Understanding clinical and immunological features associated with Pseudomonas and Staphylococcus keratitis

Pseudomonas aeruginosa and Staphylococcus aureus are the two dominant Gram-negative and -positive species, respectively, isolated from patients with contact lens-related bacterial keratitis. The clinical features of bacterial keratitis vary, such that timely differential diagnosis can be challenging, which may cause a delay in diagnosis resulting in poorer outcome. This review aims to explore the current understanding of clinical and immunological features associated with contact lens-related P. aeruginosa and S. aureus keratitis based on currently available evidence.Firstly, the review characterises contact lens-related P. aeruginosa and S. aureus keratitis, based on clinical features and prognostic factors. Secondly, the review describes the primary immune response associated with a bacterial infection in in-vivo non-scratch contact lens-wearing animal models, colonised by bacteria on contact lens and topical administration of bacteria on the cornea. Finally, the review discusses the role of macrophage inflammatory protein-2 (MIP-2) and intercellular adhesion molecule (ICAM-1) in neutrophil recruitment based on both in-vivo scratch models of bacterial keratitis and bacterial challenged in cell culture models.     

Prevalence and antimicrobial susceptibility of gram-negative bacteria isolated from bovine mastitis between 2003 and 2008 in Korea

The objective of this study was to assess trends in the prevalence and distribution of gram-negative bacteria isolated from bovine mastitis and their antimicrobial susceptibilities during a 6-yr period between 2003 and 2008 in Korea. Escherichia coli, Pseudomonas fluorescens, Klebsiella pneumoniae, Enterobacter cloacae, Acinetobacter lwoffi/junii, Pseudomonas aeruginosa, and Serratia marcescens were the most commonly observed pathogens during this period. Generally, gram-negative bacteria showed low susceptibilities to most of the antimicrobials tested in this study, except amikacin and gentamicin. Although these 2 aminoglycosides were broadly active against gram-negative bacteria, less than half of those bacteria showed susceptibilities to streptomycin. The β-lactams, except piperacillin, had the lowest activity among antimicrobials tested in this study. Susceptibilities to chloramphenicol and trimethoprim were fairy high in all genera of gram-negative bacteria, except Acinetobacter spp. and Pseudomonas spp., whereas relatively high resistance to tetracycline was observed uniformly among gram-negative bacteria. There was no significant change in the prevalence of bacterial and the proportion of antimicrobial resistance among gram-negative bacteria isolates during a 6-yr period.     

Prevalence,Antimicrobial Resistance,and Virulence Characteristics of mecA‐Encoding Coagulase‐Negative Staphylococci Isolated from Soft Cheese in Brazil

Coagulase‐negative staphylococci (CoNS), which are generally neglected as foodborne bacteria, are emerging as significant opportunistic pathogens that may be highly resistant to available antimicrobial drugs. In this study, antimicrobial susceptibility patterns, mecA gene occurrence, and virulence‐associated characteristics were evaluated in CoNS isolated from soft cheese in Brazil. A total of 227 bacterial isolates were recovered from 35 cheese samples belonging to 5 batches with 7 different trademarks. The CoNS counts ranged from 10⁶ to 10⁷ CFU/g. High antimicrobial resistance percentages were observed for oxacillin (76.2%), penicillin (78.5%), erythromycin (67.8%), gentamicin (47.2%), clindamycin (35.7%), rifampicin (26.8%), azithromycin (14.7%), tetracycline (14.7%), levofloxacin (14.2%), and sulfamethoxazole‐trimethoprim (11.9%). A low antimicrobial resistance percentage was observed for chloramphenicol (2.3%), and all of the tested bacteria were susceptible to vancomycin and linezolid. In total, a multiple antibiotic resistance (MAR) index of >0.2 was observed for 80.6% of the isolated CoNS. However, the MAR index ranged from 50% to 92.6% when only bacterial cheese isolates belonging to the same trademark were considered. Regarding to the prevalence of CoNS carrying mecA gene, 81.5% of the isolated strains were mecA⁺, and 76.2% of these were phenotypically resistant to oxacillin. Three isolates carried the enterotoxin A gene (sea), 29.5% produced biofilm in a laboratory test, and α‐ or ß‐hemolysis were observed for 3% and 5.2%, respectively. This study highlights the extent of the antimicrobial resistance phenomenon in neglected foodborne microorganisms and the potential public health risks that are related to the consumption of CoNS‐contaminated soft cheese.     

