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[Current aspects of the fecal flora of the newborn without antibiotherapy during the first 7 days of life: Enterobacteriaceae, enterococci, staphylococci].

Last years, il became obvious that the colonization pattern described in 1976-1978 was no more valid: early colonization by Enterobacteriaceae at the 2-3 rd day of life in all newborns, with constant presence of antibioresistant strainseven in non treated newborns. To establish the new pattern of colonization, the same quantitative method of dilution and culture on selective media was used daily from day 1 to day 7 (5 days only for M). The number of Enterobacteriaceae, enterococci and staphylococci was determined in the stools of 10 newborns in the Maternity unit (= M) (term 40 weeks +/- 1, birth weight 3,356 g +/- 383), 10 in the Premature nursery (= P) (term 34.9 weeks +/- 1, birth weight 2,457 g +/- 676), and 14 in the Neonatal intensive care unit (= R) (term 35.2 weeks +/- 3.8, birth weight 2,457 g +/- 763). The results establish that colonization by Enterobacteriaceae is no more constant at D3. It could be demonstrated only in 8/10 M, 1/10 P, and 6/14 R (statistically different - p < 0.01 - between M and P). At D5, 9/10 M, 5/10 P, 10/14 R, and at D7, 6/10 P and 10/14 R were colonized. Resistant Enterobacteriaceae (Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae) could be found in only 3/10 M, 4/10 P and 6/14 R. Enterococci could be found in 1 newborn M, 2 P and 7 newborns R. Staphylococci appeared earlier: all newborns M, P and R were colonized at D2, 4 and 5 respectively. These bacteria were coagulase negative, associated with Staphylococcus aureus in 3 P. Our hypothesis is that late colonization with Enterobacteriaceae and enterococci is due to the improvement of hygiene procedures and due to the decontaminating effect of antibiotics in other treated newborns (Enterobacteriaceae by 3 rd generation cephalosporin and enterococci by pharyngeal vancomycin).

Anti-Bacterial Agents↗

Inhibition of Salmonella typhimurium attachment to chicken cecal mucus by intestinal isolates of Enterobacteriaceae and lactobacilli.

The ability of selected strains of Enterobacteriaceae or lactobacilli isolated from the intestines of adult chickens to inhibit in vitro attachment of Salmonella typhimurium 3333/O to cecal mucus in the presence or absence of D-mannose was determined. Attachment in the absence of mannose was reduced by prior exposure of mucus to cultures of two isolates of Enterobacteriaceae, an Escherichia coli and a Hafnia alvei strain, but not to a third isolate, an Enterobacter agglomerans strain. Attachment of S. typhimurium was not inhibited when mannose was present in the blocking or attachment step. Formation of fimbriae by the two inhibitory Enterobacteriaceae strains and the S. typhimurium strain, as indicated by titers of mannose-sensitive hemagglutination of guinea pig erythrocytes was optimal in Z biphasic medium (consisting of tryptone, yeast extract, dextrose, and NaCl) incubated anaerobically at 42 C. Fimbriae of each of three strains prepared from these cultures also inhibited attachment. These are characteristics consistent with attachment and inhibition of attachment mediated by a mannose-sensitive adhesin associated with type 1 fimbriae on bacterial cells of Enterobacteriaceae strains. Attachment in the presence of mannose was significantly reduced by prior exposure of mucus to cultures of a Lactobacillus salivarius strain and a Lactobacillus delbrueckii delbrueckii strain but not to a strain of Lactobacillus for which the species had not been determined. Washed cells or spent culture supernatant fluid from brain-heart infusion broth, Z broth, or Z biphasic cultures of the inhibitory strains of lactobacilli incubated at 37 or 42 C inhibited this form of attachment. Of 27 intestinal isolates of Enterobacteriaceae and 21 of lactobacilli, the lactobacilli strains were generally more hydrophobic than the Enterobacteriaceae as determined by adherence to hexadecane. The lactobacilli isolates did not agglutinate guinea pig erythrocytes. The data suggest more than one mechanism for mediating attachment of inhibitory bacterial strains and for subsequent attachment of S. typhimurium.

Adhesins, Bacterial↗

Longitudinal trends in fluoroquinolone resistance among Enterobacteriaceae isolates from inpatients and outpatients, 1989-2000: differences in the emergence and epidemiology of resistance across organisms.

