Average Colony Count Research Articles

Effectivity of Irrigation Solution from Stevia rebaudiana Bertoni Leaf Extract on the Growth of Enterococcus faecalis Bacteries

Disinfection of Enterococcus faecalis bacteria in dental root canal treatment using NaOCL 0.5-2.5% is still a challenge. There is still very little data on the effectvity of Stevia rebaudiana Bertoni leaf extract irrigation solution against the growth of Enterococcus faecalis. This study evaluated the effectiveness of Stevia rebaudiana Bertoni leaf extract irrigation solution against Enterococcus faecalis ATCC 29212 through MIC (Minimal Inhibitory Concentration) and MBC (Minimal Bactericide Concentration) values. A number of 100 µl of E. faecalis suspension was inoculated into 96 microtitration wells, each containing 100 µl of 100%, 50%, 25%, 12.5%, 6.25%, 3.125%, 1.56% of Stevia leaf extract irrigation solutions, positive (NaOCL 2.5%) and negative (distilled water) control. Incubation was carried out for 24 hours at 37°C then optical density was measured with a microplate reader at 600 nm. Five µl of 10-9 dilution of each group was dripped and spread onto nutrients agar plate. The number of Enterococcus faecalis colonies was counting manually. There was no significant difference in average colony count of Enterococcus faecalis between the 100% and 50% Stevia leaf extract irrigation solution groups and the positive control (p>0.05). However, the 25%, 12.5%, 6.25%, 3.125%, and 1.56% Stevia leaf extract irrigation solution groups showed significant differences (p<0.05). Stevia leaf extract irrigation solution suppressed growth at MIC 6.25% by turbidimetry and MIC 1.56% by total plate count method using nutrient agar and than killed Enterococcus faecalis at MBC 50%. The bactericidal ability of 100% and 50% Stevia leaf extract irrigation solution is equal to NaOCL 2.5%.

Bacterial contamination of endoscopist and assistant face visors during gastrointestinal endoscopy: a pilot study

Background and aimDuring the COVID-19 pandemic, health workers’ facial exposure to pathogens has been brought into focus. In this study, we aimed to determine the occurrence and degree of facial...

Detection of Biofilm Layers in Water Plumbings and Determination of its Effect on Water Quality

Water quality deteriorations in the plumbing are mainly resulted from accumulations and biofilm layer formation on the pipe surfaces. Biofilms may cause significant health risks on human health due to its pathogen content. In this study, biofilm formation on the plumbing, the presence of pathogenic microorganisms in biofilms and their effects on water quality were examined by membrane filtration with dehydrated medium (NPS) method in 22 different buildings in a facility’s water network. A relation with biofilm formation and water quality deterioration with the age of the plumbing and the pipe material was also studied. The results were expressed as colony forming units (CFU). According to the results of the galvanized metal pipes, the average colony count was determined as >200 CFU/250 mL, average pathogen count was calculated as 107 CFU/250 mL and the highest deterioration in the water quality were observed. For the PE (Polyethylene) pipes, the average colony count was found as >200 CFU/250 mL and the average pathogen count was found as 145 CFU/250 mL. No physical and chemical changes in water quality were observed. For the PPRC (Polypropylene Random Copolymer) pipes, neither physical/chemical change in water quality nor pathogenic growth were observed. The total colony count was found as 34 CFU/250 mL. Biofilm formation was detected at 15 points in the network. Pseudomonas aeruginosa was the most common detected pathogens in plumbing as 12 points from 22 buildings. The highest colony formation was Escherichia coli, which was detected in four of 22 plumbing as 600 CFU/250 mL colonies. It has been observed that more accumulations occurred in galvanized metal pipe surfaces, and microbiological growth was higher than PE and PPRC pipes.

Vegetable fermentations brined with low salt for reclaiming food waste.

