Glyphosate perturbs the gut microbiota of honey bees.

Glyphosate, the primary herbicide used globally for weed control, targets the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) enzyme in the shikimate pathway found in plants and some microorganisms. Thus, glyphosate may affect bacterial symbionts of animals living near agricultural sites, including pollinators such as bees. The honey bee gut microbiota is dominated by eight bacterial species that promote weight gain and reduce pathogen susceptibility. The gene encoding EPSPS is present in almost all sequenced genomes of bee gut bacteria, indicating that they are potentially susceptible to glyphosate. We demonstrated that the relative and absolute abundances of dominant gut microbiota species are decreased in bees exposed to glyphosate at concentrations documented in the environment. Glyphosate exposure of young workers increased mortality of bees subsequently exposed to the opportunistic pathogen Serratia marcescens Members of the bee gut microbiota varied in susceptibility to glyphosate, largely corresponding to whether they possessed an EPSPS of class I (sensitive to glyphosate) or class II (insensitive to glyphosate). This basis for differences in sensitivity was confirmed using in vitro experiments in which the EPSPS gene from bee gut bacteria was cloned into Escherichia coli All strains of the core bee gut species, Snodgrassella alvi, encode a sensitive class I EPSPS, and reduction in S. alvi levels was a consistent experimental result. However, some S. alvi strains appear to possess an alternative mechanism of glyphosate resistance. Thus, exposure of bees to glyphosate can perturb their beneficial gut microbiota, potentially affecting bee health and their effectiveness as pollinators.

Detection and Characterization of Streptomycin Resistance (strA-strB) in a Honeybee Gut Symbiont (Snodgrassella alvi) and the Associated Risk of Antibiotic Resistance Transfer.

Use of antibiotics in medicine and farming contributes to increasing numbers of antibiotic-resistant bacteria in diverse environments. The ability of antibiotic resistance genes (ARG) to transfer between bacteria genera contributes to this spread. It is difficult to directly link antibiotic exposure to the spread of ARG in a natural environment where environmental settings and study populations cannot be fully controlled. We used managed honeybees in environments with contrasting streptomycin exposure (USA: high exposure, Norway: low exposure) and mapped the prevalence and spread of transferrable streptomycin resistance genes. We found a high prevalence of strA-strB genes in the USA compared to Norway with 17/90 and 1/90 positive samples, respectively (p < 0.00007). We identified strA-strB genes on a transferrable transposon Tn5393 in the honeybee gut symbiont Snodgrassella alvi. Such transfer of resistance genes increases the risk of the spread to new environments as honeybees are moved to new pollination sites.

Emergence and genomic analysis of MDR Laribacter hongkongensis strain HLGZ1 from Guangzhou, China.

Background: Laribacter hongkongensis is a facultative anaerobic, non-fermentative, Gram-negative bacillus associated with community-acquired gastroenteritis and traveller's diarrhoea. No clinical MDR L. hongkongensis isolate has been reported yet. Methods: We performed WGS (PacBio and Illumina) on a clinical L. hongkongensis strain HLGZ1 with an MDR phenotype. Results: HLGZ1 was resistant to eight classes of commonly used antibiotics. Its complete genome was a single circular chromosome of 3 424 272 bp with a G + C content of 62.29%. In comparison with the reference strain HLHK9, HLGZ1 had a higher abundance of genes associated with DNA metabolism and recombination. Several inserts including two acquired resistance gene clusters (RC1 and RC2) were also identified. RC1 carried two resistance gene cassette arrays, aac(6')-Ib-cr-aadA2-Δqac-Δsul1-floR-tetR-tetG and arr-3-dfrA32-ereA2-Δqac-sul1, which shared significant nucleotide sequence identities with the MDR region of Salmonella Genomic Island 1 from Salmonella enterica serovar Typhimurium DT104. There was also an integron-like structure, intl1-arr3-dfrA27-Δqac-sul1-aph(3')-Ic, and a tetR-tetA operon located on RC2. MLST analysis identified HLGZ1 as ST167, a novel ST clustered with two strains previously isolated from frogs. Conclusions: This study provides insight into the genomic characteristics of MDR L. hongkongensis and highlights the possibilities of horizontal resistance gene transfer in this bacterium with other pathogens.

Antibiotics reduce genetic diversity of core species in the honeybee gut microbiome.