We conducted a 12-year study to identify and compare trends in annual prevalence of fluoroquinolone (FQ) resistance among Enterobacteriaceae isolates obtained from inpatients and outpatients in our health care system. A total of 46,070 clinical Enterobacteriaceae isolates underwent susceptibility testing. Although there were significant increases in inpatient FQ resistance for all Enterobacteriaceae, FQ resistance trends differed significantly across Enterobacteriaceae (P<.001). For isolates obtained from outpatients, only Escherichia coli and Proteus mirabilis demonstrated significant increases in FQ resistance (P<.001 for each). Trends in outpatient FQ resistance also differed significantly across Enterobacteriaceae (P<.001). There were significant differences between inpatient and outpatient FQ resistance trends for all Enterobacteriaceae except P. mirabilis and Enterobacter cloacae. Although hospital-wide use of certain antibiotics correlated significantly with inpatient FQ resistance, these correlations differed substantially across organisms. Efforts to elucidate the epidemiology of FQ resistance and identify targets for intervention must recognize and account for the variability of FQ resistance across organisms and clinical settings.

Anti-Infective Agents↗

Activity of ciprofloxacin (BAYo 9867) against Pseudomonas aeruginosa and ampicillin-resistant Enterobacteriaceae.

We studied the in vitro activity of ciprofloxacin against 570 strains of ampicillin-resistant Enterobacteriaceae and 286 Pseudomonas aeruginosa strains. 95.26% of the Enterobacteriaceae and 53.45% of the P. aeruginosa were inhibited by 0.1 mg/l of ciprofloxacin. 2 mg/l of ciprofloxacin inhibited all of the Enterobacteriaceae strains and 4 mg/l all of the P. aeruginosa. We compared the activity of ciprofloxacin with that of temocillin in the Enterobacteriaceae strains. In the P. aeruginosa strains, classified according to their susceptibility to carbenicillin and gentamicin, we compared the activity of ciprofloxacin with that of ceftazidime. In the strains studied, the in vitro activity of ciprofloxacin is superior to that of temocillin against the Enterobacteriaceae and to that of ceftazidime against the P. aeruginosa strains.

Ampicillin↗

Control of Enterobacteriaceae producing extended-spectrum beta-lactamase in intensive care units: rectal screening may not be needed in non-epidemic situations.

OBJECTIVE: To evaluate the usefulness of screening cultures in the control of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae in intensive care units (ICUs). DESIGN: A 4-year retrospective study. SETTING: Two adult ICUs of a university-affiliated public hospital in France. RESULTS: A total of 7,777 specimens were analyzed and 28 (0.97%) of 2,883 screened patients had a positive result on a screening test, among the 3,678 admitted patients. Thirteen of these 28 patients were only carriers; 4 were carriers and then were colonized or infected 2, 2, 3, and 8 days later, respectively; and 11 were colonized or infected before a screening test was positive. Cluster analysis showed that the occurrence of ESBL-producing Enterobacteriaceae cross-transmission within both ICUs was limited to 9 cases. Thus, most cases (19 of 28) were probably imported. Surveillance cultures failed to detect 9 of the 19 cases. CONCLUSION: The low prevalence of ESBL-producing Enterobacteriaceae carriers on admission (0.45%) and the relative ineffectiveness of our screening test to detect imported cases suggest that systematic detection of ESBL-producing Enterobacteriaceae in ICU patients is not cost-effective and that the use of clinical cultures may be sufficient to control ESBL-producing Enterobacteriaceae in non-epidemic situations.

Adult↗

Extended-spectrum beta-lactamase-producing Enterobacteriaceae strains in community-acquired bacteremia in Southern Israel.

