The in vitro antifungal activity of ophiobolins was determined in a 96-well microtiter plate bioassay by measuring the

absorbance of the fungal cultures at 620 nm. The wells contained SPEC medium supplemented with ophiobolin A or B and inoculated with the appropriate sporangiospore suspension (105 spores mL–1). The drug concentrations applied were 100, 50, 25, 12.5, 6.25, 3.175 and 1.5875 μg mL–1, respectively. The plates were incubated for 72 h at 24 or 37 °C depending on the culturing requirements of the strains. Absorbances were measured using an ASYS Jupiter HD microplate reader (ASYS Hitech) every 24 h. Each test plate contained a sterile control (containing medium alone), a growth control (containing inoculated medium without the drugs) and a drug-free control (containing inoculated medium and methanol in the appropriate dilution without the ophiobolins). The uninoculated medium was used as the background Selleck Nutlin 3a for the calibration of the spectrophotometry. Absorbance of the untreated control cultures was referred to 100% of growth in each case. To decide whether the antifungal effect was fungistatic or fungicidic, 10 μL of each suspension in the microdilution plates was dropped onto YEG plates. After incubation for 24 h, the plates were checked visually. If colony formation was observed, the antifungal effect was considered to be fungistatic; otherwise, it was

fungicidic. Each experiment was repeated three times. For morphological examinations, the Mucor circinelloides strain ATCC 1216b was cultured Talazoparib ic50 on a solid and in a liquid YEG medium containing different concentrations of the drug (1.6, 3.2, 6.25 or 12.5 μg mL–1) at 24 °C. If the fungus was cultured on

a solid medium, microscopy was performed after incubation for 24 h. In the case of the liquid cultures, ophiobolin A was added to the fungus either at the time of spore inoculation (0 h) or 4 h postinoculation, and cells were examined microscopically 5 h after the addition of the inhibitor (5 or 9 h postinoculation, respectively). Treated cells were stained to using the annexin V-fluorescein isothiocyanate (FITC) Apoptosis Detection Kit (Sigma) according to the manufacturer’s instructions. For nuclear staining, cells were resuspended in 1 mL of 0.1 μg mL–1 4′-6-diamidino-2-phenylindole (DAPI) staining solution and were allowed to stain for 30 min at room temperature. Stained spores were collected, washed twice with distilled water (dw), and resuspended in 50 μL dw. Microscopic examinations were performed with a Zeiss Jenalumar fluorescence microscope using an excitation filter U 205 g, a barrier filter G-244 and a 510 nm dichroic splitter. The susceptibility to ophiobolins A and B of 17 fungal isolates representing six different genera (Micromucor, Mortierella, Mucor, Rhizomucor, Rhizopus and Gilbertella) was tested and their MIC values were determined (Table 1).

The sublethal concentration of zoocin A determined for each strain is given in Table 1. The growth assay proved simple and highly reproducible. Although somewhat arbitrary, setting the lag phase cut off point at initial OD+0.1 yielded highly reproducible experimental data. Determining the growth rate constant did not allow us to reliably distinguish treated from untreated cultures. Once treated cultures reached log phase, they grew as fast as untreated cultures, suggesting that once the cells have repaired their peptidoglycan and degraded any remaining intracellular PS-ODN, there were no remaining constraints to cellular growth. The addition of 0.1 μg mL−1

VX-809 mouse zoocin A and 10 μM of either FABM or FBA to S. IGF-1R inhibitor mutans OMZ175 resulted in a lag phase that was significantly longer (P=0.001) than that observed for the addition of zoocin A alone (Fig. 1). The effect of zoocin A and FABM on S. mutans OMZ175 growth was dose dependent. In the absence of zoocin A, FABM (1–20 μM) had no significant

effect on S. mutans OMZ175 growth (Table 2). When combined with 0.1 μg mL−1 zoocin A, the lag phase increased proportionally (R2=0.9928) with increasing FABM concentration. Similarly, using a fixed concentration of FABM (10 μM), the increase in lag phase was proportional to the zoocin A concentration (Table 2) both in the presence (R2=0.9919) and in the absence (R2=0.9069) of the PS-ODN. Growth inhibition was target specific. Only S. mutans strains were severely inhibited in the presence of FABM, whereas all streptococcal strains except S. oralis were severely inhibited by FBA (Table 3). Streptococcus

