Some further observations on synergism between chloramphenicol and polymyxin B in relation toSalmonella bacteria

Summary The synergism between chloramphenicol and polymyxin B sulphate, earlier noticed using a replica technique, has been reexamined applying quantitative methods. Vital count experiments in suspensions ofSalmonella typhimurium in broth have revealed that addition of bacteriostatic concentrations of chloramphenicol in a range of 5–100 μg/ml to bacteriostatic concentrations of polymyxin (0.1–0.9 μg/ml) produce a strong bactericidal effect (synergism). Combinations of bactericidal concentrations of polymyxin (1 μg/ml and higher) with chloramphenicol also exert a very high degree of bactericidal activity, though no more than with polymyxin alone (no synergism). Chloramphenicol therapy sustained with polymyxin, may be of therapeutic value in the eradication of stubborn enteralSalmonella infections. With the technical assistance of MissM. J. Wisse.     

Detection of Salmonella enterica serovar Typhimurium by selective amplification of fliC, fljB, iroB, invA, rfbJ, STM2755, STM4497 genes by polymerase chain reaction in a monoplex and multiplex format

The aim of the work was to specifically differentiate S. typhimurium from other closely related Salmonella serovars by monoplex or multiplex PCR and to detect it from water and food samples. Genes targeted were invA, iroB, STM4497, STM2755, fliC, fljB and rfbJ and evaluated on 58 Salmonella standard serovars/strains including 9 S. typhimurium strains, 7 suspected Salmonella isolates and 8 other organisms as negative controls. Both invA and iroB showed a uniform amplification with all serovars of S. enterica group. STM2755 and STM4497 gene based PCR’s specifically exhibited amplification in all the nine confirmed S. typhimurium strains. The rfbJ PCR produced amplification with confirmed S. typhimurium strains, in addition showed reaction with S. abony. Both STM4497, STM2755 PCR’s and rfbJ could identify two of the seven biochemically suspected Salmonella isolates that were later confirmed to be S. typhimurium on the basis of sequence data. PCR for fliC genes had amplification exhibited by a large number of serovars of the S. enterica group, including S. typhimurium strains but not to S. brunei, S. newporti, S. abony and S. weltevreden. fljB was detected in all strains of S. enterica and E. coli with the exception of S. typhi. fljB and fliC were amplified in 6/7 and 5/7 presumptive Salmonella isolates. The same PCR’s were converted into two multiplex formats for simultaneous identification of the Salmonella genus, S. enterica group and S. typhimurium as a species. The first multiplex set comprised on invA, iroB, STM4497, STM2755 and the IAC. The second multiplex set comprised of invA, iroB, fljB, fliC, rfbJ along with IAC. The detection limit for S. typhimurium in the two multiplex PCR sets was in the range of 350–400 cfu/PCR reaction and that of DNA around 2 pg. The multiplex PCR (format 1) was first evaluated on spiked water, chicken and mutton samples and the detection limit for S. typhimurium was in the range of 100 cfu/100 ml, <60 and <50 cfu/gm, respectively. Further evaluation of multiplex PCR (format 1) was undertaken on 50 natural samples of chicken, eggs, litter, soil etc. and the comparison done with conventional culture isolation and identification procedure. The multiplex PCR could identify the presence of Salmonellla in three samples and the same three samples also yielded Salmonella by the conventional method. Therefore, the presently described multiplex PCR can serve as an alternative to the tedious time-consuming procedure of Salmonella culture and identification in food safety laboratories.     

Bioconversion of L-tyrosine to L-DOPA by a novel bacterium Bacillus sp. JPJ

l-DOPA is an amino acid derivative and most potent drug used against Parkinson’s disease, generally obtained from Mucuna pruriens seeds. In present communication, we have studied the in vitro production of l-DOPA from l-tyrosine by novel bacterium Bacillus sp. JPJ. This bacterium produced 99.4% of l-DOPA from l-tyrosine in buffer (pH 8) containing 1 mg ml⁻¹ cell mass incubated at 40°C for 60 min. The combination of CuSO₄ and l-ascorbic acid showed the inducing effect at concentrations of 0.06 and 0.04 mg ml⁻¹, respectively. The activated charcoal 2 mg ml⁻¹ was essential for maximum bioconversion of l-tyrosine to l-DOPA and the crude tyrosinase activity was 2.7 U mg⁻¹ of tyrosinase. Kinetic studies showed significant values of Y _p/s (0.994), Q _s (0.500) and q _s (0.994) after optimization of the process. The production of l-DOPA was confirmed by analytical techniques such as HPTLC, HPLC and GC–MS. This is the first report on rapid and efficient production of l-DOPA from l-tyrosine by bacterial source which is more effective than the plant, fungal and yeast systems.     