Fermentation of eight vegetables was studied as an alternative for reclamation of surplus volumes. Fermentation performance was predicted by comparing the amounts of acid that could be produced from the intrinsic sugar content with that buffered by the fresh vegetable matrices prior to reaching an inhibitory pH for fermentative microbes (3.30). Native fermentations were brined with 345.0mM sodium chloride, 40.0mM calcium chloride, 6.0mM potassium sorbate, and vinegar to adjust the initial pH to 4.70. High-performance liquid chromatography analysis, pH, and carbon dioxide measurements and spiral plating on selective media were employed to monitor the progress of fermentations. The average colony counts for yeast and/or molds and Enterobacteriaceae declined to undetectable levels from 3.6± 1.5 log CFU/ml within 7 days of fermentation. The fermentation of sugars produced lactic, acetic, succinic, and/or malic acids, and ethanol. As predicted, the fermentation of vegetables with low sugar content, such as broccoli, green leaf lettuce, and green pea proceeded to completion. The fermentation of vegetables with a moderate sugar content, such as green bell pepper, red ripened tomato, and green bean were incomplete at pH 3.1± 0.2. The fermentation of high sugar vegetables including sweet potato and corn were expected and observed to be incomplete. It is concluded that the intrinsic sugar content and buffer capacity of surplus vegetables are relevant parameters in obtaining complete fermentations. PRACTICAL APPLICATION: Vegetables are the second most wasted commodity in the United States and a substantial constituent of the global food waste. Development of fermentation to reclaim surplus vegetables from farms, grocery stores, and farmer's markets offers opportunities to ameliorate economic losses and environmental impact and add value to waste. The research described here suggests that a fraction of vegetables could be fermented in cover brines while others, with high sugar content, need specialized handling. Evidently, optimization of vegetable fermentation with starter cultures and added buffers represent an opportunity to stimulate complete bioconversions useful for reclaiming surplus volumes.

Flood induced changes in the diversity of yeasts from the mangroves of Valanthakadu, Kerala, India

This is the first report on the influence of the devastating August 2018 floods of Kerala on yeast communities associated with the mangrove ecosystem of Valanthakadu, 'Kochi’s oxygen parlour’. Samples were collected for the study from six stations in Valanthakadu during three different periods with regard to floods i.e. April – May 2018: Pre flood (PRF), September – October 2018: Just after floods (JAF), and March 2019: Post floods (POF) and plated onto Malt-yeast-glucose-peptone agar for isolation and enumeration. Morphologically distinct colonies were picked, clustered on biochemical basis, representative strains identified on molecular basis or using API 20 C AUX strip. On enumeration, the average colony counts of yeasts in the sediments were found to be greater than in water samples. Out of 89 morphologically distinct yeast strains isolated, the majority were non-pigmented (73 %) and belonged to the phyla Ascomycetes (58 %). Six different yeast genera: Candida spp., Cryptococcus spp., Hortaea sp., Rhodotorula spp., Rhodosporidiobolus sp., and Trichosporon sp. were recorded in this study. C. tropicalis was noted to be not only the predominant species but also resilient to the effect of floods. Flood was found to influence the diversity of yeasts, with the introduction of certain genera (Hortaea wernekii (first report)) and the loss of certain others (C. gatti). Analysis of diversity indices evidently revealed the influence of floods, with an increase in yeast diversity just after floods (1.62) when compared to other periods of sampling. A change in the hydrolytic profile of isolates was also noticed in relation to floods. This study clearly indicated that floods do affect the diversity of yeast communities and their function in the ecosystems, at least temporarily. Nevertheless, the long-term impacts remain unknown and deserve further studies.

Potential sources of operating room air contamination: a preliminary study

The Neptune® surgical suction system (NSSS) and the Bair Hugger® (BH) forced-air warmer both discharge filtered exhaust or heated air into the operating room (OR), often in close proximity to a surgical site. To assess the effectiveness of this filtration, we examined the quantity and identity of microbial colonies emitted from their output ports compared with those obtained from circulating air entering the OR. Air samples were collected from each device using industry-standard sampling devices in which a measured volume of air is impacted on to a blood agar plate at a controlled flow rate. Twelve ORs were studied. Sample plates were incubated for one week per study protocol, then interpreted for colony counts and sent for species identification. The average colony count from the NSSS exhaust was not significantly different from that obtained from room air samples, however the average count from the BH output was significantly higher (P=0.0086) than room air. Genetic identification profiles revealed the presence of environmental or commensal organisms that differed depending on the source. High variability in colony counts from both devices suggests that certain NSSS and BH devices could be significant sources of OR air contamination. Our study showed that the BH patient warming device could be a source of airborne microbial contamination in the OR and that individual BH and NSSS units exhibit a higher output of microbial cfu than would be expected compared with incoming room air. We make simple suggestions on ways to mitigate these risks.

The effects of radiofrequency on the bacteriological and histological characteristics of tonsils in patients with chronic and persistent tonsillitis.

