Susceptibility Testing Of Bacteria Research Articles

Antimicrobial susceptibility in bacterial isolates from water and agricultural products of the Chanchán river, Chimborazo Province, Ecuador

Introduction: river contamination with pathogenic and multidrug-resistant microorganisms is a public health issue due to its impact on health and its potential to transmit infectious diseases. Objective: To determine the presence of multidrug-resistant bacteria in irrigation water samples and agricultural products from the Chachán River. Method: a descriptive, non-experimental study with a quantitative and cross-sectional approach was conducted. Water samples were collected from six different points, and pH and temperature were measured. Additionally, 13 agricultural product samples from the same points were analyzed. The identification and antimicrobial susceptibility testing of bacteria were performed using the VITEK2 system and amplification and sequencing of the 16S rRNA gene. Results: a total of 17 bacterial strains were isolated from the water samples, 16 of which (94 %) belonged to the order Enterobacterales, including Proteus mirabilis, Proteus vulgaris, Morganella morganii, and Citrobacter diversus. From the agricultural products, 24 bacteria were isolated, including species of Klebsiella, Stenotrophomonas maltophilia, and Enterococcus faecalis. Antimicrobial resistance was observed in various strains, notably P. mirabilis, C. diversus, and P. vulgaris with multiple resistances. Resistance was also identified in pathogenic strains such as Yersinia enterocolitica, Plesiomonas shigelloides, and Aeromonas hydrophila. Conclusions: the Chachan River and the agricultural products irrigated with its waters contain multidrug-resistant bacteria, posing a significant risk to public health. It is crucial to implement control and monitoring measures to reduce the spread of these pathogens and ensure food safety

A simplified version of rapid susceptibility testing of bacteria and yeasts using optical nanomotion detection.

We present a novel optical nanomotion-based rapid antibiotic and antifungal susceptibility test. The technique consisted of studying the effects of antibiotics or antifungals on the nanometric scale displacements of bacteria or yeasts to assess their sensitivity or resistance to drugs. The technique relies on a traditional optical microscope, a video camera, and custom-made image analysis software. It provides reliable results in a time frame of 2-4 h and can be applied to motile, non-motile, fast, and slowly growing microorganisms. Due to its extreme simplicity and low cost, the technique can be easily implemented in laboratories and medical centers in developing countries.

Glucose-metabolism-triggered colorimetric sensor array for point-of-care differentiation and antibiotic susceptibility testing of bacteria

Simple and sensitive discrimination of multiple bacteria and antimicrobial susceptibility test (AST) are significant for food safety, clinical diagnosis and treatment. Herein, based on different metabolic ability of bacteria on glucose, we presented a colorimetric sensor array for point-of-care testing (POCT) of multiple bacteria with methyl red (MER), bromothymol blue (BTB) and bromocresol green (BCG) as probes. Different bacteria resulted in different color changes of three probes, which was converted to RGB (Red (R)/Green (G)/Blue (B)) signals by the color recognizer APP loaded on smartphone. The sensor array performed differentiation of eleven species of bacteria, achieving the quantitative analysis of individual bacteria in tap water and differentiation of bacterial mixtures. Interestingly, the sensor array can be used for AST and evaluating minimal inhibitory concentration (MIC) of antibiotics to bacteria. The research provided meaningful guidance for distinguishing multiple bacteria and evaluating MIC, presenting great potential in practical application.

The first study on antimicrobial resistance of Staphylococcus aureus isolated from raw goat milk associated with subclinical mastitis in Siliragung Subdistrict, East Java, Indonesia.