Antimicrobial susceptibility of bacterial isolates from the conjunctiva,storage cases and mobile phones of university students using contact lenses

Purpose: To investigate the presence of bacterial pathogens on the palpebral conjunctiva, mobile phones, and storage cases of contact lens wearers to study any possible correlation between types of bacteria isolated from the 3 sites and to determine their antibiotic profiles.Methods: One hundred and eighty nine swabs from the conjunctiva, mobile phones, and storage cases were collected from 63 contact lens wearing university students. The swabs were collected and transported to the microbiology laboratory within one hour and inoculated on nutrient agar, MacConkey agar, blood agar and mannitol salt agar. The subsequent bacterial isolates were identified by their cultural, morphological and biochemical characteristics.Results: Nine bacterial species were isolated and identified in the current study namely: Staphylococcus. aureus, Streptococcus. pyogenes, Enterococcus. faecalis, Shigella dysentery, Pseudomonas aeruginosa, Proteus mirabilis, Citrobacter spp., Klebsiella pneumoniae and Escherichia coli. Nine (26%) mobile phone and 7 (21%) conjunctival samples were contaminated with five different bacterial species. The highest level of contamination was detected in contact lens storage cases where 18 (52%) bacterial isolates were detected in cases.Conclusions: The storage cases and mobile phones of contact lens wearing university students were highly contaminated with pathogenic bacteria and may act as a carrier for the transmission of such bacteria to the eye causing eye infections which can be controlled by proper hygiene and using effective disinfectant for storage cases. Pathogenic bacteria were detected with multiple antibiotic resistance indices.     

Spoilage potentials and antimicrobial resistance of Pseudomonas spp. isolated from cheeses

Pseudomonas spp. are aerobic, gram-negative bacteria that are recognized as major food spoilage microorganisms. A total of 32 (22.9%) Pseudomonas spp. from 140 homemade white cheese samples collected from the open-air public bazaar were isolated and characterized. The aim of the present study was to investigate the biochemical characteristics, the production of extracellular enzymes, slime and β-lactamase, and antimicrobial susceptibility of Pseudomonas spp. isolated from cheeses. The identified isolates including Pseudomonas pseudoalcaligenes, Pseudomonas alcaligenes, Pseudomonas aeruginosa, Pseudomonas fluorescens biovar V, and P. pseudoalcaligenes ssp. citrulli were found to produce extracellular enzymes, respectively: protease and lecithinase production (100%), and lipase activity (85.7, 42.9, 100, and 100%, and nonlipolytic, respectively). The isolates did not produce slime and had no detectable β-lactamase activity. The antimicrobial susceptibility of the isolates was tested using the disk diffusion method. Pseudomonas spp. had the highest resistance to penicillin G (100%), then sulphamethoxazole/trimethoprim (28.1%). However, all Pseudomonas spp. isolates were 100% susceptible to ceftazidime, ciprofloxacin, amikacin, gentamicin, and imipenem. Multidrug-resistance patterns were not observed among these isolates. In this study, Pseudomonas spp., exhibiting spoilage features, were isolated mainly from cheeses. Isolation of this organism from processed milk highlights the need to improve the hygienic practices. All of the stages in the milk processing chain during manufacturing have to be under control to achieve the quality and safety of dairy products.     

