Amies Transport Medium Research Articles

Mycotic flora characteristics of the endontic flora in patients affected by COVID-19

BACKGROUND: Owing to the fact that, presently, at a dental appointment, it is possible to observe a wide range of consequences arising after COVID-19, among which are observed lesions of the pulp and periodontium of the tooth, additional diagnostic methods should be introduced before choosing treatment tactics. In our previous study, we analyzed the contents of root canals of teeth in patients who had COVID-19 using bacterioscopy. Coccus flora was detected in all cases and elements of Candida yeast-like fungi were recorded in 89 cases (76.1%). Based on the obtained data, it was advantageous to perform microbiological examination of the endodontium in this group of patients to further study their physical and chemical properties and correct the treatment regimen in gangrenous form of chronic pulpitis and aggravated forms of periodontitis, which determined the relevance of the present study. AIM: To reveal the characteristics of the mycotic flora of the endodontium in patients who previously had COVID-19. MATERIALS AND METHODS: A bacteriologic study of root canal contents in patients who had previously undergone a new coronavirus infection at different times, diagnosed as “exacerbation of chronic periodontitis”, “chronic gangrenous pulpitis”, and “acute suppurative periodontitis”, was conducted to detect fungi of the genus Candida in the tooth root canal system, which was obtained during mechanical treatment with a sterile endodontic instrument. The patients (n=49) were divided into groups according to their final diagnosis: group 1, chronic gangrenous pulpitis (27 patients); group 2, acute purulent periodontitis (9 patients); and group 3, exacerbation of chronic periodontitis (13 patients). Then, the collected material was placed in a tube with Amies transport medium and sent to the laboratory. Seeding of the material was performed in sterile Petri dishes on Sabouraud agar by rubbing with a plastic spatula. Further, Petri dishes were placed in the thermostat for 24–48 hours of incubation at 37±10 °C. Then, the study was carried out according to the generally accepted scheme: the obtained cultures were identified to species by the character of growth on dense medium. RESULTS: The given bacteriological study of the patients’ endontic contents confirmed the data of bacterioscopy and concretized the previously obtained results. On final examination, black colonies of Candida albicans were observed in 95.92% of cases. No growth of Candida albicans colonies was observed in two cases (4.08%). CONCLUSION: The bacteriologic study performed after bacterioscopy of diagnosis confirmed the presence of fungi of the genus Candida in the root canals of teeth in patients who previously had COVID-19. The study showed that when treating similar patients, the high risk of infection of periodontal complex tissues by Candida fungi should be considered. It is advisable to supplement the standard protocols of endodontic treatment with physical exposure, for example, laser radiation, or additional medication with antifungal drugs, such as fluconazole.

Nasal carriage rate, associated factors, and antimicrobial susceptibility patterns of methicillin resistance Staphylococcus aureus among pre-clinical undergraduate students at the College of Health and Medical Sciences, Haramaya University, Ethiopia

BackgroundStaphylococcus aureus nasal carriage has been linked to higher rates of infection and morbidity. People with Methicillin-resistant Staphylococcus aureus can be a potential source of infection for others. University students living together in crowded conditions increase their risk of acquiring infections. The prevalence of S. aureus, particularly Methicillin-resistant Staphylococcus aureus nasal carriage, in Ethiopian university students is sparse.ObjectiveThis study aimed to determine the nasal carriage rate, associated factors, and antimicrobial susceptibility patterns of methicillin-resistant Staphylococcus aureus among pre-clinical students at the College of Health and Medical Sciences, Haramaya University, Ethiopia, from 1 July to 30 August 2022.MethodsAn institutional-based cross-sectional study was conducted among 270 randomly selected pre-clinical Health and Medical Sciences students. Data on associated factors were collected using pre-tested, structured questionnaires. A nasal swab was taken from each participant and sent to the microbiology laboratory via Amies transport media in a cold chain. There, it was cultivated using conventional techniques. The isolated colonies were found to be S. aureus, and its antimicrobial susceptibility was performed using the Kirby–Bauer disk diffusion method on Muller–Hinton agar. Methicillin-resistant Staphylococcus aureus expressing using cefoxitin based on CLSI breakpoint. Data were entered into Epi-Data version 4.4.2.1 and exported to the Statistical Package for Social Sciences (SPSS) software version 25 for analysis. Pearson’s chi-square test was performed to predict the associations between variables. A p-value less than 0.05 was regarded as statistically significant.ResultMethicillin-resistant Staphylococcus aureus nasal carriage was 5.9% (95% CI: 3.09–8.7) of cases of S. aureus nasal colonization, which was found to be 12.96% (95% CI: 8.85–16.96). Methicillin-resistant Staphylococcus aureus nasal colonization was significantly associated with the history of cigarette smoking (p = 0.000), intake of khat (p = 0.042), nose-picking habit (p = 0.003), history of sharing personal goods (p = 0.021), and history of hospitalizations (p = 0.00). All of the Methicillin-resistant Staphylococcus aureus isolates were resistant to ampicillin and cefoxitin.ConclusionBased on the findings, a considerable proportion of healthy students harbored Methicillin-resistant Staphylococcus aureus strains associated with behavioral factors. Furthermore, these isolates showed high resistance to cefoxitin and ampicillin. Hence, it is crucial to regularly test pre-clinical students to prevent endogenous infections and the spread of Methicillin-resistant Staphylococcus aureus.