oralis 34 does contain the FBA target sequence within its genome but is not sensitive to zoocin A. Compared with growth in the presence of zoocin A, there were large increases in the lag phase (between 25% and >134%) for all S. mutans strains Adenosine triphosphate grown in the presence of zoocin A plus FABM. With the exception of S. oralis 34, compared with growth in the presence of zoocin A, there were large increases in the lag phase (between 30% and >134%) for all streptococcal strains grown in the presence of zoocin A plus FBA. Streptococcus sobrinus 6715 and S. sanguinis K11 showed no response to either FABM or FBA used at concentrations of 10 μM, but both showed significant (P=0.001) increases in lag phase in the presence of zoocin A plus 50 μM FBA. There were some strains that showed a small (<11%) but statistically significant increase in lag phase when incubated with the ATS control, suggesting a degree of nonspecific toxicity by these constructs. As a consequence of their high GC content, negative charge and or sulphur group, PS-ODN have been reported to interact with cellular proteins Brown et al., 1994), resulting in nonspecific toxicity (Chrisey et al., 1995; Stein, 1996).

There was a difference in the rate of drug resistance favouring ATV/r (RR 3.94, 95% CI 2.37–6.56; P < 0.00001) but the overall rate of emergent drug resistance was low for both treatments. This difference is a class effect and has previously been reported for other NNRTIs and PI/r. Differences were also identified in the rate of grade 3/4 central nervous this website system (CNS) events and the rate of lipid abnormalities favouring both ATV/r and RAL. These differences may well influence the choice between preferred third agents for individual patients. There are no RCTs comparing DRV/r vs. EFV directly. Thus an indirect comparison was undertaken using data from studies comparing DVR/r

vs. LPV/r [35-37] and LPV/r vs. EFV [17, 18] to assess

outcomes between the two treatment options. Some differences between these studies were identified in terms of comparability and are outlined in Appendix 3. Overall, these differences were judged insufficient to invalidate an indirect comparison between EFV and DRV/r. Comparing DRV/r and LPV/r there were clinically significant differences in the critical outcomes virological suppression, discontinuation due to adverse events and serious adverse events in favour of DRV/r but no differences in the critical outcomes virological failure and drug resistance. Comparing EFV and LPV/r there were clinically significant differences in the critical outcomes virological failure and suppression at 96 weeks Neratinib price in favour of EFV but no differences in the critical outcomes drug resistance and discontinuation due to adverse events. In addition, there were significant differences in some adverse events favouring EFV over LPV/r. RPV has been compared directly with EFV in RCTs [30-32]. With respect to critical

virological outcomes there was no difference in virological suppression but there were differences in drug resistance (RR 0.38, 95% CI 0.20–0.72; P = 0.003) and virological failure (RR 0.55, 95% CI 0.29–1.02; P = 0.06), both in favour of EFV. Pooled analyses by the investigators of the two RCTs showed the risk of virological failure PtdIns(3,4)P2 with RPV was highest in patients with a baseline VL >100 000 copies/mL [32]. For critical safety outcomes there was a difference in the proportion discontinuing for adverse events in favour of RPV (RR 2.29, 95% CI 1.15–4.57; P = 0.02) but no difference in serious adverse events. RPV also had better lipid profile outcomes. The StAR study showed overall noninferiority of the fixed-dose combination of TDF/FTC/RPV to fixed-dose TDF/FTC/EFV at 48 weeks. In a subgroup analysis in patients with baseline viral load less than 100 000 copies/mL, superiority of the RPV-based regimen was demonstrated. Similarly to ECHO and THRIVE, StAR confirmed higher rates of virological failure on RPV at high viral loads (greater than 100 000 copies/mL) but not at lower baseline viral load (less than 100 000 copies/mL).

