High Inoculum Concentration Research Articles

Characterization of defense responses in the ‘green ring’ and ‘red ring’ on jujube fruit upon postharvest infection by Alternaria alternata and the activation by the elicitor treatment

Alternaria alternata is the major threat to postharvest storage of jujube (Ziziphus jujuba Mill.) fruit. A typical phenotype that a ‘red ring’ circling an enclosed ‘green ring’ area surrounding the spot of Alternaria rot on the jujube peel was first found and investigated in this study. Results indicated that the pathogenic infection is essential for the onset and development of the phenotype on jujubes, with high inoculum concentration and low incubation temperature leading to the enlarged ‘green ring’. The boundary line between the two rings seemed to be pre-fixed on the A. alternata-infected jujubes once the phenotype appeared. The ‘green ring’ was invaded gradually and ‘eaten-up’ eventually by the expansion of the surrounded rot spot. Biochemical measurements showed that such successful infection on jujubes might attribute to the great loss of individual phenolic compounds, including (+)-catechin, chlorogenic acid, (−)-epicatechin and p-coumaric acid, in the peel tissue of the ‘green ring’ and ‘red ring’, while some antifungal compounds increased in the flesh beneath the peel tissue in response to the infection. The elicitor salicylic acid (SA) triggered the early and rapid occurrence of the ‘green ring’ and ‘red ring’ on the A. alternata-infected jujubes. The SA treatment activated the phenylpropanoid pathway and elevated the accumulation of total phenolic compounds in the ‘green ring’ to combat the fungal attack and thereby restrict the expansion of disease lesion. Characterization of defense responses in the ‘green ring’ and ‘red ring’ on jujubes against Alternaria infection is helpful for unravelling the underlying mechanisms of the formation of defense resistance in fruit upon postharvest infection.

Identification and Characterization of Fungal Pathogens Causing Fruit Rot of Deciduous Holly.

Cut branches of deciduous holly (Ilex spp. L.) harboring colorful berries are traditionally used as ornaments in holiday decorations. Since 2012, a fruit rot of unspecified cause has resulted in significant yield reduction and economic losses across Midwestern and Eastern U.S. nurseries. In this study, symptomatic fruit samples collected from nine different locations over five years were analyzed, and several fungal species were isolated. A combination of morphological characterization, multilocus phylogenetic analyses, and pathogenicity assays revealed that Alternaria alternata and Diaporthe ilicicola sp. nov. were the primary pathogens associated with symptomatic fruit. Other fungi including A. arborescens, Colletotrichum fioriniae, C. nymphaeae, Epicoccum nigrum, and species in the D. eres species complex appeared to be minor pathogens in this disease complex. In detached fruit pathogenicity assays testing the role of wounding and inoculum concentration on disease development, disease incidence and severity increased when fruit was wounded and inoculated with a higher inoculum concentration. These findings indicate that management strategies that can protect fruit from injury or reduce inoculum may lower disease levels in the field. This research established the basis for further studies on this emerging disease and the design of research-based management strategies. To our knowledge, it also represents the first report of species of Alternaria, Colletotrichum, Diaporthe, and Epicoccum causing fruit rot of deciduous holly.

Combined use of the entomopathogenic fungus, Metarhizium brunneum, and the mosquito predator, Toxorhynchites brevipalpis, for control of mosquito larvae: Is this a risky biocontrol strategy?