The gut microbiome plays a key role in animal health, and perturbing it can have detrimental effects. One major source of perturbation to microbiomes, in humans and human-associated animals, is exposure to antibiotics. Most studies of how antibiotics affect the microbiome have used amplicon sequencing of highly conserved 16S rRNA sequences, as in a recent study showing that antibiotic treatment severely alters the species-level composition of the honeybee gut microbiome. But because the standard 16S rRNA-based methods cannot resolve closely related strains, strain-level changes could not be evaluated. To address this gap, we used amplicon sequencing of protein-coding genes to assess effects of antibiotics on fine-scale genetic diversity of the honeybee gut microbiota. We followed the population dynamics of alleles within two dominant core species of the bee gut community, Gilliamella apicola and Snodgrassella alvi, following antibiotic perturbation. Whereas we observed a large reduction in genetic diversity in G. apicola, S. alvi diversity was mostly unaffected. The reduction in G. apicola diversity accompanied an increase in the frequency of several alleles, suggesting resistance to antibiotic treatment. We find that antibiotic perturbation can cause major shifts in diversity and that the extent of these shifts can vary substantially across species. Thus, antibiotics impact not only species composition, but also allelic diversity within species, potentially affecting hosts if variants with particular functions are reduced or eliminated. Overall, we show that amplicon sequencing of protein-coding genes, without clustering into operational taxonomic units, provides an accurate picture of the fine-scale dynamics of microbial communities over time.

Laribacter hongkongensis: clinical presentation, epidemiology and treatment. A review of the literature and report of the first case in Denmark.

BACKGROUND: Laribacter hongkongensis is an emerging pathogen related to gastroenteritis that can cause invasive and even fatal disease. The aim of this review is to describe the clinical presentation, epidemiology, treatment options and implications for the clinical microbiology laboratory. METHODS: We searched Pubmed using the term Laribacter hongkongensis with limitations human and language English, and identified 35 publications with eight reports on human cases. RESULTS: We describe our first case of prolonged, travel-related gastroenteritis where Laribacter hongkongensis was isolated as the sole pathogen. Our review suggests that L. hongkongensis causes non-bloody acute diarrhoea with potential for invasive disease, since three cases of bacteraemia and one case of dialysis related peritonitis have been described previously. L. hongkongensis has primarily been described in Asia, but reports from Europe, North America and Australia suggests a worldwide distribution. Broad culturing with subsequent identification by the MALDI-TOF is the current strategy for detection of L. hongkongensis. Phenotypic susceptibility testing is necessary to guide the treatment choice. Few resistance genes have been described in L. hongkongensis. CONCLUSION: L. hongkongensis should be considered a potential cause of acute and prolonged diarrhoea. Clinicians must be aware of the test methods in the local clinical microbiology laboratory, since L. hongkongensis is difficult to detect and easily overlooked.

Prevalence and characterization of quinolone resistance in Laribacter hongkongensis from grass carp and Chinese tiger frog.

Laribacter hongkongensis is a food-borne bacterium associated with community-acquired gastroenteritis and diarrhoea. Quinolone resistance was recently reported in bacterial isolates from aquatic products, but the molecular mechanisms for resistance were still unknown. In this study, a total of 157 L. hongkongensis strains were isolated from grass carps (n = 443) and Chinese tiger frogs (n = 171). Twenty-one ciprofloxacin-resistant strains were analysed for mutations in quinolone resistance-determining regions (QRDR), acquired quinolone resistance (AQR) genes and the role of efflux pumps in resistance. All QRDR mutations in gyrA (codons 85 and 89) and parC (codons 83 and 231) were found to be closely associated with ciprofloxacin resistance. The AQR gene aac(6')-Ib-cr was found in 42.9% (9/21) of the resistant strains, but qnrA, qnrB, qnrC, qnrD, qnrS and qepA were not detected. No significant change of MICs to ciprofloxacin was observed in the presence of an efflux pump inhibitor, indicating the role of efflux pump was probably absent. All 21 ciprofloxacin-resistant strains showed different electrophoretic patterns, which suggested they were not genetically related. These data highlight the importance of QRDR mutations and the AQR gene aac(6')-Ib-cr during the development of quinolone resistance in a heterogeneous population of L. hongkongensis.

The prevalence, antimicrobial resistance and PFGE profiles of Laribacter hongkongensis in retail freshwater fish and edible frogs of southern China.

