text:
Mechanisms of Resistance
Identification of vat(E) in Enterococcus faecalis Isolates from Retail Poultry and Its Transferability to Enterococcus faecium
S. Simjee, D. G. White, D. D. Wagner, J. Meng, S. Qaiyumi, S. Zhao, P. F. McDermott
S. Simjee
U.S. Food and Drug Administration Center for Veterinary Medicine, Laurel, Maryland 20708
For correspondence: ssimjee@cvm.fda.gov
D. G. White
D. D. Wagner
J. Meng
Department of Nutrition and Food Science, University of Maryland, College Park, Maryland 20742
S. Qaiyumi
S. Zhao
P. F. McDermott
DOI: 10.1128/AAC.46.12.3823-3828.2002
Sixteen isolates of Enterococcus faecalis were recovered from retail poultry samples (seven chickens and nine turkeys) purchased from grocery stores in the greater Washington, D.C., area. PCR for known streptogramin resistance genes identified vat(E) in five E. faecalis isolates (three isolates from chickens and two isolates from turkeys). The vat(E) gene was transmissible on a ca. 70-kb plasmid, along with resistance to erythromycin, tetracycline, and streptomycin, by conjugation to E. faecalis and Enterococcus faecium recipient strains. DNA sequencing showed little variation between E. faecalis vat(E) genes from the chicken samples; however, one E. faecalis vat(E) gene from a turkey sample possessed 5 nucleotide changes that resulted in four amino acid substitutions. None of these substitutions in the vat(E) allele have previously been described. This is the first report of vat(E) in E. faecalis and its transferability to E. faecium, which indicates that E. faecalis can act as a reservoir for the dissemination of vat(E)-mediated streptogramin resistance to E. faecium.
Enterococci form part of the normal host flora of both the human and the animal gastrointestinal tract, where they seldom cause serious infections. However, it has been well documented that enterococci are etiological agents of endocarditis and urinary tract sepsis in humans (9). Of increasing concern is the rapid emergence of enterococci as the third leading cause of nosocomial infections in the hospital setting, surpassed only by Staphylococcus aureus and coagulase-negative staphylococci (15, 16). As a result, enterococci have eloquently been referred to as the "pathogens of the 90s" (7). Approximately 85 to 90% of enterococcal infections are attributed to Enterococcus faecalis and 5 to 10% are attributed to Enterococcus faecium. Infections caused by other Enterococcus species (E. durans, E. avium, E. raffinosus, E. gallinarum, and E. casseliflavus) occasionally emerge and have warranted attention (16).
Infections caused by multidrug-resistant enterococci are usually treated with the glycopeptide vancomycin. However, as the use of vancomycin has steadily increased, so, consequently, has the incidence of vancomycin-resistant enterococci (21). Recent reports suggest that almost one-quarter of enterococci isolated from patients in intensive care units in the United States are vancomycin resistant (11, 12). Quinupristin-dalfopristin (Synercid), a semisynthetic mixture of the compounds streptogramins A and B, was recently approved for use for the treatment of vancomycin-resistant enterococci in both the United States and Europe (18, 19). Virginiamycin, another mixture of the compounds streptogramins A and B, has been used as a growth promoter in animal production for over two decades. The use of antimicrobials in the animal production environment has the potential of selecting for resistant zoonotic bacterial pathogens, and it has been speculated that the extensive use of virginiamycin in animal husbandry may have contributed to the emergence of quinupristin-dalfopristin resistance among human gram-positive pathogens (5, 6; L. B. Jensen, L. B., A. M. Hammerum, F. M. Aerestrup, A. E. van den Bogaard, and E. E. Stobberingh, Letter, Antimicrob. Agents Chemother. 42:3330-3331, 1998). Consequently, the use of virginiamycin has been banned in the European Union since July 1999 (1). However, despite the 1998 ban of virginiamycin in Denmark, a recent report showed a 22.5% prevalence of virginiamycin-resistant E. faecium isolates from pigs in 2000 (1). Although there is no clear explanation for this, it has been suggested that the presence of resistance to other antimicrobial agents such as erythromycin, tetracycline, and streptomycin may coselect for virginiamycin resistance (1).
Resistance to streptogramins was first reported in staphylococci in 1980 (4). Only resistance to the A component is required; however, resistance to both the A component and the B component may result in higher MICs (4). To date a number of genes that confer streptogramin A resistance in both staphylococci and E. faecium have been reported (2, 4, 18, 22, 23; K. V. Singh, B. M. Jonas, G. M. Weinstock, and B. M. Murray, Abstr. 41st Intersci. Conf. Antimicrob. Agents Chemother., p. 103, 2001). E. faecalis is intrinsically resistant to streptogramin antibiotics, although the mechanism(s) underlying this resistance has yet to be fully described (13; Singh et al., 41st ICAAC). However, preliminary data suggest that an efflux pump, designated ABC23, may play a role in conferring quinupristin-dalfopristin resistance in E. faecalis (Singh et al., 41st ICAAC). The amino acid sequence of the ABC23 efflux pump appears to show high degrees of similarity (41 to 64%) to those of the ABC proteins of Lactococcus lactis; Bacillus subtilis; Streptococcus pneumoniae; Msr(C) of E. faecium; and Vga(A), Vga(B), and Msr(A) of Staphylococcus aureus (Singh et al., 41st ICAAC). To date, none of the streptogramin resistance genes found in staphylococci or E. faecium have been identified in E. faecalis.
The aim of the present study was to determine if any of the known staphylococci or E. faecium streptogramin resistance genes are present in E. faecalis isolated from retail poultry samples.
Sample collection and isolation of enterococci.Prepackaged retail chicken and turkey samples were collected from four supermarket chains in the Maryland suburbs of Washington, D.C., in June 1999. The packaging was aseptically opened, and the meat samples were rinsed in 50 ml of sterile buffered peptone water. One milliliter of the rinse suspension was placed in 10 ml of Enterococcosel (BBL) broth, and the mixture was incubated at 45°C for up to 48 h. Esculin-positive cultures were streaked onto Enterococcosel agar, and the plates were incubated at 35°C for 24 h. A single colony characteristic of Enterococcus spp. was picked from each plate and streaked onto Trypticase soy agar with 5% sheep blood to ensure purity and to check for hemolysis. Single colonies were picked from blood agar plates and streaked onto brain heart infusion agar. Suspect enterococci were Gram stained and tested for catalase and pyrimidase production. Catalase-negative, gram-positive, pyrimidase-positive isolates were confirmed as Enterococcus spp. with the AccuProbe Enterococcus identification test (Gen-Probe, Inc.). AccuProbe-positive isolates were identified to the species level with the Automated Microbial Identification system (bioMerieux Vitek, Inc.).
Antimicrobial susceptibilities of enterococci.The MICs of antimicrobials for enterococci were determined with the Sensititre Automated Antimicrobial Susceptibility system (Trek Diagnostic Systems, Westlake, Ohio) and were interpreted according to National Committee for Clinical Laboratory Standards (NCCLS) guidelines for broth microdilution methods (10). Escherichia coli ATCC 25922, S. aureus ATCC 29213, Pseudomonas aeruginosa ATCC 27853, and E. faecalis ATCC 29212 were used as quality control microorganisms.
DNA extraction, PCR studies, and DNA sequencing.Total bacterial DNA was extracted by the guanidium thiocyanate method described previously (16) for use as the template for PCR. Known streptogramin resistance genes as well as the macrolide resistance genes ermA, ermB, and ermC were amplified by PCR by using the oligonucleotide primers, PCR conditions, and control strains described previously (18, 20). The PCRs were performed with the AmpliTaq Gold PCR system (Perkin-Elmer, Norwalk, Conn.).
As the vat(E) primers amplified a 512-bp internal region of the vat(E) allele representing 80% of the coding region, the primers described by Soltani et al. (19) were used to amplify the 3′ end of the vat(E) allele. For amplification of the 5′ end of the vat(E) allele, we used the PCR primers and conditions described previously (17). The amplification gave overlapping PCR products which spanned the entire vat(E) gene and extended into both the upstream and downstream regions flanking the vat(E) gene. The DNA sequence of each overlapping amplicon was determined in both directions (SeqWright, Houston, Tex.) and compared to that of vat(E-1) (GenBank accession no. AF242872) with the BLAST program of the National Center for Biotechnology Information (3).
Recently, Jensen et al. (L. B. Jensen, A. M. Hammerum, and F. M. Aarestrup, Letter, Antimicrob. Agents Chemother. 44:2231-2232, 2000) described the linkage of vat(E)-ermB on a transposon-like element residing on a plasmid in E. faecium. We also examined the possibility of vat(E)-ermB gene linkage in E. faecalis using the PCR primers and conditions described previously (Jensen et al., Letter, Antimicrob. Agents Chemother. 44:2231-2232, 2000).
PFGE.Pulsed-field gel electrophoresis (PFGE) was performed after digestion of the DNA with SmaI as described previously (16). To analyze the PFGE results to determine the relatedness of the E. faecium isolates, we used the interpretive criteria described elsewhere (21). The PFGE fingerprints were compared by computer-assisted analysis (BioNumerics; Applied Maths, Austin, Tex.).