Tonsillotomy with radiofrequency (RF) is one of the newest treatments for chronic tonsillitis, but the mechanism of RF effects and complications are still pending. The aim of this study was to evaluate the effects of RF on the histological and bacteriological characteristics of the tonsils (Case-control study). In fifty-two patients with chronic tonsillitis in 2017-2018, immediately after tonsillectomy, the tonsils were divided into 2sections; one sample treated with RF, and the other one considered as control, without intervention. All tonsil samples sent for histological and bacteriological study: morphometric assays made by Digitizer software, and type of bacterial colonies identified by microbiological and biochemical tests. Willcoxon and McNemar tests were used for statistical analysis and level of significance was p≤.05. Tonsil mucosal thickness (2202.98±323.09 vs. 2463.94±357.61μm) and size of the tonsil nodule (28,000.42±9608.75 vs. 36,692.81±7040.74μm2) were significantly lower in the RF+ group than other group (p=.001 and p=.01, respectively). There was no significant differences in thickness of the tonsil epithelium (p=.075), number of lymphoid nodules (p=.860), and the number of reticular tonsil epithelium (p=.813) between the two groups. Bacterial growth in RF- and RF+ groups had no statistically significant difference (p=.06), however, the average colony count of S. aureus in RF+ tonsils were significantly lower, and total number of bacterial colonies were significantly lower in RF+ group(1405±156 vs. 2471±156), (p=.001). RF surgery has significant effects on size of the nodules, thickness of the mucous layer and bacteriological characteristics of tonsil tissue. Especially S. aureus seems to be more sensitive to RF effects.

A randomized control trial evaluating efficacy of antimicrobial impregnated hospital privacy curtains in an intensive care setting

Acquisition of pathogens into health care settings from prior room occupants has been documented. Hospital room privacy curtains are at high risk for pathogenic bacterial contamination. Antimicrobial impregnated curtains could be effective in reducing contamination. Rooms within an intensive care unit at The University of Iowa Hospitals and Clinics were randomized to 3 arms. The 2 intervention arms: (1) halamine antimicrobial curtains (BioSmart curtain [BSC]) and (2) halamine antimicrobial curtains sprayed twice weekly with a sodium hypochlorite-based disinfecting spray (BSC-pre and BSC-post) and a third control arm (standard curtain [SC]). Samples were collected twice weekly for 3 weeks to assess pathogenic bacterial contamination. The likelihood of remaining uncontaminated was 38% for SC, 37% for BSC, and 60% for the BSC-pre group. Time to event (contamination) analysis found no statistically significant difference between pathogenic contamination between the SC, BSC, and BSC-pre groups (P value = .1921). There was a decrease in average colony count for BSC curtains compared with control, however, this difference was not statistically significant. Hypochlorite spray was found to transiently decontaminate curtains, but effects dissipated after 72 hours. BSC did not show a significant reduction in pathogenic contamination compared with control. Antimicrobial curtains could have a role in reducing environmental contamination in the health care setting. Future studies should be done to determine the long-term effects of using antimicrobial curtains in health care.

Studies on Predominant Epiphytic Micro-flora as Antagonists to Postharvest Pathogens of Apple

Apple (Malus domestica Borkh) is an important horticultural crop that is affected by the number of diseases round the year. The fruit is particularly susceptible to a number of pathogens both pre- and post-harvest. Management of these diseases is based mostly on the application of synthetic fungicides with obvious disadvantages of environmental pollution, health hazards, pathogen resistance, etc. In the present study, eleven epiphytes were isolated using potato dextrose agar, nutrient agar and yeast maltose agar media. Amongst them, five fungal isolates viz., Aspergillus sp. (I1), Penicillium sp. (I2), Fusarium sp. (I3), Rhizopus sp. (I4) and Alternaria sp. (I5) and six bacterial isolates viz., Pseudomonas sp. (I6), Pseudomonas sp. (I7), Bacillus sp. (I8), Bacillus sp. (I9), Staphylococcus sp. (I10) and Micrococcus sp. (I11) were predominantly noticed under all the three methods (leaf impression, serial dilution and fruit washing) and were hence, used for further studies. The highest average colony count of 3.62 colonies/cm2 was recorded in fruit washing method followed by leaf impression (3.17) and lowest in serial dilution method (2.12). The in vitro screening of various bacterial and fungal epiphytes revealed that isolates of Pseudomonas sp. (I6) and Bacillus (I8 and I9) were the only bacterial strains capable of inhibiting the growth of all the test pathogens using dual culture method. Assays on wounded apples revealed that Pseudomonas sp. I6 at 107 cfu/ml was effective antagonist against Penicillium sp. and Fusarium sp., Bacillus sp. I9 at 107 cfu/ml was effective antagonist against Alternaria sp., whereas, Bacillus sp. I8 at 107 cfu/ml was most effective antagonist against Diplodia sp. The present study revealed that the antagonists were more or less efficient towards each pathogen and can be utilized for the management of post-harvest diseases of apple.