Raw goat milk is a highly nutritious dairy product and a suitable medium for the growth of antimicrobial-resistant Staphylococcus aureus, the leading cause of subclinical mastitis. This study aimed to investigate the resistance status of S. aureus which isolated from goat milk associated with subclinical mastitis cases in Siliragung Subdistrict, Banyuwangi District, East Java, Indonesia. The S. aureus isolates were recovered from 258 raw goat milk samples from seven different dairy goat farms. Preliminary screening of subclinical mastitis using the California mastitis test, then samples with score +3 and +4 were taken for further isolation and identification, followed by a biochemical test to determine the S. aureus. Moreover, the bacteria susceptibility test against several antimicrobials was done using the disk diffusion method. Based on our findings, a total of 66 (25.58%) raw goat milk samples were tested positive for S. aureus, of which 36.36% were identified as multidrug-resistant. Moreover, S. aureus were also identified as resistant to penicillin (81.82%), ampicillin (65.15%), erythromycin (50.52%), and gentamicin (36.09%). The prevalence of S. aureus isolated from raw goat milk associated with subclinical mastitis in Siliragung Subdistrict, Banyuwangi District, Indonesia, was recorded at 25.58%. Moreover, 36.36% of S. aureus isolates were categorized as resistant to three or more classes of antibiotics. The biosafety and biosecurity procedures during the milking process should be strengthened in dairy goat farms to prevent the transmission of antimicrobial resistance among animals, humans, and environments.

Trends of Antibiotic Resistance in Multidrug-Resistant Pathogens Isolated from Blood Cultures in a Four-Year Period.

Multidrug-resistant organisms cause serious infections with significant morbidity and mortality in the worldwide. These organisms have been identified as urgent and serious threats by CDC. The aim of this study was to determine the prevalence and changes of antibiotic resistance of multidrug-resistant pathogens isolated from blood cultures over a four-year period in a tertiary-care hospital. Blood cultures were incubated in a blood culture system. Positive signalling blood cultures were subcultured on 5% sheep-blood agar. Identification of isolated bacteria was performed using conventional or automated identification systems. Antibiotic susceptibility tests were performed by disc diffusion and/or gradient test methods, if necessary, by automated systems. The CLSI guidelines were used for interpretation of antibiotic susceptibility testing of bacteria. The most frequently isolated Gram-negative bacteria was Escherichia coli (33.4%) followed by Klebsiella pneumoniae (21.5%). ESBL positivity was 47% for E. coli, 66% for K. pneumoniae. Among E. coli, K. pneumoniae, Pseudomonas aeruginosa, and Acinetobacter baumannii isolates, carbapenem resistance was 4%, 41%, 37%, and 62%, respectively. Carbapenem resistance of K. pneumoniae isolates has increased from 25% to 57% over the years, and the highest rate (57%) occured during the pandemic period. It is noteworthy that the aminoglycoside resistance in E. coli isolates gradually increased from 2017 to 2021. The rate of methicillin-resistant S. aureus (MRSA) was found to be 35.5%. Increased carbapenem resistance in K. pneumoniae and A. baumannii isolates is noteworthy, but carbapenem resistance in P. aeruginosa decreased. It is of great importance for each hospital to monitor the increase in resistance in clinically important bacteria, especially isolated from invasive samples, in order to take the necessary precautions in a timely manner. Future studies involving clinical data of patients and bacterial resistance genes are warranted.

Methicillin-Resistant Staphyloccocus aureus Contamination in Hospital Door Handles of Pokhara, Nepal

