Sterile Solution Research Articles

799. A Pseudo-Outbreak of Pseudomonas fluorescens Infections

Background Pseudomonas fluorescens is a water-borne pathogen that has been associated with outbreaks from transfusion of contaminated blood products or medical equipment. Our institution had a cluster of cultures that grew an uncommonly encountered microbe P. fluorescens within a period of one week. This prompted an internal investigation. We summarize the investigational process that led to the resolution of this pseudo-outbreak.MethodsWe conducted a retrospective chart review of surgical and non-surgical patients with cultures positive for P. fluorescens from July 2nd to July 8th 2020. Baseline patient characteristics, clinical course, laboratory data, use of blood-associated products, and microbiology cultures were analyzed.ResultsEight patients were identified with positive tissue cultures for P. fluorescens. Among those, 5 specimens (62.5%) were from osteoarticular sites (1 prosthetic hip, 1 prosthetic knee, 1 right foot, 1 sternum, and 1 vertebral source). One culture (12.5%) was obtained from a sacral soft tissue wound. Two tissue specimens (25%) were collected from respiratory sites (1 lung tissue and 1 bronchoalveolar lavage). No association with specific surgical personnel or operating room was identified. During routine specimen processing, a small amount of sterile normal saline is added to the conical grinder prior to culture preparation. It was discovered that a non-sterile normal saline had been inadvertently utilized during that step. These eight tissue specimens were subsequently reprocessed with sterile solution; P. fluorescens was not re-isolated. Specimen processing protocols were reinforced. Adjustment of antimicrobial therapy was made accordingly without reported subsequent adverse clinical outcomes.ConclusionA multi-faceted team approach in collaboration with Infection Prevention, Infectious Diseases, Surgery, operating room personnel, and Microbiology identified an unintended breakdown in sterile laboratory protocols which resulted in a cluster of falsely positive cultures. An increased incidence of infection with an uncommon pathogen initiated a prompt investigation that resulted in the identification of a pseudo-outbreak event.Disclosures All Authors: No reported disclosures

805. Outbreak of Ralstonia pickettii Bacteremia Caused by Contaminated Hydromorphone in a Universitary Hospital in Bogota, Colombia

Background Ralstonia pickettii are aerobic non fermenter gram negative bacilli isolated in water and soil. It is related to nosocomial infection outbreaks and considered an opportunistic pathogen. There have been outbreaks reports due to contaminated water systems and sterile drug solutions which mainly occurs during manufacturing. We present the report of an outbreak of R. pickettii bacteremia secondary to a contamination of hydromorphone vials.MethodsIn February 2021 an outbreak of R. pickettii bacteremia was identified. All isolates were from blood cultures with slow growth, thus indicating the culturing of liquid inputs, intravenous administration solutions and commonly used drugs among patients including hydromorphone. Mass spectrometry (MALDI-TOF) was used for the identification and automated microdilution to determine sensitivity to antimicrobials of the isolates and clonality analysis of genetic relationships was carried out using the DICE coefficient, UPGMA algorithm ResultsDuring the outbreak, 19 patients with R. pickettii bacteremia were identified The global attack rate was 1,9%. 11/19 (58%) were women and 13/19 (68%) of the isolations were from inward patients and 6/19 (32%) were from intensive care unit. Factors that could contribute to the appearance of the outbreak were underlying pathology, 2 patients with a diagnosis of diabetes mellitus, 10 patients with a diagnosis of arterial hypertension, 5 patients with obesity, 6 patients with heart disease, additionally 7 patients with a diagnosis of SARS COV 2 and 6 patients with the use of corticosteroids. The global attack rate was 1,9% and mortality was 31.5% (6 patients). R. pickettii was identified from two batches of hydromorphone by MALDI-TOF and the clonality study concluded that the isolates analyzed, were clonal with a 100% similarity. The associated mortality rate was 5/29 (26.3%). ConclusionWe confirmed an outbreak of R. pickettii due to the contamination of two hydromorphone badges in Colombia. It is crucial to acknowledge the importance of infection control and surveillance during the COVID-19 pandemic as well as maintaining adequate quality control of medication production in order to avoid presenting this kind of outbreaks. Disclosures Sandra Liliana. Valderrama-Beltrán, MD, MSc, Biotoscana (Speaker’s Bureau)MSD (Grant/Research Support, Scientific Research Study Investigator, Research Grant or Support, Speaker’s Bureau)Pfiezer (Speaker’s Bureau)

ОЦЕНКА КУМУЛЯТИВНЫХ СВОЙСТВ КОМПЛЕКСНОГО СРЕДСТВА З-88

This present work is devoted to the study of cumulative properties of the complex means Z-88. The studies were carried out on intact mongrel laboratory rats that had previously passed quarantine measures for 12 days. Two equivalent groups were created. The animals of the experimental group were injected intramuscularly in the thigh area with Z-88, and the control group was similarly injected with 0.9% sterile sodium chloride solution. On the first day of the experiment, the rats were injected with test drugs at a dose of 0.5 ml per animal, which was 1/10 of the maximum volume used in experiments to determine the parameters of acute toxicity. Both Z-88 and sterile saline solution were administered once a day throughout the experiment. Z-88 was administered once a day throughout the experiment. Starting from the fifth day, in every subsequent four days, the dose of the drug was increased by 1.5 times. The duration of the experiment was 24 days. During the experimental period of visible changes on the part of the studied indicators.