Clp proteases

including ClpA act as molecular chaperones with a similar function as DnaK/DnaJ (Wickner et al., 1994). It is likely that the up-regulation of DnaJ-class molecular chaperone in CSM2 mutant is due to the substitution of the partial Clp protease function by DnaJ when Clp protease is dysfunctional. Further, our metabolomic analysis of the Clp protease mutant identified down-regulation of amino acid and oligosaccharide transporters that are part of ABC transporter pathways (Table 2). The mechanism is likely to be similar to that observed in Mycobacterium tuberculosis under hypoxia, where Clp proteases degrade the factors that inhibit DNA replication and transcription to initiate the synthesis of amino acids during stressful conditions (Sherrid et al., 2010). Changes in the levels of TCA cycle enzymes in response to copper identified by the metabolomic analysis (Table 2) were confirmed by the Ceritinib purchase proteomic analysis with up-regulation of ketol-acid reductoisomerase, which participates

in the production of CoA (Table 1). In addition, down-regulation of MalR, a transcriptional regulatory protein for malate and citrate metabolism (Table 1) under copper stress would result in the accumulation of malate and citrate, the intermediate products in TCA cycle (Papa et al., 2009). Higher levels of malate allow the organism to cope with oxidative stress caused selleck chemical by copper toxicity, by producing more NADPH, an important antioxidant (Singh et al., 2007). Citrate is a metabolite involved in the sequestration of aluminum and the increase of

citrate accumulation Glutathione peroxidase was previously shown in P. fluorescens grown under aluminum stress (Mailloux et al., 2008). Our results suggest that citrate is involved in the sequestration of copper. Based on our results, we propose a model for the response to toxic levels of copper in Pseudomonas sp. TLC6-6.5-4 (Fig. 4). High copper concentrations reduce its cell size, which decreases the amount of copper bound on the cell surface. In addition, smaller cells need less energy for maintenance under copper stress. CopA and lipoprotein mediate sequestration and efflux of copper outside the cytoplasm. Heat shock proteins including Clp protease and DnaJ-class molecular chaperone either degrade the damaged proteins or prevent their irreversible aggregation under copper stress. Furthermore, Clp protease is directly involved in copper resistance by up-regulation of amino acid transporters, proteins related to oxidative stress and proline accumulation. This organism maintains a fine metabolic balance to enable the cells to survive in environment with high copper concentration by increasing amino acid production and regulating TCA cycle. The authors gratefully thank J. Lutz and W. He for technical assistance with proteomic experiments and GC-MS analysis. We also thank R. Shaik for his help on proteomic and metabolomic data analysis. “
“Salmonella enterica serovar Typhi and Typhimurium are closely related serovars. However, S.

What is more important is the pre-deployment

education or orientation of each traveler with regards to the characteristics of the vector anopheles and the proper use of individual personnel protective equipment such as long-acting insect repellent lotion containing N,N-Diethyl-3-methylbenzamide (DEET), its reapplication when needed, PS-341 molecular weight and proper use of insecticide impregnated bed nets. Health education sessions are organized for servicepersons not only before leaving or upon arrival overseas but also just before returning home. It is unfortunately a well-known fact that disseminating information, even if it is of high quality, does not automatically lead to modification of risk behavior.9 Regular assessment of the impact of health education campaigns has, therefore, been implemented by the French Military Health Service to assess how the transmitted Metformin concentration message is perceived and if necessary adapt it to increase its effectiveness. The authors state they have no conflicts of interest to declare. “
“We report the case of an immunocompetent traveler returning from Morocco who presented with a giant splenic abscess, revealing an infection by Salmonella enterica serovar enteritidis.

Salmonellae are an important cause of food-borne infections in returning travelers. In immunocompetent hosts Salmonella typhi and Salmonella paratyphi cause enteric fever whereas other Salmonellae are commonly diagnosed in returning travelers with diarrhea.1 These Salmonella usually cause self-limited gastroenteritis but many other sites may be involved, particularly in patients with preexistent disease.2 In addition,

invasive infections may occur in infants, adults over the age of 65, and patients with debilitating or underlying illnesses.3 We report an uncommon complication revealing a disseminated Salmonella enteritidis infection, in a young and immunocompetent traveler. A 17-year-old man was admitted to our hospital with high-grade fever, anorexia, nausea, and abdominal pain lasting for 8 days. This French native student had returned 1 month earlier from Morocco where he had been vacationing Calpain for 5 weeks. He recalled symptoms of intermittent left abdominal and shoulder pain during the last 3 years, but denied any history of trauma. Eight days before admission, severe left upper abdominal and left shoulder pain appeared suddenly, together with nausea and high-grade fever. He initially received ofloxacin (200 mg bid) for 2 days and then co-amoxicillin (1 g tid) for 4 days without any improvement. On admission, the patient appeared ill and pale and complained of severe pain in the left upper abdominal quadrant. Physical examination revealed fever (39.2°C), tachycardia (pulse rate : 120/min), normal blood pressure, and a painful, large, and tender mass in the left upper abdominal quadrant. Laboratory tests revealed a white blood cell count at 20,000/mL (including 83% neutrophils).