BACKGROUND: In recent years, extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae have emerged in many hospitals worldwide. The increasing dissemination and long-term carriage of these organisms within the community carry tremendous implications on the empirical therapy of community-acquired infection. MATERIAL/METHODS: To evaluate the prevalence and clinical features of community-acquired bacteremia involving ESBL-producing Enterobacteriaceae in southern Israel (ESBL-P) we retrospectively studied all Enterobacteriaceae bacteremias during an 8-month period in the Negev region using medical and laboratory records. Antibiotic susceptibility was determined using the disk-diffusion method. ESBL production was determined using an E-test ESBL strip. Cases involving ESBL-P were compared to those involving non-producing strains (ESBL-NP) using the chi-square test. RESULTS: In all, 187 Enterobacteriaceae bacteremias were detected, of which 119 were community-acquired (63.6%). ESBL-P were found in six cases (5%) which are described herein in greater detail. Patients with ESBL-P bacteremia were older, and were more likely to stay in the intensive-care unit. Urinary catheterization and bed-ridden conditions were significant risk factors for ESBL-P. ESBL-P strains were significantly resistant to nearly all antibiotic agents except for imipienem and piperacillin-tazobactam as opposing to ESBL-NP. Patients with ESBL-P bacteremia were more likely to suffer from complications and had a higher mortality. CONCLUSIONS: This paper is the first to describe community-acquired Enterobacteriaceae bacteremia involving ESBL-P strains in Israel. Although the exact prevalence of these organisms in Israel is currently unknown, our findings suggest that ESBL-producers have already begun to disseminate in our community.

Adult↗

[Patterns of susceptibility to antibiotics of Enterobacteriaceae causing intra-abdominal infection in Spain: SMART 2003 study outcomes].

SMART (Study for Monitoring Antimicrobial Resistance Trends) is an ongoing global antimicrobial surveillance program focused on clinical isolates from intra-abdominal infections. The objective of this subanalysis was to assess antimicrobial susceptibility patterns among Entero-bacteriaceae recovered at 13 participating Spanish sites during 2003. Antimicrobial susceptibility testing was performed using broth microdilution techniques according to the CLSI (formerly NCCLS) guidelines for MIC testing. The presence of extended-spectrum beta-lactamases (ESBL) was confirmed in isolates with a MIC of ceftriaxone, ceftazidime, or cefepime>or=2 mg/l by comparing cefepime MICs with and with-out clavulanate. A total of 981 Enterobacteriaceae recovered from 840 patients were tested, of which 398 (41%) were community-acquired. Escherichia coli was the most common isolate (571 isolates; 58%), followed by Klebsiella spp. (153; 16% Enterobacter spp. (97; 10%), and Proteus spp. (63; 6%). A total of 191 isolates (19%) from 176 patients produced inducible beta-lactamases. The carbapenems and amikacin were the most consistently active agents against the Enterobacteriaceae (susceptibility>or=99%). Resistance rates for ceftazidime, cipro-floxacin, and levofloxacin exceeded 10%. ESBLs were detected phenotypically in 61 (6%) isolates, being the most common E. coli (61%), Klebsiella spp. (20%), and Enterobacter spp. (8%). Antimicrobial resistance among Enterobacteriaceae isolated from intra-abdominal infections is a problem in Spain. A significant proportion of inducible beta-lactamase and ESBL-producing Enterobacteriaceae causing intra-abdominal infection were acquired in the community. The carbapenems ertapenem, imipenem and meropenem and the aminoglycoside amikacin were highly active in vitro against Enterobacteriaceae isolated from intra-abdominal sites, including ESBL-producing organisms.

Abdomen↗

Enterobacteriaceae peritonitis complicating peritoneal dialysis: a review of 210 consecutive cases.

Enterobacteriaceae peritonitis is a serious complication in peritoneal dialysis (PD), but the clinical course of PD-related Enterobacteriaceae peritonitis remains unclear. We reviewed all Enterobacteriaceae peritonitis in our dialysis unit from 1995 to 2004. During this period, there were 1748 episodes of peritonitis recorded; 210 episodes (12.0%) in 123 patients were caused by Enterobacteriaceae. The most common species was Escherichia coli, accounting for 111 episodes. The primary response rate was 84.8% and complete cure rate was 58.1%. The presence of exit site infection was associated with a lower complete cure rate (43.2 versus 61.3%, P = 0.034). A total of 82 episodes (39.0%) did not respond to single antibiotic treatment despite sensitivity in vitro, and a second antibiotic was added. Patients treated with two antibiotics had a marginally lower risk of relapse and recurrence than those with one antibiotic (21.4 versus 36.1%, P = 0.051). The episodes that had recent antibiotic therapy had a marginally lower complete cure rate (49.3 versus 62.8%, P = 0.06). There was a gradual increase in the prevalence of resistance to several commonly used antibiotics over the years. Recent antibiotic therapy was associated with resistance to cefotaxime, ceftazidime, cefoperazone/sulbactam, and piperacillin/tazobactam. We conclude that Enterobacteriaceae peritonitis is a serious complication of PD. Recent antibiotic therapy is the major risk factor of antibiotic resistance. Exit site infection, and probably recent antibiotic therapy, is associated with poor therapeutic response. Contrary to the current recommendation, treatment with two antibiotics may reduce the risk of relapse and recurrence.

