. Together, these data provide further support for the hypothesis that ghrelin plays a role in the food-entrainable clock, but also suggest that there may be an interaction between the effect of light and ghrelin that extends beyond a simple deficit in the ability of GHSR-KO animals to entrain to scheduled feeding.

Further research is required to explore the detailed mechanisms

Further research is required to explore the detailed mechanisms. “
“l-Asparaginase-producing microbes are conventionally screened on phenol red l-asparagine-containing plates. However, sometimes the contrast of the zone obtained (between yellow and pink) is not very sharp and distinct. In the present investigation, an improved method for screening of the microorganisms producing extracellular l-asparaginase is reported wherein bromothymol blue (BTB) is incorporated as pH indicator in l-asparagine-containing medium instead of phenol red. Plates containing BTB at acidic pH are yellow and turn dark blue at alkaline selleck products pH. Thus, a dense dark blue zone is formed around microbial colonies producing l-asparaginase, differentiating between enzyme

producers and non-producers. The present method is more sensitive and accurate than the conventional method for screening of both fungi and bacteria producing extracellular l-asparaginase. Furthermore, BTB gives a transient green colour at neutral pH (7.0) and dark blue colour at higher pH 8.0–9.0, indicating the potency of the microorganism for l-asparaginase production. ERK inhibitors library
“Studies of enterohemorrhagic Escherichia

coli (EHEC) infection mechanisms using mammals require large numbers of animals and are both costly and associated with ethical problems. Here, we evaluated the pathogenic mechanisms of EHEC in the silkworm model. Injection of a clinically isolated EHEC O157:H7 Sakai into either the silkworm hemolymph or intraperitoneal fluid of mice killed the host animals. EHEC O157:H7 Sakai deletion mutants of the rfbE gene, which encodes perosamine synthetase, a monosaccharide

component synthetase of the O-antigen, or deletion mutants of the waaL gene, which encodes O-antigen ligase against the lipid A-core region of lipopolysaccharide (LPS), had attenuated killing ability in both silkworms and mice. Introduction of the rfbE gene or the waaL gene into the respective mutants pheromone restored the killing ability in silkworms. Growth of both mutants was inhibited by a major antimicrobial peptide in the silkworm hemolymph, moricin. The viability of both mutants was decreased in swine serum. The bactericidal effect of swine serum against both mutants was inactivated by heat treatment. These findings suggest that the LPS O-antigen of EHEC O157:H7 plays an important defensive role against antimicrobial factors in the host body fluid and is thus essential to the lethal effects of EHEC in animals. Infectious diseases caused by enterohemorrhagic Escherichia coli (EHEC) O157:H7 are a serious clinical problem and are associated with encephalopathy and nephropathy (Tarr, 1995; Law, 2000). An understanding of the molecular mechanisms of EHEC O157:H7 virulence is important for establishing effective therapeutic strategies. Unlike other E. coli strains, EHEC produces Shiga toxins and hemolysins. Shiga toxins are encoded by the stx1 and stx2 genes on the phage DNA that is integrated into the EHEC genome (Sato et al.

5, lane 12) An extensive mutagenesis of the E coli ArgR was per

5, lane 12). An extensive mutagenesis of the E. coli ArgR was performed in order to determine its precise role in cer site-specific recombination. The ArgR protein binds DNA at ARG boxes localized in promoter regions of several genes of the arginine regulon and at the cer site, which

contains half an ARG box. It is quite likely that the DNA-binding activity of this protein is an important contributor to its role as an accessory factor in cer recombination by bringing two cer sites together. However, ArgR by itself cannot support cer recombination in the presence of the XerCD recombinases; PepA is also required for this reaction. Alén et al. (1997) have demonstrated that PepA and ArgR interact directly with cer, forming a complex in which the accessory sequences of two cer sites are interwrapped approximately three times in a right-handed fashion. Using pentapeptide scanning mutagenesis, we isolated a series of ArgR mutants that showed an approximate 90% reduction in cer recombination, but were still able to repress an argA∷lacZ fusion effectively in vivo (Figs 1 and 2). The mutant proteins also displayed