Staphylococcus aureus Bacteria Resistant to Methicillin in Raw Milk

The presence of antibiotic-resistant strains of Staphylococcus aureus (particularly methicillin-resistant strains) in food of animal origin is considered as a severe threat to human health due to numerous clinical complications. This study tended to determine the prevalence of methicillin-resistant S. aureus in samples of raw milk distributed in Tehran using antibiotic susceptibility testing methods. In the present study, 100 raw milk samples were taken from the centres of production and purchase of milk and its products in Tehran; the samples were evaluated by culture in terms of infection with coagulase-positive S. aureus. Finally, antibiotic resistance pattern of isolates was studied using disk diffusion agar. The average colony count was estimated. Raw milk cultures were estimated at 2×105-4×105 cfu/ml. Based on the results of culture, 36 samples of raw milk tested were icnfected with positive-coagulase S. aureus. The highest susceptibility was observed for ciprofloxacin and gentamicin (100%) and the highest resistance was observed to penicillin, tobramycin, oxacillin and ceftazidime. The results showed the prevalence of infection of raw milk with S. aureus. Moreover, prevalence of S. aureus resistant to a wide range of antibiotics, more importantly, methicillin resistant, was significant in the tested samples. Therefore, adherence to and control of sanitation in different stages of production, supply and consumption of milk can prevent human infection.

Abstract NTOC-095: THE SYNERGISTIC EFFECTS OF CARBOPLATIN AND PF–477736, A SMALL MOLECULE CHECKPOINT KINASE 1 INHIBITOR ON A PLATINUM RESISTANT OVARIAN CANCER CELL LINE

Abstract PURPOSE: To investigate the effects of PF-477736, a small molecular checkpoint kinase 1 (Chk1) inhibitor with and without carboplatin on cell apoptosis and proliferation in a p53-mutated platinum resistant ovarian cancer cell line. METHODS: OVCAR3 cells were treated with PF-477736 alone and in combination with carboplatin. Western blot analyses were performed to assess the protein expression of Chk1, phosphorylated Chk1, H2AX, and phosphorylated H2AX. Cell proliferation was evaluated with colony formation assays 10 days following treatment and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) proliferation assays over the course of 5 days following treatment. Cell apoptosis was measured with flow cytometry after staining with FITC Annexin V and propidium iodide. Paired t-tests were performed for comparative analyses. RESULTS: PF-477736 decreased the expression of phosphorylated Chk1 on serine 249 within 24 hours following treatment as seen on Western blot assay. Total Chk1 expression was unchanged after treatment with PF-477736. The combination treatment of PF-477736 and carboplatin effectively decreased OVCAR3 cell proliferation. Colony formation assays revealed fewer colonies after dual treatment with PF-477736 and carboplatin compared to PF-477736 alone (average colony count 1.5 vs 13.6, p = 0.03) and compared to carboplatin alone (average colony count 1.5 vs 9.7, p < 0.001). MTT assays demonstrated a synergistic effect with PF-477736 and carboplatin: dual treated cells had lower mean dye absorbance values than PF-477736 alone (mean absorbance fold change 1.7 vs 3.2, p = 0.009) and carboplatin alone (mean absorbance fold change 1.7 vs 5.6, p < 0.001). OVCAR3 cells after treatment with PF-477736 and carboplatin had higher rates of apoptosis. On flow cytometry, a higher proportion of cells stained with the apoptotic markers Annexin V and propidium iodide after treatment with PF-477736 and carboplatin compared to PF-477736 alone (percentage of apoptotic cells 34.9% vs 13.0%, p = 0.004) and carboplatin alone (percentage of apoptotic cells 34.9% vs 16.0%, p = 0.008). In addition, western blot analysis of the cell lysates following dual treatment of PF-477736 and carboplatin showed increased expression of phosphorylated H2AX, a marker of double-stranded DNA damage while there was minimal expression with PF-477736 alone and carboplatin alone. CONCLUSIONS: Carboplatin and PF-477736, a small molecule Chk1 inhibitor, synergistically decreased cell proliferation and increased cell apoptosis in OVCAR3 cells, an in vitro model of platinum resistant ovarian cancer. PF-477736 may be a potential new therapeutic strategy for the resensitization of platinum resistant ovarian cancer. Citation Format: Jessica Lee MD, Stephanie V. Blank MD, Robert J. Schneider MD. THE SYNERGISTIC EFFECTS OF CARBOPLATIN AND PF–477736, A SMALL MOLECULE CHECKPOINT KINASE 1 INHIBITOR ON A PLATINUM RESISTANT OVARIAN CANCER CELL LINE [abstract]. In: Proceedings of the 11th Biennial Ovarian Cancer Research Symposium; Sep 12-13, 2016; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(11 Suppl):Abstract nr NTOC-095.