Investigation of hydrophilic interaction liquid chromatography coupled with charged aerosol detector for the analysis of tromethamine

Tromethamine (TMM), often encountered in a final drug product, exhibits interesting chemical properties as a counter ion, buffer, or active ingredient. European and US pharmacopeias propose titration against hydrogen chloride for TMM assays. However, this method can be a hindrance when using drugs containing low concentrations of TMM in complex buffered formulations. Due to the lack of chromophores and the high hydrophilicity of TMM, we performed a simple and reliable hydrophilic interaction chromatography coupled with a charged aerosol detector (HILIC-CAD) separation approach as an alternative for TMM analysis. An amide stationary phase and a mobile phase consisting of a binary mixture of acetonitrile and 10 mM ammonium formate, pH 3 (80/20, V/V) were used. As the CAD response deeply depends on parameters such as stationary phases and pH buffer, we investigated their impact and explored the optimal signal conditions. Including TMM analogs such as tris(hydroxymethyl) nitromethane and 2-amino-2-ethyl-1,3-propanediol allowed us to select these parameters appropriately. The effects of the evaporation temperature, flow rate, and power function value (PFV) on the CAD signal response were also studied and optimized. The method was validated according to the ICH Q2 R1 guidelines. A linear response (mean R2 > 0.997) covering the range for low TMM concentrations (170–520 μg/mL) was achieved. Satisfactory intra-day and inter-day precisions were obtained with RSDs lower than 1.9% and 2.8%, respectively. The trueness ranged from 99.6% to 101.2%, and the LOD was found to be 1.1 μg/mL. The HILIC-CAD method has been applied to a sterile TMM solution for injection.

EVALUATION OF PATHOGENICITY OF NODULE BACTERIA STRAIN RHIZOBIUM LEGUMINOSARUM G 222

Objective. Investigate pathogenic (non-pathogenic) properties of a new strain of nodule bacteria Rhizobium leguminosarum G 222. Methods. Microbiological (obtaining a suspension of R. leI guminosarum G 222, determination of its titre by opacity optical standard and by seeding on a digest medium; microscopy of stained imprint smears of internal organs of experimental animals and seeding of tissue samples on a digest medium), pathoanatomical (to determine possible invasiveness and dissemination of bacteria in the tissues of internal organs of animals — in the model of outbred white mice after administration of a suspension of live bacterial cells (oral doses from 0.5 × 109 to 5 × 109 CFU in 0.5 cm3 of sterile isotonic sodium chloride solution per mouse and intraperitoneal doses from 1 × 109 to 5 × 109 CFU/0.5 cm3) and statistical. Results. Over 15 days of observations after administration of a suspension of live bacterial cells, no death of experimental animals was registered. The strain did not lead to any changes in the general condition of the mice. No changes in their behaviour were reported. Fifteen days after the start of the study, it was found that this strain is non-infective (non-invasive), does not disseminate and does not reproduce in the body of experimental animals. Oral and intraperitoneal doses of a suspension of live bacterial cells did not result in bacterial invasion of animal internal organs. No retrocultures were registered. Gross examination did not find characteristic changes in the internal organs of the experimental animals. The obtained results indicate the avirulence of the strain for the studied warm-blooded animals (intraperitoneal LD50 > 5 billion cells/mouse, oral LD50 > 5 billion cells/mouse). Conclusion: According to the results regarding the lack of virulence and according to regulations, the new strain R. leguminosarum G 222 belongs to the group of avirulent microorganisms that are not able to invade the internal organs of studied warm-blooded laboratory animals and can be considered nonpathogenic and used as a basis for microbial preparations to increase crop yields.