Laribacter hongkongensis is a novel emerging pathogen associated with human gastroenteritis. We aimed to investigate the prevalence, antimicrobial resistance and genotypic relationship of 199 L. hongkongensis isolates from 690 intestinal samples of fish and frogs. These samples were collected from retail markets in the city of Guangzhou in southern China from October 2008 to September 2009. L. hongkongensis was detected in from 80 (16.3%) out of 490 freshwater fish, and this number included 76 (32.3%) out of 235 grass carp and 4 (14.8%) out of 27 bighead carp. A higher isolation rate of 59.5% (119 out of 200) was observed in edible frogs. The isolation rate was highest in the spring in comparison with other seasons. Notably, 63.8% of the isolates were resistant to at least one class of antimicrobial agents. Analysis by pulsed-field gel electrophoresis (PFGE) revealed that the isolates could be grouped into three clusters. Isolates from fish intestines were grouped into two clusters: cluster I and II. Isolates of frog-origin and several fish-origin isolates were grouped into cluster III. Two patient-derived strains could be classed into cluster III. Extensive genetic heterogeneity among the isolates was observed. The results indicate that L. hongkongensis isolates exhibits host tropism, extensive resistance to widely used antimicrobials and diverse biological evolution in an aquatic environment. The frog is more likely than the freshwater fish to be the potential source for human infection with L. hongkongensis.

Identification and characterization of integron-associated antibiotic resistant Laribacter hongkongensis isolated from aquatic products in China.

Laribacter hongkongensis is a recently discovered bacterium associated with gastroenteritis. In this study, a total of 199 isolates of this species obtained from aquatic products (n=462) in Guangzhou City, China, were examined for their susceptibility to 19 antimicrobial agents and the presence of antimicrobial resistance integrons. The genetic relatedness of the isolates with integrons was also evaluated. A PCR-based method was used to screen integrons and found that 13 (6.5%) of the isolates harbored class 1 integrons. The antimicrobial resistance rates of integron-positive isolates were significantly higher than integron-negative ones for cefepime, ciprofloxacin, gentamicin, tetracycline, trimethoprim-sulfamethoxazole and rifampicin. Genetic sequence analysis revealed that these integrons contained various antimicrobial-resistance genes (dfrA1, dfrA14, dfrA17, dfrA32, aadA1, aadA2, aadA5, cmlA5, arr2, ereA and orfC) organized into different gene cassettes arrangements including a novel array of dfrA14-arr2-cmlA5. Pulsed-field gel electrophoresis (PFGE) yielded 13 different patterns among 13 integron-positive isolates, which could be grouped into four clusters. These indicate the dispersal of multi-resistant integrons among different molecular types. To the best of our knowledge, this is the first report showing distribution and characterization of class 1 integrons among L. hongkongensis isolates.

[Prevalence and antimicrobial susceptibility of Laribacter hongkongensis and enterotoxigenic Escherichia coli isolated from patients with diarrhea in Guangzhou].

OBJECTIVE: To survey the prevalence of enterotoxigenic Escherichia coli (ETEC) and Laribacter hongkongensis (LH) and their drug resistance in diarrhea patients in Guangzhou. METHODS: We detected 646 fecal cases collected between Sep 2008 and Oct 2009 from the out-patient and emergency departments in a hospital. EC enriched culture medium was used for enrichment. MAC- and CMAC-specific culture media were used to isolate ETEC and LH from the specimens. The biochemical agents API20NE and API20E were employed for biochemical identification, and PCR was used for genetic identification. K-B disk diffusion method was used for antimicrobial susceptibility testing. RESULTS: No LH was detected in the total 646 patients, and 38 patients were positive for ETEC, with a detection rate of 6%. Antibiotics resistance test showed that 38 strains of ETEC had a high resistance rate to penicillin, tetracycline and sulfa, but remained sensitive to cephalosporins. CONCLUSIONS: LH may have a low prevalence in Guangzhou. The incidence of diarrhea caused by ETEC tends to decrease as compared with that a decade ago, and further multi-center survey is needed for confirmation. Consumption of aquatic products may be one of the major risk factors for ETEC infection. Cephalosporins can be used for ETEC-induced diarrhea.

Current status and future directions for Laribacter hongkongensis, a novel bacterium associated with gastroenteritis and traveller's diarrhoea.