Conjugation of streptogramin resistance determinants.The frequencies of transfer of the streptogramin resistance determinants from E. faecalis to E. faecalis and from E. faecalis to E. faecium were examined by the filter mating method (14). The recipient strains used for the conjugation studies were E. faecalis CVM3463 (quinupristin-dalfopristin MIC, 8 μg/ml; gentamicin MIC, 1,028 μg/ml) and E. faecium GE-1 (quinupristin-dalfopristin MIC, <1 μg/ml; rifampin MIC, 100 μg/ml; fusidic acid MIC, 50 μg/ml). Transconjugants were selected on blood agar base supplemented with 5% defibrinated sheep blood containing either 16 μg of quinupristin-dalfopristin per ml and 500 μg of gentamicin per ml for the selection of E. faecalis transconjugants or 8 μg of quinupristin-dalfopristin per ml, 50 μg of rifampin per ml, and 25 μg of fusidic acid per ml for the selection of E. faecium transconjugants. Following incubation at 37°C for 48 h, 10 transconjugants were selected from each mating, and the transconjugants were subjected to susceptibility testing and analyzed for the presence of plasmids.
Plasmid analysis.Plasmid extraction was done by the alkaline lysis method described previously (14) to detect plasmids and to determine the number of plasmids in each strain and the transconjugants. Two strains of E. coli, strains V517 (8) and 39R861 (22), harboring plasmids of known sizes were used as plasmid size markers. Purified plasmid preparations were electrophoresed through 0.8% agarose gels at 90 V for 2 h with 1× TBE (Tris-borate-EDTA) electrophoresis buffer. The plasmids were visualized under UV light following ethidium bromide staining.
Antimicrobial susceptibility phenotypes of enterococcal isolates.Sixteen independent E. faecalis isolates were recovered from 16 independent retail meat samples (seven chicken samples and nine turkey samples) and further examined for their antimicrobial susceptibilities. As E. faecalis is intrinsically resistant to the streptogramins, it was no surprise to find that all 16 E. faecalis isolates exhibited reduced susceptibilities to quinupristin-dalfopristin and the related veterinary streptogramin, virginiamycin (Table 1). In addition, 66% of the turkey sample isolates and 85% of the chicken sample isolates showed resistance to erythromycin (NCCLS breakpoint, >8 μg/ml). All 16 isolates were resistant to tetracycline (NCCLS breakpoint, >16 μg/ml). None of the isolates were resistant to penicillin (NCCLS breakpoint, >16 μg/ml). Twenty-two percent of the turkey sample E. faecalis isolates and 14% of the chicken sample E. faecalis isolates showed high-level resistance to gentamicin (NCCLS breakpoint, >500 μg/ml). All of the turkey sample E. faecalis isolates and 42% of the chicken sample E. faecalis isolates were resistant to streptomycin. The susceptibility profiles of the 16 E. faecalis isolates are shown in Table 1.
Antimicrobial susceptibility profiles, PCR data, and PFGE grouping of the 16 E. faecalis isolates recovered from retail meatsa
PCR studies and DNA sequencing.vat(E) was amplified by PCR from 5 of the 16 (31%) E. faecalis isolates. Three vat(E)-positive isolates (isolates CVM 3478, CVM 3972, and CVM 3973) were recovered from chicken sample E. faecalis isolates, and two vat(E)-positive isolates (isolates CVM 3476 and CVM 3477) were recovered from turkey sample E. faecalis isolates. No other known streptogramin resistance determinants [vat, vat(B), vat(C), vat(D), vga, vga(B), vgb, vgb(B)] were amplified by PCR. Additionally, ermA was detected in 33% of chicken sample E. faecalis isolates and 43% of turkey sample E. faecalis isolates. Two of the chicken sample E. faecalis isolates and two of the turkey sample E. faecalis isolates were also vat(E) positive by PCR (Table 1). In contrast, ermB was detected in 100% of the vat(E)-positive chicken sample E. faecalis isolates but was not detected in any of the vat(E)-positive turkey sample isolates. Among the E. faecalis isolates negative for vat(E) by PCR, ermB was detected in 75% of the chicken sample E. faecalis isolates and 85% of the turkey sample E. faecalis isolates. One chicken sample E. faecalis isolate was positive for ermA, ermB, and vat(E). Additionally, one turkey sample E. faecalis isolate had both ermA and ermB; however, this isolate was negative for vat(E) by PCR. None of the isolates tested positive for ermC by PCR (Table 1).
Even though ermB and vat(E) reside in the same isolate in two chicken sample E. faecalis isolates that were PCR positive for vat(E) and ermB (isolates CVM 3972 and CVM 3973), we were unable to show vat(E)-ermB linkage on a transposon-like element, as described by Jensen et al. (Letter, Antimicrob. Agents Chemother. 44:2231-2232, 2000), in these two isolates. These findings indicate that vat(E) and ermB may not be close to each other on the plasmids found in these two isolates.
DNA sequencing confirmed the presence of vat(E) PCR products; however, interestingly, several variations between the vat(E) DNA sequences indicating allelic variation were detected. The vat(E) sequences of the two turkey sample E. faecalis isolates varied from the original vat(E-1) sequence originally described (GenBank accession no. AF242872). Strain CVM 3476 had a single vat(E) mutation, A43→G (Ile15→Val). In contrast, turkey sample E. faecalis strain CVM 3477 possessed five point mutations resulting in four amino acid substitutions and one silent mutation. These changes were G40→T (Ala14→Ser), A46→C (Lys16→Gln), G51→T (Glu17→Asp), T56→A (Val19→Asp), and G273→T (Ser89→Ser). Less sequence divergence was observed between the vat(E) genes from chicken sample E. faecalis isolates. The vat(E) sequences of two of the chicken sample E. faecalis isolates were identical to the vat(E-1) sequence originally described; however, the sequence of vat(E) from E. faecalis CVM 3478 varied by a single base, resulting in a single amino acid change, A43→G (Ile15→Val). This was the same base change seen with turkey sample E. faecalis isolate CVM 3476. All the amino acid changes are outlined in Fig. 1.
Variations in vat(E) alleles of streptogramin A acetyltransferase found in chicken and turkey sample isolates.
PFGE.PFGE of the 16 E. faecalis isolates identified a total of 10 different PFGE patterns, which were arbitrarily assigned to groups A through J (Table 1). The nine turkey sample E. faecalis isolates were distinguishable from the seven chicken sample E. faecalis isolates by PFGE. The two vat(E)-positive turkey sample isolates were indistinguishable from each other by PFGE (PFGE group E), even though they came from different meat samples from different stores. Of the three vat(E)-positive chicken sample isolates, two belonged to PFGE group J, while one belonged to PFGE group I.
Plasmid analysis.Analysis of the five E. faecalis isolates positive for vat(E) by PCR confirmed the presence of either three to four plasmids ranging in size from ca. 5 to ca. 70 kb. Turkey sample isolates CVM 3476 and CVM 3477 and chicken sample isolate CVM 3478 each had three plasmids that appeared to be of the same size, ca. 70, 15, and 5 kb, respectively. The remaining two chicken sample E. faecalis isolates (isolates CVM 3972 and CVM 3973) appeared to have the same plasmid profile outlined above; in addition, they also carried a ca. 7-kb plasmid.
Conjugation studies.Conjugation studies demonstrated that all five vat(E)-positive E. faecalis isolates were able to transfer the vat(E) streptogramin resistance determinant to both E. faecalis and E. faecium recipient strains. Conjugation frequencies for the transfer of vat(E) varied from 1 × 10−2 to 8 × 10−3 per recipient for both E. faecalis and E. faecium recipients.
Ten transconjugants per mating were selected and subjected to plasmid analysis and antimicrobial susceptibility testing. PCR analysis confirmed the presence of the vat(E) genes in the transconjugants. The plasmid profiles among the transconjugants varied, the ca. 70-kb plasmid was present in all transconjugants examined; however, the full complement of plasmids identified in donor strains was never observed in any of the transconjugants analyzed (data not shown). In each case, the MICs of quinupristin-dalfopristin, erythromycin, tetracycline, and streptomycin were the same for the transconjugants and the original parent strain.
Enterococci are an increasing problem in human medicine, as the array of antibiotics available for the treatment of enterococcal infections is diminishing. This is compounded by the fact that enterococci are intrinsically resistant to a large number of antibiotics and can rapidly acquire and disseminate resistance genes. Quinupristin-dalfopristin was approved for use in the United States in 1999 for the treatment of glycopeptide-resistant E. faecium infections. However, it is not efficacious against E. faecalis, as this species of Enterococcus is intrinsically resistant to the streptogramins.