Експериментальне моделювання туберкульозного спондиліту

Актуальність. Туберкульозний спондиліт (ТС) в структурі кістково­суглобового туберкульозу у дорослих займає провідне місце і досягає 40–61,5 %. Метою даної публікації є висвітлення процесу моделювання туберкульозного спондиліту і його особливостей у морських свинок для подальшого використання результатів експерименту у лікувальній та науковій практиці. Матеріали та методи. Експеримент ставився на 40 статевозрілих морських свинках. Моделювання туберкульозного спондиліту проводилось на основі розробленого нами способу (патент № 112423 (UА) Україна). Усі тварини були розподілені на чотири рівні групи: 1­ша — 3­тя групи (основні) — виконувалась ін’єкція 0,5 мл суспензії М.tuberculosis (0,1 мг сухої маси в 1 мл) в тіло хребця згідно з методикою; 4­та група — контрольна. Тваринам робилась ін’єкція стерильного фізіологічного розчину (0,9% — 0,5 мл) в тіло хребця. Група 1 (10 свинок) — лікування специфічними антибактеріальними препаратами (АБП) першого ряду (ізоніазид, стрептоміцин, рифампіцин). Група 2 (10 свинок) — лікування специфічними антибактеріальними препаратами другого ряду (амікацин, рифабутин, офлоксацин). Група 3 (10 свинок) — не проводилося лікування. За всіма тваринами проводилося динамічне спостереження з клінічними, рентгенологічними, патоморфологічними і лабораторними дослідженнями. Виведення ­експериментальних тварин з досліду проходило за раніше розробленим графіком — при виявленні ознак досліджуваних стадій туберкульозного процесу. Всім виведеним з експерименту тваринам проводилося патоморфологічне дослідження. При цьому основна увага приділялася вивченню патоморфології уражених специфічним процесом хребців. Результати. У результаті морфологічного дослідження встановлено наявність активного туберкульозного процесу в тілах хребців і паравертебральних тканинах у тварин із модельованим туберкульозом і лікуванням специфічним АБП першого ряду, а також у тварин, які не приймали специфічного лікування. Важливо зазначити, що ступінь вираженості деструктивних змін у вражених хребцях у тварин нелікованих і тих, які приймали АБП першого ряду, практично однаковий. У тварин, яким моделювали туберкульоз і лікували специфічним АБП другого ряду, виявлено пригнічення патологічного процесу з утворенням молодої кісткової та сполучної тканини різного ступеня зрілості та наявністю зони, що відмежовує вогнище запалення від здорової тканини в ранні терміни захворювання (один місяць). В результаті виконаної роботи вдалося простежити стадійність розвитку туберкульозного спондиліту у морської свинки та сучасні особливості клінічного, рентгенологічного та патоморфологічного перебігу. Виявлена ідентичність моделі основних клінічних форм туберкульозного спондиліту у морської свинки та людини. Висновки. Дане дослідження показало, що проведення сучасної інтенсивної специфічної антибактеріальної терапії в умовах експерименту дозволяє досягти відмежування деструктивного процесу в порівняно ранні терміни розвитку захворювання (4–5 тижнів). Отримані нові знання про патоморфологічні особливості перебігу туберкульозного спондиліту дозволяють проводити радикальні оперативні втручання на хребті без ризику генералізації туберкульозного процесу в більш ранні терміни.

Biological control of Listeria monocytogenes in soil model systems by Enterococcus mundtii strains expressing mundticin KS production

The agricultural practices applied in pre-harvest greatly influence the presence and the levels of microorganisms in fresh produce. Among these, Listeria monocytogenes represents one of the most lethal foodborne pathogens associated with vegetables. The main hypothesis of this work is that bacteriocin producer Enterococcus mundtii strains can be effective against L. monocytogenes in soil. To this purpose, bacteriocin production by E. mundtii WFE3, WFE20 and WFE31, three strains showing a strong bacteriocin activity in terms of inhibitory power and inhibition spectra, was evaluated in sterile extracts from agricultural soil and peat moss, in organic nutrient solution (ONS) and mineral nutrient solution (MNS). ONS supernatants from E. mundtii WFE3 showed the highest inhibition of the strain L. monocytogenes ATCC 19114. Thus, this strain [104 colony forming units (CFU) g−1 dw] was co-inoculated with E. mundtii WFE3 (106 CFU g−1 dw) in the sterile extracts and solutions. A general increase of E. mundtii cell densities and the contemporary decrease of the levels of L. monocytogenes were observed, particularly in ONS. This solution was used to amend autoclaved soil to test in vivo the competition among bacteriocin producer and sensitive strain. L. monocytogenes decreased of almost 1.5 log CFU when E. mundtii was added to soil. The test was also carried out with basil plants showing that the anti-Listeria effect of E. mundtii is exerted during the very first days from inoculation. After 2 days, the levels of NO3−-N in soil increased for all trials, while the concentrations of ammonium and α-amino N decreased. The lowest concentrations of NH4+-N were found in presence of L. monocytogenes. At harvest, the plants were analysed for the presence of E. mundtii and L. monocytogenes, but none of the two bacterial species deliberately added to soil was transferred to plants. Although a biocontrol based on bacteriocin application in soil has to be properly set, this study provides useful insight for the future development of chemical-free agricultural strategies.