The presence of contaminants on the door handles surfaces of hospitals may be a common means of transfer of potentially pathogenic bacteria among users and can act as vehicles of diseases transmission. The aim of this study was to isolate, identify and evaluate the presence of bacterial contaminants and Methicillin Resistant Staphylococci (MRSA) with their antimicrobial susceptibility pattern present on the door handles of selected hospitals of Pokhara Metropolitan City, Western Nepal to take the necessary remedial measures. Isolation, identification and antimicrobial susceptibility test of bacteria were done using standard microbiological procedures. Further screening and confirmation of MRSA were done according to Clinical Laboratory Standard Institutes (CLSI) guidelines. Out of the 100 swab samples cultured, 96 (96%) showed bacterial growth. 62.86% (88/140) of the isolates were found to be Gram positive. Percentage distribution of the isolates showed that the most prevalent Gram-positive bacteria identified was Staphylococcus aureus (43.18%), followed by Coagulase negative Staphylococci (15.91%), Bacillus spp (11.36%), Diptheroides (10.23%) but considerable number of Micrococci (7.95%), Streptococcus pneumoniae (6.82%), Enterococci (4.55%). Prevalence of MRSA in this study was 39.47%. Significant difference in antibiotic resistance pattern was found among MRSA and MSSA isolates (P<0.05) reflecting increased ability of MRSA to develop resistance against various antimicrobials. Drugs like clindamycin, cefazoline and amikacin were found quite effective against MRSA in the present study would be better options for the management of such infections. MRSA strains’ contaminating on the door handles of hospitals may cause threat of infections to patients, hospital staffs, visitors to hospitals and people in the community. Thus, regular surveillance and disinfection with appropriate agent at regular interval would minimize the settlement and transmission of various pathogens including MRSA.Keywords: Contaminants; disinfection; door handles; hospital; MRSA; pathogen

Phase Contrast Image-Based Rapid Antimicrobial Susceptibility Testing of Bacteria in Liquid Culture Media

Currently, the world is facing the problem of bacterial resistance, which threatens public health, and bacterial antimicrobial susceptibility testing (AST) plays an important role in biomedicine, dietary safety and aquaculture. Traditional AST methods take a long time, usually 16-24 h, and cannot meet the demand for rapid diagnosis in the clinic, so rapid AST methods are needed to shorten the detection time. In this study, by using an in-house built centrifuge to centrifuge bacteria in a liquid medium onto the inner wall of the bottom surface of a counting plate, and using a phase contrast microscope to track bacterial growth under the effect of different antibiotic concentrations, the results of the minimum inhibitory concentration (MIC) of bacteria under the effect of antibiotics can be obtained in as early as 4 h. We used a combination of E. coli and tigecycline and obtained MIC results that were consistent with those obtained using the gold standard broth micro-dilution method, demonstrating the validity of our method; due to the time advantage, the complete set can be used in the future for point of care and clinical applications, helping physicians to quickly obtain the MIC used to inhibit bacterial growth.

Antibiotics Usage and Resistance among Patients with Severe Acute Respiratory Syndrome Coronavirus 2 in the Intensive Care Unit in Makkah, Saudi Arabia.

Antibiotic resistance is a global health and development threat, especially during the Severe Acute Respiratory Syndrome Coronavirus 2 (COVID-19) pandemic. Therefore, the current study was conducted to describe antibiotic usage and resistance among patients with COVID-19 in the intensive care unit (ICU) in Makkah, Saudi Arabia. In this cross-sectional study, only patients with positive COVID-19 status (42 patients) admitted to the ICU at the King Faisal Hospital were selected using a census sampling method. The susceptibility test of bacteria was carried out according to the standard protocol. The identified strains were tested in-vitro against several antibiotics drugs. Statistical analysis was performed using SPSS version 24. A total of 42 patients were included, with a mean age of 59.35 ± 18 years. Of them, 38.1% were males, and 61.9% were females. 35.7% have blood group O +. For age and blood groups, statistically significant associations were found between males and females, with p-values = 0.037 and 0.031, respectively. A large percentage (42.7%) of the obtained samples contained Klebsiella Pneumoniae; all bacteria were multidrug-resistance bacteria. Furthermore, 76.2% of bacteria were resistant to Ampicillin, 66.7% were resistant to Ciprofloxacin, 64.3% were resistant to Levofloxacin, 57.1% were resistant to Imipenem, and 57.1% were resistant to Moxifloxacin. On the contrary, among the 40 examined antibiotics, the effective antibiotics were Daptomycin, Linezolid, Mupirocin, Synercid, Teicoplanin, Vancomycin, and Nitrofurantoin. Our study demonstrates that antibiotic resistance is highly prevalent among ICU patients with COVID-19 at the King Faisal Hospital. Additionally, all bacteria were multidrug-resistance bacteria. Therefore, this high prevalence should be seriously discussed and urgently considered.