To our knowledge the present study is the first to report the presence of the streptogramin resistance gene vat(E) in E. faecalis. This is surprising since one might expect that a species already intrinsically resistant to an antibiotic class would not acquire additional resistance determinants against that class of antibiotics. In the present study, besides vat(E), no other streptogramin resistance determinants were identified in E. faecalis. The fact that no other streptogramin resistance determinants were identified is an important observation, as reports from Europe have identified vat(D) and vgb, in addition to vat(E), in E. faecium (19; Jensen et al., Letter, Antimicrob. Agents Chemother. 42:3330-3331, 1998). As the present study evaluated only 16 E. faecalis isolates, the ability to detect other possible resistance determinants was limited.
In addition, the three vat(E)-positive chicken sample E. faecalis isolates also harbored the ermB gene; however, the turkey sample vat(E)-positive E. faecalis isolate tested negative for ermB by PCR. Of the vat(E)-negative E. faecalis turkey isolates, 85% were positive for ermB by PCR. Jensen et al. (Letter, Antimicrob. Agents Chemother. 44:2231-2232, 2000) previously demonstrated that vat(E) and ermB are linked on the same plasmid in 70% of E. faecium poultry isolates from Denmark. However, in the present study we did not find linkage of vat(E)-ermB as part of a possible transposon-like element. Despite this, the fact that both vat(E) and ermB are present in the same strain, and in some cases on the same plasmid, may help to explain the acquisition of vat(E) by E. faecalis as a result of selective pressure from macrolide use in poultry.
Examination of the vat(E) DNA sequences obtained from E. faecalis isolates of chicken origin revealed that two of the vat(E) sequences were identical to that of vat(E-1); however, the sequence of strain CVM 3478 showed a single base change from that of the vat(E-1) allele originally described. In contrast, much more variation was observed between the vat(E) sequences of the two turkey sample E. faecalis isolates, which exhibited 3% sequence divergence, resulting in 93% amino acid identity. However, in one chicken sample isolate (CVM 3478) and one turkey sample isolate (CVM 3476), vat(E) showed the same base change, resulting in the same amino acid substitution (Ile15→Val). The MICs for these two isolates were almost identical (only differences in the MICs of erythromycin were seen), and the plasmid profiles of the two isolates were identical. We could therefore predict either that these two isolates are clones or that they share common plasmids. However, these two isolates were distinguishable by PFGE, indicating that perhaps they may have only acquired vat(E) from a common reservoir. The DNA sequence data obtained from this study suggest that the vat(E) genes in the chicken and turkey sample isolates may not, in all cases, have originated from the same source. There appears to be greater homology among the chicken E. faecalis vat(E) sequences, whereas heterogeneity may exist between the turkey E. faecalis vat(E) sequences. To confirm this hypothesis, a larger number of isolates would need to be studied.
Eight allelic variations of vat(E) from E. faecium, designated vat(E-1) through vat(E-8), have been deposited in GenBank (17, 19). However, the vat(E) allelic variations observed from E. faecalis isolates in this study have not previously been observed in E. faecium. It is of interest that the substitutions found in the vat(E) alleles, including those described in the present study, are all focused around a very small region of the vat(E) allele, namely, between base positions 34 and 57 (amino acids 11 to 19). How these amino acid substitutions affect the activity of the Vat(E) protein is not known at present, as the three-dimensional crystal structure of the Vat(E) protein has not been determined. However, we suspect that this small region (amino acids 11 to 19) does not play an important role in streptogramin binding either directly or indirectly, as amino acid variations in this region do not seem to have any effect on the level of resistance to quinupristin-dalfopristin.
PFGE analysis showed that the two vat(E)-positive turkey sample isolates were indistinguishable from each other. Similarly, two vat(E)-positive chicken sample E. faecalis isolates were assigned to PFGE group J and one vat(E)-positive chicken sample isolate was assigned to PFGE group I. These results suggest that the vat(E) genes are not confined to a single clone in E. faecalis, as determined by PFGE.
It was interesting that the two vat(E)-positive turkey sample E. faecalis isolates belonged to the same PFGE group; however, there was 3% DNA sequence divergence in the vat(E)-coding region, suggesting that a clone of E. faecalis may have acquired vat(E) from different sources.
Conjugation studies demonstrated that the vat(E) gene could be transferred not only from an E. faecalis donor to an E. faecalis recipient but also from an E. faecalis donor to a streptogramin-susceptible E. faecium recipient. This implicates E. faecalis as a reservoir for vat(E) genes that can be transferred to E. faecalis or E. faecium. This is not surprising, as previous studies have shown that plasmids and transposons readily move between enterococcal species (15, 16). Susceptibility to quinupristin-dalfopristin, erythromycin, tetracycline, and streptomycin was detected following the transfer of the 70-kb plasmid. This is based on the observation that not all transconjugants received the full complement of plasmids that were contained within the donor strain; however, each transconjugant received the 70-kb plasmid. Although the linkage of vat(E) and ermB as part of a possible transposon-like element has recently been reported (Jensen et al., Letter, Antimicrob. Agents Chemother. 44:2231-2232, 2000), we could not establish a vat(E)-ermB linkage. It is possible that vat(E) may have appeared in E. faecalis by virtue of gene linkage on the same plasmid as the resistance determinants for either erythromycin, tetracycline, or streptomycin resistance. Thus, the vat(E) gene may have been coselected by the use of either macrolides, tetracyclines, or the aminoglycosides in the poultry production environment. It is therefore feasible that an organism such as E. faecalis that is intrinsically resistant to a given antimicrobial may, under certain conditions, harbor resistance genes as a consequence of coselection. This notion is supported by reports from Denmark, which showed that in 1998, 55.6% of E. faecium isolates recovered from pigs were resistant to virginiamycin. Following the 1998 ban on the use of virginiamycin in feeds, this prevalence decreased to 8.0% in 1999 and then increased again to 22.5% in 2000. Although it was unclear why resistance to virginiamycin increased in 2000, the investigators suggested that the increase was due to the emergence of isolates that were simultaneously resistant to erythromycin, kanamycin, penicillin, streptomycin, and tetracycline (1). This may explain to some degree the appearance of vat(E) in our isolates of E. faecalis. All five E. faecalis isolates that were vat(E) positive by PCR showed reduced susceptibilities to erythromycin, streptomycin, and tetracycline. It is plausible that the use of these antibiotics, either singly or in combination, may have coselected for the appearance and maintenance of vat(E) in a population of E. faecalis isolates.
In conclusion, we have detected vat(E) in E. faecalis, a species intrinsically resistant to quinupristin-dalfopristin, probably by virtue of efflux mechanisms, and found that this gene could readily be transferred to other E. faecalis and E. faecium isolates. The recovery of vat(E)-containing E. faecalis isolates from retail chickens and turkey samples (42 and 22% of samples, respectively) indicates a reservoir of streptogramin resistance genes that can potentially be transferred to human gram-positive pathogens via consumption of contaminated food. Even though the study cohort comprised a small number of randomly selected isolates, the presence of vat(E) in approximately 30% of the E. faecalis isolates suggests that the prevalence of vat(E) in the larger environment is significant. Until we fully understand the mechanisms of streptogramin resistance in enterococci and quantify the risk of gene transfer from nonhuman isolates to human isolates, the occurrence of streptogramin resistance genes in enterococci of animal origin remains a potential public health hazard.
Received 27 February 2002.
Returned for modification 1 July 2002.
Accepted 29 August 2002.
American Society for Microbiology
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Tenover, F. C., R. D. Arbeit, R. V. Goering, P. A. Mickelsen, B. E. Murray, D. H. Persing, and B. Swaminathan. 1995. Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis criteria for bacterial strain typing. J. Clin. Microbiol. 33:2233-2239.
Threlfall, E. J., B. Rowe, J. L. Ferguson, and L. R. Ward. 1986. Characterization of plasmids conferring resistance to gentamicin and apramycin in strains of Salmonella typhimurium phage type 204c isolated in Britain. J. Hyg. (London) 97:419-426.
Werner, G., I. Klare, H. Heier, K. H. Hinz, G. Bohme, M. Wendt, and W. Witte. 2000. Quinupristin/dalfopristin-resistant enterococci of the satA (vatD) and satG (vatE) genotypes from different ecological origins in Germany. Microb. Drug Resist. 6:37-47.
Antimicrobial Agents and Chemotherapy Dec 2002, 46 (12) 3823-3828; DOI: 10.1128/AAC.46.12.3823-3828.2002
You are going to email the following Identification of vat(E) in Enterococcus faecalis Isolates from Retail Poultry and Its Transferability to Enterococcus faecium


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Arts + Commerce
Essays + Humor
Land + Sea
Gardens + Landscapes
Best of the Vineyard
Cook the Book
It's a birder! It's a painter! Look, it's Lanny McDowell!
The Accidental Ornithologist
Nelson Sigelman
Artist Lanny McDowell has always been fascinated by birds. But it was the appearance of three unusual winged visitors to Menemsha Pond in early summer 2001 that set him on the path to become one of Martha's Vineyard's foremost birders and avian photographers. At the time the former art history major turned Island carpenter and part-time artist was living in West Tisbury. His friend Richard Cohen, who regularly raced his Herreshoff 12.5 on Menemsha Pond, knew that McDowell was knowledgeable about birds.