Facultative thermophilic microorganisms in potato products in retort packaging

Thermally treated potato products in retort packaging with an extended shelf-life (up to 90 days before opening) at room temperature (20 ± 1 °C) from two Latvian vegetable processing companies (A and B) were analysed in this research. The aim of this research was to determine the presence of thermotolerant microbial contamination in thermally treated potato products in retort packaging, to ensure their safety and suitability for immediate consumption. Samples from three production batches were used. The total plate count was determined, and a microscopic examination of potato product samples was performed. Microscopic examination of microorganisms showed the presence of microbial vegetative forms in potato products from company B, but the standard investigation did not register them; therefore, Tryptone Glucose Yeast Extract Agar (TGYA), TGYA enriched with 1 % starch solution and TGYA enriched with 5 % starch solution were used for potential microorganism isolation. Samples were incubated at 25 °C; 37 °C; 41.5 °C; 50 °C; 55 °C and 60 °C temperatures under aerobic and anaerobic conditions, incubation time 12, 24 and 48 h. Facultative thermophilic Bacillus licheniformis bacteria which produce thermotolerant spores were isolated from thermally treated potato products in retort packaging from company B. The average colony count in company B samples was 1.27´100 CFU/g (SD = 78.25). The presence of microorganisms was not detected in potato products produced at company A, while facultative thermophilic and thermotolerant microorganisms were found in potato products produced at company B. DOI: http://dx.doi.org/10.5755/j01.ct.66.1.12363

Diversity of organotrophic bacteria, activity of dehydrogenases and urease as well as seed germination and root growth Lepidium sativum, Sorghum saccharatum and Sinapis alba under the influence of polycyclic aromatic hydrocarbons.

Polycyclic aromatic hydrocarbons are organic compounds with highly toxic, carcinogenic, and mutagenic properties, which adversely affect the basic biological parameters of the soil, including the count of microorganisms, and the enzymatic activity. In addition to disturbances to the biological activity of the soil, PAHs may also exhibit toxic effects on plants. In view of the above, the study involved testing aimed at the determination of the effects of polycyclic aromatic hydrocarbons in a form of naphthalene, phenanthrene, anthracene and pyrene on the count, colony development (CD) index, ecophysiological (EP) diversity index of organotrophic bacteria, and the activity of soil dehydrogenases and soil urease. Moreover, an attempt was made to determine the soil’s resistance based on the activity of the above-listed enzymes, and the effect of polycyclic aromatic hydrocarbons on seed germination and root growth was assessed by Lepidium sativum, Sorghum saccharatum, and Sinapis alba. In addition, the species of bacteria found in a soil subjected to strong pressure of polycyclic aromatic hydrocarbons were isolated. The experiment was performed in a laboratory on samples of loamy sand. Polycyclic aromatic hydrocarbons were introduced into the soil in an amount of 0, 1000, 2000, and 4000 mg kg−1 of soil dry matter. Germination and growth of cress (L. sativum), white mustard (S. alba), and sweet sorghum (S. saccharatum) were determined using Phytotoxkit tests. It was found that the tested PAHs increased the average colony counts of organotrophic soil bacteria; pyrene did so to the greatest extent (2.2-fold relative to non-contaminated soil), phenanthrene to the smallest extent (1.4-fold relative to non-contaminated soil). None of the PAHs changed the value of the bacterial colony development (CD) index, while anthracene and pyrene increased the value of the eco-physiological (EP) diversity indicator. PAHs lowered the activity of the tested enzymes. The activity of dehydrogenases was dependent on a greater extent by the type of hydrocarbon (54.56 %) rather than by the dose (10.64 %), while for the activity of urease, it was the opposite. The greater extent was dependent on dose (95.42 %) rather than by type (0.21 %). Dehydrogenases are characterised by greater resistance to the action of PAHs than urease. Based on seed germination and root growth, it has shown that S. alba is best suited, being the most vulnerable plant, while S. saccharatum is the least suited. Subjecting a soil to strong pressure of PAHs leads to disturbances to the biological parameters of the soil, seed germination, and root growth L. sativum, S. saccharatum, and S. alba.