Interaction between tobramycin and CSA-13 on clinical isolates of Pseudomonas aeruginosa in a model of young and mature biofilms

The bactericidal activity of a cholic acid antimicrobial derivative, CSA-13, was tested against eight strains of Pseudomonas aeruginosa (both reference and clinical strains) and compared with the response to tobramycin. In planktonic cultures, the minimal inhibitory and minimal bactericidal concentrations of CSA-13 and tobramycin were in the 1–25 mg/L range except for one mucoid clinical strain which was much less sensitive to tobramycin (minimal bactericidal concentration, 65–125 mg/L). In young (24 h) biofilms, the sensitivity to CSA-13 was reduced (half-maximal concentration CSA-13 averaged 88 mg/L) and varied among the eight strains. The sensitivity to tobramycin was also very variable among the strains and some were fully resistant to the aminoglycoside. The combination of tobramycin with CSA-13 was synergistic in five strains. Only one strain showed antagonism between the two drugs at low concentrations of CSA-13. One reference and five clinical strains were tested in mature (12 days) biofilms. The effect of CSA-13 was delayed, some strains requiring 9 days exposure to the drug to observe a bactericidal effect. All the strains were tolerant to tobramycin but the addition of CSA-13 with tobramycin was synergistic in three strains. CSA-13 permeabilized the outer membrane of the bacteria (half-maximal concentration, 4.4 mg/L). At concentrations higher than 20 mg/L, it also permeabilized the plasma membrane of human umbilical vein endothelial cells. In conclusion, CSA-13 has bactericidal activity against P. aeruginosa even in mature biofilms and cationic steroid antibiotics can thus be considered as potential candidates for the treatment of chronic pulmonary infections of patients with cystic fibrosis. Considering its interaction with the plasma membrane of eukaryotic cells, less toxic derivatives of CSA-13 should be developed.     

Sialic Acid-Containing Lipopolysaccharides of Salmonella O48 Strains—Potential Role in Camouflage and Susceptibility to the Bactericidal Effect of Normal Human Serum

Sialic acid (N-acetylneuraminic acid, NeuAc) plays an essential role in protecting gram-negative bacteria against the bactericidal activity of serum and may contribute to the pathogenicity of bacteria by mimicking epitopes that resemble host tissue components (molecular mimicry). The role of sialic acid (NeuAc)-containing lipopolysaccharides (LPS) of Salmonella O48 strains in the complement activation of normal human serum (NHS) was investigated. NeuAc-containing lipooligosaccharides cause a downregulation of complement activation and may serve to camouflage the bacterial surface from the immunological response of the host. Serotype O48 Salmonella strains have the O-antigen structure containing NeuAc while its serovars differ in outer membrane protein composition. In this study, the mechanisms of complement activation responsible for killing Salmonella O48 serum-sensitive rods by NHS were established. Four of such mechanisms involving pathways, which are important in the bactericidal mechanism of complement activation, were distinguished: only the classical/lectin pathways, independent activation of the classical/lectin or alternative pathway, parallel activation of the classical/lectin and alternative pathways, and only the alternative pathway important in the bactericidal action of human serum. To further study the role of NeuAc, its content in bacterial cells was determined by gas-liquid chromatography-mass spectrometry in relation to 3-deoxy-D-manno-2-octulosonic acid (Kdo), an inherent constituent of LPS. The results indicate that neither the presence of sialic acid in LPS nor the length of the O-specific part of LPS containing NeuAc plays a decisive role in determining bacterial resistance to the bactericidal activity of complement and that the presence of sialic acid in the structure of LPS is not sufficient to block the activation of the alternative pathway of complement. We observed that for three strains with a very high NeuAc/Kdo ratio the alternative pathways were decisive in the bactericidal action of human serum. The results indicated that those strains are not capable of inhibiting the alternative pathway very effectively. As the pathogenicity of most Salmonella serotypes remains undefined, research into the interactions between these bacterial cells and host organisms is indispensable.     