"He called to tell me that he'd seen some black-and-white duck-like birds that he hadn't seen before and I ought to check them out," McDowell recalled recently. McDowell hopped in his kayak. "I found these guys and I had my little camera with me and I shot over the bow and I got some pictures."
It was a family of razorbills. Not uncommon to see in the winter off our coast, but very unusual for a species that was not known to nest farther south than on islands off the coast of northern Maine. McDowell called his friend, the late Vineyard Gazette bird columnist E. Vernon Laux. He was exuberant.
"In the long ornithological history of Massachusetts and Martha's Vineyard, there has never been an occurrence like this," Laux wrote in a story that ran on the front page of the Vineyard Gazette. Soon The New York Times got wind of the surprise visitors. Not for the first time, and certainly not the last, the national editorial hook was the Island's reputation as a haven for celebrities – human versus avian.
In 2001 The New York Times published one of Lanny McDowell's photos of a razorbill, launching his bird photography career.
Lanny McDowell
"Rare Birds Eclipse Vineyard's Celebrities" was published on August 26, 2001. Reporter Sara Rimer wrote, "To outsiders, this paradisiacal island off the coast of Massachusetts is known as a place to spot celebrities. This summer, the lineup has included Bill and Hillary Rodham Clinton, James Taylor, Carly Simon, Alan Dershowitz and Sarah Jessica Parker…To birders, the human species is the least of the excitement."
The New York Times had sent a photographer along to the Vineyard, but a day sailing on the pond in search of razorbills came up empty. A photo was needed. McDowell had it, and The New York Times bought it.
With the sale of that one photo, McDowell saw an opportunity to do something that would be fun and satisfying: "As silly as it sounds after the fact, I thought to myself coyly, 'This is a great way to make a living.'"
Which is not to say he immediately quit his various day jobs, of course. Or creating his paintings, which are mostly brightly colored, often geometric works of acrylic on canvas.
McDowell's artistic creativity and appreciation of nature springs from a family tree rooted in outdoor pursuits and art. He grew up in the rural confines of Kent, Connecticut, one of a generation of family members – Harts, Moores, Lows, Youngs, and Peases – that summered in the Oak Bluffs family enclave on Nantucket Sound known as Harthaven, founded by William H. Hart, president of the Stanley Works in New Britain, Connecticut. There were so many artists among the various members of the clan in the middle of the twentieth century in Harthaven that informal family art shows were a regular feature of summer.
Twilight Cedars, acrylic on canvas, 24 x 36 inches.
In 1970, McDowell and his wife were living in New Haven, where he had attended Yale University, when they decided to move to the Island to live year-round. What brought him back? "Everything I knew about the Vineyard from the past," McDowell said.
Chilmark builder Emmett Carroll gave him his first job. He joined a crew building a house off Lighthouse Road in Gay Head (now Aquinnah). "I didn't know the first thing about carpentry and Emmett just put up with my ignorance," McDowell said. "It was a really cold winter with northwest exposure and that was my baptism to Island life in the wintertime."
It was an introduction to year-round Island culture working with men only a few years older than he. Names familiar to a now aging generation of Islanders – Huey Taylor, Danny Larsen, Richard Greene – were part of the crew.
Soon, he used his newly acquired building skills to construct his own house – "hippie funk with some taste" – on family land at the end of Middle Point on Tisbury Great Pond, where his Harthaven relatives kept a rustic duck hunting camp. "I was definitely off the grid out there," McDowell said.
Oriole, Through the Temple Wall, acrylic on canvas, 36 x 36 inches.
Following the death of both his parents in the summer of 1983, he and his wife moved back to the family home in Kent overlooking the Housatonic Valley. He was working in real estate and hadn't painted since college when his artistic genes stirred at the sight of deer against the landscape. "I would sit there, either with a spotting scope or binoculars, and watch the deer against the snow in the wintertime. And I said to myself, 'I'm going to paint that scene.'" He's been painting off and on ever since.
In 1995, recently divorced, he returned to the Island and carpentry. At the same time, his interest in birds, which had never waned, was amplified when he renewed his friendship with Laux. "All of a sudden I was off and running, tagging along with Vern, who was a moving force."
Then, The New York Times flew in. "Instead of having a seasonal, very local audience, I thought I could have a year-round national audience for bird photographs, so I embarked on that project," he said. "Unfortunately, my timing was terrible….There are a lot of birders out there with digital cameras."
Their level of expertise became apparent in August 2004 when a wayward red-footed falcon visited the grass strip airfield in Katama. It was the first sighting of its species in North America, and it set the bird-watching world on fire. Birders flocked to the Island. Stalking the falcon with other photographers was when McDowell realized he would have to up his game. "I knew that to be a professional and have a satisfying result I would have to take it more seriously. I got a good camera and a good lens and birded more and shot more, and if I traveled, I took the camera with me."
Hollow Man, sculpture in half-inch steel, 40 x 42 inches.
McDowell is low key about the photographs and paintings he sells off his website, lannymcdowellart.com, but he speaks with pride about Martha's Vineyard Bird Alert, the "wildly successful" Facebook group he started in November 2010 that now boasts more than 3,200 members. "So I get around in the birding world, I spend a lot of time doing what I like to do," he said. "I get a lot of exposure and I give away a lot of photographs to any Massachusetts nonprofit that wants them. It's very satisfying. It takes up most of my so-called work time – it takes away from painting, for sure – but my exposure is large for being in a little pond."
McDowell and his wife Melissa Keeler, whom he married in 2019, live on the west side of Tashmoo Pond in Vineyard Haven in a delightful rambling home with abundant nooks and crannies that house the couple's eclectic collection of paintings, books, sculpture, decoys, and knick-knacks. They share this space with three cats, a Labrador retriever, a beagle, a boxer, and two French bulldogs that vie for the attention of any visitor.
On the walls hang McDowell's artistic work, which includes a photo series of dolphins, paintings of waves, and a variety of abstract and representational paintings. Most recently, he has created a series of bird portraits he calls "Double Takes," which he describes as "fanciful symmetries" in which the photo is flipped around a vertical axis to create a mirror image.
He has also stretched his wings and created a painted metal sculpture titled Hollow Man that sits outside the house welcoming visitors. The piece, which he said represents "the spirit of hope and encouragement," started out as a painting until he thought to himself, "Why stop there?"
He said he knows a lot of people, whether it's dealing with a camera or picking up paints, who say they've always wanted to do something creative but don't know where to start. "They're clearly afraid; they're just afraid of making a mistake. But being afraid is like a dead-end street. What could you possibly be afraid of? I'm sure you could tell me, but does it have value enough to stand between you and the satisfaction of at least saying: I gave it a shot? It's a little bit quirky but it's mine."
An uncompleted painting sat on the easel in his studio, waiting for a spell of bad weather, which is when McDowell said he gets most of his painting done. "And it better be bad weather that excludes going out to shoot birds, because I'm pretty much addicted to going out and shooting birds."
He described his paintings as "about feeling good, projecting interesting creativity, and inspiring people, but they're basically decoration. It's décor." In contrast, he said his photographs of birds and his ability to reach people on social media are intended to help a wider audience to understand the beauty and fragility of the natural world that is outside their window.
Clockwise from top left: Yellow warbler, American oystercatcher, yellow-crowned night heron, eastern bluebird.
He is also passionate about a collaboration with his friend, pianist, and inventor David Stanwood of West Tisbury. The show, "Avian Improv: Martha's Vineyard Edition," features a sequence of 234 bird photos and an improvisational background track by Stanwood. "Avian Improv is sort of the current project that has the most promise of turning people on to birds, healing them, putting them at ease – any number of desirable adjectives that I want in my life and that I can make available," he said.
The slideshow, available at mvmuseum.org/learn/
avianimprov, is intended as a way to make the photos and music available to a wider audience. He speaks enthusiastically about his hope that the project will someday supplant the visual version of elevator music: waiting room monitors tuned to cable news. It's ideal for places such as hospitals and nursing homes, anyplace where people may not have the opportunity to be exposed to the "healing and contemplative power of nature," he said.
For those more fortunate, the Island provides many wonderful opportunities to observe birds in their natural environment. One tip, he said, is to go out early, preferably at first light. His favorite birding locations include the Katama plains ("Specifically around the FARM Institute"), Norton Point Beach ("I never fail to have a good time just being out there"), the Sheriff's Meadow Sanctuary off Plantingfield Way in Edgartown ("A great little bird mecca"), Aquinnah ("It has so much to offer in the fall in terms of being a funnel for migrating birds"), and Cape Pogue.
Long-tailed Encounter, Double Take, 12 x 30 inches.
Being able to approach a bird or a flock without disrupting them and acquire a connection with what you're photographing is a challenge, McDowell said.
"But when you're done shooting, and you're able to turn around and walk away almost as stealthily as you were when you approached the bird, that's a kick. So you take home a great sense of your time being well-spent and respect for your subject matter. But it's actually the experience of being there that is so gratifying.
"Being outdoors and being aware, it's great medicine."