MYCOLOGICAL ANALYSIS OF CYSTIC CONTENT IN PATIENTS WITH POLYMICROCYSTIC CHANGES OF THE MAXILLARY SINUS MUCOSA

The prevalence of mycotic infections of the paranasal sinuses is constantly increasing worldwide. Pathogens can enter the sinuses through the respiratory tract that leads to the colonization of microorganisms in both the lungs and paranasal sinuses as part of the normal microflora. The presence of filamentous fungi in the maxillary sinus may result from endodontic treatment of molars directly adjacent to the sinus, increasing the risk of fungal foci development. Fungal paranasal sinusitis manifests in invasive or non-invasive forms. While mycological culture remains the traditional gold standard for mycosis diagnosis, its sensitivity is reportedly low. Paranasal sinus cysts comprise 4-8% of general ENT pathologies and approximately 4-16% of chronic paranasal sinus diseases, with a predominant presence in the maxillary sinus. Factors that contribute to the development of sinusitis and create favorable conditions for fungal spore adhesion and growth include impaired ventilation and transport function of the sinus mucosa, inadequate use of broad-spectrum antibacterial drugs and steroid therapy, as well as household factors. Recently, there has been an increase in the number of cases of multiple small cysts in maxillary sinuses. Clinical manifestations range from asymptomatic incidental findings to varying degrees of discomfort and/or pain localized in the maxillary sinus area. The aim of this study is to identify the involvement of a fungal agent and to develop a set of therapeutic and prophylactic measures for polymicrocystic changes in the mucous membrane of the maxillary sinuses. The study enrolled 37 patients aged 18 to 65 years. Atypical cystic content was collected and cultured in Sabouraud's nutrient broth and Amies transport medium at 35-37°C for 5-7 days, with final measurements on day 21. Treatment involved a 14-day regimen of itraconazole (100 mg once daily) alongside topical miramistin (0.01% solution, 2 ml) sinus injections. The findings obtained demonstrate the following: positive fungal growth was observed in 26 cases (70.3%), predominantly Penicillium (48.6%) and Aspergillus (21.6%). Absence of growth was noted in 11 cases (29.7%). Follow-up examinations in 6 months and in a year revealed no crusts in the nasal cavity, moderate mucous discharge in the sinuses, and no polymicrocystic changes. Conclusion. The analysis has demonstrated the involvement of fungal flora in the development and recurrence of cystic changes in the maxillary sinus mucosa, advocating for mycological examination alongside bacteriological analysis. Itraconazole for systemic therapy and miramistin for topical therapy demonstrate efficacy in treatment and recurrence prevention.