De la Sensibilitéin vitro de Quelques Microorganismes D'origine Animale à la Viomycine

Summary The results of the study of the action of Viomycin on several microorganisms of animal origin isolated in the Belgian Congo and in the Ruanda-Urundi proved that the majority of the strains ofB. anthracis were inhibited by this antibiotic. All the other strains under study were not significantly affected by Viomycin with the exception of some strains ofSalmonella.      

Antigenic Determinants of Salmonella for Production of “Clearance” Factor in Mouse Typhoid

The “clearance” factor produced in the peritoneal cavity of mice immunized with killed vaccines prepared from Salmonella typhimurium or S. enteritidis was identified as the specific antibodies elicited by the O side chain of the cell wall polysaccharides in the organisms used as immunogens. After immunization of mice with vaccines prepared from virulent Salmonella strains, complement-dependent antibacterial antibodies in the serum and “clearance” factors in the peritoneal cavity were found to appear coincidentally, to last for more than one year, and to have the same specificity against the virulent bacterial strains. The relationship between the complement-dependent antibacterial antibodies and “clearance” factor, and the mechanisms of bactericidal action of these antibacterial agents in experimental typhoid were discussed.     

Presence of polar type granulations in E. coli bacterial cells. Coli and B. subtilis under the influence of detergents.

Résumé L’action de détersifs des groupes anioniques, cationiques et nonioniques à la concentration d’ 1 g 0/00, permet, dans certains cas, la mise en évidence de granulations dans les cellules bactériennes d’E. coli et duB. subtilis. Ces granulations se voient sans coloration, mais elles sont généralement plus visibles au contraste de phase ou par l’emploi de colorants basiques tels que fuchsine basique, bleu de Nil, vert Janus et brun Bismarck. Elles peuvent également être vues par l’action du bleu de Nil en l’absence de détersifs. Ces granulations sont du type polaire et peuvent être assimilées aux granulations polaires deBondarenko-Zozulina (6). Elles résistent à l’action de ribonucléase, de la désoxyribonucléase et de l’hyaluronidase. L’amylase seule fait dispara?tre les granulations observées dans les cellules duB. subtilis, tandis qu’il faut l’action simultanée et prolongée d’un mélange de désoxyribonucléase, ribonucléase, hyaluronidase et amylase pour faire dispara?tre la plupart des granulations d’E. coli.     

Reptiles as a Source of Salmonella O48��Clinically Important Bacteria for Children: The Relationship Between Resistance to Normal Cord Serum and Outer Membrane Protein Patterns

Bacteria of the Salmonella O48 somatic antigen group are clinically important strains causing intestinal dysfunction and diarrhoea, especially in children. The susceptibility of Salmonella O48 strains containing sialic acid (N-acetylneuraminic acid (NeuAc)) in lipopolysaccharide (LPS) to the bactericidal action of normal cord serum (NCS) was determined. The authors' previous results published in Microbial Ecology in 2010 indicated that neither the presence of NeuAc in LPS nor the length of the O-specific part of LPS containing NeuAc plays a decisive role in determining bacterial resistance to the bactericidal activity of normal human serum (NHS), and that the presence of NeuAc in the LPS structure is not sufficient to block the activation of the alternative pathway of complement in NHS. The current results showed that the tested strains showed various sensitivities also to the bactericidal action of NCS. The authors postulate that the presence of certain outer membrane proteins (OMPs) are characteristic of the resistant and sensitive phenotypes of Salmonella O48 strains. To establish a possible relationship between resistance to NCS and OMPs band patterns, ten Salmonella O48 strains were studied as follows: susceptibility to the bactericidal effect of NCS, the mechanisms of NCS activation and OMP band patterns obtained by sodium dodecyl sulphate-polyacrylamide gel electrophoresis.     

Killing of Gram-Negative Bacteria with Normal Human Serum and Normal Bovine Serum: Use of Lysozyme and Complement Proteins in the Death of Salmonella Strains O48