Linda carnegie
Oakbluffs
Great article about of a one of a kind friend of many talents❤
House of the Dragonfly
Fourteenth Annual Photography Contest
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Catawba County has a population of 154,810, according to the 2010 census. Its county seat is Newton and its largest city is Hickory.
Catawba County is part of the data center corridor. Maiden is home to the Apple iCloud data center, and Conover is home to the ncDataCampus and the largest privately owned solar farm in the United States operated by Apple.
The county was formed in 1842 from Lincoln County. It was named for the Catawba tribe of Native Americans who once inhabited the area. German Lutheran farmers settled the area in the 1700's.
The county has a total area of 414 square miles, of which 400 square miles is land and 14 square miles (3.27%) is water.
With approximately twenty freight trains a day, Catawba County is a freight railroad transportation center. This is largely due to the area's strong manufacturing based economy, and its placement along the Norfolk Southern Railway line. The Caldwell County Railroad also serves the county and interchanges with Norfolk Southern in Hickory.
The county is divided into eight townships.
Iredell County had a population of 159,437 according to the 2010 census. Its county seat is Statesville; its largest town is Mooresville. It is named for James Iredell, a justice of the Supreme Court of the United States.
According to the U.S. Census Bureau, the county has a total area of 597 square miles, of which 576 square miles is land and 21 square miles (3.59%) is water.
Iredell County is located within the Piedmont Region of central North Carolina. The northwestern section of the county contains the Brushy Mountains, a deeply eroded spur of the Blue Ridge Mountains far to the west. The highest point in Iredell County, Fox Mountain, is in the Brushies; it rises to 1,760 feet. Although the "Brushies", as they are often called locally, are not high in the normal sense, they do rise prominently above the surrounding countryside. The remainder of Iredell County consists of gently rolling countryside occasionally broken by low hills and small river valleys. The county's largest river, the Catawba, forms much of its western border. Lake Norman, North Carolina's largest man-made lake, is the most prominent geographic feature of southern Iredell County; it is often called North Carolina's "inland sea".
Iredell County is one of the longest counties in the state and stretches for nearly fifty miles north to south from Yadkin County in the north to Mecklenburg in the south. The county is divided into seventeen townships.
Iredell County is a major hub of NASCAR racing, with many race shops located in the county (mostly around Mooresville). Many NASCAR drivers live around Mooresville and Lake Norman. Although northern Iredell County has retained much of its rural character, the southern half of the county is experiencing rapid growth, largely due to the immense popularity of the Lake Norman area for residents of nearby Charlotte, North Carolina's largest city.
Lincoln County, North Carolina has an estimated population of 79,740. The county seat is located in Lincolnton. The county was formed in 1779 from the eastern part of Tryon County and was named for Benjamin Lincoln, a general in the American Revolutionary War.
In 1782, the southeastern part of Burke County was annexed to Lincoln County. In 1841, parts of Lincoln County and Rutherford County were combined to form Cleveland County. In 1842, the northern third of Lincoln County became Catawba County and in 1846, the southern half of what was left of Lincoln County became Gaston County.
According to the U.S. Census Bureau, Lincoln County has a total area of 307 square miles, of which, 299 square miles of it is land and 8 square miles of it (2.68%) is water. The population density is 213.5 people per square mile.
Mecklenburg County is both the most populated and densely populated county in North Carolina. On September 12, 2013, the county welcomed its one millionth resident.
The county seat and largest city is Charlotte.
Mecklenburg County was formed in 1762 from the western part of Anson County. It was named in commemoration of the marriage of King George III to Charlotte of Mecklenburg-Strelitz. Due to unsure boundaries, a large part of south and western Mecklenburg County extended into areas that would later form part of the state of South Carolina. In 1768, most of this area (the part of Mecklenburg County west of the Catawba River) was designated Tryon County, North Carolina. Determining the final boundaries of these "western" areas between North and South Carolina would become a decades- long process. In 1792 the northeastern part of Mecklenburg County became Cabarrus County. Finally, in 1842 the southeastern part of Mecklenburg County was combined with the western part of Anson County to become Union County.
The Mecklenburg Declaration of Independence was allegedly signed on May 20, 1775; if the document is indeed genuine, Mecklenburg County was the first part of the Thirteen Colonies to declare independence from Great Britain.
According to the U.S. Census Bureau, the county has a total area of 546 square miles of which 526 square miles is land and 20 square miles (3.65%) is water.
The county's primary commercial aviation airport is Charlotte Douglas International Airport in Charlotte.
Light rail service in Mecklenburg County runs from Uptown to Pineville. Currently a 9.6-mile line, build-out is expected to be complete by 2034.
Mecklenburg's manufacturing base, its central location on the Eastern Seaboard and the intersection of two major interstates in the county have made it a hub for the trucking industry.
The major industries of Mecklenburg County are banking, manufacturing and professional services, especially those supporting banking and medicine. Mecklenburg County is home to six Fortune 500 companies, including 21st-ranked Bank of America.


Neither option is very good for you, your customers, or your company. There is another industry that used to produce their products this way that we should learn a lot from: the auto industry.
Cars used to get produced in huge batches as it was seen as inefficient to produce a variety of cars at the same time. Large caches of parts were held in inventory every step of the way to minimize fluctuation in supply and demand. This resulted in greatly reduced flexibility. In fact the Model T famously came in any color you wanted "so long as it is black." This was the Ford way, and Ford was seen as the epitome of success with their assembly line production.
TPS actually does the exact opposite thing to minimize fluctuations in production. Even though this minimizes fluctuations it actually improves responsiveness to customer and market demands. Most cars are available in at least a dozen colors and have so many possible options (sunroof, satellite radio, GPS, power seats, engines, transmissions, etc) that there are potentially hundreds of thousands or even millions of possible versions of a very common car such as a Toyota Camry.
Of the many advantages to Scrum which are commonly cited the most important benefit is implied but never really articulated and that is reducing what I call "intellectual inventory". Software development can have many forms of this type of inventory. It can be any artifact (requirement/PRD, design document, untested code, etc) that is completely created before being passed off to another functional role.
The key steps of the waterfall process are actually all forms of Intellectual Inventory: requirement definition, design, implementation, and verification. In fact the waterfall model is basically building up inventories in large batches.
Creating requirements takes a lot of "non-development" time from team members like the Product Owner/Manager, User Experience, Design, etc. The more complete the requirements are the more investment you are making. It takes development time to implement something that doesn't get tested. Even if it does get tested but the feature is too buggy to pass QA you have still invested in something with zero return; even though your engineers were "done" with their phase of the waterfall. Make no mistake about it, intellectual inventory is something you spend investment on.
In an Agile/Scrum model intellectual inventory = waste as well. Requirements that are twelve months from being implemented don't need to be refined to the point that the feature could be implemented today. A lot can change in a year and that feature could get dropped, expanded on, or maybe it will be delivered in a different way (iPad app instead of desktop client, HTML5 instead of Flash, etc) because users and the market change. Clearly any time spent refining the requirements that are discarded would have been a complete waste. Of course in practice it isn't possible to completely eliminate inventory, but it can be greatly reduced.
I have personally seen feedback from our user experience (UX) team make it into a shipped product in less than a month from when they did user testing on the product that was in development. That type of responsiveness is impossible in a waterfall model because the UX would be done in a batch that would then lead to a batch of requirements, and so on. Just as in automobile production, reducing batch production of intellectual inventory allows much more responsiveness to consumer demand and reduces waste.
Waste is probably actually worse in software development than the auto industry. Even if a batch of cars is built that ends up having low demand they probably won't be sold for less than the marginal cost to produce them, so the inventory still retains much of its value. If a requirement or design is never built because consumers don't want it then the value of that inventory is zero. All of that investment was lost.
Self-organizing teams with daily status meetings and weekly retrospectives are just a form of Kaizen.
Ability for any team member to bring any issue to light at any time in the development process is also a similar to the role of an Andon system.
Using a "pull" system (as opposed to waterfall's "push") is basically Kanban.
So the next time you are trying to convince someone in your company of the virtues of adopting a "radical" process like Scrum point out how well these same principles worked for a plucky up-start named Toyota.


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Jon Waterhouse and Rob Rosenfeld provide an overview of the work accomplished by the Yukon River Inter-Tribal Watershed Council, demonstrating the benefits of Native nations who have common cultures and challenges to band together to solve issues of mutual concern.
Rosenfeld, Rob and Jon Waterhouse. "The Yukon River Inter-Tribal Watershed Council." Honoring Nations symposium. Harvard Project on American Indian Economic Development, John F. Kennedy School of Government, Harvard University. Cambridge, Massachusetts. September 27-28, 2007. Presentation.
"Good morning. My name's Jon Waterhouse and I work with the Yukon River Inter-Tribal Watershed Council. I'll start with a quick introduction. I'm S'Klallam, Chippewa, Cree and Scottish. I always kind of like to think I got the best parts. My wife has let me know that's my own private delusion. That's okay.