Mosquitoes transmit several diseases, which are of global significance (malaria, dengue, yellow fever, Zika). The geographic range of mosquitoes is increasing due to climate change, tourism and trade. Both conidial and blastospore formulations of the entomopathogenic fungus, Metarhizium brunneum ARSEF 4556, are being investigated as mosquito larvicides. However, concerns have been raised over possible non-target impacts to arthropod mosquito predators such as larvae of Toxorhynchites brevipalpis which feed on larvae of mosquito vector species. Laboratory-based, small container bioassays showed, that T. bevipalpis larvae are susceptible to relatively high concentrations (i.e. ≥107 spores ml-1) of inoculum with blastospores being significantly more virulent than conidia. At lower concentrations (e.g. <107 spores ml-1), it appears that M. brunneum complements T. brevipalpis resulting in higher control than if either agent was used alone. At a concentration of 105 spores ml-1, the LT50 of for conidia and blastospores alone was 5.64 days (95% CI: 4.79-6.49 days) and 3.89 days (95% CI: 3.53-4.25 days), respectively. In combination with T. brevipalpis, this was reduced to 3.15 days (95% CI: 2.82-3.48 days) and 2.82 days (95% CI: 2.55-3.08 days). Here, combined treatment with the fungus and predator was beneficial but weaker than additive. At 107 and 108 blastospores ml-1, mosquito larval mortality was mostly due to the fungal pathogen when the predator was combined with blastospores. However, with conidia, the effects of combined treatment were additive/synergistic at these high concentrations. Optimisation of fungal concentration and formulation will reduce: (1) risk to the predator and (2) application rates and costs of M. brunneum for control of mosquito larvae.

The Distribution of Listeria in Pasture-Raised Broiler Farm Soils Is Potentially Related to University of Vermont Medium Enrichment Bias toward Listeria innocua over Listeria monocytogenes.

The occurrence of Listeria monocytogenes has been widely investigated in the poultry production chain from the processing plant to the final product. However, limited data are available on Listeria species, including Listeria monocytogenes, in the poultry farm environment. Therefore, fecal and soil samples from 37 pastured poultry flocks from 10 all-natural farms over 3 years were assessed to determine the prevalence and diversity of Listeria within these alternative poultry farm environments using standard cultural and molecular methods. Listeria species were isolated in 15% of poultry farm samples and included Listeria innocua (65.7%), L. monocytogenes (17.4%), and Listeria welshimeri (15.1%). Additional multiplex PCR serotyping showed group 1/2a-3a to be the most dominant L. monocytogenes serovar group. Based on these results, monoculture growth experiments were conducted on four Listeria soil isolates (three L. monocytogenes isolates representing the three recovered serovar groups and one L. innocua isolate) to determine if culture medium [tripticase soy broth (TSB) and University of Vermont modified Listeria enrichment broth (UVM)], inoculum concentration (102 or 105 CFU/ml), or incubation temperature (20, 30, and 42°C) differentially affected these Listeria species. Overall, very few significant growth differences were observed between the behavior of the three L. monocytogenes isolates (representing the three recovered serovar groups) under the growth conditions tested. Alternatively, at 30°C in UVM with the lower inoculum concentration, the L. innocua isolate had a significantly shorter lag phase than the L. monocytogenes isolates. In coculture growth studies under these same incubation conditions, the lag phase of L. innocua and L. monocytogenes was similar, but the final concentration of L. innocua was significantly higher than L. monocytogenes. However, cocultures in UVM for high inoculum concentration did not show preferential growth of L. innocua over L. monocytogenes. These results indicate that the use of UVM as an enrichment medium may preferentially allow L. innocua to outcompete L. monocytogenes at low concentrations, biasing the Listeria prevalence from these farm samples toward L. innocua and potentially underreporting the presence of L. monocytogenes in these environments.

Valorisation of the liquid fraction from hydrothermal carbonisation of sewage sludge by anaerobic digestion

AbstractBACKGROUNDThe mesophilic anaerobic digestion of the liquid fraction from hydrothermal carbonisation (208°C, 1 h) of dehydrated sewage sludge has been studied. Two initial inoculum concentrations (IC) (10 and 25 g COD L‐1) and four inoculum to substrate ratios (ISR) (2, 1, 0.5 and 0.4 on a COD basis) have been selected to analyse their influence on the evolution of the anaerobic digestion process.RESULTSThe substrate is characterised by a high COD (95.5 g L‐1) and TKN (8.7 g N L‐1) values. High inoculum concentration (25 g COD L‐1) and/or low ISR (≤ 0.5) inhibited methanogenesis due to the high ammonia nitrogen (1.4 g TAN L‐1) and VFA (&gt;4 g COD L‐1) released. For the inhibited samples final COD removals lower than 15% and IA/TA ratios higher than 0.3 were found. The greatest methane yield (177±5 mL CH4 STP g‐1 CODadded) was achieved at 25 g COD L‐1 of IC and at an ISR of 2.CONCLUSIONDuring anaerobic digestion of the liquid fraction from the hydrothermal carbonisation of sewage sludge, the IC and ISR must be adequately selected for proper operation of the process and successful valorisation. According to the results, working at an ISR ≥ 1 is recommended. © 2017 Society of Chemical Industry