CHARACTERISTICS OF ROOT CANAL MICROBIOTA IN ACUTE PULPITIS OF TEMPORARY TEETH

Aims. To determine the representatives of the microbiota of the root canals seeded in acute forms of pulpitis of temporary teeth among children of the Transcarpathian region, taking into account the endemic features of the Transcarpathian region and its influence on the course of the carious process.
 Materials and methods. Collection of biomaterial was carried out using transport tubes with Amies transport medium. The biomaterial was delivered to the laboratory within 2-4 hours after selection. In order to evaluate the microbiota of the root canals, cultivation of biomaterial was carried out on differential diagnostic nutrient media. The material was sown by the method of sector sowing according to Gold: Saburo for the cultivation of microscopic mushrooms; blood agar (MPA + 5% blood) Streptococcus and Neisseria bacteria; bacteria of the family Enterobacteriaceae Endo and Levin; yolk-salt agar with mannitol bacteria of the genus Staphylococcus; enterococcus agar for isolation of enterococci (Farmaktiv, Ukraine). Additionally, the biomaterial was sown on a chromogenic medium (bioMerieux, France). Microorganisms were identified by cultural, morphological and biochemical characteristics using API-test systems (bioMerieux). The results of the quantitative and qualitative evaluation of the microbiota of the oral cavity were expressed in CFU/ml.
 Results and discussion. Today, in the daily practice of a dentist, among issues of diagnosis, prevention and treatment of dental diseases, the study of the microbiota of the oral cavity, which is a collection of different taxonomic groups of microbes that inhabit the oral cavity and enter into biochemical, immunological and other relationships with macroorganism and each other.
 At the same time, the microbial landscape of the oral cavity is represented by two types of microbiota: obligate, that is, one that is constantly present in the oral cavity and is mainly saprophytic, and provides metabolic processes and the function of protecting the body from virulent infectious agents, and facultative, which is represented opportunistic microorganisms, which, when immune protection is reduced, acquire aggressive properties and contribute to the development of diseases [1].
 Conclusion. Opportunistic bacteria play a significant role in the development of inflammatory diseases. Considering the fact that pulpitis is an inflammatory reaction caused by the activity of opportunistic microflora, the study of the characteristics of the microbiota of root canals is relevant, which will help to improve the selection of antiseptic agents for medicinal treatment of root canals in order to obtain positive delayed treatment results.

Bacterial Profile, Antimicrobial Susceptibility Patterns, and Associated Factors of Puerperal Sepsis in Asella, Central Ethiopia: A Cross-sectional Study

Sepsis, caused by various bacterial pathogens, is a significant contributor to maternal mortality worldwide. In many developing countries, including Ethiopia, empirical or syndromic treatment is commonly employed for puerperal sepsis, which may promote antimicrobial resistance (AMR). We conducted a cross-sectional study to investigate bacterial pathogens, their antimicrobial susceptibility patterns, and associated factors among women with suspected puerperal sepsis attending Asella Referral and Teaching Hospital from September 2020 to August 2021. A total of 174 participants were enrolled, and the sociodemographic and obstetric data were collected using a pretested structured questionnaire and checklist, respectively. Blood samples (approximately 20 ml) were collected from all study participants and incubated in BacT/ALERT® 3D automated blood culture system. In addition, endocervical swabs were collected in Amies transport media. Bacterial isolation and identification were performed following standard bacteriological methods. Antimicrobial susceptibility profiles of bacterial isolates were determined using the disc diffusion method. Data were entered into EpiData version 4.6 and analyzed using SPSS version 25.0. The overall positivity rate of bacterial isolates among puerperal sepsis-suspected women was 48.9%. Among these, 87.1% of the isolates were Gram-negative bacteria. The most common isolates were Escherichia coli (54.1%), followed by Klebsiella spp. (23.5%) and Staphylococci aureus (10.6%). High resistance rates were observed in E. coli to piperacillin (87%), in Klebsiella spp. to aztreonam (65%) and ceftriaxone (65%), and in S. aureus to trimethoprim-sulfamethoxazole (66.6%). Multidrug-resistant bacterial pathogens accounted for 81.2% of the isolates in this study. Multivariate regression analysis did not reveal any statistically significant association between the presence of bacteria and the sociodemographic and obstetrics factors. Our findings emphasize the urgency of strengthening microbiology services to optimize patient management and combat AMR in puerperal sepsis.