"Podiums are difficult for me as well; I am standing up. It is a true honor to be here among so many great nations. All great nations have allies and it's been a great honor for me to be an ally of Indigenous nations throughout the Yukon River Watershed and beyond.
Jon spoke briefly about the backhaul program. It's been a very, very exciting program. And the genesis of it was at the very first meeting of leaders in 1997. The leaders identified many of the different contaminant sources, from one end of the river to the other, including mining sites, military contaminants, but they looked mostly at their own municipal solid waste. And instead of pointing fingers at polluters, they decided to focus on what they could do in their own backyards. With that the backhaul program was created and, with one of the guiding principles of the organization, it was driven. Through inclusiveness, all the different backhaul companies, 12 airlines, three barge companies, two shipping companies, Alaska Railroad, as well as battery recycling companies joined together. They go out to the villages filled with all sorts of goods and services and come back empty. Not any more. They're loaded up to the gills. And we realized that they were going above and beyond and we realized the importance of rewarding them for their efforts and encouraging corporate responsibility.
And the way that we did that was -- actually, I'll give Jon Waterhouse the credit for that. I asked Jon to come up with a very creative way to say thank you to the many different transportation companies. He walked into my office one day with an idea to do a commercial on the Super Bowl. I thought he was nuts, actually. I had no idea that was even feasible. Well, within about six months, it came to reality. And Super Bowl Alaska aired a 60-second piece that focused on the good work of the transportation companies and the tribes. And that has now been shown two years in a row, last year twice at halftime.
And one of the things I wanted to focus on is, really, how the Honoring Nations award program has helped amplify the work of the Yukon River [Inter-Tribal] Watershed [Council]. Since the Honoring Nations Board of Directors acknowledged the good work of the Tribes on the Yukon River [Inter-Tribal] Watershed [Council], the organization has tripled in size and tripled in credibility. The money that came with the award was used for the video footage that was used for the Super Bowl commercial. It was also used for footage that was utilized to create a new documentary called Yukon Circles. That documentary is now part of the permanent archives of the National Museum of American [Indians]. It travels with the Wild and Scenic Film Festival all around the United States. It's showing in a film festival in Italy this next month in October, and it's been in nine other film festivals around the country.
I do want to switch for a moment to how not only Honoring Nations matters, but also unified voices of sovereign nations matters most of all. When 66 indigenous governments come together and take a position, that position is hard to ignore. It wasn't long ago, maybe three years ago, that the community of Galena was looking at and still is looking at a proposed nuclear reactor, being donated by Toshiba©. Now, it's kind of cute. It sounds kind of cute -- the 4S reactor. It's super, safe, small and simple. It sounds like you want to take it to bed with you. But it's actually 100 feet big; it's 60 feet in the ground and 40 feet above the ground. There's 450, or so, reactors in the world. Three of them are sodium cooled. Every single sodium-cooled reactor in the world that has ever gone online has gone on fire, blown up, and in one case killed everyone on site. Again, corporate America is looking to use Indigenous peoples as guinea pigs. And the Yukon River Inter-Tribal Watershed Council provided information to the Tribes and First Nations. It was a no brainer. It didn't take long. The Tribes and First Nations have unified and they oppose the transportation of radioactive materials on the Yukon River watershed, they stand against the storage of radioactive materials on the Yukon River watershed, the experimentation with Yukon radioactive materials on the watershed, and they opposed this particular sodium-cooled reactor. We got a phone call two nights ago from one of the city council members who once voted in favor of this proposed reactor who now, as of last night, took an official position against the reactor because he wanted to encourage all the city council members to recognize the desires of the First Nations and Tribes on the Yukon River watershed. So we hope next time to come back and let you know that there is no more talk of that reactor.
One other important thing that I wanted to mention was the unified voice that is starting to come to be with regards to a critical issue that affects all of us here in this room and beyond. There is a proposed land transfer in the Yukon Flats Wildlife Refuge, one of the biggest refuges in the United States. The Department of Interior is proposing trading some of the refuge land with a corporation. That would be the beginning of the erosion of refuges in the United States. To go along with that, in an area that has 80 percent unemployment in most places, Fish and Wildlife has put a high dollar -- and is proposing to put a high-dollar value -- on buying Native allotments. What it could amount to is yet another federal land grab of Native lands. I can say that from a meeting over three weeks ago or four weeks ago more than 50 Tribes and First Nations have expressed their opposition for this proposed land transfer. And we'll be going to Secretary Kempthorne's office in the very near future, hopefully, if he will honor the request for a meeting. And the request will be made to "˜please, not to release a draft environmental impact statement without first consulting with all the governments, all the tribal governments on the Yukon River watershed,' before a draft environmental impact statement is released, so that their concerns can be integrated, rather than being reactionary to a position that gets put out. Finally, the leaders have requested a Senate Committee on Indian Affairs hearing to look at the impacts of military contaminants throughout the Yukon River watershed.
And I'll just close with sharing the theme of this initiative here today and this symposium, is sharing successes. And since the Yukon River Inter-Tribal Watershed Council received high honors in 2005, and received incredible exposure as a result, the leaders on the Yukon River [Inter-Tribal] Watershed [Council] have since been invited to go to Bolivia, Paraguay and Argentina to help the Indigenous governments form an inter-tribal watershed effort in a similar way. Since then they have gone already to the Salish Sea Conference, where 88 First Nations and Tribes came together to also emulate and take the best from this idea of organizing on a watershed basis, a paradigm shift. In addition, the leaders have been invited to Hungary, Romania, Bulgaria, the Czech Republic and Slovakia as those countries work together on cross-border environmental cooperation. Just three days ago, we spoke at a NAFTA [North American Free Trade Agreement] conference where the people of Mexico asked for the documentary to include subtitles in Spanish, so that the people of Mexico protecting their rivers can begin to learn from this effort and invitations have already been extended. We've learned of the Three Rivers effort in England, where now there's been invitations for leadership to go to England to teach how to organize for river watershed protection. And finally, there's been invitations to speak at French universities.
Honoring Nations: Manley Begay: So You Have a Great Program...Now What?!


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Home Tea Party What Went Wrong? Everything.
What Went Wrong? Everything.
Ah… The Democratic Party. Such high ideals. Such high expectations. A couple of years ago it looked like you were going to pick up the charred remains of this country after the previous administration left it in ruins and reshape this nation into a better more noble version of itself.
You elected the first black man to be president. You controlled both houses of Congress. You were the the big cheese in the catbird set. What happened? Oh, that's right you're Democrats…
"We like to sit around the office and we have a little game called 'How will the Democrats blow it?'
– The Daily Show's Jon Stewart on Fresh Air with Terry Gross
CNN breaks down what went wrong.
The Tea Party Is Tomorrow
Grab A Friend and Go To A Tea Party Friday April 15th
It's Not Too Late For A Tea Party


The first half of April featured my thoughts on three city centre restaurants: Revolution de Cuba, Tuk Tuk Indian Street Food and Smashburger (If you haven't read it yet, it's available here). Last month, I also visited FF Finsbay, the new steak restaurant in Woodlands rd, Gnom – new cafe/restaurant in Strathbungo, and tried the new pop up Undercover Dinner Society, which was held in Finnieston. Have a read below to see what I thought of these three new Glasgow restaurants.
££: Breakfast options are around £7-9, and coffee is around £2-3 quid. Fairly standard for brunch in the area, and definitely worth it!
Good for: breakfast, brunch, lunch, coffee and cake! Also slightly larger than some of the other popular spots in the area so maybe you will find it easier to grab a table.
Glasgow's food scene right now is on fire with amazing street food, brunch, pop ups and tasting menus. This year, chef John Traynor, who has done a number of pop ups in Glasgow already (Flock Pop up) and has worked at Porter & Rye, launched Undercover Dinner Society which was held in the Drugstore Social. The menu featured the best of seasonal Scottish produce mixed in pan-Asian flavours to create six delicious courses. We also had an option to pair our food with cocktails or wine. Both me and my friend chose to go for the cocktails as we wanted to try something new and interesting.
The food was definitely interesting and very creative. Some of the flavour combinations were unusual but are ones that I still talk about, almost a month after I've been there. There were two dishes that really stood out in my opinion. The first course which featured delicious Goat's cheese mouse in a charcoal cracker, which melted in the mouth in a symphony of fantastic and subtle flavours. It was served with a vodka based watermelon cocktail which was absolutely yum! And the third course, which was a seaweed cured trout in kimchee consume with korean noodles. I was really impressed at how well the flavours balanced each other – a very salty fish with a subtle consume and noodles, which worked together extremely well. Bonus point for the wee fortune cookies that came with it!
The cocktail pairing was a bit hit and miss for me, but that's more because I'm very fussy with my spirits (i.e. not a fan of gin and really dislike whisky). Since more than half of the cocktails featured quite strong on the gin and whisky I couldn't really drink them, but it's my fault for not really looking at the ingredients properly. My friend thought they were delicious. The menu also featured two desserts, which I was initially slightly worried about as I'm not really a dessert person. However, I have to say that the Mango, coconut & Yogurt sorbet (Course No 5) was so light and sweet, and very balanced in it's flavours. The second dessert was slightly sweeter but delicious and once again, featured and balanced delicious flavours. Also.. who can say no to rainbow meringues?!