Effect of growth conditions on advantages of hup− strain for H2 photoproduction by Rubrivivax gelatinosus

H2 photoproduction by growing cultures of hup− mutant and parental strain RL2 of Rubrivivax gelatinosus was compared. We checked the influence of different substrates, presence of air and N2, culture shaking, inoculum concentrations. At low inoculum concentration, hup− strain demonstrated significant advantage over the parental strain in microaerobic conditions, while under N2–Ar atmosphere it was lower and vanished in anaerobic conditions (Ar only). This advantage was evident when using substrates with low degree of reduction (malate and succinate). Culture shaking under microaerobic conditions and in presence of N2 completely prevented H2 production by both strains. The high inoculum concentration inhibited H2 production under microaerobic conditions and in presence of N2, unlike to anaerobic conditions. With inoculum concentration increase, H2 production decreased not gradually but stepwise which means some metabolic shift. H2 production by hup− strain seems to be more tolerant to air traces than by parental strain.

Metabolic cost of resistance in clubroot-resistant canola and napa cabbage

Genetic resistance is widely used to manage clubroot (Plasmodiophora brassicae) in canola (Brassica napus) and other brassica crops. Replicated field trials were conducted with three clubroot-resistant cultivars and one susceptible cultivar of either canola or napa cabbage (B. rapa) in southern Ontario in 2014 and 2015. These studies indicated that plant growth and development in resistant cultivars of both crops were often reduced at high concentrations of inoculum (resting spores) of the pathogen. In napa cabbage, leaf length was reduced by 31% in 2014 and 22% in 2015 at a site with high spore concentration relative to a site with lower concentration. In canola, height of resistant plants was reduced by 30%, biomass by 43%, and maturity was delayed at a site with high spore concentration relative to an adjacent site with lower spore concentration in 2014. However, there were no differences among sites in 2015. A growth room study demonstrated that plant height of a clubroot-resistant canola cultivar declined by 12–14% at high spore concentrations. Biomass of canola was also lower and maturity was delayed with increasing concentration of spores in some, but not all, field and laboratory studies. We suggest that the expression of resistance to clubroot in resistant cultivars can result in reduced plant growth and development in both B. rapa and B. napus, but that substantial reductions in plant growth and development occur only at high concentrations (> 106 g−1) of resting spores in soil.

Biorremediación de organofosforados por hongos y bacterias en suelos agrícolas: revisión sistemática

&lt;p&gt;Los organofosforados son un tipo de plaguicidas ampliamente utilizados en el sector agrícola para el control de plagas. Dado que estos son compuestos químicos altamente tóxicos, su uso excesivo ha causado gran deterioro en los suelos cultivables, así como graves daños para los ecosistemas y la salud humana. La biorremediación surge como una alternativa para transformar los plaguicidas en compuestos más simples y poco contaminantes mediante el uso del potencial metabólico de los microorganismos. Por lo anterior, el objetivo de esta investigación fue describir los hongos y bacterias involucrados en la biorremediación de los principales plaguicidas organofosforados empleados en suelos agrícolas por medio de una revisión sistemática de la literatura científica, con el fin de aportar información útil para la realización de estudios posteriores. Se obtuvo información científica de las bases de datos ScienceDirect y Springer Link, y también información no indexada del buscador Google Scholar. Se encontró que el organofosforado que más se ha estudiado es el clorpirifós (categoría toxicológica III) y los microorganismos que más se utilizan como biorremediadores de organofosforados son los géneros Serratia, Bacillus y Pseudomonas. Se concluye que el éxito de la biorremediación depende de la capacidad competitiva de los microorganismos, de la biodisponibilidad y la concentración del organofosforado, del pH, la temperatura y el tipo de suelo, así como de la presencia de suplementos nutricionales y de la concentración alta del inóculo.&lt;/p&gt;