Antimicrobial resistance profiles for bacteria isolated from rectal swabs in patients candidate for prostate biopsy

Objective: Infection is one of the common complications of post-prostate biopsy. Therefore, prophylaxis with fluoroquinolones and/or cephalosporins is recommended. We aimed to evaluate the prevalence of antibiotic-resistant bacteria isolated from rectal swabs in candidates for transrectal ultrasound-guided (TRUS) biopsy. The possible patient’s related risk factors attributing to resistance to antibiotics were also assessed. Methods: This cross-sectional study was performed on 126 male patients who were candidates for TRUS biopsy. Rectal swabs were collected and the samples were transferred to the laboratory in Amies transport medium during 2 hours and cultured on MacConkey agar with ciprofloxacin 1 mg/L and ciprofloxacin-resistant strains were identified. Kirby-Bauer disc diffusion method was used to determine the antibiotic susceptibility of the isolates. Results: In total, 59 bacterial isolates were obtained, which were Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa, respectively, in terms of frequency. They showed the lowest resistance to levofloxacin. Smoking was associated with positive culture results. Age was a factor with a significant effect on carrying ciprofloxacin-resistant strains. Conclusion: Ciprofloxacin resistance was high in almost all strains, but post-biopsy infectious complications were very low. Level of evidence: Not applicable

Portrait of Microbiology Profile in Obstructive Jaundice Cases: A Single Center Experience

Introduction: Physiologically bile is aseptic, however bacteria may be present in bile in cases of biliary infections. Portrait of biliary microbiology in patients with obstructive jaundice in Indonesia have never been reported before. Methods: Data of obstructive jaundice patients in digestive surgery division, Fatmawati General Hospital were collected prospectively from November 2020 to June 2021. Data collected included age and gender, diagnosis, causes of jaundice, preoperative total and direct bilirubin, preoperative leucocyte and culture results. Bile was aspirated, using a 3cc syringe, directly from the biliary tree above the obstruction and placed into amies transport medium and sent to microbiology laboratory. Results: Bile samples were obtained in 28 out of 39 cases of obstructive jaundice. All samples were obtained during an open surgical procedure without prior intervention. There was equal distribution between male and female female patients with the mean age of 51.8 (± 12.7). The cause of obstructive jaundice were 57% biliary stones and 39% of malignant causes. Positive culture was found in 25% of cases (n=7), the most common pathogen found was Escherichia coli (57%). Fungal infection, candida tropicalis, was found in one case (14%). Conclusion: Although bile is essentially sterile, pathogens can grow in selected cases. The frequency and spectrum of bacteria that infect bile, including their sensitivity to antibiotics are important in guiding patient management. This is a preliminary result from an ongoing study in our centre to determine the best choice of antibiotics given to obstructive jaundice patients.

Improved Blastocystis spp. detection method using swabs with Amies transport medium and charcoal.

Blastocystis is one of the most frequently detected protozoa in the human large intestine. One of the most effective and cheap methods for detecting Blastocystis in faeces is culture on a special medium in anaerobic conditions. Sampling faeces using traditional containers and their transport to the laboratory has certain limitations: a sample taken in this way should reach the laboratory relatively quickly, moreover, some patients are uncomfortable during sampling and protection of material in this way. We propose utilizing a swab for sampling and transportation of the faeces samples to be examined for Blastocystis instead of using traditional containers. We believe this is an excellent method allowing the material to be transported over longer distances without additional, and sometimes expensive, safety measures, and at the same time permitting the possibility of obtaining living cells after a relatively long period of storage.

Validation and implementation of a commercial real-time PCR assay for direct detection of Candida auris from surveillance samples.

Rapid and reliable laboratory methods are required for detecting the nosocomial yeast Candida auris. AurisID® (Olm Diagnostics) is a real-time PCR assay approved for detecting C. auris in fungal cultures and directly from blood samples, involving a nucleic acid extraction as a prior step. The purpose of this study is to validate the AurisID® kit for direct detection of C. auris from surveillance samples without prior DNA extraction and to analyse the results of implementing this methodology to our daily laboratory routine protocol for C. auris surveillance studies. Our PCR method using the AurisID® kit was compared with our routine protocol, consisting of culture in CHROMagar® Candida and identification by mass spectrometry. A total of 113 swabs were used for validation and 136 pair of surveillance samples were tested. Limit of detection (LOD) was determined by using swabs in Amies transport medium, which were spiked in a series of dilutions of a C. auris standardised suspension (0.5 McFarland). The PCR method showed high sensitivity, specificity, predictive positive value and predictive negative value (96.6%, 100%, 100% and 98.2%, respectively) when compared with the routine protocol. LOD was 500CFU/ml, which corresponds to approximately 1CFU/PCR. Our PCR method using the AurisID® kit allows a reduction in the turnaround time for surveillance of C. auris compared with other methods. These results are expected to contribute to control C. auris outbreaks, allowing isolation of patients and cleaning of environmental surfaces in advance.