Effect of sanitizers and Lactobacillus rhamnosus R23 on the growth of Salmonella spp. in raw chicken fillets during temperature abuse storage

One of the food products commonly contaminated by Salmonella is raw chicken. In wet markets in Southeast Asian countries, chicken carcasses frequently were handled and sold at abused temperatures, above the refrigeration temperatures (>5°C), thus could support the growth of Salmonella. One way to extend the shelf life of raw chicken carcasses at room temperature is by reducing the initial contamination using sanitisers such as ozone micro-bubble water (OMBW) or hypochlorite (NaOCl) solution. The other option is by adding bio-preservative agents such as lactic acid bacteria. This study aimed to evaluate the effect of sanitisers in reducing the initial contamination and the potential of lactic acid bacteria in inhibiting the growth of Salmonella in raw chicken fillets stored at abused temperatures. Chicken fillets were artificially inoculated with Salmonella (~5 log CFU/g) and rinsed for 5 minutes with sterile water, OMBW (1 and 2 ppm), or NaOCl solution (50 and 100 ppm). The results showed that washing the chicken fillets with NaOCl 100 ppm gave the most reduction of Salmonella counts. However, there were no significant effects regarding the inhibition of Salmonella growth during temperature abuse between those previously washed with OMBW or sterile water. The addition of L. rhamnosus R23 (6 log CFU/g and 8 log CFU/g) did not significantly inhibit the growth of Salmonella as compared to the control.

A microdosing framework for absolute bioavailability assessment of poorly soluble drugs: A case study on cold-labeled venetoclax, from chemistry to the clinic.

This work presents an end‐to‐end approach for assessing the absolute bioavailability of highly hydrophobic, poorly water‐soluble compounds that exhibit high nonspecific binding using venetoclax as a model drug. The approach utilizes a stable labeled i.v. microdose and requires fewer resources compared with traditional approaches that use radioactive 14C‐labeled compounds. The stable labeled venetoclax and internal standard were synthesized, then an i.v. formulation was developed. In the clinical study, female subjects received a single oral dose of venetoclax 100 mg followed by a 100‐µg i.v. dose of cold‐labeled 13C‐venetoclax at the oral time of maximum concentration (Tmax). The i.v. microdose was prepared as an extemporaneous, sterile compounded solution on the dosing day by pharmacists at the clinical site. Several measures were taken to ensure the sterility and safety of the i.v. preparation. A sensitive liquid chromatography‐tandem mass spectrometry method was developed to allow the detection of plasma levels from the i.v. microdose. Plasma samples were collected through 72 h, and pharmacokinetic parameters were estimated using noncompartmental methods. Postdosing sample analysis demonstrated the consistency of the preparations and allowed the precise calculation of the pharmacokinetic parameters based on the actual injected dose. The absolute bioavailability of venetoclax was estimated at 5.4% under fasting conditions. Venetoclax extraction ratio was estimated to be 0.06 suggesting that the fraction transferred from the enterocytes into the liver is limiting venetoclax bioavailability. The proposed framework can be applied to other highly hydrophobic, poorly water‐soluble compounds that exhibit high nonspecific binding to support the understanding of their absorption and disposition mechanisms and guide formulation development.

Effect of Cinnamon Nanoparticles in Presence of HAMLET on the Healing of Wounds Infected with Pseudomonas aeruginosa: An Animal Model Study

The objective was to evaluate the ability of cinnamon nanoparticles (CNPs) in the healing of wounds with Pseudomonas aeruginosa (PAE) infection as well as HAMLET sensitization in rats. Fifty healthy male Wistar rats were used in this study. The rats were divided into 5 groups (n=10), randomly. In the Normal group, no infected wounds were treated with a sterile solution of saline 0.9% (0.1 ml). In the PAE group, the wounds with Pseudomonas aeruginosa infection were only treated with a sterile solution of saline 0.9% (0.1 ml). In the PAE-HMLT group, HAMLET (100 µg) was used to treat infected wounds. In the PAE-CNM group, 1 mg/ml CNPs (0.1 ml) was applied topically to treat PAE-infected wounds. In the PAE-HMLT-CNM group, HAMLET (100 µg) and 1 mg/ml CNPs (0.1 ml) were applied topically to treat PAE-infected wounds. Microbiological examination, planimetric and biochemical showed significant differences between rats in the PAE-HMT-CNM group in comparison with other groups (p < 0.05). In conclusion, CNPs could offer potential to pay more attention to this harmless and easily available agent to be topically applied in wounds with infection.

