Commonly associated aerobic microbial pathogens and their antibiotic susceptibility profile in diabetic foot ulcers in tertiary care centre in Western Maharashtra.

PURPOSE: Diabetes mellitus (DM-II) is a metabolic disorder either due to reduced insulin production or reduced insulin sensitivity. Diabetic foot ulcer (DFU) is one of the most devastating complications of DM-II. This study was performed to assess commonly isolated micro-organisms and their anti-microbial sensitivity pattern in diabetic foot ulcers in a tertiary care centre in Western Maharashtra. METHODS: Adult patients with a known case of DM-II with foot lesions, suspected to be a Diabetic Foot Infections (DFIs) at the tertiary care hospital from Aug 2022 to Sept 2022 were included in the study. After obtaining informed written consent, pus sample was collected with sterile swab from lesion's base and submitted to Microbiology Laboratory for aerobic culture and sensitivity. RESULTS: Out of 56 enrolled patients, 47 (83.9%) patients tested positive for bacteriological growth and there was 'no growth' in 9 (16.07%) patients at the end of 48 h of aerobic incubation. There was male preponderance and patients were in age group of 35-85 years. The most commonly isolated micro-organisms were P. aeruginosa (17.8%), followed by S. aureus (14.2%), K. pneumonia and P. mirabilis (12.5% each). The resistance markers observed was ESBL producer, AmpC producer, MBL producer, Methicillin resistance and Inducible Clindamycin Resistance (ICR). CONCLUSION: Due to the injudicious use of antibiotics, antibiotic resistance has been increased in all types of soft tissue infections. The empirical formula for the treatment of DFIs should be decided for given geographical reasons according to antimicrobial susceptibility profile from particular geographical area or health care institute.

Effect of Antibiotic Exposure on Upper Respiratory Tract Bacterial Flora.

BACKGROUND The human microbiota modulates the immune system and forms the surface flora. Antibiotic administration causes dysbiosis in the intestinal flora. It is not clear if antibiotic administration in the community effects the upper airway flora in the mid-term or long-term. This study aims to define long-term influence of antibiotics on upper airway flora. MATERIAL AND METHODS In this prospective study, aerobic microbiological analysis of nasal and nasopharyngeal surfaces was performed. Antibiotic administration history of the last 6 months was retrieved using the social insurance database. Culture results of antibiotic-treated and antibiotic-naïve subjects were compared by Pearson's chi-square test or Fisher's exact test. RESULTS A total of 210 subjects were included in the study. Normal flora were documented in 86 nasal swabs and 99 nasopharyngeal swabs. Most of the remaining cases demonstrated gram-positive bacterial overgrowth. There were 113 subjects who did not receive any antibiotic, and 93% of the remaining 97 patients received broad-spectrum antibiotics. Statistical analysis showed that nasal and nasopharyngeal flora did not change upon antibiotic administration, but antibiotic administration during the last month caused increased methicillin resistance development of coagulase-negative Staphylococcus and Staphylococcus aureus microorganisms. CONCLUSIONS Antibiotic exposure did not lead to perturbations in general composition of upper airway flora within 6 months, although the incidence of methicillin resistance in coagulase-positive and -negative Staphylococci demonstrated significant increases when patients received antibiotic during the last month. This should be considered in case of broad-spectrum antibiotic administration, since methicillin resistance increases the morbidity and mortality of nosocomial Staphylococcus infections.

Effectiveness of Quaternary Ammonium in Reducing Microbial Load on Eggs.

Table eggs are an affordable yet nutritious protein source for humans. Unfortunately, eggs are a vector for bacteria that could cause foodborne illness. This study aimed to investigate the effectiveness of a quaternary ammonium compound (quat) sanitizer against aerobic mesophilic bacteria, yeast, and mold load on the eggshell surface of free-range and commercial farms and the post-treatment effect on microbial load during storage. Total aerobic mesophilic bacteria, yeast, and molds were enumerated using plate count techniques. The efficacy of the quaternary ammonium sanitizer (quat) was tested using two levels: full factorial with two replicates for corner points, factor A (maximum: 200 ppm, minimum: 100 ppm) and factor B (maximum: 15 min, minimum: 5 min). Quat sanitizer significantly (p < 0.05) reduced approximately 4 log10 CFU/cm2 of the aerobic mesophilic bacteria, 1.5 to 2.5 log10 CFU/cm2 of the mold population, and 1.5 to 2 log10 CFU/cm2 of the yeast population. However, there was no significant (p ≥ 0.05) response observed between individual factor levels (maximum and minimum), and two-way interaction terms were also not statistically significant (p ≥ 0.05). A low (<1 log10 CFU/cm2) aerobic mesophilic bacteria trend was observed when shell eggs were stored in a cold environment up to the production expiry date. No internal microbial load was observed; thus, it was postulated that washing with quat sanitizer discreetly (without physically damaging the eggshell) does not facilitate microbial penetration during storage at either room temperature or cold storage. Current study findings demonstrated that the quat sanitizer effectively reduced the microbial population on eggshells without promoting internal microbial growth.