Molecular Mechanism of Action of 5‐Azacytidine and its Resistance in MDA‐MB 435 Cells

DNA methyltransferase inhibitors such as 5‐Azacytidine (5‐Aza) have shown promise in cancer treatment because of their ability to allow re‐expression of tumor suppressor genes in cancer cells. Preliminary studies showed that high doses of 5‐Aza are clearly cytotoxic to MDA‐MB 435 tumor cells. However, we have begun treating the MDA‐MB 435 cells with a low dose so that the majority of the cells survive. A resistant cell line has been developed through continuous growth in the presence of 5‐Aza. Tests were done to determine if the resistance inhibited growth in a methylcellulose assay. The number of colonies decreased in the 5‐Aza resistant cell line compared to the control. Control 435 cells were found to have an average colony count of 969±332 but when 5‐Aza was added the colony count dropped to 464±31. In order to look at the mechanism of 5‐Aza and its resistance, quantitative reverse transcriptase PCR was done to measure the expression of various genes. Between the control cells and the control cells plus 5‐Aza there was a five‐fold decreased expression of Mammaglobin B, a member of the secretoglobin family associated with chemoresistance, while the other genes tested did not have any change (TP53, RB1, BCL‐2, Myc). When the same tests were done on the resistant cell line the colony counts showed little change, 287±129 and 357±46, respectively, nor was there a difference between the resistant and resistant plus 5‐Aza in terms of Mammaglobin B expression. However, the resistant and the resistant plus 5‐aza both showed a six‐fold decrease in Mammaglobin B expression compared to the control cells. Future experiments will investigate a phenotypic role for this gene in the resistance.

058 Cryopreservation of non-manipulated stem cell products; clinical implication

Cryopreservation is of critical importance to the successful clinical use of stem cell products since, except for freshly harvested marrow, most products are cryopreserved months to years before infusion into patients. Despite the importance, data regarding the optimization of cryopreservation of stem cells remains rather limited. Questions remaining that are of interest to the field include defining a shelf life for stem cell products, and determining how to assess successful freezing and thawing procedures. We recently performed two studies to help address these questions. In the first, we assessed the viability of peripheral blood stem cells (PBSC) cryopreserved for 16–18 years (average 17.5 years). Post-thaw cell recovery, viability and colony forming ability were assessed in vitro. To evaluate engraftment capabilities in vivo, four units were thawed, CD34+ cells isolated, and 2 ×105 CD34+ cells were infused into sublethally irradiated NOD/SCID/IL-2R γ null (NSG) mice. Average % cell recovery after thawing the seven units was 92% ± 11.4 with average % post-thaw viability of 72% ± 19. Average colony counts from the seven thawed units were 3.4 ± 3 CFU-GM and 3.1 ± 3 BFU-E. Significantly, at twelve weeks, NSG mice transplanted with two of the units showed evidence of long-term engraftment with the presence of human CD45+ cells by flow cytometry (hCD45+ range 26-81%). Mice transplanted with the other two units are in progress. This study demonstrates PBSC can be cryopreserved for over 16 years and retain colony forming ability, engraft into NSG mice and likely lead to successful clinical transplantation. In the second study, we examined the recovery of nucleated cells from cord blood units and correlated recovery with engraftment kinetics. Retrospective analysis was performed on 59 cord blood units transplanted into 52 patients at Indiana University (average age 23.6 years) with malignant and non-malignant diseases from January 2010 to October 2012. Cord blood units were obtained from 15 different cord blood banks and thawed using the Rubenstein thaw-wash method. Pre-freeze and post-thaw cell dose and viability with trypan blue were analyzed. The corrected recovery was calculated by (pre-freeze – post-thaw cell count) ×viability. This was correlated with neutrophil and platelet engraftment data. Average cell recovery was 81 ± 9.8% with post-thaw viability of 82 ± 6.4% for a corrected recovery of 66 ± 9.9%. Time to engraftment significantly correlated with infused cell dose/kg corrected for viability ( p = 0.0056). Platelet engraftment was significantly predicted by both corrected and uncorrected cell dose/kg ( p < 0.0005). In this study, infused cell dose/kg corrected for post-thaw viability was a stronger predictor of delayed neutrophil engraftment than cell dose/kg alone. In our patient cohort, we were surprised to find that this population averaged a corrected recovery of 66%, suggesting that it is essential to consider a loss of up to a third of the reported cell dose in cord blood unit selection practices. Along with HLA match, optimizing infused cell dose by accounting for additional cell losses using corrected recovery could impact cord selection and patient outcomes. These studies illustrate that while cryopreservation of hematopoietic stem cell products has been successful clinically for many years, work remains to optimize the freezing and thawing process to improve clinical results. Moreover, development of cryopreservation techniques for manipulated stem cell populations such as MSC are in their infancy, and established techniques used for hematopoietic stem cells may not necessarily be optimal for MSC. Some data from the studies in this abstract were presented at the Tandem ASPHO/PBMTC meeting in Miami FL April 2013, and accepted for a poster presentation at the annual AABB meeting in Denver CO, October 2013. Source of funding: None declared. Conflict of interest: None declared. sgoebel2@iupui.edu