SUMO1 modification of IGF-1R combining with SNAI2 inhibited osteogenic differentiation of PDLSCs stimulated by high glucose

BackgroundPeriodontal disease, an oral disease characterized by loss of alveolar bone and progressive teeth loss, is the sixth major complication of diabetes. It is spreading worldwide as it is difficult to be cured. The insulin-like growth factor 1 receptor (IGF-1R) plays an important role in regulating functional impairment associated with diabetes. However, it is unclear whether IGF-1R expression in periodontal tissue is related to alveolar bone destruction in diabetic patients. SUMO modification has been reported in various diseases and is associated with an increasing number of biological processes, but previous studies have not focused on diabetic periodontitis. This study aimed to explore the role of IGF-1R in osteogenic differentiation of periodontal ligament stem cells (PDLSCs) in high glucose and control the multiple downstream damage signal factors.MethodsPDLSCs were isolated and cultured after extraction of impacted teeth from healthy donors or subtractive orthodontic extraction in adolescents. PDLSCs were cultured in the osteogenic medium with different glucose concentrations prepared by medical 5% sterile glucose solution. The effects of different glucose concentrations on the osteogenic differentiation ability of PDLSCs were studied at the genetic and cellular levels by staining assay, Western Blot, RT-PCR, Co-IP and cytofluorescence.ResultsWe found that SNAI2, RUNX2 expression decreased in PDLSCs cultured in high glucose osteogenic medium compared with that in normal glucose osteogenic medium, which were osteogenesis-related marker. In addition, the IGF-1R expression, sumoylation of IGF-1R and osteogenic differentiation in PDLSCs cultured in high glucose osteogenic medium were not consistent with those cultured in normal glucose osteogenic medium. However, osteogenic differentiation of PDLCSs enhanced after adding IGF-1R inhibitors to high glucose osteogenic medium.ConclusionOur data demonstrated that SUMO1 modification of IGF-1R inhibited osteogenic differentiation of PDLSCs by binding to SNAI2 in high glucose environment, a key factor leading to alveolar bone loss in diabetic patients. Thus we could maximize the control of multiple downstream damage signaling factors and bring new hope for alveolar bone regeneration in diabetic patients.

The Involvement of Macrophage Colony Stimulating Factor on Protein Hydrolysate Injection Mediated Hematopoietic Function Improvement.

Protein hydrolysate injection (PH) is a sterile solution of hydrolyzed protein and sorbitol that contains 17 amino acids and has a molecular mass of 185.0–622.0 g/mol. This study investigated the effect of PH on hematopoietic function in K562 cells and mice with cyclophosphamide (CTX)-induced hematopoietic dysfunction. In these myelosuppressed mice, PH increased the number of hematopoietic cells in the bone marrow (BM) and regulated the concentration of several factors related to hematopoietic function. PH restored peripheral blood cell concentrations and increased the numbers of hematopoietic stem cells and progenitor cells (HSPCs), B lymphocytes, macrophages, and granulocytes in the BM of CTX-treated mice. Moreover, PH regulated the concentrations of macrophage colony stimulating factor (M-CSF), interleukin (IL)-2, and other hematopoiesis-related cytokines in the serum, spleen, femoral condyle, and sternum. In K562 cells, the PH-induced upregulation of hematopoiesis-related proteins was inhibited by transfection with M-CSF siRNA. Therefore, PH might benefit the BM hematopoietic system via the regulation of M-CSF expression, suggesting a potential role for PH in the treatment of hematopoietic dysfunction caused by cancer therapy.

PSXVI-3 Supplemental effects of Bacillus sp. in prevention of post-weaning diarrhea caused by enterotoxigenic F18+ Escherichia coli in nursery pigs