Comparison of two methods for obtaining and transporting corneal samples in suspected infectious keratitis

The purpose of this study is to compare two alternative methods of collecting and transporting media for the diagnosis of corneal ulcers, as not all clinical settings have conventional culture materials and transport media available. In this open-label, prospective, comparative, and randomized study, patients with clinical suspicion of infectious keratitis with high risk of loss of vision had corneal specimens collected using two methods and transport media: Eswab scraping with Amies transport medium and 23-gauge needle scraping in BACTEC Peds broth. The order of each collection method was randomized. The samples were processed by standard methods, comparing the positivity frequencies for both by parametric and nonparametric tests, according to normality criteria. Corneal infiltrates from 40 eyes of 40 patients were analyzed. Culture positivity rate was 50% for Eswab and 35% for 23-gauge needle (P=0.258). The overall growth rate of the two methods combined was not higher than with the swab alone. The results obtained with a swab were not influenced by the collection sequence (P=0.112); however, the positivity rate was significantly higher when the sample taken with the needle was performed first (P=0.046). The single sample Eswab method of collection and transportation for the diagnosis of high risk corneal ulcers is a valid alternative and can be used in cases in which, for various reasons, there is no access to the full set of traditional culture materials.

The Potential Sources of Transmitting of Hospital Acquired Infection by Routine Devices in Adult ICU in Alrass General Hospital

Hospital-acquired infections (HAI) reflect as a major global safety concern for both patients and health-care professionals. These infections could be in the form of cross-infection, endogenous infection and environmental Infection. Over 80% of these infections are related to devices’ utilization needed for patients’ life support. Methods show this is an observational and cross-sectional study, to identify the microorganism and determine the potential source of transmitting of hospital acquired infection by routine devices in adult ICU. The samples were collected using Amies transport media; three swabs were taken from the surfaces of indwelling urinary catheter, mechanical ventilation device and central venous catheter used from every twelve patients. The samples were cultured and analyzed by using microbiologic technique. Finally, all samples analyzed by MicroScan WalkAway 96 pulse. Results showing the most bacteria isolated are “Klebsiella pneumonia” (18.37%), “Acinetobacter baumannii” (11.48%), “Staphylococcus epidermidis” (4.59%), “Staphylococcus haemolyticus” (4.59%), “E. coli” (4.59%), “Serratia marcescens” (2.3%), “Pseudomonas luteola” (2.3%), “Kocurio kristinae” (2.3%) and “Photorhabdus luminscens” (2.3%). This study detects a high contamination of routine devices and resistant organisms. In the end it is recommended that effective infection control practices and effective strategies to control antibiotic-resistant bacteria should be applied.

Antimicrobial resistance of Neisseria gonorrhoeae in Jakarta, Indonesia: a cross-sectional study.

A cross-sectional study was conducted in Jakarta, Indonesia, from September to November 2018. In all, 98 high-risk males and females who fulfilled the study criteria were included. Specimens were collected from urethral or endocervical swabs, put into Amies transport medium and then transported to the Laboratory of Clinical Microbiology, Universitas Indonesia for culture and identification. Proven gonococcal isolates were examined for susceptibility to various antimicrobials using the disk diffusion method according to Clinical and Laboratory Standard Institute guidelines. Of the 98 specimens, 35 were confirmed to be N. gonorrhoeae. The proportion of N. gonorrhoeae specimens resistant to penicillin, tetracycline, levofloxacin, cefixime and ceftriaxone among the high-risk population was 97.1%, 97.1%, 34.3%, 0% and 0% respectively. The possible factors associated with resistance could only be analysed for levofloxacin. Age, sexual orientation and a history of orogenital sexual activity during the past month were not associated with N. gonorrhoeae resistance to levofloxacin. This study detected no resistance of N. gonorrhoeae to cefixime and ceftriaxone. Further studies with larger samples are needed to obtain more representative results of N. gonorrhoeae resistance and the possible factors associated with resistance.