Bacterial load in expressed and stored breast milk of lactating mothers in Abia state, Nigeria

The use of expressed breast milk has been advocated as an effective way of encouraging and maintaining lactation when the mother is separated from the baby for a while. However, prospects of storage of expressed breast milk for any considerable period of time is hindered by the possibility of bacterial contamination and growth of infectious pathogens in the stored milk. Previous research worldwide has focused on optimal conditions for storing expressed breast milk. This study investigated bacterial growth in expressed breast milk stored at two different conditions and time periods. A total of 240 randomly selected lactating mothers participated in a survey used to elicit information on their practices concerning expressed and stored breast milk. Out of this, 20 lactating mothers were willing to donate their breast milk , which was used for microbial studies. Using antiseptic procedures, breast milk samples were collected from these 20 lactating mothers by manual expression and put into sterile containers. One half was stored at room temperature (30 o C), while the other half was immersed in a container of water (26 o C) for 0, 3, 6 and 9hrs. The samples at different time periods in both storage conditions were plated on three different culture media (Nutrient agar , MacConkey agar and Chocolate agar) and incubated for 24hrs. Bacterial load was enumerated and isolates were characterized and identified. Data was analyzed using one way ANOVA and presented as frequencies, percentages means and standard deviations. Results showed that in all the culture media, bacteria l load was higher in breast milk samples stored at room temperature for up to 9hrs compared to that immersed in water (p&lt;0 .05). However, the average colony counts were within acceptable limits (&lt;10 4 CFU/ml). Enterococcus faecalis , Escherichia coli and Staphylococcus aureus were the predominant bacteria isolated in the breast milk samples under both storage conditions. The findings from this study revealed that storage of breast milk either at room temperature or immersed in a container of water appeared to be safe for up to 9 hours of storage in a tropical environment . Generally, the number of microbes was within levels considered acceptable in expressed breast milk .

In vitro bactericidal activity of jinghua weikang capsule (荆花胃康胶丸) and its individual herb Chenopodium Ambrosioides L. against antibiotic-resistant Helicobacter Pylori

To investigate the bactericidal effects of Jinghua Weikang Capsule and its major component Chenopodium ambrosioides L. on antibiotic-resistant Helicobacter pylori. Four clinical antibiotic-resistant H. pylori strains were isolated and incubated in liquid medium containing Jinghua Weikang Capsule or Chenopodium ambrosioides L. By means of time-kill curve method, the average colony counts and bactericidal rate were calculated at time points of 0, 4, 8 and 24 h after the incubation and the time-kill curves were charted. Both Jinghua Weikang Capsule and Chenopodium ambrosioides L. at a concentration of 0.64 g/L showed obvious bactericidal effect against antibiotic-resistant H. pylori after 4 h of incubation. Jinghua Weikang Capsule and Chenopodium ambrosioides L. are considered to be active against antibiotic-resistant H. pylori in vitro.