Abstract This study investigated the effects of Bacillus sp. on intestinal health and prevention of diarrhea by F18+ Escherichia coli (ETEC) in nursery pigs. Forty-eight weaned pigs (7.9 ± 0.5 kg BW) were allotted to 4 treatments with 12 replicates (NC: no-challenge; PC: challenge/no-treat; BMD: challenge/bacitracin; BAC: challenge/Bacillus sp. at 109 CFU/kg feed). All pigs were fed diets for 28 d. At d 7, challenged groups were orally inoculated with ETEC (1.8 × 1010 CFU) and NC group received sterile solution. Growth performance was analyzed weekly and pigs were euthanized on d 28 to measure intestinal health. Data were analyzed using SAS 9.4. There were no difference on growth performance among treatments during pre-challenge period. From d 7 to 14, PC tended to reduce (P = 0.067) ADG (352 to 440 g/d), whereas no effect was observed during the overall period. The PC increased (P &amp;lt; 0.05) fecal score at d 7 (post-challenge) (4.1 to 3.3) and d 8 (4.1 to 3.3), whereas BMD decreased (P &amp;lt; 0.05) it at d 9 (4.3 to 3.5) and d 11 (3.8 to 3.3). The PC increased (P &amp;lt; 0.05) MDA (0.56 to 0.30 μmol/mg protein) and tended to increase TNFα (P = 0.084; 0.89 to 1.04 pg/mg protein), whereas BMD and BAC reduced (P &amp;lt; 0.05) MDA (0.56 to 0.31, 0.29 μmol/mg protein) and TNFα (1.04 to 0.81, 0.77 pg/mg protein) in jejunal mucosa. The PC reduced (P &amp;lt; 0.05) α-diversity (Chao1: 32.3 to 19.4), whereas BAC increased it (Chao1: 19.4 to 30.7) whereas BMD increased (P &amp;lt; 0.05) it (Shannon: 1.85 to 2.44; Simpson: 0.55 to 0.70) of jejunal mucosa-associated microbiota. In conclusion, ETEC challenge caused diarrhea, increased immune response, and oxidative stress by disrupting mucosa-associated microbiota. The BMD and BAC reduced jejunal immune response and oxidative stress by restoring mucosa-associated microbiota.

PSXVI-7 Supplemental effects of Lactobacillus extract postbiotics in prevention of post-weaning diarrhea caused by enterotoxigenic F18+ Escherichia coli in nursery pigs

Abstract This study determined the supplemental effects of Lactobacillus extract (LBE) postbiotics on intestinal health and prevention of postweaning diarrhea caused by F18+ Escherichia coli (ETEC) in nursery pigs. Sixty-four weaned pigs (6.6 ± 0.7 kg BW) were allotted in a RCBD to 4 dietary treatments (NC: no-challenge; PC: challenge/no-treat; BMD: challenge/bacitracin; LBE: challenge/LBE 0.2%) and fed diets for 28 d. At d 7, challenged groups were orally inoculated with ETEC (2.4 x 1010 CFU) and NC group received sterile solution. Growth performance was analyzed weekly and pigs were euthanized on d 28 to measure intestinal health. Data were analyzed using the SAS 9.4. During post-challenge period, PC tended to decrease (P = 0.067) ADG (373 to 284 g/d), whereas BMD increased (P &amp;lt; 0.05) ADG (284 to 408 g/d); LBE tended to increase (P = 0.081) ADG (284 to 370 g/d). PC increased fecal score (P &amp;lt; 0.05, 3.4 to 3.9) on d 14, whereas BMD decreased it (P &amp;lt; 0.05, 3.9 to 3.5) on d 21. PC increased (P &amp;lt; 0.05) protein carbonyl (0.76 to 1.12 nmol/mg protein), crypt cell proliferation (28 to 36%), and Helicobacter rodentium (0.4 to 3.7%). However, BMD decreased (P &amp;lt; 0.05) crypt cell proliferation (36 to 32%) and Helicobacter spp. (15.0 to 1.4%); and increased (P &amp;lt; 0.05) villus height (309 to 377 µm), Bifidobacterium boum (0.04 to 2.0%), Pelomonas spp. (1.5 to 8.5%), and Microbacterium ginsengisoli (0.5 to 3.0%). LBE reduced (P &amp;lt; 0.05) crypt cell proliferation (36 to 27%) and Helicobacter rodentium (3.7 to 0.04%); and increased (P &amp;lt; 0.05) Lactobacillus salivarius (0.3 to 4.1%) and Propionibacterium acnes (0.4 to 7.4%). Collectively, ETEC reduced growth performance by adversely affecting microbiota and intestinal health. BMD and LBE improved growth performance by enhancing intestinal health and increasing beneficial microbiota in ETEC challenged pigs.

Corrosion behavior of Q345R steel influenced by Thiobacillus ferrooxidans

Here, the corrosion weight-loss method, surface analysis technology, and electrochemical test methods were used to study the corrosion behavior and electrochemical characteristics of experimental samples of Q345R steel in a sterile solution (pH 2.0) and a solution containing T. ferrooxidans. The growth cycle of T. ferrooxidans was determined to be approximately 8 days. The corrosion weight-loss method showed that the corrosion rate of Q345R carbon steel coupons decreased with time in the T. ferrooxidans system and the sterile system; the corrosion rate was approximately two times higher in the T. ferrooxidans system than in the sterile system. The corrosion morphology results showed that the presence of T. ferrooxidans promotes the corrosion of Q345R steel and increases the local corrosion of the matrix material. The electrochemical test results showed that after 5 days of corrosion, the polarization resistance of the T. ferrooxidans system was approximately 50% of that of the sterile system, and the corrosion current density of the T. ferrooxidans system was approximately twice as high as that of the sterile system. Therefore, T. ferrooxidans can accelerate the corrosion of Q345R steel two-fold.