PURPOSE OF REVIEW: Despite extensive investigations, a microbiological cause cannot be found in about half of the patients with infectious disease. Throughout the years, scientists have spent tremendous efforts in looking for microorganisms associated with these "unexplained infectious disease syndromes". Recently, a novel bacterium, Laribacter hongkongensis, was discovered and shown to be associated with gastroenteritis and traveller's diarrhoea. This review summarizes the current status, and shares with the readers the authors' experience in the microbiology, classification, epidemiology, clinical disease, laboratory diagnosis, antibiotic resistance and treatment of L. hongkongensis. It also discusses the importance and perspective of describing novel pathogenic bacterial species. RECENT FINDINGS: L. hongkongensis was shown to be associated with gastroenteritis and traveller's diarrhoea. Consumption of fish was associated with recovery of L. hongkongensis. Freshwater fish was a reservoir of L. hongkongensis. Genotypic typing revealed the possibility of virulent clones of L. hongkongensis. The class C beta-lactamase of L. hongkongensis has been cloned and characterized. SUMMARY: In 2001, L. hongkongensis, a novel genus and species, was first discovered in Hong Kong from the blood and empyema pus of a patient with alcoholic cirrhosis. Subsequently, it was isolated from patients in other parts of the world. Recently, this bacterium was found to be associated with community-acquired gastroenteritis and traveller's diarrhoea using cefoperazone MacConkey agar as the selective medium. Further studies, including setting up of animal and tissue culture models and characterization of virulence factors, should be performed. For pathogenic microbes, even one strain of a novel species should be described, so that global concerted efforts can be drawn to look for more cases associated with such a pathogen.

Use of cefoperazone MacConkey agar for selective isolation of Laribacter hongkongensis.

A new selective medium, cefoperazone MacConkey agar (CMA), was developed for primary isolation of Laribacter hongkongensis from stool. Its performance in quantitative recovery and in a clinical evaluation of 4,741 human diarrheal stool specimens was superior to that of charcoal cefoperazone deoxycholate agar. In addition, with CMA, Arcobacter butzleri was unexpectedly isolated from the stools of six patients.

Effects of Gypsophila saponins on bacterial growth kinetics and on selection of subterranean clover rhizosphere bacteria.

Plant secondary metabolites, such as saponins, have a considerable impact in agriculture because of their allelopathic effects. They also affect the growth of soil microorganisms, especially fungi. We investigated the influence of saponins on rhizosphere bacteria in vitro and in soil conditions. The effects of gypsophila saponins on the growth kinetics of rhizosphere bacteria were studied by monitoring the absorbance of the cultures in microtiter plates. Gypsophila saponins (1%) increased the lag phase of bacterial growth. The impact of gypsophila saponins on subterranean clover rhizosphere was also investigated in a pot experiment. The addition of gypsophila saponins did not modify clover biomass but significantly increased (twofold with 1% saponins) the weight of adhering soil. The number of culturable heterotrophic bacteria of the clover rhizosphere was not affected by the addition of gypsophila saponins. Nevertheless, the phenotypical characterization of the dominant Gram-negative strains of the clover rhizosphere, using the Biolog system, showed qualitative and quantitative differences induced by 1% saponins. With the addition of saponins, the populations of Chryseomonas spp. and Acinetobacter spp., the two dominant culturable genera of control clover, were no longer detectable or were significantly decreased, while that of Aquaspirillum dispar increased and Aquaspirillum spp. became the major genus. Aquaspirillum dispar and Aquaspirillum spp. were also the dominant rhizosphere bacteria of Gypsophila paniculata, which greatly accumulates these saponins in its roots. These results suggest that saponins may control rhizosphere bacteria in soil through rhizodeposition mechanisms.

[Dog bite infections associated with CDC group EF-4a. Report of 2 cases]. / Infecciones por mordedura de perro asociadas a CDC grupo EF-4a. Comunicación de 2 casos.