Effect Of Abdominal Insufflation On Bacterial Growth In An Experimental Model Of Peritonitis - A Randomized Controlled Trial.

Background: Despite the advantages of laparoscopy, studies suggest that gases used for insufflation may adversely affect patients with peritonitis by bacterial proliferation and impair the clearance of bacteria from the peritoneal cavity.Methods: This was an experimental study conducted on 33 albino-rats from February 2004 to September 2004 in Department of Surgery N.S.C.B, Government Medical College, Jabalpur (MP), India. This study was done to assess the effect of pneumoperitoneum on peritonitis. The rats were divided in three groups (A, B, and C). In group A (control group) 0.5ml of normal saline (5%) was instilled intraperitoneally. In group B and C, a single infusion of 0.5ml suspension containing 1×10 Escherichia coli/ml was instilled intraperitoneally. In group A and C, an additional 5ml of atmospheric air was instilled. The colony counts in each group were noted after 72 hours after laparotomy. Results: In the control group there was no peritoneal contamination, no bacterial growth, none of the rats had died and there was no evidence of pus or pus pockets seen. In the experimental group the mean colony count was 659.91 (±680.34). There were no macroscopic pus pockets or adhesions seen but there was frank pus in one of the rats of this group. The t-value for comparison of colony count was tA/B =3.22 (p<.05). In group C there were no macroscopic pus pockets seen, no adhesions observed and a mean colony count of 840.09 (±681.21) was observed.Conclusion: There is an increase in average colony count in the pneumoperitoneum group as compared to nonpneumoperitoneum group.

Ecological health of the Usuthu and Mbuluzi rivers in Swaziland based onselected biological indicators

Biological monitoring, using coliform bacteria and macroinvertebrate populations and diversity, was carried out monthly from August 2004 to January 2005 to determine the ecological health of the Usuthu and Mbuluzi rivers in Swaziland. Water temperature and pH were not significantly different between sites, but differed significantly between months. Bacterial species identified from both rivers were Escherichia coli, Klebsiella pneumoniae and Proteus vulgaris, with no significant differences in mean colony counts between the two rivers. Average colony counts were significantly different between months. A total of 926 macroinvertebrates, representing 28 families, were collected from both rivers and were significantly different between sites, rivers and months. Macroinvertebrate populations and diversity were negatively correlated with bacterial colony counts. Compliance with regulated water quality standards did not necessarily indicate acceptable water quality for biological communities, and thus there is a need for an integrated approach in water quality monitoring.

Effect of face washing with soap and water and cleaning with antiseptics on upper-lid bacteria of surgical eye patients

Objective: To determine the effect of face washing with soap and water and cleaning with povidone iodine and cetrimide/chlorhexidine gluconate (Savlon) on upper-lid bacteria. Design: Prospective, nonrandomized clinical trial. Participants: Eighty patients attending the Eye Clinic, University College Hospital, Ibadan, Nigeria. Methods: Eighty patients assigned to 4 groups had swabs of the upper eyelid skin taken before and after face wash with soap and water, and cleansing with Savlon and 5% povidone iodine. Specimens were cultured and Gram stained. Bacterial counts were carried out using standard techniques. Results: Face washing with soap and water increased the proportion of patients with bacterial isolates from 80.0% to 87.5%. The average colony count increased from 187.1 to 318.5 colony units per mL ( p = 0.02). Application of 5% povidone iodine without face washing with soap and water reduced the proportion of patients with bacterial isolates from 82.6% (mean count 196.5) to 28.6% (mean count 34.1) ( p = 0.001); in comparison, the application of 5% povidone iodine after face washing with soap and water reduced the proportion from 71.4% (mean count 133.9) to 40.0% (mean count 69.0) ( p = 0.01). Application of Savlon without face washing with soap and water reduced the proportion of patients with bacterial isolates from 100% (mean count 3 10.9) to 41.2% (mean count 19.8) ( p = 0.004) compared with the application after face washing, which reduced the proportion from 89.5% (mean count 240.3) to 41.2% (mean count 82.9) ( p = 0.02). Conclusions: Both povidone and Savlon are effective in reducing periocular bacteria in an African setting. Prior face washing with soap and water had no added benefit in reducing bacterial colony count.