Selective pressure of biphenyl/polychlorinated biphenyls on the formation of aerobic bacterial associations and their biodegradative potential.

Unique bacterial associations were formed in the polluted soils from territory of the industrial factories Open Joint Stock Company "The Middle Volga Chemical Plant," Chapaevsk, Russia and Open Joint Stock Company "Lubricant Producing Plant," Perm, Russia. This study evaluates the influence of the biphenyl/polychlorinated biphenyls (PCB) on the formation of aerobic bacterial associations and their biodegradative potential. Enrichment cultivation of the soil samples from the territories of these industrial factories with PCB (commercial mixture Sovol) was lead for forming aerobic bacterial enrichment cultures showing a unique composition. The dominating in these bacterial cultures was the phylum Proteobacteria (Beta- and Gammaproteobacteria). Using biphenyl as a carbon source led to decrease of biodiversity in the final stable bacterial associations. Periodic cultivation experiments demonstrated that the association PN2-B has a high degradative potential among the six studied bacterial associations. PN2-B degraded 100% mono-chlorobiphenyls (94.5 mg/L), 86.2% di-chlorobiphenyls (22.3 mg/L), 50.9% Sovol, and 38.4% Delor 103 (13.8 mg/L). Qualitative analysis of metabolites showed that association performed transformation of chlorobenzoic acids (PCB degradation intermediates) into metabolites of citrate cycle. Twelve individual strain-destructors were isolated. The strains were found to degrade 17.7-100% PCB1, 36.2-100% PCB2, 18.8-100% PCB3 (94.5 mg/L), and 15.7-78.2% PCB8 (22.3 mg/L). The strains were shown to metabolize chlorobenzoic acids formed during degradation of chlorobiphenyls. A unique ability of strains Micrococcus sp. PNS1 and Stenotrophomonas sp. PNS6 to degrade ortho-, meta-, and para-monosubstituted chlorobenzoic acids was revealed. Our results suggest that PN2-B and individual bacterial strains will be perspective for cleaning of the environment from polychlorinated biphenyls.

Antimicrobial coating is associated with significantly lower aerobic colony counts in high-touch areas in an orthopedic ward environment.

BACKGROUND: Hospital acquired infections (HAI) are the most common complication found in the hospital environment. The aim of the study was to examine whether the use of an antimicrobial coating in high-touch areas in an orthopedic ward could reduce bacterial growth and HAI. METHODS: From December 2017 to February 2018, HAI were registered on two orthopedic wards. A second registration was performed from December 2018 to February 2019. On the second occasion, an antimicrobial organosilane coating was applied just before the study period and thereafter weekly on one ward, while the other ward served as a control. Twenty defined high-touch areas on each ward were cultured before treatment and after 1, 2, 4, 8, 12, 14 and 16 weeks. Samples were cultured for aerobic colony counts, Staphylococcus aureus and E. coli. RESULTS: The total aerobic colony counts were 47% lower on the treated ward compared with the non-treated ward over the study period (p = 0.02). The colony counts for Staphylococcus aureus and E. coli were low on both wards. During the first registration period, the incidence of HAI was 22.7% and 20.0% on the non-treated and subsequently treated ward respectively. On the second occasion, after treatment, the incidence was 25.0% and 12.5% (treated ward) respectively (p = 0.0001). CONCLUSIONS: The use of a long-lasting antimicrobial organosilane coating appears to reduce the bioburden and reduce HAI. Since the incidence of HAI varies substantially over time, longer observation times are needed.

Composition of semen and foreskin mucosa aerobic microbiota and its impact on sperm parameters of captive collared peccaries (Pecari tajacu).