Aged↗

Caseinolysis in cheese by Enterobacteriaceae strains of dairy origin.

AIMS: To study the effect of Enterobacteriaceae strains of dairy origin on caseins under cheese manufacture and ripening conditions. METHODS AND RESULTS: Strains belonging to the genera Enterobacter, Escherichia, Hafnia and Serratia were isolated from fresh raw milk cheeses. Residual caseins in cheeses made from milk individually inoculated with 10 strains of Enterobacteriaceae were determined by capillary electrophoresis. Hierarchical cluster analysis of strains based on data of residual caseins grouped together strains from the same genus, excepting Hafnia strains, which were separated into two groups. Serratia was the most proteolytic genus in our study. Preferences for degradation of casein fractions differed among the four genera studied. CONCLUSIONS: Enterobacteriaceae strains posses proteolytic systems active on all casein fractions under cheese manufacture and ripening conditions. The effects on caseins were similar for strains belonging to the same genus. SIGNIFICANCE AND IMPACT OF THE STUDY: The presence of Enterobacteriaceae in cheeses may affect proteolysis during ripening. Assays of Enterobacteriaceae proteolytic activity on milk agar plates may underestimate their caseinolytic activity in cheese.

Animals↗

Rapid identification of Enterobacteriaceae using a novel 23S rRNA-targeted oligonucleotide probe.

The aim of this study was to rapidly identify bacteria of the family of Enterobacteriaceae using fluorescent in situ hybridization (FISH). A comparative sequence analysis was carried out and a 23S rRNA signature sequence for Enterobacteriaceae was identified. A 23S rRNA-targeted oligonucleotide probe (EBAC1790) was constructed and subsequently tested against 40 reference strains. Nearly all of the Enterobacteriaceae used in this study yielded positive results with EBAC1790, except for Edwardsiella tarda (ATCC 15947). None of the non-Enterobacteriaceae reference strains gave positive signals with the probe. The possibility of a rapid detection of Enterobacteriaceae in groundwater was demonstrated using colony hybridization.

Enterobacteriaceae↗

Molecular epidemiology of extended-spectrum beta-lactamase-producing Enterobacteriaceae isolated from environmental and clinical specimens in a cardiac surgery intensive care unit.

OBJECTIVES: To investigate environmental contamination by extended-spectrum beta-lactamase-producing Enterobacteriaceae and to perform a comparative molecular analysis of clinical and environmental strains. SETTING: A 17-bed cardiac surgery intensive care unit of a 480-bed university teaching hospital. METHODS: Following an outbreak of extended-spectrum beta-lactamase-producing Enterobacteriaceae, an environmental survey revealed extensive contamination of the environment (particularly faucets, sink drains, and the joints of the countertops) by extended-spectrum beta-lactamase-producing Enterobacteriaceae. Environmental strains were compared with clinical strains by pulsed-field gel electrophoresis and randomly amplified polymorphic DNA. RESULTS: A total of 62 environmental strains belonging to 4 species of extended-spectrum beta-lactamase-producing Enterobacteriaceae were analyzed and compared with 43 clinical strains obtained from 34 patients. Comparative molecular analysis revealed 4 identical or closely related patterns (3 from Klebsiella oxytoca and 1 from Enterobacter cloacae) between environmental and clinical strains. CONCLUSIONS: Moist surfaces may serve as sources of multiply resistant Enterobacteriaceae in the intensive care unit. Identification and disinfection of such sources may therefore be helpful in prevention and control of outbreaks.

Cardiac Surgical Procedures↗

The enumeration of thermotrophic types amongst the Enterobacteriaceae colonizing perishable foods.