Characterization of bacterial susceptibility isolates in sixteen dairy farms in Taiwan

Bacteria were isolated from dairy cows, dairy farm environments, and dairy workers in 2 geographically different areas of eastern and northern Taiwan. Isolates were evaluated for antimicrobial susceptibility and the phylogenetics of isolated Escherichia coli O157:H7 were characterized. A total of 1,346 bacteria were identified, including 226 E. coli, 30 Pseudomonas spp. (7 Pseudomonas aeruginosa), 259 other gram-negative bacteria, 271 Enterococcus spp., 314 Staphylococcus spp., 195 Streptococcus spp., and 51 other gram-positive bacteria. Among them, 88% (1,184/1,346) of the isolates were resistant to sulfadimethoxine. The percentages of gram-negative bacteria resistant to oxy-tetracycline and streptomycin were 48% (249/515) and 78% (404/515), respectively. Gram-positive bacteria isolated from eastern Taiwan, the least polluted region of Taiwan, were found to have greater antimicrobial resistance than those isolated from northern Taiwan. Two E. coli O157:H7 from 2 different geographical areas were isolated. Both were vt2-positive but vt1-negative and had phylogenetic similarities of 82 and 67%, respectively, compared with previous isolates. Information on antimicrobial susceptibility revealed from this dairy farm survey may serve as a baseline for future studies and may also highlight the need to formulate better regulation strategies for the safe use of antimicrobials on food-producing farms.     

Microbial background flora in small-scale cheese production facilities does not inhibit growth and surface attachment of Listeria monocytogenes

The background microbiota of 5 Norwegian small-scale cheese production sites was examined and the effect of the isolated strains on the growth and survival of Listeria monocytogenes was investigated. Samples were taken from the air, food contact surfaces (storage surfaces, cheese molds, and brine) and noncontact surfaces (floor, drains, and doors) and all isolates were identified by sequencing and morphology (mold). A total of 1,314 isolates were identified and found to belong to 55 bacterial genera, 1 species of yeast, and 6 species of mold. Lactococcus spp. (all of which were Lactococcus lactis), Staphylococcus spp., Microbacterium spp., and Psychrobacter sp. were isolated from all 5 sites and Rhodococcus spp. and Chryseobacterium spp. from 4 sites. Thirty-two genera were only found in 1 out of 5 facilities each. Great variations were observed in the microbial background flora both between the 5 producers, and also within the various production sites. The greatest diversity of bacteria was found in drains and on rubber seals of doors. The flora on cheese storage shelves and in salt brines was less varied. A total of 62 bacterial isolates and 1 yeast isolate were tested for antilisterial activity in an overlay assay and a spot-on-lawn assay, but none showed significant inhibitory effects. Listeria monocytogenes was also co-cultured on ceramic tiles with bacteria dominating in the cheese production plants: Lactococcus lactis, Pseudomonas putida, Staphylococcus equorum, Rhodococcus spp., or Psychrobacter spp. None of the tested isolates altered the survival of L. monocytogenes on ceramic tiles. The conclusion of the study was that no common background flora exists in cheese production environments. None of the tested isolates inhibited the growth of L. monocytogenes. Hence, this study does not support the hypothesis that the natural background flora in cheese production environments inhibits the growth or survival of L. monocytogenes.     

Short communication: β-Lactam resistance and vancomycin heteroresistance in Staphylococcus spp. isolated from bovine subclinical mastitis

The use of antimicrobial agents has led to the emergence of resistant bacterial strains over a relatively short period. Furthermore, Staphylococcus spp. can produce β-lactamase, which explains the survival of these strains in a focus of infection despite the use of a β-lactam antibiotic. The aim of this study was to evaluate the resistance of Staphylococcus spp. isolated from bovine subclinical mastitis to oxacillin and vancomycin (by minimum inhibitory concentration) and to detect vancomycin heteroresistance by a screening method. We also evaluated β-lactamase production and resistance due to hyperproduction of this enzyme and investigated the mecA and mecC genes and performed staphylococcal cassette chromosome mec typing. For this purpose, 181 Staphylococcus spp. isolated from mastitis subclinical bovine were analyzed. Using the phenotypic method, 33 (18.2%) of Staphylococcus spp. were resistant to oxacillin. In contrast, all isolates were susceptible to vancomycin, and heteroresistance was detected by the screening method in 13 isolates. Production of β-lactamase was observed in 174 (96%) of the Staphylococcus spp. isolates. The mecA gene was detected in 8 isolates, all of them belonging to the species Staphylococcus epidermidis, and staphylococcal cassette chromosome mec typing revealed the presence of type I and type IV isolates.     