££: £30 for a six course menu, £20 for cocktail/wine pairings.
Good for: those who like to eat interesting food and flavours and fusion food.
FF Finsbay opened in the West End only a couple of months ago, and since, it has gained huge popularity with Glasgow foodies. They specialise in flatiron steaks and have a banging deal during the week – £8 for a flatiron steak between Sun-Thurs, £10 Fri-Sat. It's great price for a steak in any case, and they also have a really good selection of interesting sides (Parmesan and Hollandaise Sauce topped chunky chips, black pudding bon bons and more).
A few weeks ago, a friend and I headed there for some steaks and cocktails. We both had the flatiron steak, medium rare as recommended by FF's team, with garlic butter and had some chips and black pudding bon bons to share. First things first, the staff are nice and friendly and explained the menu options and steaks well (there are sometimes other steaks on offer as specials). The cocktails were also really tasty. However, when it came to the steak… it wasn't so great. The steaks arrived and looked absolutely delicious, however when I dug in I was disappointed to discover a well-done, chewy steak. When you have a steak restaurant which doesn't cook it's steak right, it's always disappointing. Even more so when they specialise in one particular cut AND recommend it medium rare. When I highlighted this to the staff they took it away and served another steak a little while later which was on the rare side – so still not quite right, but an improvement.
The restaurant is only opened at the end of February so still quite new, and perhaps the chef had a bad night that night. But even so – it was a Friday night (so perhaps a busy night for them) and not doing well what they specialise in was disappointing. Everything else was pretty good, so I might pop in again for another try.
££: Our bill came to just over £40 for the two steaks, sides and drinks.
Good for: midweek dinner, cocktails. Specialises in steak.
I hope you will go and try some of these places, and if you do, please pop your thoughts in the comments below!


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FBI arrests alleged white supremacist accused of planning to bomb a Pueblo synagogue
Posted 7:45 PM, November 4, 2019, by CNN Wire
A 27-year-old man who allegedly espoused white supremacist ideology online was arrested by the FBI and accused of plotting to bomb a synagogue in Colorado.
Richard Holzer had brought a knife and a mask to a motel room and was examining inert pipe bombs prepared by undercover agents moments before he was arrested late Friday, according to a criminal complaint.
Holzer is charged with attempting to obstruct the free exercise of religious beliefs with the attempted use of explosives — a hate crime. In court documents, officials called him a domestic terrorist and described his alleged anti-Semitic ideology and desires for a racial holy war.
According to the complaint, Holzer talked about killing Jews in forums online and shared video of himself casing a synagogue in Pueblo. He described himself as a skinhead, and shared pictures of himself with other social media users with guns and knives alongside white supremacist symbols, the complaint says.
Holzer, according to prosecutors, wrote on one Facebook account: "I wish the Holocaust really did happen …. they need to die."
He also said he paid a "witch doctor" $70 to put arsenic in the water pipes of the synagogue and "hex" the place last year, according to the complaint.
Holzer made his first appearance in Denver federal court on Monday afternoon. A preliminary hearing and a detention hearing are scheduled for Thursday morning, according to US Attorney Jason R. Dunn.
The suspect, who does not yet have an attorney listed on his case, faces a maximum 20-year sentence if convicted of a hate crime, officials said.
How suspect came to FBI's attention
Domestic terrorism and hate crimes have become a growing concern for law enforcement in recent years as a number of high-profile attacks have left scores dead across the country. Eleven worshippers were killed in October last year when a white supremacist opened fire at the Tree of Life synagogue in Pittsburgh.
Holzer first came on the FBI's radar after a tip in late September regarding comments he'd made online "of concern indicating a possible threat to the community," FBI Denver Special Agent in Charge Dean Phillips said at a news conference.
"This case emphasizes our continued efforts to aggressively and promptly address threats to our community to include violence against places of worship. I cannot stress enough the importance of reporting threats in our neighborhoods," Phillips said.
Undercover FBI agents approached Holzer online soon after the tip and were interacting with him as he allegedly plotted the attack on Temple Emanuel. In a meeting with three undercover agents at a restaurant in Colorado Springs last month, Holzer proposed using Molotov cocktails to destroy the Pueblo synagogue, according to the complaint.
"I want something that tells them they are not welcome in this town," Holzer allegedly said. "Let's get that place off the map."
Authorities say they have video evidence
Holzer allegedly visited Temple Emanuel multiple times after he hatched his plot with the undercover agents. On October 19, Holzer sent one agent a video of himself walking around the exterior of the synagogue and "commenting on various features of the building," the complaint says.
On the night of November 1, Holzer donned a Nazi armband and drove with an undercover agent to a motel. There, he toasted a "move for our race" and commented that the pipe bombs, which authorities said had been prepared with simulated black powder at the FBI lab in Quantico, Virginia, looked "absolutely gorgeous," according to the complaint.
The FBI arrested him soon afterward and took him to a local police station, where he admitted to planning to blow up the synagogue, the complaint says.
Florida man wanted ISIS to bomb deans at colleges that suspended or expelled him, feds say
Police say man posed for sketch, then robbed artist – but left portrait behind
Pittsburgh Pirates pitcher Felipe Vázquez hit with more charges in alleged sex assault case
A runner who smacked a reporter's backside on air now faces charges
3 people arrested, charged with child pornography
Police believe drug deal gone wrong led to shooting of 17-year-old Dover HS student in York
He was abused as an 11-year-old boy. A new law in California is making it possible to sue more than 40 years later
Chambersburg man accused of sexually assaulting 9-year-old girl
Man accused of exposing himself near play area at York Chick-Fil-A
Murder charge dropped against one of two men allegedly involved in Regal Cinemas shooting on Dec. 2
Man accused of raping teen at park in Chambersburg
Safety Kameron Kelly waived by Steelers after he was arrested for alleged altercation with police


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How to Calm Your Bladder
Reduce that "gotta go" feeling with these four small lifestyle changes
By Beth Howard and Grandparents.com
(This article appeared previously on Grandparents.com)
Do you often feel the urge to pee and feel like you just can't hold it? Do you head to the bathroom far more than you think is normal? You may have overactive bladder (OAB), a condition that affects about 33 million Americans. With OAB, people experience sudden urges to urinate that they can't control, as well as the need to urinate frequently. People with OAB may also leak when they experience the urge.
Who Gets OAB?
According to the American Urological Association, up to 30 percent of men and 40 percent of women in the U.S. experience OAB symptoms. Women who have gone through menopause and men who have had prostate problems are at greater risk. And risk also increases with age.
In addition, some medications may cause or worsen urinary symptoms. These include blood pressure drugs — diuretics and a class called alpha blockers (such as Cardura and Minipress) — antidepressants and sleeping medications.
Doctors diagnose OAB based on a physical exam, symptoms of frequency and urgency, and other test results. Urinary frequency is defined as urinating at least eight times in a 24-hour period. You may also wake up to urinate two or three times a night. In about 40 percent of patients, OAB resolves on its own within a year, but most people suffer from symptoms for years.
Luckily there is a lot you can do to alleviate symptoms and overcome overactive bladder.
OAB Treatments
Lifestyle changes are a first step in overcoming overactive bladder. "What you eat, how you exercise, and how well you take care of yourself all factor in your overall bladder health and your success at alleviating overactive bladder symptoms," says Karen Sebastian, a physical therapist in Honolulu who specializes in bladder control issues. Here, a primer on four lifestyle practices that can help:
1. Change your diet What you eat and don't eat can affect your symptoms.
Reduce: Diuretics such as alcohol and caffeine. They stimulate muscles in the bladder, increasing urgency. Limit caffeine to 100 mg a day (about half a cup of coffee). Irritants, including acidic fruits and juices, carbonated beverages, spicy foods and artificial sweeteners like saccharine and aspartame should also be reduced.
Eat more: Fiber. "Because the bladder and rectum are in close proximity, being constipated puts constant pressure on the bladder," says Sebastian. In fact, women with OAB have higher rates of constipation than those without the condition. Load up on fruits, vegetables and whole grains.
2. Exercise While Kegels are the best exercise for women with urinary health (see how to do them below), for men and women, having a strong back and core are also important. "We're finding more and more in research that our muscles work together in groups," says Sebastian. "Your abs and adductors — hip muscles — attach directly to the pubic bone. And muscle fibers in the hip feed into the pelvic floor. They all help each other activate."
To ease bladder symptoms, try three moves once a day, while lying down on your back:
Inner thigh squeeze: With your knees bent and feet on the floor, place a pillow between your knees and lightly squeeze to work your adductors. Inhale, and as you exhale, clench your stomach muscles to tilt your pelvis up, while keeping your tailbone on the floor. (It should feel like you're pulling your pelvis towards your belly button.) Hold for 10 seconds while breathing normally. Relax completely. Repeat 10 times.