AIM: To evaluate the bacterial composition of collared peccary semen and foreskin mucosa, and to verify the sensitivity of isolates to antimicrobials used in semen conservation and to Aloe vera gel, which is an alternative external cryoprotectant. METHODS AND RESULTS: Nine foreskin mucosa and ejaculate samples from adult animals were used. Sperm characteristics and bacterial load were evaluated in fresh semen. The preputial mucosa and semen bacterial isolates were identified and tested against five concentrations of each antimicrobial (streptomycin-penicillin and gentamicin) and A. vera gel. Corynebacterium sp. and Staphylococcus sp. were isolated in greater numbers than others in both semen (64·10 and 20·51%, respectively) and the foreskin mucosa (60·60 and 24·25%, respectively), and ranged from 0·4 to 21 × 105 colony-forming units (CFU) per ml. The average load of Corynebacterium sp. was negatively correlated (P < 0·05) with the sperm membrane integrity (r = -0·73055) and curvilinear velocity (r = -0·69048). Streptomycin-penicillin and gentamicin inhibited most micro-organisms, and A. vera showed lower antimicrobial activity. CONCLUSION: Several Gram-positive bacteria are present in semen and foreskin mucosa of collared peccary, and the benefits of using primarily penicillin-streptomycin and gentamicin antimicrobials in the bacterial control of diluted semen of these animals are strongly indicated. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides insight into the reproductive microbiota of captive male-collared peccary. This work provides a theoretical basis to assist reproductive biotechnologies for ex situ conservation of the species.

[Aerobic bacteria from skin lesions in reptiles and their antimicrobial susceptibility]. / Aerobes Keimspektrum und Resistenzlage bei Hautläsionen von Reptilien.

OBJECTIVE: Bacterial skin infections are common in reptiles. Although many such infections are influenced by multifactorial problems, specific treatment of bacterial infections is an important consideration. The objective of this study was to evaluate the range of aerobic bacteria in skin lesions of reptiles and to determine their antimicrobial susceptibility. MATERIAL AND METHODS: Swabs of skin lesions from 219 reptiles were cultured for aerobic bacteria between January 2017 and June 2018. Isolates were identified based on growth on selective agar plates, biochemical parameters, as well as MALDI-TOF MS. Antibiotic susceptibility testing was carried out using the microdilution method. RESULTS: A total of 306 isolates were identified, mostly gram-negative, including Pseudomonas spp. (n = 48), Citrobacter spp. (n = 31, only in chelonians), aerobic spore-forming bacteria (n = 30), Aeromonas spp. (n = 20), Acinetobacter spp. (n = 20), Proteus spp. (n = 15), Staphylococcus spp. (n = 15), Klebsiella spp. (n = 13), Enterococcus spp. (n = 13), Morganella spp. (n = 11) as well as 78 other gram-negative and 12 other gram-positive bacteria. Colonization with 2 (n = 80) or more (n = 16) bacterial isolates was seen in 96 animals. Antibiotic susceptibility testing was carried out with 208 of the 306 isolated bacteria. Many isolates were sensitive (minimal inhibitory concentration [MIC] in µg/ml ≤ breakpoint) to enro- (E) and marbofloxacin (M): 86.4 % MIC ≤ 0.5 (E) and 95.5 % MIC ≤ 1 (M) for Pseudomonas spp., 86.4 % MIC ≤ 0.5 (E) and 90.9 % MIC ≤ 1 (M) for Citrobacter spp., 75.0 % MIC ≤ 0.5 (E) and 100 % MIC ≤ 1 (M) for Aeromonas spp. Trimethoprim/sulfamethoxazol proved to be effective against most of the Citrobacter spp. (90.9 % MIC ≤ 2/38) and Aeromonas spp. (75.0 % MIC ≤ 2/38). Amikacin was effective against nearly all Pseudomonas spp. (97.7 % MIC ≤ 16), Citrobacter spp. (95.5 % MIC ≤ 16) and Aeromonas spp. (93.8 % MIC ≤ 16). CONCLUSION AND CLINICAL RELEVANCE: The majority of isolates were gram-negative; the clinical relevance of individual isolates must, however, be evaluated on a case by case basis. Many of the isolated bacteria were sensitive to fluoroquinolones as well as aminoglycosides. Susceptibility testing is recommended since use of these antibiotics should be limited and for every tested group of antibiotics resistant isolates were found.

Fecal cultivable aerobic microbiota of dairy cows and calves acting as reservoir of clinically relevant antimicrobial resistance genes.

Antimicrobial resistance has become a global threat to public health since multidrug-resistant (MDR) bacteria have been reported worldwide carrying different antimicrobial resistance genes (ARGs), and animals have been described as a reservoir of ARGs. The presence of antimicrobial-resistant bacteria and ARGs in the food matrix is a risk to public health. This study aimed to research the presence of clinically relevant ARGs for important antimicrobials and genetic elements in fecal samples from dairy cows and calves on a Brazilian farm. In this study, a total of 21 fecal samples were collected, and then, the DNA of cultivable aerobic bacteria was extracted. Fifty-seven ARGs and twenty-three genetic elements were researched by PCR and confirmed by sequencing. Several ARGs that confer resistance to ß-lactams, tetracyclines, fluoroquinolones, sulphonamides, phenicols, aminoglycoside, glycopeptides, and macrolides were detected. A total of 200 amplicons from 23 ARGs (blaCTX-M-Gp2, blaCMY, blaSHV, tetA, tetB, tetC, qepA, qnrB, qnrS, oqxA, oqxB, vanC1, vanC2/3, aadA, sul1, sul2, sul3, ermB, mefAE, floR, cmlA, aadA, aph(3')-Ia, aac(3')-Ia), and 145 amplicons from 12 genetic elements (IncF, IncFIA, IncFIB, IncI1, IncY, IncU, IncK, IncP, IncR, IncHI1, ColE-like, intI1) were detected. The results presented in this study call attention to the monitoring of antimicrobial resistance in dairy farms worldwide. MDR bacteria and ARGs can spread to different sources, including milk products, which are one of the most consumed products worldwide, representing a potential risk to human health.