Antimicrobial resistance profiles of 5 common bovine mastitis pathogens in large Chinese dairy herds

The prevalence of antimicrobial resistance (AMR) is increasing in human and animal pathogens, becoming a concern worldwide. However, prevalence and characteristics of AMR of bovine mastitis pathogens in large Chinese dairy herds are still unclear. Therefore, our objective was to determine the AMR profile of bacteria isolated from clinical mastitis in large (>500 cows) Chinese dairy herds. A total of 541 isolates of the 5 most common species, Staphylococcus aureus (n = 103), non-aureus staphylococci (NAS; n = 107), Streptococcus species (n = 101), Klebsiella species (n = 130), and Escherichia coli (n = 100), isolated from bovine clinical mastitis on 45 dairy farms located in 10 provinces of China were included. Presence of AMR was determined by minimum inhibitory concentrations using the microdilution method. Prevalence of multidrug resistance (resistance to >2 antimicrobials) was 27% (148/541). A very wide distribution of minimum inhibitory concentrations was screened in all isolates, including Staph. aureus isolates, which were resistant to penicillin (66%). In addition, NAS (30%) were more resistant than Staph. aureus to oxacillin (84%), penicillin (62%), tetracycline (34%), and clindamycin (33%). Prevalence of resistance to tetracycline was high (59%) in Streptococcus spp. Additionally, prevalence of resistance of both E. coli and Klebsiella spp. was high to amoxicillin/clavulanate potassium (81 and 38%, respectively), followed by tetracycline (only Klebsiella spp. 32%). A high proportion (27%) of isolates were multidrug resistant; the most frequent combinations were clindamycin-cefalexin-tetracycline or enrofloxacin-cefalexin-penicillin patterns for Staph. aureus; enrofloxacin-oxacillin-penicillin-tetracycline patterns for NAS; clindamycin-enrofloxacin-tetracycline patterns for Streptococcus spp.; amoxicillin/clavulanate potassium-ceftiofur-polymyxin B patterns for Klebsiella spp.; and amoxicillin/clavulanate potassium-ceftiofur-polymyxin B patterns for E. coli. Resistance for 4 kinds of antimicrobials highly critical for human medicine, including daptomycin, vancomycin, imipenem, and polymyxin B, ranged from 0 to 24%. In conclusion, prevalence of AMR in mastitis pathogens was high on large Chinese dairy farms, potentially jeopardizing both antimicrobial efficacy and public health. Results of this study highlighted the need for improvements in antimicrobial stewardship and infection control programs in large Chinese dairy farms to reduce emergence of AMR.     

Presence of Campylobacter and Arcobacter species in in-line milk filters of farms authorized to produce and sell raw milk and of a water buffalo dairy farm in Italy

The objectives of this study were to investigate the presence of Campylobacter spp. and Arcobacter spp. in dairy herds authorized for the production and sale of raw milk and in a water buffalo dairy farm, and to test the antimicrobial susceptibility of the isolates. A total of 196 in-line milk filters were collected from 14 dairy farms (13 bovine and 1 water buffalo) for detection of Campylobacter spp. and Arcobacter spp. by microbiological culture. For each farm investigated, 1 isolate for each Campylobacter and Arcobacter species isolated was tested using the Etest method (AB Biodisk, Solna, Sweden) to evaluate the susceptibility to ciprofloxacin, tetracycline, chloramphenicol, ampicillin, erythromycin, and gentamicin. A total of 52 isolates were detected in 49 milk filters in 12 farms (85.7%) out of 14 and the isolates were identified as Campylobacter jejuni (6), Campylobacter hyointestinalis ssp. hyointestinalis (8), Campylobacter concisus (1), Campylobacter fetus ssp. fetus (1), Arcobacter butzleri (22), and Arcobacter cryaerophilus (14). The small number of isolates tested for antimicrobial susceptibility precludes any epidemiological consideration but highlights that all Campylobacter isolates were susceptible to macrolides, which are the first-choice drugs for the treatment of campylobacteriosis, and that resistance to fluoroquinolones and tetracycline was detected; for Arcobacter isolates, resistance to ampicillin and chloramphenicol was detected. The sale of raw milk for human consumption by self-service automatic vending machines has been allowed in Italy since 2004 and the presence of C. jejuni in in-line milk filters confirms that raw milk consumption is a significant risk factor for human infection. The high occurrence of emerging Campylobacter spp. and Arcobacter spp. discovered in dairy farms authorized for production and sale of raw milk represents an emerging hazard for human health.     