Bacterial profile and antibacterial susceptibility of otitis media among pediatric patients in Hawassa, Southern Ethiopia: cross-sectional study

BackgroundOtitis Media (OM) is the most common disease of childhood. Twenty thousand people die each year from otitis media. It is an important cause of preventable hearing loss, affects children's intellectual performance and language development. There are very small numbers of studies done in Ethiopia concerning this topic. This study aimed to identify bacterial pathogens related to ear infection and to assess antibacterial susceptibility of isolated organisms.MethodA cross-sectional study was conducted on 152 children from April 2018 to July 2018 at selected health facilities in Hawassa city, SNNPR, Ethiopia. All pediatric patients having ear discharge were included. Convenient sampling technique was used to collect clinical and demographic data using standard questionnaires after child care-takers signed the consent. Ear discharge specimens were collected using a sterile swab, and transported using Amies transport media to Hawassa University Comprehensive Specialized Hospital laboratory. Bacterial isolates were characterized based on colony appearance, Gram reaction, culture characteristics, and biochemical tests after inoculating on appropriate culture media. Antibacterial susceptibility testing was performed using the disc diffusion method according to the criteria of the Clinical and Laboratory Standards Institute (CLSI).ResultsAmong 152 children included, 115(75.6%) of them demonstrated pathogenic bacterial growth. Staphylococcus aureus 41(27%) was the most frequently isolated pathogen, followed by Proteus mirabilis 19 (12.5%). Of the total isolates, 11.2 and 7.3% were resistant to gentamicin and ciprofloxacin respectively. Over three-fourth (85.2%) of the isolates were resistant to ampicillin. More than two-third of the isolates were resistant to both penicillin (71.4%) and trimethoprim-sulphamethoxazole (72.0%).ConclusionsS. aureus is the most commonly isolated bacterial pathogen from ear discharge among children. Even though gentamicin is a parenteral drug and ciprofloxacin is rarely used in children due to concerns of bone/joint effects, these two drugs were highly effective antibiotics and thus should be considered in treating children with otitis media since most organisms were resistance or poor response to first line drugsHigh level of antibiotic resistance was observed so antimicrobial susceptibility test is needed before prescribing drugs for treatment of OM.

Comparison of postmortem airway swabs and lung tissue for detection of common porcine respiratory pathogens by bacterial culture and polymerase chain reaction assays

Objective: To compare pathogen detection from tracheobronchial swabs with lung tissue in diagnostic submissions from pigs with reported respiratory disease. Materials and methods: Individual lung samples (n = 153) from 133 laboratory submissions were included in this study. Inclusion criteria were a lung sample where the tracheal bifurcation or major bronchus was readily identifiable and a clinical report of respiratory disease symptoms. Sterile, nylon-flocked swabs were used to sample the largest available airway before the lung tissue was routinely processed for diagnostic testing. Swabs were placed in Amies transport medium and tested in blinded parallel with the lung tissue by bacterial culture and polymerase chain reaction (PCR) for common swine respiratory pathogens. Results: There was excellent agreement between PCR detection from lung and bronchial swab samples for porcine reproductive and respiratory syndrome virus, influenza A virus, Mycoplasma hyopneumoniae, and porcine circovirus 2 (kappa > 0.8, all assays). Agreement between bacterial culture from lung and swabs was substantial for Pasteurella multocida and Salmonella spp. and fair for Streptococcus suis. Lung tissue was culture positive more often than swabs for Haemophilus parasuis and Actinobacillus spp.; however, in these cases, PCR for the respective pathogen was 100% positive on swab samples regardless of culture status of the swab. Implications: Tracheobronchial swabs are a single, uniform sample that can be easily collected at postmortem and transported to the laboratory for detection of swine respiratory pathogens by culture and PCR. Such swabs may serve as a rapid screening tool for unexpected mortalities in a population.

Detection of Extended Spectrum Beta-Lactamases from a Pet Blue-Fronted-Amazon Parrot (Amazona aestiva)