BACKGROUND: Group EF-4 bacteria make up part of the normal flora of the oral cavity of dogs and cats. Few reports have been published on the incidence of human infections by this group of bacteria and these are associated with animal bite or scratch. Two cases of infections by CDC group EF-4 by dog bite were diagnosed in 1996 by the Bacteriology Laboratory of the authors' hospital. These cases are herein described and the biochemical analysis and profile of sensitivity of this little known group of bacteria evaluated. METHODS: Two clinical cases of infection by CDC group EF-4a by dog bite are described. Identification of the bacteria was performed by conventional biochemical tests and quantitative antibiotic sensitivity to 12 antibiotics was carried out by the seried broth macrodilution method. RESULTS: The two strains isolated corresponded to biovar "a" of group EF-4 being sensitive to: ampicillin, ceftriaxone, aminoglycosides, chloramphenicol, rifamipicin, TMS and ciprofloxacin, intermediate sensitivity to erythromycin and were resistant to cefalotine, oxacillin and vancomycin. With respect to penicillin, one of the strains was sensitive and the other presented intermediate sensitivity. Neither of the strains produced beta lactamase. CONCLUSIONS: Although Pasteurella sp. is usually considered in dog bite wounds, the possible presence of group EF-4 should be taken into account since the sensitivity of both microorganisms against penicillin and cefalotin, which are effective against Pasteurella but less active against group EF-4 bacteria differ.

Timed kill kinetic studies of levofloxacin, ofloxacin, and ciprofloxacin against Moraxella catarrhalis.

Levofloxacin bactericidal activity was compared to ciprofloxacin and ofloxacin against 10 strains of Moraxella catarrhalis. The cidal action (by kill-curve analysis) was slightly more rapid for levofloxacin, but all tested fluoroquinolones were considered bactericidal for all strains tested, including those producing BRO-1 and 2 beta-lactamases.

Prediction of bacterial susceptibility to cefpodoxime by using the ceftriaxone minimum inhibitory concentration result.

The cross-resistance or cross-susceptibility of cefpodoxime and ceftriaxone for 3700 strains of Enterobacteriaceae, oxacillin-susceptible staphylococci, Streptococcus spp., Haemophilus influenzae, Moraxella catarrhalis, and Neisseria gonorrhoeae was evaluated. With the exception of tests with Enterobacter spp. and Morganella morganii, the ceftriaxone minimum inhibitory concentration (MIC) result interpretive criteria predicted bacterial susceptibility (or resistance) to cefpodoxime with an acceptable rate of serious interpretive errors (1.5%) and an absolute categorical agreement > 92%. By using cefpodoxime interpretive criteria for ceftriaxone MICs, the interpretive errors for testing enteric bacilli were reduced from 2.1% to 1.5%.

[In vitro antibacterial activity of RU 51746 (sodium salt of cefpodoxime). Results of a multicenter study]. / Activité antibactérienne in vitro du RU 51746 (sel de sodium du cefpodoxime). Résultats d'une étude multicentrique.

Cefpodoxime proxetil, a new oral cephalosporin, is the prodrug ester of cefpodoxime. Minimal inhibitory concentrations (MIC) of RU 51746 (sodium salt of cefpodoxime: CPD) were evaluated by agar dilution for 1 696 bacterial strains isolated in 5 hospitals. For Enterobacteriaceae, MIC 50 and 90% were respectively (micrograms/ml): (1) naturally non bêtalactamase producing species: E. coli, Shigella and Salmonella 0.25-0.5; P. mirabilis 0.06-0.12. (II) chromosomal penicillinase producing species: Klebsiella 0.12-1. (III) chromosomal cephalosporinase producing species: E. cloacae and C. freundii 2-greater than 128; S. marcescens 2-64; indole + Proteus 0.25-64; P. stuartii 0.25-16. Activity of CPD was not modified on plasmid mediated penicillinase producing strains, but CPD was inactive on cephalosporinase hyperproducing strains, and on broad spectrum bêtalactamases producing strains. CPD was inactive on P. aeruginosa (MIC greater than or equal to 64) and on A. baumannii (16-pi 128). Haemophilus, regardless on bêtalactamase production status, were very susceptible to CPD (MIC less than or equal to 0.25) and B. catarrhalis was generally inhibited by 0.12 to 1. CPD was poorly active on methicillin susceptible Staphylococci (MIC 50 and 90%: 2-4) and inactive on methicillin resistant strains. Enterococci and Listeria monocytogenes were generally resistant; Streptococci A, B, C, G and Pneumococci were inhibited by low concentration: 0.002 to 0.25 (MIC 50 and 90%: 0.016-0.032) whereas MIC for other Streptococci were 0.004 to 32 (MIC 50 and 90%: 0.25-4). These antibacterial properties placed CPD in excellent position among oral cephalosporins.