Inhibitory effect of ethanol and thiamine dilaurylsulfate against loosely, intermediately, and tightly attached mesophilic aerobic bacteria, coliforms, and Salmonella Typhimurium in chicken skin.

The effects of 3 ethanol levels (30, 50, and 70%) with and without thiamine dilaurylsulfate (TDS; 1,000 ppm) were evaluated for the reduction of natural mesophilic aerobic bacteria (MAB), coliforms, and inoculated Salmonella Typhimurium (S. Typhimurium) in chicken skin. The chicken skin was inoculated with a 7 log cfu/mL suspension of S. Typhimurium. Loosely, intermediately, and tightly attached cells were recovered from chicken skin through shaking at 200 rpm for 5 min, stomaching for 1 min, and blending for 1 min, respectively. Increasing the ethanol concentration reduced the number of MAB, coliforms, and S. Typhimurium on the chicken skin, whereas TDS treatment without ethanol was not effective. Intermediately and tightly attached microorganisms (total MAB, coliforms, and S. Typhimurium) were more resistant to chemical disinfectants than loosely attached microorganisms. The combination of 70% ethanol with TDS was most effective than the combination of TDS with lower concentrations of ethanol in reducing populations of loosely, intermediately, and tightly attached MAB (by 1.88 log cfu/g, 1.21 log cfu/g, and 0.84 log cfu/g, respectively), coliforms (by 1.14 log cfu/g, 1.04 log cfu/g, and 0.67 log cfu/g, respectively), and S. Typhimurium (by 1.62 log cfu/g, 1.72 log cfu/g, and 1.27 log cfu/g, respectively). However, the chicken skin treated with higher concentrations of ethanol was tougher (P < 0.05) and more yellow and less red (P < 0.05) than that treated with lower concentrations of ethanol or with water (control). On the other hand, a combination of 30% ethanol and TDS yielded the best results, showing the reduction greater than 0.5 log cfu/g in S. Typhimurium, with no negative effect on chicken skin color or texture. Thus, a combination of 30% ethanol and TDS appears to be the optimal treatment for reducing microbial contamination of skin-on chicken products to enhance poultry safety without decreasing food quality, and this treatment could be applied in the poultry industry.

Fortified hyperbranched PEGylated chitosan-based nano-in-micro composites for treatment of multiple bacterial infections.

Bacterial resistance is a real threat to human health. One of the most common strategies used to overcome this problem is the combination therapy. This study proposes a new chitosan-based nano-in-microparticles (NIMs) antibacterial platform that can deliver multiple antibacterial therapeutics at the same time. Chitosan (CS) was PEGylated to overcome its limited water solubility. Then, the antibacterial activity of the resulting PEG-CS was fortified via conjugation with dendritic polyamidoamine hyperbranches (HB) as well as in-situ immobilization of silver nanoparticles (AgNPs) to be efficient against multiple bacterial strains. Montmorillonite nanoclay (MMT) was prepared and used to encapsulate ibuprofen (IBU) as anti-inflammatory drug to reduce any concomitant inflammatory response during bacterial infection. The successful synthesis of PEG-HBCS-AgNPs as well as IBU-MMT nanocomplex was confirmed using FTIR, 1H NMR, DSC, TGA and EDX. SEM micrographs showed a complete formation of NIM spherical particles with a size around 13 µm. Besides, the newly developed drugs-loaded CS-based NIM formulation showed a better widespread activity on the tested aerobic and anaerobic bacterial species, and it may represent, after further optimization, a promising approach for overcoming multiple-bacterial infection.