Background: The indiscriminate use of antibiotics both in humans and in animals, has contributed to the development of bacterial resistance. One of the key mechanisms in the resistance of enterobacteria to antibiotics is the production of extendedspectrum β-lactamases (ESBLs), which reduce the therapeutic options available. Several studies have been performed in different animal species in order to isolate and identify multidrug-resistant strains and determine their antibiotic sensitivity profile. The purpose of this study was to detect ESBL-producing enterobacteria in isolates from a pet blue-fronted amazon parrot.Case: A 16-year-old pet blue-fronted-amazon parrot (Amazona aestiva) weighing 0.445 kg, from the city of Presidente Castelo Branco, Paraná, Brazil, was admitted into a specialized veterinary clinic in the city of Maringá. This parrot was presented with agitation and aggressive behavior. It was fed with sunflower seeds, and its cage was small and unsanitary especially in the feeding and drinking areas, which were heavily contaminated with feces. The parrot had been diagnosed with acute sinusitis approximately one year prior, with a history of treatment with tylosin, thuya (a herbal remedy popularly used for fowl pox), and a mineral-vitamin complex. The clinical symptoms included dyspnea and bilateral increase of facial swelling, with a mass in the peri-nasal region. A membranous red tissue not related to the nictitating membrane was observed in the lower left eyelid. To obtain a better clinical evaluation, the bird was anesthetized with an intramuscular injection of dextroketamine 50 mg/kg (KetaminS+™). Subsequently, physical examination, head radiographic examinations, and an attempt at sinus draining were performed. The radiographic examinations showed a decreased amount of air in the nasal sinuses. For treatment, oral itraconazole and vitamin A were prescribed, and a dietary improvement, prioritizing the offering of fruits and industrialized food for parrots was proposed. Samples were collected from the cloaca and choanae by making rotating movements with compressed sterile swabs in the corresponding locations. All samples were kept in Amies transport media with activated coal and forwarded under refrigeration to the Laboratory of Preventive Veterinary Medicine and Public Health in the Post-Graduation Program in Animal Sciences with Emphasis in Bioactive Products at Universidade Paranaense (UNIPAR).Discussion: The samples were subjected to phenotypic antimicrobial sensitivity tests and phenotypic testing for detecting ESBL-producing strains. Escherichia coli was identified and isolated from the cloacal sample. Phenotypic tests for antimicrobial sensitivity, detected resistance to the following antimicrobials: ceftriaxone, ceftiofur, cefotaxime, cefepime, ampicillin, amoxicillin, amoxicillin + clavulanate, and tetracycline. The phenotypic test for detecting ESBL-producing strains was positive. The findings in this study had no relation with the clinical symptoms presented by the parrot. However, in the samples collected, it was possible to detect the presence of ESBL-producing Escherichia coli, indicating that this parrot had strains in its cloacal microbiota that were either multidrug-resistant or at a subclinical stage of an infection. This knowledge is important, since the presence of this bacteria in a pet represents an important factor in the dissemination of multidrug-resistant strains into the environment, as well as being a source of contamination for both humans and other animals.

Impact of sample processing on human airways microbial metagenomes

Whole metagenome shotgun sequencing provides information about the gene content and the composition of microbial communities provided that the processing of the samples does not introduce a methodology-driven bias. We tested the impact of DNA isolation and storage period on the metagenome profile. Deep throat swabs were collected from healthy adults and an infected infant. DNA was isolated by sonification or enzymatic lysis either immediately or after 24h storage in agar gel Amies transport medium at room temperature. Disruption of cells and subsequent fragmentation of DNA by sonification was as suitable as the common enzymatic lysis to generate high-quality metagenomes particularly for low total DNA input of less than ten nanograms. Conversely, storage of samples for 24h produced severely distorted metagenomes. The majority of species became less abundant or even extinct, whereas a few Streptococcus, Neisseria and Haemophilus spp. proliferated so that the total number of bacterial reads increased at the expense of human reads. We recommend that samples for metagenome analysis should be immediately processed or frozen at −80°C.

Rapid screening for group B Streptococcus in near-term pregnant women by Granada™ biphasic broth

Background: Prenatal screening for group B Streptococcus (GBS) colonization can reduce the incidence of neonatal GBS infections. We aimed to improve the screening-based approach of GBS in a limited resources antenatal care clinic by using Strep B Granada™ Biphasic Broth.Methods: This study included 80 pregnant women between 35 and 37 weeks of gestation, who attended the antenatal care clinic of Kasr El-Aini University Hospital from November 2013 to January 2014. Two high vaginal swabs were collected, then transported using Amies transport medium. One vaginal swab was processed by conventional culture-based methods on 5% sheep blood agar plates. The other swab was immersed in 3 mL selective enrichment broth (Granada™ Biphasic Broth bioMérieux).Results: Among 80 pregnant women, GBS was detected in 9 (11.25%) of the studied cases within 18–24 hours. Detection of orange-red colonies in GBS Granada broth was 100% specific for the presence of beta-hemolytic group B streptococci.Conclusion: Using Granada biphasic broth media was easy, affordable and shortened the turnaround time needed for the detection of GBS by conventional culture methods. Routine screening of pregnant women for vaginal GBS colonization by Granada™ Biphasic broth would allow properly timed prenatal antimicrobial prophylaxis to prevent possible neonatal infections.