Bacteriological assessment of stethoscope used by health care personnel in attat hospital, SNNP, Gurage Zone, Ethiopia

Abstract Stethoscope has always been a part of the physicians’ basic tool for examining patients. Universal use of stethoscope for examination of patients by health care personnel makes it a potential source for spread of nosocomial infection. This study was designed to assess the potential for bacterial transmission by stethoscopes used by different health-care workers in Attat hospital. A cross sectional study was conducted from April to June 2018 in university of Wolkite, department of biotechnology and biology in molecular laboratory. During the study period there were a total of 26 stethoscopes from health professionals who had direct contact with patients are collected. Sample was collected by sterile cotton-tipped applicator moistened in a sterile solution of physiologic saline (0.85% sodium chloride) that was used to swab the entire surface of the diaphragm of the stethoscope and then inoculated into macconkey agar, tryptone soya agar and blood agar media. 18(69.2%) stethoscopes out of total collected stethoscopes had bacterial growth and 12 bacterial isolates were selected and characterized to genus level. Isolates include staphylococcus aureus(37.5%), coagulase negative staphylococci (28.12%), Streptococcussp. (21.88%), and Bacillus sp.(12.5%). All isolates were susceptible to the co-trimoxazole and ciprofloxacine, while resistant to cifoxitine. They showed intermediate growth against vancomycine. All except streptococcus were found resistant against penicillin. Both S. aureus and CoNS were sensitive to the chloramphenicol; Streptococcus was intermediate while bacillus was resistant to the chloramphenicol All stethoscopes (42.2%) that had never cleaned and last cleaned a week agowere highly contaminated while lower levels of contamination (27%) were found on those cleaned several times a day and cleaned between each patient before the examination of the patients.

Ameliorative Effect of Neem Leaf and Pomegranate Peel Extracts in Coccidial Infections in New Zealand and V-Line Rabbits: Performance, Intestinal Health, Oocyst Shedding, Carcass Traits, and Effect on Economic Measures.

Simple SummaryCoccidiosis, one of the most contagious diseases among domestic rabbits, negatively affects production and results in massive economic losses. This study evaluates the therapeutic efficacy of treatment with aqueous neem leaf extract and ethanolic pomegranate peel extract (PPE) individually and in combination on the intestinal coccidiosis caused by Eimeria spp. in New Zealand white and V-line (VL) rabbits. Rabbits from two breeds were divided into ten equal groups (five groups each for NZ and VL). All rabbits were inoculated with Eimeria spp. oocysts except for the rabbits in the first group (G1) (negative control). The remaining groups were: G2, positive control, G3, treated with neem leaf extract, G4, treated with pomegranate peel extract (PPE), and G5, treated with a combination of neem leaf extract and PPE. Our results showed that the use of neem leaf and/or pomegranate peel extract for both breeds resulted in improved growth performance, a significant reduction in mean oocyst count, no mortalities, an anticoccidial index > 120, and significantly improved economic efficiency measures when compared to the positive control group.Healthy, weaned, coccidial-free male rabbits from two breeds (New Zealand white (NZ) and V-line (VL)) were divided into 10 equal groups (5 groups each for NZ and VL) (3 replicates/group, 6 rabbits/replicate, 18 rabbits/group). All rabbits were inoculated with 5 × 104 Eimeria spp. oocysts (E. intestinalis (67%), E. magna (22%), and E. media (11%)) except for the rabbits in the first group (G1), which were inoculated with a sterile solution and served as a negative control. The remaining four groups were treated as follows: G2, no treatment/positive control, G3, treated with neem leaf extract, G4, treated with pomegranate peel extract (PPE), and G5, treated with a combination of neem leaf extract and PPE. For both breeds, our results showed that the use of neem leaf and/or pomegranate peel extract resulted in improved growth performance, with a significant improvement in relative feed conversion ratio (FCR) compared to the positive control groups, which recorded the worst values, as well as a significant (p ≤ 0.05) reduction in mean oocyst count compared to the positive control groups. We also observed downregulation of mRNA levels of IL-1βα, IL6, and TNF-α in the herbal treatment groups compared with the mRNA levels of these genes in the positive control groups. Herbal treatment with neem leaf and/or pomegranate peel extracts had positive effects on the NZ and VL rabbits experimentally infected with mixed Eimeria species, as evidenced by their healthy appearance, good appetite, no mortalities, an anticoccidial index > 120, and a significantly higher total return and net profit when compared to the positive control groups of both breeds. In NZ rabbits, the treatment with neem leaf extract alone (G3) or in combination with PPE (G5) recorded the most efficient economic anticoccidial activity.