Recovery of aerobic and anaerobic bacteria from patients with acute appendicitis using blood culture bottles / Recuperación de bacterias aerobias y anaerobias de pacientes con apendicitis aguda mediante botellas de hemocultivo

Introduction: Acute appendicitis is the first cause of acute abdomen, however, there is a little information about the associated bacteria and its sensibility profile. Objetive: To identify and to determine the resistance pattern of aerobic and anaerobic bacteria isolated in periapendicular fluid cultures taken in patients with acute appendicitis and to establish the proportions of isolates according to the clinical phase. Materials and methods: A descriptive and prospective study was undertaken at the Hospital Universitario de San José (Bogotá, Colombia) of patients older than sixteen years of age, undergoing an open appendectomy. A sample of periappendiceal fluid was taken, which was deposited directly into aerobic and anaerobic blood culture bottles. Results: One hundred and fifty-four patients were included. The overall positivity of cultures was 87% (n=1344); 77% (n=118) for aerobes and 51% (n=79) for anaerobes. The proportion of positive cultures was lower in the uncomplicated appendicitis cases as compared to the complicated ones (80% (66/83) vs. 95%(67/71), p = 0.003). The microorganisms isolated most frequently were: Escherichia coli (53%) (n=84); Bacteroides spp. (25%) (n=25); Propionibacterium acnes (21%) (n=21); coagulase negative Staphylococci (17%) (n=27); Enterococcus spp. (11%) (n=15), and Fusobacterium spp. (11%) (n=11). The sensitivity of E. coli to ampicillin/sulbactam was 30%. The sensitivity of Bacteroides spp. to clindamycin and ampicillin/sulbactam was 91%. All anaerobe isolates were sensitive to piperacillin/tazobactam, ertapenem, meropenem and metronidazole. Conclusions: Intraoperative cultures in acute appendicits are relevant in order to determine the local epidemiological pattern and to establish prophylactic and therapeutic antibiotics for this pathology; direct inoculation in blood culture bottles allows a high recovery of microorganisms.

Introduccción. La apendicitis aguda es la primera causa de abdomen agudo; sin embargo, poco se conoce sobre las bacterias asociadas y su perfil de sensibilidad. Objetivo. Identificar y determinar el patrón de resistencia de las bacterias aerobias y anaerobias aisladas en cultivo de líquido periapendicular tomado de los pacientes con apendicitis aguda, y establecer la proporción de bacterias según la fase clínica. Materiales y métodos. Se llevó a cabo un estudio descriptivo y prospectivo en el Hospital Universitario de San José de Bogotá (Colombia), en pacientes mayores de 16 años sometidos a apendicectomía abierta. Se tomaron muestras de líquido periapendicular, las cuales se sembraron directamente en botellas de hemocultivos para aerobios y anaerobios. Resultados. Se incluyeron 154 pacientes. Del total de cultivos, el 87 % (n=134) fueron positivos: 77 % (n=118) para aerobios y 51 % (n=79) para anaerobios. La proporción de cultivos positivos fue inferior en los casos de apendicitis no complicada, en comparación con aquellos de apendicitis complicada (80 % (66/83) Vs. 95 % (67/71); p=0,003). Los microorganismos aislados con mayor frecuencia fueron: Escherichia coli (53 %) (n=84), Bacteroides sp. (25 %) (n=25), Propionibacterium acnes (21 %) (n=21), Staphylococci coagulasa negativo (17 %) (n=27), Enterococcus sp. (10 %) (n=15) y Fusobacterium sp. (11 %) (n=11). La sensibilidad de E. coli a la amplicilina sulbactam fue de 30 %. La sensibilidad de Bacteroides spp. a la clindamicina y la ampicilina sulbactam fue de 91 %. El 100 % de los anaerobios fueron sensibles a piperacilina tazobactam, ertapenem, meropenem y metronidazol. Conclusiones. Los cultivos intraoperatorios son pertinentes en la apendicitis para determinar el patrón epidemiológico local, y establecer los antibióticos profilácticos y terapéuticos para esta enfermedad. Su siembra directa en botellas de hemocultivo permite una gran recuperación de microorganismos.

Antimicrobial resistance in pathogenic aerobic bacteria causing surgical site infections in Mbarara regional referral hospital, Southwestern Uganda.

Surgical site infections (SSI) remain a common postoperative complication despite use of prophylactic antibiotics and other preventive measures, mainly due to increasing antimicrobial resistance. Here, we present antimicrobial resistance rate of bacteria isolated in clinical cases of SSI. A hospital based descriptive cross sectional study was conducted on 83 consented postoperative patients with clinical SSI. Data on patients was obtained using structured data collection form. Two swabs were collected aseptically from each patient. Bacteriological culture examination and identification was done following standard microbiological techniques. Antibiotic susceptibility test was done by Kirby-Bauer disc diffusion method. Gram negative bacteria (GNB) were predominant (65.59%) with the dominant being Klebsiella species (29.03%). Overall 86% of aerobic bacteria isolated were multidrug resistant (MDR) where 65.63% and 96.72% of Gram positive and Gram negative isolates were MDR respectively. All the isolates with exception of Enterococci species were resistant to ampicillin. GNB showed high resistance to ceftriaxone, sulfamethoxazole/trimethoprim and gentamicin. All the isolated Klebsiella spp were MDR. S. aureus were all resistant to oxacillin. The isolation rate was higher in emergency, males and dirty wounds in relation to nature of surgery, gender and class of surgical wound respectively. These findings necessitate judicious antibiotic use and calls for surveillance of SSIs periodically as well as strict adherence to good sanitation practice to reduce spread of drug-resistant pathogens.