Serum is an environment in which bacterial cells should not exist. The serum complement system provides innate defense against microbial infections. It consists of at least 35 proteins, mostly in pre-activated enzymatic forms. The activation of complement is achieved through three major pathways: the classical, alternative, and lectin. Lysozyme, widely present in body fluids, catalyzes the hydrolysis of β 1,4 linkage between N-acetyloglucosamine and N-acetylmuramic acid in the bacterial cell wall and cooperates with the complement system in the bactericidal action of serum. In this study, ten strains of serotype O48 Salmonella, mainly associated with warm-blooded vertebrates and clinically important causing diarrhea in infants and children, were tested. The results demonstrated that the most efficient killing of Salmonella O48 occurred when all the components of normal bovine serum (NBS) and normal human serum (NHS) cooperated. To prove the role of lysozyme in the bactericidal activity of bovine and human serum, the method of serum adsorption onto bentonite (montmorillonite, MMT) was used. In order to investigate structural transitions accompanying the adsorption of serum components, we applied X-ray diffraction methods. The results of this investigation suggested that apart from lysozyme, other proteins (as, e.g., C3 protein or IgG immunoglobulin) were adsorbed on MMT particles. It was also shown that Ca²⁺ cations can be adsorbed on bentonite. This may explain the different sensitivities of the serovars belonging to the same O48 Salmonella serotype to NBS and NHS devoid of lysozyme.     

The estimation of a bivariate fitness function from two samples taken from a population

Summary The fitness of animals subjected to natural selection can be defined as the probability of surviving selection for a given interval of time, or some convenient multiple of this. If the fitness is related to some measurable variablesX, Y, Z,… then the relationship is expressed mathematically in the fitness functionw(x, y, z,…) and this function can be estimated by comparing the joint distribution ofX, Y, Z,… in samples taken before and after selection. In an earlier paper (Manly, 1975) the problems involved in estimating a fitness function of one variable were discussed. In the present paper various methods for estimating a bivariate fitness function are proposed and compared on some semiartificial sample data. It is concluded that either a generalized version ofO’Donald’s (1968) method of moments or a weighted multiple regression method will be most satisfactory. Alternative methods involving assumptions of normality will need to be used with great care.     

A study of the influence of polymyxine combined with chloramphenicol onSalmonella in vitro and in vivo in carriers ofS. paratyphi B

Summary A study was made of the effect of polymyxine and of chloramphenicol on Salmonellae in vitro, and also in vivo in a number of carriers ofS. paratyphi B. The findings of other investigators concerning synergistic action of polymyxine and chloramphenicol in vitro could be confirmed. Excreters who had had a course of treatment with polymyxine and chloramphenicol in most of the cases were still found positive one or more times forS. paratyphi B in the feces. Most excreters became negative spontaneously. The question whether combined antibiotic treatment is indicated for chronic excreters ofSalmonella is not answered. The differences between the results of in vitro and in vivo experiments, and the divergences in the results obtained after in vivo application by these and other authors are possibly to be atributed to an insufficient or absent penetration of polymyxine into the biliary ducts and the gall bladder.     

Chemical Communication of Antibiotic Resistance by a Highly Resistant Subpopulation of Bacterial Cells

The overall antibiotic resistance of a bacterial population results from the combination of a wide range of susceptibilities displayed by subsets of bacterial cells. Bacterial heteroresistance to antibiotics has been documented for several opportunistic Gram-negative bacteria, but the mechanism of heteroresistance is unclear. We use Burkholderia cenocepacia as a model opportunistic bacterium to investigate the implications of heterogeneity in the response to the antimicrobial peptide polymyxin B (PmB) and also other bactericidal antibiotics. Here, we report that B. cenocepacia is heteroresistant to PmB. Population analysis profiling also identified B. cenocepacia subpopulations arising from a seemingly homogenous culture that are resistant to higher levels of polymyxin B than the rest of the cells in the culture, and can protect the more sensitive cells from killing, as well as sensitive bacteria from other species, such as Pseudomonas aeruginosa and Escherichia coli. Communication of resistance depended on upregulation of putrescine synthesis and YceI, a widely conserved low-molecular weight secreted protein. Deletion of genes for the synthesis of putrescine and YceI abrogate protection, while pharmacologic inhibition of putrescine synthesis reduced resistance to polymyxin B. Polyamines and YceI were also required for heteroresistance of B. cenocepacia to various bactericidal antibiotics. We propose that putrescine and YceI resemble "danger" infochemicals whose increased production by a bacterial subpopulation, becoming more resistant to bactericidal antibiotics, communicates higher level of resistance to more sensitive members of the population of the same or different species.     

Rôles présumés de la membrane embryonnaire d’un parasite ovo-larvaire dePyralidae phanerotoma flavitestace [Hym.: Braconidae]

Résumé Après l’éclosion de la larve du parasitePhanerotoma flavitestacea Fisch., la membrane embryonnaire persiste pendant 3,5 jours. Différents r?les de cette membrane sont envisagés et discutés. Trois d’entre eux sont retenus (alimentation et protection de la larve parasite et action sur le développement ultérieur de l’h?te). Ce choix est basé sur nos propres observations biologiques et physiologiques et celles de divers auteurs concernant d’autres insectes entomophages.