Caracterização epidemiológica e fosfitos no manejo da podridão por Aspergillus niger em uva de mesa

The aim of this study was to evaluate the influence of epidemiological characteristics on the development of postharvest rot in table grape caused by Aspergillus niger Tiegh. and the effect of phosphite salts in vitro and in vivo. The inoculum concentration (102 to 107 conidios mL-1), wetness period (0, 12, 24, 36 and 48 h) and temperature (2, 5, 10, 15, 20 and 25 °C) of two A. niger isolates were evaluated. At postharvest management were used Ca, K, Cu, Mg e Zn phosphites at the concentrations 0.3; 0.9; 1.25; 1.7 and 2 g L-1 by dipping the grape clusters. After 12 and 24 h, the berries were wounded and inoculated with the fungus. The favorable conditions for the establishment of disease are high inoculum concentration (106 and 107 conidios mL-1), 48 h of wetness period and temperature about 25 °C. Phosphites shown to have a direct effect on the fungus by inhibiting the mycelial growth, especially zinc phosphite. There were no significant difference between phosphites for conidia germination. Just calcium phosphite is efficient in reducing disease severity, on the berries inoculated 12 hours after treatment. The treatments do not influence the chemical characteristics of grapes.

Association between Type A blaZ Gene Polymorphism and Cefazolin Inoculum Effect in Methicillin-Susceptible Staphylococcus aureus.

Some proportion of type A blaZ gene-positive methicillin-susceptible Staphylococcus aureus strains exhibit the cefazolin inoculum effect (CIE). The type A blaZ gene was divided into two groups by single nucleotide polymorphisms (SNPs) at Ser226Pro and Cys229Tyr. The median cefazolin MICs at a high inoculum concentration were 5.69 μg/ml for the Ser-Cys group and 40.32 μg/ml for the Pro-Tyr group (P = 0.01). The SNPs at codons 226 and 229 in the amino acid sequence encoded by the blaZ gene were closely associated with the CIE.

Combined effect of carvacrol and citral on the growth of Listeria monocytogenes and Listeria innocua and on the occurrence of damaged cells

The aim of this study was to evaluate the growth kinetics of Listeria innocua Serovar 6a (CECT 910) and Listeria monocytogenes Serovar 4b (CECT 4032) exposed to combinations of carvacrol and citral (0.0 μL/mL (control), 0.050 μL/mL of carvacrol and 0.075 μL/mL of citral, 0.050 μL/mL of carvacrol and 0.125 μL/mL of citral, 0.085 μL/mL of carvacrol and 0.075 μL/mL of citral, and 0.085 μL/mL of carvacrol and 0.125 μL/mL of citral), with two initial inoculum concentrations, and also the occurrence of sublethal damage in these cell populations. The terpene combinations exhibited antibacterial activity against L. innocua and L. monocytogenes and the effects were dependent on the concentration of terpenes present in the culture medium (p ≤ 0.05). When terpene-treated L. innocua and L. monocytogenes were incubated in TSB, significant differences in lag phase and growth rate were observed between low and high inoculum concentrations (p ≤ 0.05), indicating that the inoculum level should be taken into account in modeling studies. When bacterial cells were exposed to terpenes the proportion of sublethally injured cells increased with the increase in the terpene dose (p ≤ 0.05). In conclusion, all of these results show that carvacrol and citral can be used in combination at 25% of the MIC in order to control Listeria growth.

Effect on nursery and field performance of Pinus patula seedlings after inoculation with Fusarium circinatum

Fusarium circinatum is an important fungal pathogen of Pinus species. In South Africa, it is the most significant pathogen of Pinus patula seedlings in forestry nurseries where it presents a substantial constraint to productivity and can continue to cause mortality in-field for up to two years after establishment. This study describes the results from two trials where P. patula seedlings were inoculated with F. circinatum to determine the impact of the pathogen on nursery and field performance. Seedlings were also subjected to water stress treatments to ascertain whether this would trigger the onset of disease symptoms. Inoculum load and timing of inoculation had significant effects on seedling survival in both the nursery and field. High inoculum concentrations caused greater levels of mortality and, where seedlings were inoculated at a young age, they showed higher levels of susceptibility to F. circinatum. Temporary water-stress in the nursery produced smaller plants and improved in-field survival, but this treatment did not trigger higher mortality in inoculated treatments. On the other hand, transplant stress was a major contributor to the higher levels of mortality observed in inoculated treatments. Overall, these studies confirmed that infection in the nursery leads to the disease problems observed during early plant establishment in the field.