A total of 41 pure cultures of Enterobacteriaceae, comprising 32 thermotrophic and nine psychrotrophic strains, pathogens or marker organisms, were examined for numbers of colony forming units obtained at 37 degrees and 42.5 degrees C (thermotrophs) and 30 degrees C (psychrotrophs), when surface-plated on a rich infusion agar and violet red bile agar. In addition 42 food and water samples, collected in a rural area of the Philippines, were examined by surface inoculating violet red bile AIPC (agar immersion plating and contact; 'dip') slides and incubating at 37 degrees and 42.5 degrees C. At 42.5 degrees C there was almost total recovery of the thermotrophic Enterobacteriaceae, whereas the psychrotrophic strains were completely suppressed. At 37 degrees C the psychrotrophs were only slightly inhibited. The Philippine foods, predominantly cooked meals, milk and drinking water, appeared to be significantly colonized by thermotrophic Enterobacteriaceae. It is concluded that incubation at 42.5 degrees C satisfactorily selects enteropathogenic and other enteric Enterobacteriaceae while suppressing the psychrotrophic types which are mainly of vegetable origin. It is emphasized that, regardless of the temperature used, a resuscitation procedure for Enterobacteriaceae populations that have incurred sublethal injury in food has to precede counts on or in the usual selective media.

Animals↗

Oral Enterobacteriaceae in patients with HIV infection.

160 oral swabs of 149 HIV-infected patients and 168 swabs of 166 controls were cultured to detect a possible colonization of the oral cavity with yeasts and Enterobacteriaceae. In 5.0% of the HIV-infected patients and in 4.8% of the controls different species of Enterobacteriaceae were found. Yeasts were always associated with Enterobacteriaceae in the group of HIV-infected patients. Clinically, different oral lesions were seen in the study group. A correlation between the nature of these lesions and the presence of Enterobacteriaceae could not be found. A possible pathogenic role of the Enterobacteriaceae associated with these oral lesions is yet uncertain and remains to be established by further studies.

Acquired Immunodeficiency Syndrome↗

Trends in antimicrobial susceptibilities among Enterobacteriaceae isolated from hospitalized patients in the United States from 1998 to 2001.

Longitudinal surveillance of Enterobacteriaceae for antimicrobial susceptibility is important because species of this family are among the most significant and prevalent human pathogens. To estimate rates of in vitro antimicrobial susceptibility among hospitalized patients in the United States, data from The Surveillance Network were studied for 14 agents tested against 10 species of Enterobacteriaceae (n = 384,279) isolated from intensive-care-unit (ICU) patients and non-ICU inpatients from 1998 to 2001. Cumulative susceptibility (percent) data for all species of Enterobacteriaceae isolated from ICU patients and non-ICU inpatients, respectively, were ranked as follows: ampicillin-sulbactam (45.5 and 57.2) << ticarcillin-clavulanate (74.8 and 83.5) < trimethoprim-sulfamethoxazole (87.0 and 84.5) congruent with cefotaxime (82.9 and 92.6) = ceftazidime (82.3 and 91.0) = ceftriaxone (86.5 and 93.9) = piperacillin-tazobactam (83.5 and 90.5) < levofloxacin (89.3 and 90.6) = ciprofloxacin (91.0 and 91.7) < gentamicin (91.8 and 94.3) < cefepime (95.0 and 97.9) < amikacin (98.5 and 99.2) < imipenem (100 and 100) = meropenem (100 and 100). Of those agents studied only susceptibilities to ciprofloxacin (94 to 89%) and levofloxacin (93 to 89%) decreased in a stepwise manner from 1998 to 2001. Decreased fluoroquinolone susceptibility was most pronounced for Escherichia coli, Proteus mirabilis, and Enterobacter cloacae. For all species of Enterobacteriaceae, trimethoprim-sulfamethoxazole resistance was more commonly observed in isolates with a single-drug resistance phenotype while gentamicin and fluoroquinolone resistances were more common in isolates resistant to at least one additional class of antimicrobial agent. Ongoing surveillance of Enterobacteriaceae will be particularly important to monitor changes in fluoroquinolone susceptibility, as well as changes in the prevalence of isolates resistant to multiple classes of antimicrobial agents.

Ampicillin↗

Accurate automated identification of selected Enterobacteriaceae at four hours.