Hip flexors: Position yourself as you did for the "inner thigh squeeze" exercise, but without the pillow. Inhale, and as you exhale, clench your stomach muscles to tilt your pelvis up, while keeping your tailbone on the floor. Then lift up your hips to create a "bridge," and slowly open and close your knees 10 times. Lower your hips and relax completely. Repeat two or three times for a total of 20 to 30 hip rotations.
3. Meditate Some people find their bladder symptoms are eased when they tap into the mind-body connection. Women who listened to a 15-minute audio recording using deep breathing and visualization exercises twice a day for two weeks cut the average number of their urge incontinence episodes from 42 to 16 per week, Loyola University researchers have reported. The women also boosted their continuous "dry time" from 32 to 100 hours per week.
Try this deep-breathing exercise: Lie down and get comfortable. Allow yourself 10 undisturbed minutes. Take a few slow, deep breaths, allowing your belly to rise and fall. With each exhale let your muscles go, allowing yourself to "melt" into your supporting surface.
Concentrate on your breath: Inhale to a silent count of 3, allowing your stomach to rise, hold for a second at the top of your inhale, then slowly exhale to a silent count of 3. Repeat this pattern five times. Next, continue breathing slowly and deeply, but now to a count of 4. Progress to a count of 5, then 6. Re-focus on your surroundings. Slowly bring your attention back to the here and now.
4. Women: Do Kegel exercises Pelvic floor muscles support a woman's urethra and bladder, so keeping them strong is crucial to preventing leaks. Research shows that Kegel exercises, which involve contracting and releasing the pelvic floor muscles, can be very helpful. "Kegels are a good starting point for anyone with a bladder problem," says Dr. Lauri Romanzi, a uro-gynecologist in New York City.
Try this Kegel routine recommended by the American Urogynecologic Society (AUS), three times a day:
Start by pulling in and holding a pelvic muscle squeeze (as if you were trying to stop yourself from urinating) for three seconds and then relax for an equal amount of time. Do this for 10 repetitions.
Increase your contraction hold by one second each week until you are contracting for a 10-second squeeze.
Remember to rest and breathe between contractions.
When you start, do the exercises while lying down. As you get stronger; do an exercise set sitting and standing.
If self-help tactics aren't enough to get symptoms under control, your doctor may suggest bladder training, which involves scheduling times for urinating instead of waiting for the urge, for instance, and other behavioral measures. A medication that relaxes the bladder may also be prescribed. Together these measures often improve the problem — and your quality of life.
Beth Howard A former magazine editor, Beth Howard specializes in health and medicine. She also writes for U.S. News & World Report; Reader's Digest; O, The Oprah Magazine; The Washington Post; and The Wall Street Journal. She is based in Charlotte, N.C. Read More
By Grandparents.com
Grandparents.com is a lifestyle website, social media community & peer group that unites & connects America's 70 million Grandparents to the best information and premier products & services just for them. Our goal is to promote well-being and give timely information on what really matters to you, from health and money to family and relationships to travel and retirement.


Israeli police have said they have found evidence to recommend pursuing bribery and corruption charges against Prime Minister Benjamin Netanyahu and his wife.
Authorities claimed on Sunday that Netanyahu gave favourable contracts to Bezeq Telecom in exchange for better coverage of the prime minister and his wife on its news website.
They also recommended his wife, Sara Netanyahu, face charges of bribery, fraud, breach of trust and obstruction of evidence.


In Ayurveda I am a so called "tri dosha" (vata-pitta-kapha type), which is very rare. It gives me a very sturdy constitution and well balanced personality. People say that my presence makes them feel relaxed. I can also feel those different components inside of me and sometimes the fight between letting the mind or letting the heart decide. The last ten years I have been living in Berlin/Germany, however feeling home at many places in the world. I try to live a slow life, which does not necessarily mean going slower but making more intentional decisions about my life and what I want to do with it.
As my aunt and uncle lived in the country side, I think I spent most of my childhood there. Strolling through the woods & fields, swimming and hanging out with the local kids and of course ice skating in the winter. We also had some horses, dogs, geese and a sheep called Robert. I am still so grateful that I had an opportunity to experience this as a child.
"je ne regrette riens" – there is nothing I would change in life if I could. I believe that everything happens for a reason. We are living our lives forward and only understanding it in retrospect. Why some things have happened and others not at this point in time it might difficult to see but later on it makes sense. I have also learned to let go and trust the universe that all the things that are meant to will happen.
After working close to 20 years in Human Resources for big international companies, I decided to turn my passion for photography into a living, leaving an excellent career behind. Since earlier this year I work full time as a freelancer and I have never looked back a single day. I think how fortunate I am, being able to do what I love doing most; showing others the world through my eyes and telling stories with my images. Will I be doing this the next 10 years? I do not know and do not want to think about it but truly enjoying the moment and this experience.
Three is my lucky number so that is already good news!
A piece of amber – I fairly big piece of amber I found at the Baltic Sea on the morning of birthday. I just love looking at it.
A book with my favourite images printed on matte paper – some images feel particular dear to me like being my own children.
Coffee beans – I simply love that smell of fresh ground coffee, I close my eyes, pretending being back at Bondi Beach.
Officially it is the name of my portfolio page where I present my work as a photographer; sharing images of food, lifestyle and travel photography. Unofficially it is place I invite people to let their mind and eyes wander. Everything is kept very minimalistic in order not to distract the mind.
I really love a good flat white if one is available and in the morning before I go out for my run I usually enjoy an quick espresso. Nowadays the coffee blends seem to be very fruity which does not go well with a milky coffee. I prefer coffee tasting and smelling of chocolate, hazelnut or vanilla which seems to be the perfect fit with an Arabica blend.
Going for my morning run at 6am and having a quick shower. Making a flat white with freshly ground espresso beans, ideally a blend I just brought back from my travel. Then sneaking back to bed for half an hour enjoying my coffee, chatting with my friend Sabine in Australia and reading some of my favourite magazines.
Feeling all refreshed and ready for the day I would be having a photoshoot with one of my favourite customers or meeting friends. I would end my perfect day with a mediation session just before going to sleep. If I am lucky I get to spend a perfect day on Sundays every now and then.


Hi everyone!!! Today I celebrate my blog's 4th birthday! Thank you so much to all of my readers and to the publishers and authors who keep trusting me with their books for me to review. To all the new friends I've met during the book signings and other events, you guys are awesome! I can't thank you enough you are all amazing!
Well... in return, I have a present for my blog giveaway winner. I won't reveal my prizes yet though here's the clue, I have a box with me which will be filled with stuff it is book related of course, some will be random things that I think will be helpful for your reading time AND there will be a book which I won't tell what will it be.
Sorry international followers, this mystery box will be available for PHILIPPINES only. I promise I'll try to arrange a giveaway like this next time.


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Elizabeth Scopel Biography, Age, Net Worth, Movies
All you need to know about actor Elizabeth Scopel: Biography, Age, Net Worth, Movies, Wiki, House, Family and Career
Elizabeth Scopel: Profile Summary
Full Name Elizabeth Scopel
Date Of Birth 5 October 1992
Birthplace Huntingtown, Maryland, United States
Ethnicity Mixed
Sexuality: Queer
Height 5 Feet 4 Inches (1.63 M)
Weight 49 Kg (108 Lbs.)
Father's Name: Donald K. Scopel
Mother's Name: Anne Kathryn Scopel
Sibling(S): 2 (Andrew And Maggie)
Education: Vassar College
Elizabeth Scopel is an actor widely known as Sarah Fier, the character they played in the Fear Street trilogy on Netflix in 2021, they also played the part of Britt Mills in the TV show Chicago Med.
Elizabeth Scopel a well-known thespian from the United States of America has captivated the hearts of people around the world with their exceptional acting abilities. Making their professional acting debut in the television series Grave Secrets in 2017, have since appeared in a number of films and television shows, including FBI: Most Wanted (2020) and Half-Empty (2019).
Elizabeth Scopel: Net Worth
Elizabeth Scopel has an estimated net worth of $500,000 as of 2022.
Elizabeth Scopel: Age, Height and Weight
Elizabeth was born on 5 October 1992. They are 5 feet 4 inches (1.63 m) tall and weighs 49 kg (108 lbs.).
Elizabeth Scopel: Gender and Pronouns
Elizabeth is non-binary. The American actor uses pronouns like they/them, according to their Instagram profile.
Elizabeth Scopel: Career
Elizabeth Scopel made their debut onscreen appearance as a Coffee Shop Girl in the 2010 short film Local Hero. They made their television debut in the 2017 television series Grave Secrets, where they played the role Noni.
Their excellent performance gained them more opportunities in a number of films and television series. Their biggest breakthrough came in 2021, when they were cast in the Netflix series Fear Street: Part One – 1994, Fear Street: Part Two – 1978, and Fear Street: Part Three – 1666 as Sarah Fier. They collaborated on the series with other prominent actors and actresses such as Kiana Madeira, Olivia Scott Welch, and Benjamin Flores Jr.