[Antibacterial effect of cefixime]. / Comportement antibactérien du céfixime.

Cefixime (CFM) is a new hemi-synthetic orally active cephalosporin which exhibits a particular affinity for PBPs 3, 1a, 1bs. Its penetration through the Gram negative bacilli outer membrane is similar to that of third generation cephalosporins. The MICs were assessed by the agar dilution method against 2,489 bacterial strains collected in 10 hospitals. Against Enterobacteriaceae, MICs50 and 90 are respectively (mg/l): naturally non beta-lactamase-producing species: E. coli and Shigella: 0.25-0.5, Salmonella: 0.06 - 0.25, P. mirabilis: 0.008 - 0.0.32; chromosomal penicillinase producing species: Klebsiella: 0.06 - 2; chromosomal cephalosporinase producing species: E. cloacae and C. freundii: 1 - greater than 128, S. marcescens: 0.25 - 16, Proteus indole: + 0.06 - 4, P. stuartii: 0.032 - 0.5. CFM activity is not altered in strains producing an acquired penicillinase. On the other hand, CFM appears to be inactive against cephalosporinase hyperproducing mutants and its activity is variably decreased against expanded spectrum beta-lactamase producing strains. CFM is inactive against P. aeruginosa (MIC50 and 90: 64 - 128) and against A. baumannii (16 - 128). Haemophilus and gonococci, beta-lactamase producing or not, as well as meningococci, are highly susceptible to CFM (MIC 0.008 - 0.12). B. catarrhalis is usually inhibited by 0.03 to 0.5. CFM is moderately active against meticillin-sensitive staphylococci (MIC50 and 90: 1-64), and inactive against meticillin-resistant strains. Enterococci are usually resistant, whereas streptococci and pneumococci are inhibited by low concentrations: 0.08 to 1. CFM is a bactericidal antibiotic, as shown by MBC and killing curves determination. These antibacterial properties relate CFM to the third generation cephalosporins and position the compound in an excellent place among the orally active cephalosporins.

High-level tetracycline resistance resulting from TetM in strains of Neisseria spp., Kingella denitrificans, and Eikenella corrodens.

Similar to Neisseria gonorrhoeae, tetracycline-resistant isolates of N. meningitidis, Kingella denitrificans, and Eikenella corrodens contained 25.2-megadalton plasmids carrying the TetM determinant. In contrast, tetracycline-resistant N. subflava biovar perflava-N. sicca and N. mucosa isolates carried the TetM determinant in the chromosome.

Clinical interpretation of beta-lactamase-producing strains of Branhamella catarrhalis in sputum Gram's stain and culture.

Branhamella catarrhalis has been implicated previously as a cause of bronchopulmonary infections. Sputum Gram's stain and culture results suggesting significant infection with beta-lactamase-producing strains of B. catarrhalis were correlated with a retrospective chart review of eight pediatric and ten adult patients. Preexisting pulmonary disease was observed in 12 patients; 5 had a history of aspiration; and 13 were intubated. Clinically, ten patients had pneumonia, five had bronchitis, and three manifested no disease. Only three sputum specimens grew a pure culture of B. catarrhalis, and six specimens yielded B. catarrhalis in the presence of normal upper respiratory flora. Analysis of broth microdilution susceptibility test results showed that 90% of the strains were inhibited at the following minimum inhibitory concentrations (MICs90): ampicillin, 8 micrograms/mL; cefotaxime, 0.5 microgram/mL; cefoxitin, 0.5 microgram/mL; cephalexin, 4 micrograms/mL; cephalothin, 8 micrograms/mL; chloramphenicol, 1 microgram/mL; clindamycin, 4 micrograms/mL; erythromycin, 0.25 microgram/mL; methicillin, 16 micrograms/mL; mezlocillin, 16 micrograms/mL; moxalactam less than or equal to 0.6 microgram/mL; penicillin, 16 micrograms/mL; piperacillin, 8 micrograms/mL; tetracycline, less than or equal to 0.3 microgram/mL; and trimethoprim/sulfamethoxazole, 1.6/30 micrograms/mL. Therapy may have been adequate in only eight (44%) of the cases. However, all but four of the patients, who died of unrelated causes, exhibited resolution of disease. The data indicate that Gram's stain and culture results of sputum specimens suggesting B. catarrhalis bronchopulmonary infection should be interpreted with caution by clinicians.