sequence-specific DNA-binding activity (Fig. 3). All of Pirfenidone the insertions mapped to the same amino acid, between residues 149 and 150 of ArgR (ArgR5aa). This region corresponds to the C-terminal region of ArgR, at the end of the α6-helix (Fig. 4). In order to show that the observed phenotype was due to the disruption of ArgR and was not caused by the additional five amino acids residues, we constructed an ArgR mutant that was truncated at this region. This protein lacks residues 150–156 (ArgR149), Ribonuclease T1 but displays the same properties as ArgR5aa, namely a significant reduction

in cer site-specific recombination in vivo (Fig. 1b), and the ability to bind to DNA at near wild-type levels in vivo (Fig. 2) and in vitro (Fig. 3). Moreover, we were able to detect the same level of DNA retardation as the wild-type protein, which suggests that both ArgR149 and ArgR5aa bind to DNA as hexamers (Fig. 3). In addition, crosslinking studies have shown that wild-type and mutant proteins are capable of forming higher-order structures in solution, although ArgR149 does not appear to form hexamers as efficiently as either wild-type ArgR or ArgR5aa under the crosslinking conditions used (Fig. 5). It is possible that the small C-terminal deletion in ArgR149 prevents this protein from forming a stable hexameric structure. Similar results have been observed with the α A-crystallin protein, where deletions of the terminal 11 amino acids from the C-terminus significantly decreased the oligomeric size of the protein (Thampi & Abraham, 2003). Despite this, ArgR149 can still bind DNA effectively both in vivo and in vitro; the addition of DNA and l-arginine may allow this mutant to form more stable hexamers under these conditions.

Alternative approaches proposed the utilization of MALDI-TOF for

Alternative approaches proposed the utilization of MALDI-TOF for species identification based on the sodA gene (Hinse et al., 2011). Most of these assays were developed using blood-derived clinical cultures of the SBSEC or restricted to a single species. In contrast to blood samples, raw dairy products were shown to contain a large diversity of different lactococci, enterococci, Streptococcus thermophilus, or Streptococcus

agalactiae (Delbès et al., 2007; Younan & Bornstein, 2007; Franciosi et al., 2009; Giannino et al., 2009; Jans, 2011), which increases the requirements regarding specificity of the primers. Even though the genes groESL or sodA provide improved capability to differentiate RGFP966 solubility dmso between species and subspecies, the 16S rRNA gene is still regarded as the recommended target for the initial identification of novel bacteria for which the higher degree of conservation of the 16S rRNA gene can be of advantage (Glazunova et al., 2009). This gene is one of the most important genotypic markers for bacterial taxonomy (Yarza

et al., 2008), and a large number of 16S rRNA gene sequences are available for downstream comparison and further analysis (Benson et al., 2009). It therefore represents an ideal target for the analysis of complex and I-BET-762 nmr less-studied ecological niches such as the human microbiota (Grice et al., 2008; Liu et al., 2008) or spontaneous food fermentations, e.g., the African dairy environment. The high-density and complex microbial communities in these niches could result in unexpected genetic modifications through horizontal gene transfer (HGT), which was observed for African S. infantarius subsp. infantarius as well as for S. thermophilus and other LAB (Hols et al., 2005; Terminal deoxynucleotidyl transferase Makarova et al., 2006; Jans, 2011).

HGT is affecting nearly all genes within prokaryotic genomes; some genes including the 16S rRNA gene are, however, hypothesized to be less affected by HGT (Jain et al., 1999). Therefore, the objective was to utilize the high conservation of the 16S rRNA gene to develop an identification assay applicable to all species within the SBSEC allowing clear differentiation from other streptococci, enterococci, and lactococci regularly found in the dairy environment. The availability of large sets of nucleotide sequences from all members of the SBSEC including dairy isolates (Jans, 2011) enabled the design of a subsequent RFLP for the discrimination of SBSEC species groups. Furthermore, the primers were designed to work with Sanger sequencing for downstream sequence analysis. The assay was then evaluated against reference strains and isolated species of dairy microbial communities. Bacterial reference strains listed in Table 1 were obtained from the Culture Collection University of Gothenburg (CCUG, Gothenburg, Sweden), the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany), and the National Collection of Type Cultures (NCTC, Porton Down, UK).