Temporal efficacy of antimicrobials against aerobic bacteria isolated from equine endometritis: an Italian retrospective analysis (2010-2017).

This study aimed to describe bacteria isolated from the reproductive tract of mares and to identify changes in antimicrobial susceptibility patterns to those antibiotics commonly used for the treatment of equine endometritis. A total of 4122 equine uterine swabs were collected from mares suffering from reproductive tract disorders in the period 2010-2017. Aerobic culture and antimicrobial susceptibility testing using agar disc diffusion were performed on each sample. Aerobic bacteria were isolated from 3171 of 4122 (76.9 per cent) samples. The most frequently isolated microorganisms were Escherichia coli (885/3171, 27.9 per cent) and Streptococcus equi subspecies zooepidemicus (791/3171, 24.9 per cent), confirming previous findings from the literature. Antimicrobial susceptibility patterns of E coli, S equi subspecies zooepidemicus and Klebsiella pneumoniae changed over time. A statistically significant decrease in antimicrobial efficacy of cefquinome against E coli was observed over the years, as well as of ampicillin, cefquinome and penicillin against S equi subspecies zooepidemicus The high frequency of resistant bacteria isolated in the present work proceeds in the same way as indicated by surveillance data on the huge antibiotic use in Italy. As a result, testing and monitoring programmes of antimicrobial efficacy are crucial to consciously using antibiotics and preserving their effectiveness both for veterinary and human medicine.

Evaluation of cultivable aerobic bacterial flora from Russell's viper (Daboia russelii) oral cavity.

Snake mouths contain a wide range of bacteria. Identifying these bacteria in snakes is very important to obtain an understanding of the etiological agents of secondary infections that may result from accidents during handling and/or snake bites. The present study aims to determine the pattern of oral bacterial flora of nine healthy Russell's vipers (Daboia russelii), and their susceptibility to common antibiotics. A total of 94 isolates were obtained in pure form, which demonstrated noticeable colony characteristics and which were further studied with several biochemical tests. The strains that showed distinctive colonies, morphology and biochemical parameters were additionally subjected to phylogenetic characterization using 16S rRNA gene sequences. Furthermore, all these isolates were studied for antibiotic susceptibility. The oral cavity of the Russell's viper harbors a wide range of pathogenic bacteria, including Gram-negative genera: Proteus sp., Pseudomonas sp., Salmonella sp., Providencia sp., Alcaligenes sp., Morganella sp., as well as E. coli, and Gram-positive genera: Bacillus and Enterococcus sp., Staphylococcus sp. and Lysinobacillus sp. Most of the isolates were resistant to antibiotics viz. penicillin, Amoxyclav, oxacillin, methicillin and streptomycin while sensitive towards imipenem, amikacin, norfloxacin, gatifloxacin, ciprofloxacin, gentamicin, tetracycline, chloramphenicol and azithromycin. The present study documents diverse bacteria predominant in the oral cavity of Daboia russelii and studied their antibiotic susceptibilities.

In-situ headspace analysis of metabolic carbon dioxide of aerobic bacteria for assessing antimicrobial activity of natural products.

Headspace analysis of the volatile metabolites has been used as a good strategy for monitoring the microbial growth in several applications, but never been established for the evaluation of the activity of microbial inhibitor in the field of drug screening from natural products. We report on a new method for the determination of antimicrobial activity of drug compounds or crude extracts from natural products by measuring the amount of metabolic carbon dioxide produced in the drug-bacteria incubation system. 2 mL of medium containing bacteria and drug of interest was incubated at 37 °C for 24 h. The amount of metabolic carbon dioxide partitioned in the headspace was measured to evaluate the drug antimicrobial activity using headspace gas chromatography (HS-GC) coupled with thermal conductive detector (TCD). The principle and the standard procedure of the present method have been developed and verified. As a result, the precision of the present method was less than 4% (expressed as relative standard deviation), and an excellent agreement was found on both inhibition rate (R2 = 0.935) and the half inhibition concentration (R2 = 0.994) between the present method and a reference method (optical density method). By comparison, the present method is simpler and safer regarding the microorganism contamination due to the in-situ incubation and detection in the closed system, and suitable for the routine analysis of antimicrobial activity of natural products with high flexibility in both bacterial strains and sample properties.