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Summary After the hatching of the parasitic larva,Phanerotoma flavitestacea Fisch., the embryonic membrane persists around it during three days and a half. The different hypothetical roles of this membrane are discussed. Three are retained, on the base of our biological and physiological observations interesting this larva or different authors’ observations concerning other entomophagous insects. These roles are alimentary and protective functions for the parasite and action on the host physiology.

     

Block of a Ca<Superscript>2+</Superscript>-activated Potassium Channel by Cocaine

The primary target for cocaine is believed to be monoamine transporters because of cocaine’s high-affinity binding that prevents re-uptake of released neurotransmitter. However, direct interaction with ion channels has been shown to be important for certain pharmacological/toxicological effects of cocaine. Here I show that cocaine selectively blocks a calcium-dependent K⁺ channel in hippocampal neurons grown in culture (IC₅₀ = ∼30 μM). Single-channel recordings show that in the presence of cocaine, the channel openings are interrupted with brief closures (flicker block). As the concentration of cocaine is increased the open-time is reduced, whereas the duration of brief closures is independent of concentration. The association and dissociation rate constants of cocaine for the neuronal Ca²⁺-activated K⁺ channels are 261 ± 37 μM⁻¹s⁻¹ and 11451 ± 1467 s⁻¹. The equilibrium dissociation constant (K_B) for cocaine, determined from single-channel parameters, is 43 μM. The lack of voltage dependence of block suggests that cocaine probably binds to a site at the mouth of the pore. Block of Ca²⁺-dependent K⁺ channels by cocaine may be involved in functions that include broadening of the action potential, which would facilitate transmitter release, enhancement of smooth muscle contraction particularly in blood vessels, and modulation of repetitive neuronal firing by altering the repolarization and afterhyperpolarization phases of the action potential.     

IL-10 suppresses bactericidal response of macrophages against Salmonella Typhimurium

We report, herein, an attempt to determine whether an IL-10-induced immunological state affects the response of macrophages against Salmonella Typhimurium (ST). Pretreatment with mrIL-10 induced the intracellular invasion of ST into macrophages in a dose-dependent manner. It also activated AKT phosphorylation, cyclin D1, Bcl-X_L, and COX-2 upon ST infection, which may correlate with Salmonella’s survival within the macrophages. However, I-κB phosphorylation was shown to be inhibited, along with the expression of TNF-α and MIP-2α mRNA. Therefore, IL-10 not only suppresses the bactericidal response of macrophages against ST, but also ultimately causes infected macrophages to function as hosts for ST replication.     

酸枣果提取物在体内外对抗生素的增效作用及机制研究

作者研究团队先前从酸枣果的氯仿提取物中精制得到其低极性范围的活性组合物Fr.2a,发现Fr.2a与多种抗生素联用显示出广泛的协同抗菌作用。该研究在Fr.2a的基础上利用硅胶柱层析对酸枣果氯仿提取物中其他极性范围内的活性成分进行了分离纯化,得到精制物Fr.B,并对精制物Fr.B进行GC MS、核磁共振氢谱、红外光谱分析,以确定Fr.B的组成成分;通过抗菌谱分析和细胞通透性分析,以明确Fr.B的抗菌增效谱和抗菌增效机制;采用熔和法将精制物Fr.B制备成软膏,通过小鼠伤口感染模型评价该软膏对抗生素的增效效果。结果表明：(1)由酸枣果氯仿提取物进一步精制得到的Fr.B组分,主要包含反油酸、油酸、顺 10 十六碳烯醇、棕榈酸等脂肪酸类化合物。(2)Fr.B分别与庆大霉素、妥布霉素、氨苄青霉素、氯霉素、红霉素、夫西地酸、制霉菌素、酮康唑和两性霉素B等多种抗生素联用时显示出广泛的协同抗菌作用。(3)Fr.B可破坏细胞膜和细胞壁的完整性而增强细菌细胞的通透性。(4)在体内和体外Fr.B均能显著增强红霉素对耐甲氧西林金黄色葡萄球菌(MRSA)的杀菌作用,从而提高红霉素对MRSA菌株引起的伤口感染的治疗效果。研究表明,本研究所得到的Fr.B具有广谱的抗菌增效活性,能够增强红霉素对伤口耐药菌感染的治疗效果。该研究结果为克服微生物对抗生素的耐药性提供了新的思路和解决方案。     