Antimicrobial activity of crude extracts from actinomycetes against mastitis pathogens

The emergence of antimicrobial resistance to commonly used antibiotics has necessitated the development of new antimicrobial products. Crude extracts produced by actinomycetes contain antimicrobial metabolites that can inhibit bacterial growth. The objective of our study was to evaluate the antimicrobial activity of crude extracts (Caat1–54 and CaatP5–8) produced by actinomycetes against isolates of Staphylococcus aureus, Staphylococcus chromogenes, Streptococcus dysgalactiae, and Streptococcus uberis, which were obtained from the milk of cows affected by mastitis in 23 dairy herds. Twenty isolates of each bacterial species were used to define minimum inhibitory concentrations (MIC) of both crude extracts and ceftiofur (positive control). The MIC₅₀ and MIC₉₀ were defined at the concentration required to inhibit the growth of 50 and 90% of bacterial isolates tested, respectively. The MIC results were evaluated by survival analysis. Staphylococcus aureus isolates presented MIC₉₀ of Caat 1–54 ≥6.25 µg/mL, ceftiofur ≥12.5 µg/mL, and Caat P5–8 ≥100 µg/mL. Streptococcus uberis presented MIC₉₀ of ceftiofur ≥0.39 µg/mL, Caat 1–54 ≥50 µg/mL, and Caat P5–8 ≥100 µg/mL. Staphylococcus chromogenes isolated from subclinical mastitis presented MIC₉₀ of Caat 1–54 ≥0.78 µg/mL and ceftiofur and Caat P5–8 of ≥6.25 and ≥100 µg/mL, respectively. Streptococcus dysgalactiae isolated from clinical mastitis presented similar MIC₉₀ values between antimicrobials tested (ceftiofur, Caat 1–54, and Caat P-58), but these values (≥100 µg/mL) were higher than the values obtained from other pathogens evaluated in the present study. Our results indicate that Caat 1–54 and Caat P5–8 crude extracts present in vitro antimicrobial activity against isolates of Staph. aureus, Staph. chromogenes, Strep. dysgalactiae, and Strep. uberis isolated from clinical and subclinical mastitis.     

In vitro antimicrobial resistance profiles of Streptococcus uberis,Lactococcus spp., and Enterococcus spp. from quarter milk samples of cows between 2015 and 2019 in Southern Germany

The objective was to describe and compare antimicrobial resistance patters of esculin-hydrolyzing streptococci and streptococcal-like organisms (Streptococcus uberis, Enterococcus faecium, Enterococcus faecalis, Lactococcus garvieae, Lactococcus lactis) from routine diagnostic samples of the udder health laboratory of the Bavarian Animal Health Services between 2015 and 2019. All routine diagnostic samples of the udder health laboratory of the Bavarian Animal Health Services, that were tested with a standard microbroth dilution, were eligible to be included in this retrospective case series. A California Mastitis Test result was available for all samples. Most Strep. uberis and L. lactis were susceptible to all antibiotics tested. Enterococcus faecium had consistently the highest minimum inhibitory concentration required to inhibit the growth of 90% of tested isolates. The resistance patterns of Lactococcus garvieae were positioned between enterococci and L. lactis. The minimum inhibitory concentration for various antibiotics and pathogens tended to decrease over the 5-yr period. Regardless of the pathogen, isolates of clinical cases were less likely to express in vitro resistance than isolates of healthy or subclinical cases. Streptococcus uberis or L. lactis showed hardly any in vitro resistance to tested antibiotic groups. Penicillin should remain the first-choice antimicrobial for the therapy of Strep. uberis and Lactococcus spp. However, a success of any antimicrobial treatment of enterococcal infections seems questionable.     