Using whole-cell voltage-clamp recording, HCN channels in the neurones were activated

in response to isoprenaline and exogenous cAMP but only occasionally did they respond to NO, although exogenous cGMP was routinely effective. With the less invasive sharp microelectrode recording technique, however, exogenous NO modulated the channels reproducibly, as measured by the size of the HCN channel-mediated voltage sag following hyperpolarization. Moreover, NO also blunted the subsequent rebound depolarizing potentials, consistent with it Protease Inhibitor Library chemical structure increasing the hyperpolarization-activated current. Optimizing the whole-cell solution to improve the functioning of NO-activated guanylyl cyclase failed to restore NO sensitivity. Minimizing cellular dialysis by using the perforated-patch technique, however, was successful. The results provide evidence that HCN channels are potential downstream mediators of NO signalling in deep cerebellar nuclei neurones and suggest that the more general importance of this transduction pathway may have been overlooked

previously because of unsuitable recording methods. “
“Although we can generate movements whenever we feel like doing so, the way in which neuronal signals regulate the timing of self-initiated movements remains elusive. There is evidence that the dorsomedial frontal cortex, including Selleck PR171 the supplementary eye field (SEF), is involved in the self-triggering of movements. Because the gradual evolution of cortical activity over the dorsomedial frontal cortex is known to reflect the temporal prediction of an upcoming event, we postulated that the timing of self-initiated movements is regulated by the time course of neuronal Diflunisal activity in the SEF. To test the causal role, we applied electrical microstimulation to the SEF when monkeys prepared for memory-guided saccades. Stimulation delayed the initiation of saccades when animals were required to make saccades 1200 ± 300 ms following the cue (self-timed task), but not when they generated memory-guided saccades in response

to the offset of the fixation point (conventional task). As well as the increment in median saccade latencies, stimulation at ∼24% of sites also increased the occurrence of early erroneous saccades. Saccades facilitated by stimulation were always directed toward the cue, even when the cue was located away from the movement field. In contrast, stimulation to the frontal eye fields during saccade preparation exerted no effects in either task. These results suggest that the preparatory signals in the SEF may play a causal role in regulating the timing rather than the direction of self-initiated saccades. “
“The mammalian main olfactory bulb (MOB) receives a significant noradrenergic input from the locus coeruleus.

A focus-group interview was carried out with 15 HIV-positive patients (both homosexual and heterosexual men and women of various ethnicities) to identify relevant questions. The responses were transcribed and analysed. The final questionnaire was validated by a pilot

study with 12 HIV-positive patients. The participants in the pilot study were not eligible to participate in the study. The following LBH589 mouse variables were recorded: gender, age, educational level, ethnicity, current job, marital status, current adherence [17], current financial situation, route of infection, HIV exposure group, unsafe sex and psychological factors (guilt, shame, anxiety, concern, stress, loneliness, influence of HIV on life situation, constant thoughts about HIV, living a double life with HIV as a secret, the feeling that HIV limits way of living and stigmatization). The Beck Depression Inventory

II (BDI-II) [18] was used to assess the prevalence and severity of depressive symptoms. The BDI-II has shown high validity and reliability in measuring depressive symptoms. Respondents Silmitasertib research buy were required to rate 21 items from 0 to 3 according to how they had felt during the previous 2 weeks. The BDI-II focuses on both the cognitive-affective symptoms of depression, such as pessimism and diminished self-esteem, and the somatic symptoms of depression such as weight loss. A score of 14 or more is widely accepted as a cut-off point indicating depression on the 21-item BDI-II. The cut-off scores were: 0–13, minimal depression; 14–19, mild depression; 20–28, moderate depression; and 29–63, major depression. A cut-off point of 20 was chosen for validation using the Hamilton Depression Scale [19]. All patients with a BDI