Physiological description of multivariate interdependencies between process parameters, morphology and physiology during fed‐batch penicillin production

Optimization of productivity and economics of industrial bioprocesses requires characterization of interdependencies between process parameters and process performance. In the case of penicillin production, as in other processes, process performance is often closely interlinked with the physiology and morphology of the organism used for production. This study presents a systematic approach to efficiently characterize the physiological effects of multivariate interdependencies between bioprocess design parameters (spore inoculum concentration, pO2 control level and substrate feed rate), morphology, and physiology. Method development and application was performed using the industrial model process of penicillin production. Applying traditional, statistical bioprocess analysis, multivariate correlations of raw bioprocess design parameters (high spore inoculum concentration, low pO2 control as well as reduced glucose feeding) and pellet morphology were identified. A major drawback of raw design parameter correlation models; however, is the lack of transferability across different process scales and regimes. In this context, morphological and physiological bioprocess modeling based on scalable physiological parameters is introduced. In this study, raw parameter effects on pellet morphology were efficiently summarized by the physiological parameter of the biomass yield per substrate. Finally, for the first time to our knowledge, the specific growth rate per spore was described as time-independent determinant for switching from pellet to disperse growth during penicillin production and thus introduced as a novel, scalable key process parameter for pellet morphology and process performance.

Biological Control of the Weed Hemp Sesbania (Sesbania exaltata) in Rice (Oryza sativa) by the Fungus Myrothecium verrucaria

In greenhouse and field experiments, a mycelial formulation of the fungus Myrothecium verrucaria (IMI 361690) containing 0.20% Silwet L-77 surfactant exhibited high bioherbicidal efficacy against the problematic weed hemp sesbania. Infection and mortality levels of 100% of hemp sesbania seedlings occurred within 48 h after fungal application in the greenhouse. In rice field tests conducted over a three year period, M. verrucaria at an inoculum concentration of 50 g L−1 (dry mycelium equivalent) controlled 95% of ≤20 cm tall hemp sesbania plants. M. verrucaria also controlled larger plants (≥60 cm tall) using this high inoculum concentration. This level of weed control, as well as rice yields from plots where weeds were effectively controlled, were similar to those which occurred with the herbicide acifluorfen. These results suggest that a mycelial formulation of M. verrucaria has potential as a bioherbicide for controlling hemp sesbania in rice.

Antagonistic Bacillus species as a biological control of ginseng root rot caused by Fusarium cf. incarnatum

BackgroundThis study aimed to develop a biocontrol system for ginseng root rot caused by Fusarium cf. incarnatum. MethodsIn total, 392 bacteria isolated from ginseng roots and various soils were screened for their antifungal activity against the fungal pathogen, and a bacterial isolate (B2-5) was selected as a promising candidate for the biocontrol because of the strong antagonistic activity of the bacterial cell suspension and culture filtrate against pathogen. ResultsThe bacterial isolate B2-5 displayed an enhanced inhibitory activity against the pathogen mycelial growth with a temperature increase to 25°C, produced no pectinase (related to root rotting) and no critical rot symptoms at low [106 colony-forming units (CFU)/mL] and high (108 CFU/mL) inoculum concentrations. In pot experiments, pretreatment with the bacterial isolate in the presumed optimal time for disease control reduced disease severity significantly with a higher control efficacy at an inoculum concentration of 106 CFU/mL than at 108 CFU/mL. The establishment and colonization ability of the bacterial isolates on the ginseng rhizosphere appeared to be higher when both the bacterial isolate and the pathogen were coinoculated than when the bacterial isolate was inoculated alone, suggesting its target-oriented biocontrol activity against the pathogen. Scanning electron microscopy showed that the pathogen hyphae were twisted and shriveled by the bacterial treatment, which may be a symptom of direct damage by antifungal substances. ConclusionAll of these results suggest that the bacterial isolate has good potential as a microbial agent for the biocontrol of the ginseng root rot caused by F. cf. incarnatum.