Development of a DNA microarray for detection and identification of Legionella pneumophila and ten other pathogens in drinking water

The safety and accessibility of drinking water are major concerns throughout the world. Consumption of water contaminated with infectious agents, toxic chemicals or radiological hazards represents a significant health risk and is strongly associated with mortality. Therefore, we have developed an oligonucleotide-based microarray using the sequences of 16S-23S rDNA internal transcribed spacer regions (ITS) and the gyrase subunit B gene (gyrB) found in the most prevalent and devastating waterborne pathogenic agents. This new diagnostic contains 26 specific probes and can simultaneously detect Aeromonas hydrophila, Klebsiella pneumoniae, Legionella pneumophila, Pseudomonas aeruginosa, Salmonella spp., Shigella spp., Staphylococcus aureus, Vibrio choleraeo, Vibrio parahaemolyticus, Yersinia enterocolitica and Leptospira interrogans. Testing was carried out against a total of 218 bacterial strains, including 53 representative strains, 103 clinical isolates and 62 strains of other bacterial species belonging to 10 genera and 48 species. The results were specific and reproducible, with a detection sensitivity of 0.1 ng DNA or 10⁴ CFU/ml achieved for pure cultures of each target organism. The diluted cultures and real drinking water samples were tested by the microarray with 100% accuracy. This novel diagnostic method is superior in time- and labor-efficiency to conventional bacterial culture and antiserum agglutination, and can be readily applied to epidemiological surveillance and other food safety applications.     

Clinical factors associated with positive corneal culture in suspected microbial keratitis

AimsDetermine demographic and clinical characteristics associated with positive culture in suspected microbial keratitis.MethodsRetrospective audit of patients that had corneal scrapings between October 1999-September 2004 at Princess Alexandra Hospital. Clinical information was gathered from medical records, smear and culture results from the local microbiology database. Univariate and multivariate analyses of variables associated with positive cultures and calculation of population attributable risk percentage (PAR).ResultsUnivariate analysis showed that positive cultures were associated with patients over 60 years (81% vs 55%; p < 0.001), presenting visual acuity (VA) of 6/24 or worse (74% vs 57%; p = 0.012) or contact lens-related keratitis (CLK 77% vs 62%; p = 0.08). Analysis of patients’ clinical presentation showed that positive culture was associated with a central epithelial defect (74% vs 57%; p = 0.012), anterior chamber reaction of 2 + cells or worse (73% vs 56%; p = 0.03), an epithelial defect of 2 mm or more in diameter (71% vs 50%; p = 0.006) or no prior treatment with antibiotics (68% vs 56%; p = 0.053). Multivariate analysis showed the independent variables associated with positive cultures were VA of 6/24 or worse on presentation, contact lens-related keratitis, age greater than 60 years, an anterior chamber reaction of 2 + cells or worse and no prior treatment with antibiotics. The factor with the highest attributable risk (PAR%) for a positive corneal scraping was VA of 6/24 or worse on presentation (21%).ConclusionsIn this series positive cultures were associated with poor presenting VA contact lens keratitis (CLK), older age, anterior chamber reaction and no prior treatment with antibiotics.     