The Enterobacteriaceae Biochemical Card of the AutoMicrobic system Vitek Systems, Inc., Hazelwood, Mo.) provides completely automated identification of members of this family within an 8-h test period. Identification of 776 clinical and stock isolates to species level by the Enterobacteriaceae Biochemical Card under routine operating conditions correlated at 96% with our present 18- to 24-h methods of identification. Further, utilizing a special program, we investigated presumptive identification of certain organisms within 4-h--an interval that provides greater practical clinical usefulness on a real-time rapid basis. In a single year, 1978, 97% of 23,464 Enterobacteriaceae isolated in our diagnostic laboratory belonged to 11 species of six genera. Our results suggest that, by limiting the number of the identified Enterobacteriaceae that could be actually presumptively reported to those 11 species of six genera. Our results suggest that, by limiting the number of the identified Enterobacteriaceae that could be actually presumptively reported to those 11 species with the highest frequency of occurrence, we could have correctly identified and presumptively reported 83% of these to genus or species after only 4 h. Approximately 2% of the isolates would have been presumptively identified and reported incorrectly, whereas the identification of the remaining 15% would not have been reported before the completed 8-h incubation period.

Autoanalysis↗

Clinical laboratory evaluation of the Auto-Microbic system for rapid identification of Enterobacteriaceae.

The capability of the Auto-Microbic system (Vitek Systems, Inc., Hazelwood, Mo.) has been expanded to identify members of the family Enterobacteriaceae with the use of a sealed, disposable accessory card (the Enterobacteriaceae Biochemical Card) containing 26 biochemical tests. To judge the accuracy of the AutoMicrobic system's identification in a hospital laboratory, 933 Enterobacteriaceae isolates were studied. The AutoMicrobic system provided the correct identification for 905 of the isolates (97%). For Enterobacteriaceae excluding Escherichia coli (533 isolates not E. coli), 506 isolates were correctly identified (94.9%). The AutoMicrobic system integrates well into the clinical microbiology laboratory, providing a simple, accurate, and efficient automated system for the rapid identification of Enterobacteriaceae.

Bacteriological Techniques↗

Salmonella and other Enterobacteriaceae found in commercial poultry feed.

Poultry feed (mash and pelleted) and meat and bone meal samples were collected from commercial mills. All samples were analyzed for Enterobacteriaceae count (ENT) and Salmonella. The genus and species of the various Enterobacteriaceae present were also determined. The average ENT for mash, pelleted, and meal samples was log 4.1, .8, and 1.8/g, respectively. Enterobacteriaceae were present in 100, 60, and 92% and Salmonella in 58, 0, and 92% of the mash, pelleted, and meal samples, respectively. Overall, the Enterobacteriaceae most frequently isolated from all samples were Enterobacter agglomerans, Enterobacter cloacae, and Klebsiella pneumoniae. Although no Salmonella were found in the pelleted samples, the presence of other Enterobacteriaceae suggests that commercial pelleting may not totally destroy Salmonella since their heat resistance is similar to the other organisms found.

Animal Feed↗

Effects of epiphytic Enterobacteriaceae and pseudomonads on the growth of Listeria monocytogenes in model media.

Four Enterobacteriaceae (Enterobacter agglomerans and Rhanella aquatilis) and six pseudomonads (Pseudomonas fluorescens, Pseudomonas chlororaphis, Pseudomonas putida) isolated from minimally processed green endive were coinoculated at 10 degrees C with Listeria monocytogenes in a minimal medium. Pseudomonads did not modify the growth of L. monocytogenes, whereas Enterobacteriaceae reduced its maximal population by 2 to 3 log CFU/ml. The same effect was observed in a diluted yeast extract medium supplemented with amino acids and glucose, in which L. monocytogenes grown alone reached 10(9) to 10(10) CFU/ml. In the same diluted yeast extract medium, not supplemented with glucose and amino acids, the maximal population of L. monocytogenes in the presence of both Enterobacteriaceae and pseudomonads was only slightly reduced (less than 0.5 log CFU/ml). Culture filtrates of the Enterobacteriaceae had no inhibitory activity on L. monocytogenes. The effect of the Enterobacteriaceae on L. monocytogenes growth was presumably due to a competition for glucose and/or amino acids.

Amino Acids↗