Simultaneous single-cell protein production and COD removal with characterization of residual protein and intermediate metabolites during whey fermentation by K. marxianus

Cheese whey fermentation with Kluyveromyces marxianus was carried out at 40 °C and pH 3.5 to examine simultaneous single-cell protein production and chemical oxygen demand (COD) removal, determine the fate of soluble whey protein and characterize intermediate metabolites. After 36 h of batch fermentation, the biomass concentration increased from 2.0 to 6.0 g/L with 55 % COD reduction (including protein), whereas soluble whey protein concentration decreased from 5.6 to 4.1 g/L. It was confirmed through electrophoresis (SDS-PAGE) that the fermented whey protein was different from native whey protein. HPLC and GC-MS analysis revealed a change in composition of organic compounds post-fermentation. High inoculum concentration in batch fermentation resulted in an increase in biomass concentration from 10.3 to 15.9 g/L with 80 % COD reduction (including protein) within 36 h with residual protein concentration of 4.5 g/L. In third batch fermentation, the biomass concentration increased from 7.3 to 12.4 g/L with 71 % of COD removal and residual protein concentration of 4.3 g/L after 22 h. After 22 h, the batch process was shifted to a continuous process with cell recycle, and the steady state was achieved after another 60 h with biomass yield of 0.19 g biomass/g lactose and productivity of 0.26 g/L h. COD removal efficiency was 78-79 % with residual protein concentration of 3.8-4.2 g/L. The aerobic continuous fermentation process with cell recycle could be applied to single-cell protein production with substantial COD removal at low pH and high temperature from cheese whey.

Virulence potential and genome-wide characterization of drug resistant Streptococcus pneumoniae clones selected in vivo by the 7-valent pneumococcal conjugate vaccine.

We used mouse models of pneumococcal colonization and disease combined with full genome sequencing to characterize three major drug resistant clones of S. pneumoniae that were recovered from the nasopharynx of PCV7-immunized children in Portugal. The three clones – serotype 6A (ST2191), serotype 15A (ST63) and serotype 19A (ST276) carried some of the same drug resistance determinants already identified in nasopharyngeal isolates from the pre-PCV7 era. The three clones were able to colonize efficiently the mouse nasopharyngeal mucosa where populations of these pneumococci were retained for as long as 21 days. During this period, the three clones were able to asymptomatically invade the olfactory bulbs, brain, lungs and the middle ear mucosa and established populations in these tissues. The virulence potential of the three clones was poor even at high inoculum (105 CFU per mouse) concentrations in the mouse septicemia model and was undetectable in the pneumonia model. Capsular type 3 transformants of clones 6A and 19A prepared in the laboratory produced lethal infection at low cell concentration (103 CFU per mouse) but the same transformants became impaired in their potential to colonize, indicating the importance of the capsular polysaccharide in both disease and colonization. The three clones were compared to the genomes of 56 S. pneumoniae strains for which sequence information was available in the public databank. Clone 15A (ST63) only differed from the serotype 19F clone G54 in a very few genes including serotype so that this clone may be considered the product of a capsular switch. While no strain with comparable degree of similarity to clone 19A (ST276) was found among the sequenced isolates, by MLST this clone is a single locust variant (SLV) of Denmark14-ST230 international clone. Clone 6A (ST2191) was most similar to the penicillin resistant Hungarian serotype 19A clone.