Influence of farming methods on microbiological contamination and prevalence of resistance to antimicrobial drugs in isolates from beef

The presence of Escherichia coli, Staphylococcus aureus, Listeria monocytogenes and Salmonella spp. was determined in 75 samples of conventional beef and in 75 samples of organic beef. All samples came from cattle slaughtered and processed in the same slaughterhouse and quartering room. A total of 180 E. coli, 180 S. aureus and 98 L. monocytogenes strains were analyzed by an agar disk diffusion assay for their resistance to 11 antimicrobials, for the case of E. coli and S. aureus, or 9 antimicrobials, for the case of L. monocytogenes. Salmonella spp. were not isolated from any of the beef samples. No significant differences in prevalence were obtained for any of the bacterial species tested between organic and conventional beef. E. coli isolated from organic beef exhibited significant differences in antimicrobial resistance against 5 of the 11 antimicrobials tested as compared to isolates recovered from conventional beef. In the case of S. aureus, these differences were only found for 3 of the 11 antimicrobials tested and for L. monocytogenes, no differences were obtained between isolates obtained from organic or conventional beef. Although no significant differences were obtained in microbiological contamination, E. coli and S. aureus isolates from organically farmed beef samples showed significantly lower rates of antimicrobial resistance in E. coli and S. aureus isolates.     

Bacterial biofilm in silver-impregnated contact lens cases

PurposeThis study investigated the efficacy of pre-conditioning lens cases on bacterial biofilm formation and removal.MethodsSilver impregnated (MicroBlock / ProGuard™ & i-Clean) and control storage cases were pre-conditioned for 24 h with their respective multipurpose solutions (MPDSs). Cases were then inoculated with 2 ml of 10⁶ CFU/mL of ocular isolates of either P. aeruginosa or S. aureus and incubated for 48 h. Cases were subsequently disinfected (4−6 hours) as per the manufacturer’s recommended disinfecting time (MRDT) followed by the recommended case hygiene procedures - recapping wet (MicroBlock / ProGuard™ cases only) or rinse and air-dry or rinse, tissue-wipe and air dry (mechanical disruption). Surviving bacteria were enumerated using standard techniques.ResultsPre-conditioning the MicroBlock / ProGuard™ cases with MPDS significantly reduced biofilm formation (-1.1 log₁₀ CFU, p < 0.01 for P. aeruginosa & -1.3 log₁₀, p < 0.001, CFU for S. aureus) compared to the i-Clean lens cases. Maintaining the MicroBlock / ProGuard™ lens cases wet after the MRDT resulted in partial removal of bacterial biofilms (-2.9 log₁₀ CFU, p < 0.001 for P. aeruginosa and -2.6 log₁₀ CFU, p < 0.001 for S. aureus). Air-drying of all three types of lens storage cases after MRDT significantly reduced the bacterial biofilm (-5.4 log₁₀ CFU, p < 0.001 for P. aeruginosa and -3.5 log₁₀ CFU, p < 0.001 for S. aureus). Mechanical disruption produced the greatest reduction in the levels of bacterial biofilm in all 3 types of lens cases tested (-6.8 log₁₀ CFU, p < 0.001 for P. aeruginosa and -4.5 log₁₀ CFU, p < 0.001 for S. aureus). Synergi MPDS was significantly better than AQuify MPDS in removing bacterial biofilm from all 3 lens case types for case hygiene treatments with an air-drying step.ConclusionPre-conditioning of silver-impregnated ProGuard™ lens cases inhibited initial bacterial biofilm formation. Synergi MPDS was more effective than AQuify MPDS in removing bacterial biofilm in silver impregnated cases and tissue-wiping significantly improved biofilm removal.     

Analysis of the cultivable bacterial community in jeotgal, a Korean salted and fermented seafood, and identification of its dominant bacteria