Degradation of ancient Maya carved tuff stone at Copan and its bacterial bioconservation

Much stone sculptural and architectural heritage is crumbling, especially in intense tropical environments. This is exemplified by significant losses on carvings made of tuff stone at the Classic Maya site of Copan. Here we demonstrate that Copan stone primarily decays due to stress generated by humidity-related clay swelling resulting in spalling and material loss, a damaging process that appears to be facilitated by the microbial bioweathering of the tuff stone minerals (particularly feldspars). Such a weathering process is not prevented by traditional polymer- and alkoxysilane-based consolidants applied in the past. As an alternative to such unsuccessful conservation treatments, we prove the effectiveness of a bioconservation treatment based on the application of a sterile nutritional solution that selectively activates the stone´s indigenous bacteria able to produce CaCO3 biocement. The treatment generates a bond with the original matrix to significantly strengthen areas of loss, while unexpectedly, bacterial exopolymeric substances (EPS) impart hydrophobicity and reduce clay swelling. This environmentally-friendly bioconservation treatment is able to effectively and safely preserve fragile stones in tropical conditions, opening the possibility for its widespread application in the Maya area, and elsewhere.

In vivo effects of exposure to Golden trumpet Handroanthus chrysotrichus in mice.

The Golden trumpet Handroanthus chrysotrichus is a tree that presents beneficial health properties against various diseases. Thus, this study aims to verify the toxicity of H. chrysotrichus bark extract, observing the effects of exposure to this extract in mice. For this, mice were separated in groups: saline (sterile solution .9%); H. chrysotrichus crude extract (HCCE) 10; HCCE 50, and HCCE 100mg. kg-1 (p.o.). We analyzed HCCE effects on acute (single exposure) and subchronic protocol (14 days exposure). After both exposures, acute, and subchronic, we collected samples from blood, brain, liver, and kidney tissues for biochemical evaluation. In addition, after subchronic exposure, we performed behavioral tests. Acute exposure caused an increase of lipid peroxidation in liver tissue. Moreover, we observed a significant carbonyl increase in liver and brain tissues from HCCE 50mg. kg-1. Kidneys presented carbonyl increase in mice treated with the highest concentration. Besides, creatinine increased in the group of the acute exposure at HCCE 100mg. kg-1. Total leukocyte count decreased in all concentrations tested. Sub-chronic exposure at HCCE 100mg. kg-1 caused a decrease in the number of crossing and an increase in its self-grooming frequency in the open field test. In this exposure, the brain and liver had a significant increase in carbonyl levels in all concentrations. We concluded that H. chrysotrichus cause behavioral and biochemical alterations in mice. HCCE primary targets seem to be the liver, kidneys, and white cells.

FEATURES OF THE DEVELOPMENT OF A LYOPHILIZED INJECTABLE DOSAGE FORM OF THE ORIGINAL ANTICANCER DRUG LCS-1208

Objective: Development of a lyophilized injectable dosage form LCS-1208, an original antitumor drug based on an indolocarbazole derivative.&#x0D; Methods: The prepared solution of the injectable dosage form LCS-1208 is transferred to sterilizing filtration, which is carried out under vacuum on a «Stericup» filter unit with a filter pore size of 0.22 μm. The sterile solution of the injectable dosage form LCS-1208 is poured into sterile vials using a dispenser and lyophilized in a freeze-drying chamber. At the end of drying, the preparation is corked in the chamber of a sublimation unit using a hydraulic device and transferred to crimping with aluminum caps using a seaming machine. Quantitative determination of the drug content was determined by spectrophotometry using a standard sample at λ = 320±2 nm. The pH was determined by potentiometry.&#x0D; Results: A freeze-drying regimen for the injectable dosage form LCS-1208 has been developed. The required solution freezing temperature was established taking into account the presence of 2 eutectic zones: a solution of LCS-1208 in DMSO (-35 ÷-32) °С, an aqueous solution of Kollidon 17PF (-10 ÷-8) °С. As a result of a series of experiments, the optimal lyophilization regime was chosen that does not require preliminary freezing in a low-temperature chamber, with freezing on the shelves of freeze-drying at a temperature of-47 °C without their preliminary cooling. The most acceptable vial filling volume was determined, amounting to 3 ml, and the rate of temperature rise during secondary drying of the preparation was justified. When using the developed regime of lyophilization of the LCS-1208 solution, it was shown that it can be sublimated while preserving the initial qualitative and quantitative characteristics.&#x0D; Conclusion: In this article, using the example of creating a lyophilized injectable dosage form LCS-1208 (the original antitumor drug from the indolocarbazole group), the main problems that arose during the lyophilization of the selected composition of the model solution, as well as ways to improve the process.