score of 20 or above were offered a clinical interview by a consultant psychiatrist. The consultant psychiatrist checked all questionnaires with cut-offs between 14 Rolziracetam and 19 and interviewed 10 randomly selected patients to be sure that the patients were not at risk for depression or committing suicide. The result of the BDI-II was documented in medical records so that patients who declined an interview with the consultant psychiatrist could be followed up. The Hamilton Depression Scale (HDS-17) was used to validate the results of the BDI-II. HDS-17 consists of a semi-structured interview with 17 items. Scores represent a synthesis of severity and frequency of occurrence. ‘Mild’ depression is generally defined by scores from 7 to 12; ‘moderate’ by scores from 13 to 20; and ‘severe’ by scores above 20 [19]. Data on diagnosed depression were also obtained from the medical records of all 391 HIV-positive patients. We conducted statistical analysis using STATA 8 (StataCorp LP, College Station, TX, USA). All data were double-entered. The primary endpoints at baseline were the prevalence of symptom criteria for depression (BDI).

A focus-group interview was carried out with 15 HIV-positive patients (both homosexual and heterosexual men and women of various ethnicities) to identify relevant questions. The responses were transcribed and analysed. The final questionnaire was validated by a pilot

study with 12 HIV-positive patients. The participants in the pilot study were not eligible to participate in the study. The following see more variables were recorded: gender, age, educational level, ethnicity, current job, marital status, current adherence [17], current financial situation, route of infection, HIV exposure group, unsafe sex and psychological factors (guilt, shame, anxiety, concern, stress, loneliness, influence of HIV on life situation, constant thoughts about HIV, living a double life with HIV as a secret, the feeling that HIV limits way of living and stigmatization). The Beck Depression Inventory

II (BDI-II) [18] was used to assess the prevalence and severity of depressive symptoms. The BDI-II has shown high validity and reliability in measuring depressive symptoms. Respondents GPCR & G Protein inhibitor were required to rate 21 items from 0 to 3 according to how they had felt during the previous 2 weeks. The BDI-II focuses on both the cognitive-affective symptoms of depression, such as pessimism and diminished self-esteem, and the somatic symptoms of depression such as weight loss. A score of 14 or more is widely accepted as a cut-off point indicating depression on the 21-item BDI-II. The cut-off scores were: 0–13, minimal depression; 14–19, mild depression; 20–28, moderate depression; and 29–63, major depression. A cut-off point of 20 was chosen for validation using the Hamilton Depression Scale [19]. All patients with a BDI

score of 20 or above were offered a clinical interview by a consultant psychiatrist. The consultant psychiatrist checked all questionnaires with cut-offs between 14 ADAMTS5 and 19 and interviewed 10 randomly selected patients to be sure that the patients were not at risk for depression or committing suicide. The result of the BDI-II was documented in medical records so that patients who declined an interview with the consultant psychiatrist could be followed up. The Hamilton Depression Scale (HDS-17) was used to validate the results of the BDI-II. HDS-17 consists of a semi-structured interview with 17 items. Scores represent a synthesis of severity and frequency of occurrence. ‘Mild’ depression is generally defined by scores from 7 to 12; ‘moderate’ by scores from 13 to 20; and ‘severe’ by scores above 20 [19]. Data on diagnosed depression were also obtained from the medical records of all 391 HIV-positive patients. We conducted statistical analysis using STATA 8 (StataCorp LP, College Station, TX, USA). All data were double-entered. The primary endpoints at baseline were the prevalence of symptom criteria for depression (BDI).

In our cohort, all rates of selected OSDs markedly decreased as HAART use increased. Our data support the conclusion that thrombocytopenia in children responds to HAART treatment, as has already been described Selleckchem PD0325901 in

adults [26]. Despite the scarcity of information in children, there is one report of three cases in which peripheral cytopenias improved under HAART [27]. Nevertheless, larger studies are needed to determine the effects of HAART on haemopoietic cell abnormalities in the paediatric population. A dramatic decrease in the rate of HIV-related wasting syndrome has been observed in our cohort as the use of HAART has increased. In the adult population, weight loss and wasting remain important AIDS-defining conditions independently associated with mortality, despite the advent of HAART [28]. Other authors have recently observed