Jeotgal or jeot, a traditional Korean salted and fermented food, is made by adding 20-30% (w/w) salt to various types of seafood. To develop a more complete overview of the bacterial community present in jeotgal, 610 pure colonies were isolated from Myeolchi-jeot and Saeu-jeot, the most commonly consumed varieties of jeotgal, which are made with anchovy (Engraulis japonicas) and tiny shrimp (Acetes japonicas), respectively. The bacterial isolates were identified by 16S rDNA sequence analysis. A total of 104 species comprising 47 genera and 31 previously unknown species were identified. Eleven genera were isolated from both jeotgal samples, including species in the genera Staphylococcus, Bacillus, Halomonas, and Kocuria, with Staphylococcus spp. constituting the highest number. The most populous genus detected in Myeolchi-jeot was Bacillus and its relatives, while the most populous in Saeu-jeot was Staphylococcus. These were isolated from both jeotgal samples, but their proportion in the bacterial community may be influenced by matrix composition and fermentation parameters. Among the proteolytic isolates, although Virgibacillus halodenitrificans KM2100 and Staphylococcus spp. maintained their growth in 20% NaCl, protease activities were not detected in these conditions. This suggests that bacteria are not the major source of the proteolytic enzyme involved in protein hydrolysis in high-salt-containing jeotgal. However, the Staphylococcus spp. isolated from Saeu-jeot were too numerous for us to ignore their possible role in jeotgal fermentation. Staphylococcus spp. may not be hugely involved in proteolysis, but they may play a significant role in the ripening of jeotgal. Bacteria of the genus Bacillus and its relatives and of the genus Staphylococcus may be the major organisms involved in jeotgal fermentation.     

In vivo efficacy of silver-impregnated barrel contact lens storage cases

PurposeThis study examined the rate and level of microbial contamination of silver and non-silver (control) barrel contact lens storage cases in vivo.MethodsA prospective, single-centre, randomized, double blinded, crossover study was conducted to evaluate the efficacy of silver and control barrel cases used with an Oxipol™-based disinfecting solution over a two-month period. Lens cases were collected, and the frequency, type and level of microbial contamination evaluated using viable culture and standard identification methods. Questionnaires were used to assess the contact lens-related hygiene compliance of the participants.ResultsForty eight daily contact lens wearers completed the study. Overall, 27 % of silver and 35 % control lens cases (P > 0.05) were contaminated, predominantly with Gram-positive bacteria. Only, 12 % of participants had good compliance, with 46 % having average compliance and 43 % poor compliance. Compliance score was not associated with level of microbial contamination. However, the use of hydrogel lenses in combination with silver cases compared to non-silver barrel cases resulted significantly greater numbers of bacteria colonising cases.ConclusionThe present investigation demonstrated that the use of silver barrel cases does not reduce the overall rate and level of bacterial contamination, but that using hydrogel lenses in combination with silver cases resulted in higher numbers of bacteria isolated from cases.     

Antibiotic resistance of Campylobacter jejuni and Campylobacter coli isolated from hog, beef, and chicken carcass samples from provincially inspected abattoirs in Ontario

Campylobacterjejuni is one of the most common causes of bacterial foodborne infection in the United States, and there are reports of resistance of Campylobacter spp. to antimicrobial agents used for the treatment of gastroenteritis. The purpose of this study was to determine the antimicrobial resistance patterns of Campylobacter spp. isolated from hog, beef, and chicken carcasses from provincially inspected abattoirs in Ontario. The agar dilution method was performed to measure antimicrobial resistance of the isolates. Antimicrobial resistance of Campylobacter isolates from hogs (n = 401), beef (n = 21), and chicken (n = 435) to ampicillin, azithromycin, chloramphenicol, ciprofloxacin, clindamycin, erythromycin, gentamicin, nalidixic acid, streptomycin, and tetracycline was determined. Resistance of chicken, hog, and beef isolates was 14.3, 18.2, and 9.5% to ampicillin; 17.9, 67.3, and 38.1% to azithromycin; 0, 0.5, and 0% to chloramphenicol; 3.7, 1.2, and 0% to ciprofloxacin; 2.3, 46.6, and 4.8% to clindamycin; 6.7, 43.6, and 4.8% to erythromycin; 0.2, 0, and 0% to gentamicin; 5.1, 10.7, and 0% to nalidixic acid; 13.6, 57.4, and 4.8% to streptomycin; and 52.6, 44.1, 42.9% to tetracycline, respectively. The hog isolates had the greatest resistance to seven of the ten antimicrobials tested. Results of this study confirm the existence of antimicrobial resistance of Campylobacter to various antimicrobial agents,especially ciprofloxacin and erythromycin, commonly used for treatment of campylobacteriosis in humans.