Biological characteristics of Monilinia fructicola isolates from stone fruits in eastern West Virginia

Monilinia fructicola, the causal agent of brown rot, was recovered from decayed stone fruits (peach, plum and nectarine) in 11 West Virginia orchards. There was significant variation among these isolates with respect to colony morphology, growth rate, sporulation level, sensitivity to fenbuconazole, vegetative compatibility and virulence. Species identification was confirmed using ITS sequences from the nuclear ribosomal RNA gene. The cultural phenotypes on potato dextrose agar (PDA) ranged from white to dark, melanized colonies. The growth rate of the isolates on PDA ranged from 0.3 to 3.2 mm day−1 at 4 °C, from 2.9 to 7.6 mm day−1 at 10 °C, and from 4.8 to 19 mm day−1 at 24 °C. There was a statistically significant relationship between the growth of the isolates on PDA and their aggressiveness on nectarines at 24 °C, especially at higher inoculum concentrations. Sporulation of 3-day-old cultures on peach agar at 24 °C varied from profuse to no sporulation, with some isolates sporulating only sparsely after 10 days. The EC50 for fenbuconazole ranged from 0.003 to 0.129 μg μL−1 and for two reference isolates was 0.020 and 0.016 μg μL−1. Only a few vegetative compatibility groups were identified among isolates within orchards, reflecting the lack of sexual recombination in this region.

Feasibility of anaerobic digestion from bioethanol fermentation residue

The focus of this study was the reuse of red algal ethanol fermentation residue as feedstock for anaerobic digestion. Levulinic acid and formic acid, the dilute-acid hydrolysis byproducts, inhibited methanogenesis at concentrations over 3.0 and 0.5 g/L, respectively. However, the inhibition was overcome by increasing inoculum concentration. A series of batch experiments with the fermentation residue showed increased methane yield and productivity at higher inoculum concentration. The maximum methane conversion rate of 84.8% was found at 5 g COD/L of fermentation residue at 0.25 g COD/g VSS of food-to-microorganism (F/M) ratio. The red algal ethanol fermentation residue can possibly be used as a feedstock in anaerobic digestion at appropriate concentration and F/M ratio.

Inoculum Effect on the Efficacies of Amoxicillin-Clavulanate, Piperacillin-Tazobactam, and Imipenem against Extended-Spectrum β-Lactamase (ESBL)-Producing and Non-ESBL-Producing Escherichia coli in an Experimental Murine Sepsis Model

Escherichia coli is commonly involved in infections with a heavy bacterial burden. Piperacillin-tazobactam and carbapenems are among the recommended empirical treatments for health care-associated complicated intra-abdominal infections. In contrast to amoxicillin-clavulanate, both have reduced in vitro activity in the presence of high concentrations of extended-spectrum β-lactamase (ESBL)-producing and non-ESBL-producing E. coli bacteria. Our goal was to compare the efficacy of these antimicrobials against different concentrations of two clinical E. coli strains, one an ESBL-producer and the other a non-ESBL-producer, in a murine sepsis model. An experimental sepsis model {~5.5 log10 CFU/g [low inoculum concentration (LI)] or ~7.5 log(10) CFU/g [high inoculum concentration (HI)]} using E. coli strains ATCC 25922 (non-ESBL producer) and Ec1062 (CTX-M-14 producer), which are susceptible to the three antimicrobials, was used. Amoxicillin-clavulanate (50/12.5 mg/kg given intramuscularly [i.m.]), piperacillin-tazobactam (25/3.125 mg/kg given intraperitoneally [i.p.]), and imipenem (30 mg/kg i.m.) were used. Piperacillin-tazobactam and imipenem reduced spleen ATCC 25922 strain concentrations (-2.53 and -2.14 log10 CFU/g [P < 0.05, respectively]) in the HI versus LI groups, while amoxicillin-clavulanate maintained its efficacy (-1.01 log10 CFU/g [no statistically significant difference]). Regarding the Ec1062 strain, the antimicrobials showed lower efficacy in the HI than in the LI groups: -0.73, -1.89, and -1.62 log10 CFU/g (P < 0.05, for piperacillin-tazobactam, imipenem, and amoxicillin-clavulanate, respectively, although imipenem and amoxicillin-clavulanate were more efficacious than piperacillin-tazobactam). An adapted imipenem treatment (based on the time for which the serum drug concentration remained above the MIC obtained with a HI of the ATCC 25922 strain) improved its efficacy to -1.67 log10 CFU/g (P < 0.05). These results suggest that amoxicillin-clavulanate could be an alternative to imipenem treatment of infections caused by ESBL- and non-ESBL-producing E. coli strains in patients with therapeutic failure with piperacillin-tazobactam.