Carvacrol antimicrobial wash treatments reduce Campylobacter jejuni and aerobic bacteria on broiler chicken skin.

Campylobacter jejuni, a major cause of human gastroenteritis worldwide, is often associated with the consumption of contaminated poultry products. With increasing consumer preference to natural and minimally processed foods, interventions utilizing natural antimicrobials for controlling C. jejuni on poultry products are gaining popularity. This study investigated the efficacy of the generally recognized as safe compound carvacrol (CR) as a wash treatment in reducing C. jejuni and aerobic bacteria on chicken skin. Two separate studies, each with 2 trials, were conducted. In the first study, the efficacy of CR suspension (0, 0.25, 0.5, 1, and 2%) was investigated, whereas in the second the efficacy of CR as suspension, emulsion, and nanoemulsion was studied. In both studies, skin samples were inoculated with 50 µL (∼8 log10 cfu/sample) of a cocktail of 4 wild strains of C. jejuni. After 30 min of attachment, samples were washed with the respective treatments for 1 min, drip dried for 2 min, and processed at 0, 8, 24, h post-treatment for enumeration of C. jejuni and aerobic bacterial counts (n = 5/treatment/time point). In addition, the effect of treatments on the color of chicken skin was evaluated. Data were analyzed using PROC MIXED procedure of SAS. In the first study, all the tested doses of CR suspension consistently reduced C. jejuni counts across all time points. The 2% CR suspension wash reduced C. jejuni counts by ∼2.4 to 4 log10 cfu/sample (P < 0.05). In addition, 1% and 2% CR suspensions significantly reduced aerobic counts at all the time points. The results from the second study suggest that anti-Campylobacter efficacy of CR emulsion or nanoemulsion treatments was not improved compared to CR suspension. Several CR suspension treatments were more effective than corresponding emulsion or nanoemulsion treatments. No significant differences were observed in the color of the samples between treatments (P > 0.05). The results suggest that CR could potentially be used as an antimicrobial wash treatment in postharvest poultry.

Comparative effectiveness of rapid-cycle ultraviolet decontamination to chemical decontamination on high-touch communication devices.

BACKGROUND: This quantitative, comparative-descriptive study of inpatient units in a large military medical center was designed to compare the effectiveness of compact ultraviolet (UV-C) decontamination to standard chemical decontamination in reducing the microbial burden on Vocera (San Jose, CA) communication devices and to characterize changes in staff cleaning practices following UV-C device implementation. METHODS: Aerobic and anaerobic swabs were used to collect microbial samples from Vocera devices (n = 60) before and after chemical decontamination (first sampling) and before and after UV decontamination (second sampling). Cleaning behaviors were assessed by observation and oral inquiry during the baseline sampling and surveyed 8 weeks after UV-C device implementation. Outcomes included aerobic and anaerobic colony-forming units and prevalence of methicillin-resistant Staphylococcus aureus, vancomycin-resistant enterococci, or Clostridium difficile, as determined by standard microbiological methods. RESULTS: No differences were found between the two cleaning methods in their ability to reduce aerobic bacteria; however, UV-C was significantly more effective at reducing bacteria grown anaerobically (P < .01). This study elucidated an 8.3% prevalence of methicillin-resistant Staphylococcus aureus on Vocera devices in the inpatient environment. Initially, 42% of respondents reported deviations from manufacturer's cleaning guidelines, and 16.7% reported daily or more frequent cleaning of the Vocera devices. CONCLUSIONS: After implementation, UV-C decontamination reduced average cleaning time by 43% and increased the rate of daily Vocera cleaning to 86.5%. Respondents reported an overall 98% user satisfaction with the UV-C device.

Pectin or chitosan coating fortified with eugenol reduces Campylobacter jejuni on chicken wingettes and modulates expression of critical survival genes.