Synergistic bactericidal activity of <Emphasis Type="Italic">Eremanthus erythropappus</Emphasis> oil or β-bisabolene with ampicillin against <Emphasis Type="Italic">Staphylococcus aureus</Emphasis>

The activity of Eremanthus erythropappus oil (EO) and some of its compounds and their potential synergistic interaction with ampicillin against different strains of Staphylococcus aureus were investigated. Determination of chemical composition of EO by gas chromatography-mass spectrometry (GC/MS) and bioguided chemical fractionation led to the identification of β-bisabolene as the main active compound. A synergistic bactericidal activity of EO or β-bisabolene with ampicillin against Staphylococcus aureus was observed in a time-kill assay. EO and β- bisabolene have the potential to restore the effectiveness of ampicillin against resistant S. aureus.     

A novel GH43 α-l-arabinofuranosidase from Humicola insolens: mode of action and synergy with GH51 α-l-arabinofuranosidases on wheat arabinoxylan

A novel α-l-arabinofuranosidase (α-AraF) belonging to glycoside hydrolase (GH) family 43 was cloned from Humicola insolens and expressed in Aspergillus oryzae. ¹H-NMR analysis revealed that the novel GH43 enzyme selectively hydrolysed (1→3)-α-l-arabinofuranosyl residues of doubly substituted xylopyranosyl residues in arabinoxylan and in arabinoxylan-derived oligosaccharides. The optimal activity of the cloned enzyme was at pH 6.7 and 53 °C. Two other novel α-l-arabinofuranosidases (α-AraFs), both belonging to GH family 51, were cloned from H. insolens and from the white-rot basidiomycete Meripilus giganteus. Both GH51 enzymes catalysed removal of (1→2) and (1→3)-α-l-arabinofuranosyl residues from singly substituted xylopyranosyls in arabinoxylan; the highest arabinose yields were obtained with the M. giganteus enzyme. Combinations (50:50) of the GH43 α-AraF from H. insolens and the GH51 α-AraFs from either M. giganteus or H. insolens resulted in a synergistic increase in arabinose release from water-soluble wheat arabinoxylan in extended reactions at pH 6 and 40 °C. This synergistic interaction between GH43 and GH51 α-AraFs was also evident when a GH43 α-AraF from a Bifidobacterium sp. was supplemented in combination with either of the GH51 enzymes. The synergistic effect is presumed to be a result of the GH51 α-AraFs being able to catalyse the removal of single-sitting (1→2)–α-l-arabinofuranosyls that resulted after the GH43 enzyme had catalysed the removal of (1→3)–α-l-arabinofuranosyl residues on doubly substituted xylopyranosyls in the wheat arabinoxylan.     

T’ef (<Emphasis Type="Italic">Eragrostis tef</Emphasis>) in Ancient Agricultural Systems of Highland Ethiopia

T’ef ( Eragrostis tef ) in Ancient Agricultural Systems of Highland Ethiopia. T’ef (Eragrostis tef) has been cultivated in the Horn of Africa for at least 2,000 years. The earliest known agricultural systems in this region date to the Pre-Aksumite period (800–400 b.c.) and appear to have focused on Near Eastern crops, with indigenous African species increasing in importance during Aksumite times (400 b.c.–a.d. 700). While palaeoethnobotanical data are available from Pre-Aksumite and late Aksumite periods, macroscopic botanical remains from the site of Ona Nagast, northern Ethiopia, provide a first glimpse of agricultural systems dating to Proto-Aksumite (400–50 b.c.), Early to Classic (50 b.c.–a.d. 340), and Post-Aksumite (a.d. 700–900) times. Archaeological t’ef remains from Ona Nagast are examined in detail. Guidelines are developed for the identification of t’ef grains preserved on archaeological sites, with a focus on how to differentiate them from seeds of wild Eragrostis species. Charring experiments reveal that in some cases t’ef may not survive high temperatures tolerated by larger cereal grains, such as wheat and barley. The domestication history of t’ef appears to be different from some other cereals, a factor which may explain the preponderance of indeterminate Eragrostis seeds in archaeological samples. Selection of large seed size and intensified tillage were not key factors in t’ef domestication. Early cultivators were likely selecting for increased branching and higher percentage seed set under conditions of minimal tillage.