The Influence of Corneal Collagen Crosslinking on Conjunctival Flora

ABSTRACTPurpose: To investigate the influence of corneal collagen crosslinking (CXL) on conjunctival flora.Materials and Methods: Included in this study were 42 patients, all of whom were treated with CXL for progressive keratoconus (KC). Conjunctival swab samples were collected under operating-room conditions immediately before and immediately after the procedure. Samples were quickly immersed in Amies Transport Medium (ATM), transferred to the microbiology laboratory, diluted, and inoculated into three areas each of blood agar, chocolate agar, eosin methylene blue (EMB) agar, and Saboraud–Dextrose agar plates. Inoculated plates were incubated at 35°C for 24–48 h. Preliminary species identification of isolated bacterial colonies was based on catalase and oxidase tests, Gram staining, and colony morphology. Definitive identification of isolates was made with Vitek-II fully automated identification kits.Results: Cultures were positive in 24 (57.1%) preoperative and 10 (23.8%) postoperative samples (p ˂ 0.05). Microorganisms isolated from preoperative conjunctival samples included coagulase-negative Staphylococcus (CNS) spp. in 23 (54.8%) samples, S. aureus in three (7.1%), Moraxella spp. in two (4.8%), Streptococcus spp. in three (7.1%), Corynebacterium spp, in one (2.4%), Micrococcus spp. in one (2.4%), and Candida spp. in one (2.4%). Microorganisms isolated from postoperative conjunctival samples included CNS spp. in nine (21.4%) samples, S. aureus in one (2.4%), Corynebacterium spp. in two (4.8%), and Acinetobacter spp. in one (2.4%).Conclusions: CXL appears to have a significant impact on conjunctiva flora.

The Effect of Chronic Alcoholism on the Conjunctival Flora

Aim: We aimed to investigate the effect of alcohol abuse on the conjunctival flora.Materials and methods: The cases were evaluated as two groups. The study group consisted of 55 heavy-drinking males diagnosed with alcohol abuse, while the control group consisted of 55 males without a history of alcohol abuse. Samples were taken from the inferior fornix conjunctiva with sterile cotton-tipped swabs (Amies transport medium) for culture. The samples were inoculated into blood agar, chocolate agar, eosine methylene blue agar and Saboraud-Dextrose agar (Oxoid/UK) with the dilution method.Results: The microorganisms that grew in study group subjects were Coagulase Negative Staphylococcus (CNS) in 30 (54.5%), Staphylococcus aureus in 14 (25.5%), Moraxella spp. in 3 (5.5%), Streptococcus spp. in 3 (5.5), Bacillus spp. in 3 (5.5%), Corynebacterium spp. in 3 (5.5%), Candida spp. in 3 (5.5%), Haemophilus spp. in 2 (3.6%), Acinetobacter spp. in 2 (3.6%), Neisseria spp. in 1 (1.8%) and Micrococcus spp. in 1 (1.8%). The results for control group were CNS in 31 (56.4%), Bacillus spp. in 7 (12.7%), S. aureus in 5 (9.1%), and Corynebacterium spp. in 2 (3.6%). Moraxella spp., Streptococcus spp., Candida spp., Haemophilus spp., Acinetobacter spp., Neisseria spp. and Micrococcus spp. microorganisms grew in the conjunctival flora samples of the study group but not in the control group. S. aureus colonization was significantly higher in the study group than the control group (p < 0.05).Conclusion: The S. aureus colonization rate was statistically significantly higher in the study group. Some microorganisms only grew in the conjunctival flora samples of the study group. These findings indicate that the conjunctival flora in persons with chronic alcoholism is different than the normal population.

Evaluation of Optimal Storage Temperature, Time, and Transport Medium for Detection of Group B Streptococcus in StrepB Carrot Broth

The performances of the ESwab and Amies transport media were evaluated for optimal survival of group B streptococcus (GBS) in StrepB carrot broth. ESwab was superior to Amies at all temperatures evaluated but was optimal at 21°C and 24°C, whereas recovery in Amies was significantly decreased at these temperatures.