Campylobacter jejuni infection in humans is strongly associated with the consumption of contaminated poultry products. With increasing consumer demand for minimally processed and natural product, there is a need for novel intervention strategies for controlling C. jejuni. Antimicrobial coatings are increasingly being used for preventing food contamination due to their efficacy and continuous protection of product. This study investigated the efficacy of pectin and chitosan coating fortified with eugenol to reduce C. jejuni on chicken wingettes. Pectin, chitosan, and eugenol are generally recognized as safe status compounds derived from berries, crustaceans, and cloves respectively. Each wingette was inoculated with a mixture of 4 wild-type strains of C. jejuni (approximately 107 CFU/sample) and randomly assigned to controls, pectin (3%), chitosan (2%), eugenol (0.5, 1, or 2%), or their combinations. Following 1 min of coating, wingettes were air-dried, vacuum sealed, and sampled on 0, 1, 3, 5, and 7 d of refrigerated storage for C. jejuni and aerobic counts (n = 5 wingettes/treatment/d). In addition, the effect of treatments on wingette color and expression of C. jejuni survival/virulence genes was evaluated. All 3 doses of eugenol or chitosan significantly reduced C. jejuni and aerobic bacteria from 0 d through 7 d. Incorporation of 2% eugenol in chitosan improved coating efficiency and reduced C. jejuni counts by approximately 3 Log CFU/sample at the end of 7 d of storage (P < 0.05). Similarly, the antimicrobial efficacy of pectin was improved by 2% eugenol and the coating reduced C. jejuni by approximately 2 Log CFU/sample at 7 d of storage. Chitosan coating with 2% eugenol also showed greater reductions of total aerobic counts as compared to individual treatments of eugenol and chitosan. No significant difference in the color of chicken wingettes was observed between treatments. Exposure of C. jejuni to eugenol, chitosan, or combination significantly modulated select genes encoding for motility, quorum sensing, and stress response. Results demonstrate the potential of pectin or chitosan coating fortified with eugenol as a postharvest intervention against C. jejuni contamination on poultry products.

Prospective, randomised clinical trial of four different presurgical hand antiseptic techniques in equine surgery.

BACKGROUND: Currently, the World Health Organization recommends the use of alcohol-based hand rubs (ABR) for surgical hand preparation in human surgery. When disinfecting soaps are used, a rubbing technique causes less skin irritation than brush scrubbing. Based on a recent survey, most equine surgeons still use disinfecting soap. The efficacy of scrubbing vs. rubbing and the use of sole ABR compared with chlorhexidine (CHx)- based products has not been evaluated in the equine surgical setting. OBJECTIVES: To compare four surgical hand antisepsis techniques in equine surgery for reduction of aerobic bacterial counts from pre- to post-preparation (immediate efficacy) and at the end of surgery (sustained efficacy). STUDY DESIGN: Randomised, prospective clinical trial. METHODS: A 4% CHx-based product applied with either a scrub or rub technique, one sole ABR (ET; 80% ethanol) and one CHx/alcohol-combination (CHx/ET; 1% CHx and 61% ethanol) product both applied with a rub technique were evaluated. Samples were collected by glove juice technique and cultured on 3M™ Petrifilm plates and counted using a 3M™ Petrifilm plate reader. RESULTS: Immediate mean bacterial log10 colony forming unit (CFU) reduction was 2.4 for CHx-scrub, 2.8 for CHx-rub, 3.1 for CHx/ET and 2.1 for ET. CHx/ET resulted in significantly lower bacterial counts than CHx-scrub (P<0.005) and ET (P<0.001) while CHx-rub resulted in significantly lower counts than ET (P<0.001). At the end of surgery bacterial counts were the lowest for CHx-rub, significantly lower than CHx/ET (P<0.001) and ET (P<0.001). There was no difference between CHx-rub and -scrub techniques (P = 0.7). MAIN LIMITATIONS: Bacterial counts were used as the outcome measure rather than prevalence of surgical site infection, and the effect of hand preparation on skin health was not assessed. CONCLUSIONS: ABR did not decrease bacterial log10 CFU counts more effectively than CHx products. When using CHx soaps in the equine setting, hand-rub is as effective as a hand-scrub-technique.

A clinical trial comparing antimicrobial efficacy of "essential oil of Ocimum sanctum" with triple antibiotic paste as an intracanal medicament in primary molars.

OBJECTIVES: To evaluate the aerobic and anaerobic antimicrobial efficacy of Ocimum Sanctum (Tulsi) essential oil and compare it with that of triple antibiotic paste (TAP) by collecting microbiological samples from the root canals of primary molars. STUDY DESIGN: Forty children were selected for the study and were randomly divided into two groups of twenty each, namely, TAP group and O. sanctum group (basil). Six intracanal samples were collected for every patient, comprising of two each after access opening, irrigation and after 3 days of intracanal medicament placement. These samples were cultured in aerobic and anaerobic environment and later colony-forming units (CFUs) were counted and intragroup as well as intergroup comparison was done. RESULTS: Analysis of the results showed that there was a statistically significant reduction in CFUs after using essential oil of O. sanctum as an intracanal medicament. Saline use also leads to a statistically significant reduction in CFUs irrespective of the intracanal medicament used. TAP showed better antibiotic properties in comparison with that of O. sanctum. CONCLUSION: Antibiotic use is often associated with the adverse effects and development of resistance due to injudicious use. O. sanctum can be used in cases of long-standing infection owing to its antimicrobial efficacy and anti-inflammatory potential as an intracanal medicament in primary teeth.