that the early use of HAART may prevent the development of chronic lung disease in children [29,30]. Lymphoid interstitial pneumonia has been described to improve as a result of HAART [31] or as a clinical manifestation of the immune reconstitution inflammatory syndrome [32]. This last effect was not observed in our patients, while the significant decrease in the rate of lymphoid interstitial pneumonia was attributed to the widespread use of HAART. Similarly, in our series, the decrease in cardiomyopathy may be attributed mainly to the use of HAART, as dilated cardiomyopathy was the only HIV-associated event recorded. However, in HAART-treated adult series, additional cardiovascular Selleckchem BMS-354825 consequences have been described as a result of the metabolic syndrome with a propensity for hyperlipidaemia. The involvement of the cardiovascular system is of major concern in HIV-infected children as the long-term consequences associated with atherosclerotic heart disease are unknown [33,34]. The frequency of the most severe

forms of HIV-associated encephalopathy among children has dropped dramatically since the introduction of HAART in our patients. Of concern, however, is the this website possibility that a more insidious form of this disorder, with residual neurological, cognitive and learning impairments, may currently be occurring among older vertically infected children as a result of inadequate penetration of the antiretroviral agents into the cerebrospinal fluid [35,36]. Thus, early predictive markers for the prompt and reliable identification of infants who are at risk for encephalopathy are needed [37]. Finally, our study had several limitations, such as the heterogeneous collection of data, both retrospective and prospective, and the lack of a direct relationship between HAART and clinical manifestations, CD4 cell counts and HIV viral loads in every CP.

, 2008a). Sugar analysis was carried out by acid hydrolysis of polysaccharides, followed by reduction, acetylation and quantification of alditol acetates by gas–liquid chromatography, using methods adapted from Blakeney et al. (1983). Total uronic acids were determined colorimetrically at 580 nm from a standard curve of galacturonic acid using the method of Blumenkrantz & Asboe-Hansen (1973). Galacturonic acid and APO866 order glucuronic acid are not differentiated by this method. Determination of phenolics and flavonoids of NS and BS and solid residues recovered after in vitro gastric and duodenal digestion was carried out using a Shimadzu HPLC system equipped with a UV-Vis photodiode-array detector

and a fluorescence detector (Hewlett Packard 1046A). Detection was performed at 270 nm for hydroxybenzoic acids and flavanones and at 370 nm for flavonols. The UV spectra of the different compounds were recorded from 240 to 400 nm. The wavelengths used for fluorescence detection Thiazovivin of flavan-3-ols were: λex, 276 nm; λem, 316 nm. Data acquisition was performed using class-vp5 chemstation software (Shimadzu, Japan) as reported previously (Mandalari et al., 2009). Water-jacketed fermenter

vessels (300 mL) filled with 135 mL of presterilized basal growth medium (2 g L−1 peptone water, 2 g L−1 yeast extract, 0.1 g L−1 NaCl, 0.04 g L−1 K2HPO4, 0.04 g L−1 KH2PO4, 0.01 g L−1 MgSO4·7H2O, 0.01 g L−1 CaCl2·6H2O, 2 g L−1 NaHCO3, 2 mL Tween 80, 0.02 g L−1 haemin, 10 μL vitamin K1, Adenosine 0.5 g L−1 cysteine HCl, 0.5 g L−1 bile salts, pH 7.0) were inoculated with 15 mL of faecal slurry, prepared by homogenizing 10% w/v freshly voided faecal material of one healthy donor in 0.1 M phosphate-buffered saline (PBS), pH 7.0. The almond skin extract (NS or BS postdigestion) or fructo-oligosaccharides (FOS) was added to yield a final concentration

of 1% (w/v). A negative control was performed with no addition in the fermenter vessels. Each vessel was magnetically stirred, the temperature was set at 37 °C and pH was automatically maintained at 6.8. Anaerobic conditions were maintained by sparging the vessels with oxygen-free nitrogen at 15 mL min−1. Samples (5 mL) were removed at 0, 4, 8 and 24 h for bacterial enumeration and fatty acid analysis. Fermentations were run on three separate occasions. Bacteria were counted using FISH (Rycroft et al., 2001). Duplicate fermentation samples were diluted four times in 4% w/v filtered paraformaldehyde and fixed overnight at 4 °C. Samples were then washed twice with filtered PBS (0.1 M, pH 7.0) and stored at −20 °C in PBS/ethanol (1 : 1, v/v) until further analysis. Hybridization was performed at optimal temperature using genus-specific 16S rRNA-targeted oligonucleotide probes labelled with the fluorescent dye Cy3 for the different bacterial groups or with 4′,6-diamidino-2-phenylindole for total cell counts.