Prevalence of subclinical mastitis, associated risk factors and antimicrobial susceptibility of the pathogens isolated from milk samples of dairy cows in Northwest Ethiopia

Subclinical mastitis (SCM) is a problem of dairy cattle worldwide that affects the income of dairy farmers and besides a health risk to humans. A cross-sectional study was carried out to estimate prevalence, identify risk factors of SCM and antimicrobial susceptibility test of bacteria isolated from SCM quarters. California Mastitis Test (CMT) positive milk samples were cultured and bacteria isolated. The isolated bacteria were checked for their antimicrobial susceptibility using Kirby-Bauer disk diffusion method. The study findings showed that 32.7% of the quarters and 60.1% of the cows were CMT positive. The isolated bacteria were 29.5% Staphylococcus aureus (S. aureus) and 9.3% Streptococcus agalactiae (Str. agalactae). In the risk factors study, cows from farms of poor hygiene, increased age and stage of lactation had higher odds of SCM. Of the isolated bacterial species, 75.7% and 71.4% of S. aureus isolates were resistant to the effects of penicillin and ampicillin, respectively. Streptococcus agalactiae were resistant to the effects of oxytetracycline (76.9%), streptomycin (61.5%) and cloxacillin (53.8%). Whereas the majority of S. aureus (98.6%) and Str. agalactiae (88.5%) were susceptible to the effects of gentamycin and Clindamycin respectively. In conclusion, SCM had high level of prevalence and associated risk factors with pathogens that were resistant to commonly used antimicrobials. Implementing strict preventive measures and limiting spread of pathogens is a preferred approach rather than treatment with antibiotics to control mastitis in the study area.

Metabolism-Triggered Colorimetric Sensor Array for Fingerprinting and Antibiotic Susceptibility Testing of Bacteria.

The rapid identification and antibiotic susceptibility testing (AST) of bacteria would help us to accurately identify the infectious sources as well as guide the use of antibiotics, which are crucial for improving the survival rate and antimicrobial resistance. Herein, a colorimetric sensor array for bacteria fingerprinting was constructed with d-amino acid (d-AA)-modified gold nanoparticles (AuNPs) as probes (Au/d-AA). Bacteria can metabolize the d-AA, triggering the aggregation of AuNPs. Making use of different metabolic capabilities of bacteria toward different d-AA, eight kinds of bacteria including antibiotic-resistant bacteria and strains of the same bacterial species are successfully differentiated via learning the response patterns. Meanwhile, the sensor array also performs well in quantitative analysis of single bacterium and differentiation of bacteria mixtures. More interestingly, a rapid colorimetric AST approach has been developed based on the Au/d-AA nanoprobes by monitoring the d-AA metabolic activity of bacteria toward various antibiotic treatments. In this regard, the outlined work here would promote clinical practicability and facilitate antibiotic stewardship.

Association of bacterial isolates and antimicrobial susceptibility between prostatic fluid and urine samples in canine prostatitis with concurrent cystitis

Most old, intact male dogs usually have prostate disorders, especially benign prostatic hypertrophy and prostatitis with or without abscesses, and concurrent cystitis. The successful treatment of dogs with prostatitis concurrent with cystitis has relied on choosing an appropriate antimicrobial drug based on a bacterial culture and drug sensitivity testing. The objective of the study was to compare the prevalence of bacterial species and results of drug susceptibility testing of bacteria that were isolated from the prostatic fluids and urine samples that were collected from dogs with both prostatitis and cystitis. One hundred and sixty intact male dogs, who presented with both diseases, were recruited for the study. The disease diagnoses were based on clinical history notes, physical examinations, abdominal ultrasonography, prostatic fluid cytology, urinalysis and bacterial cultures from both prostatic fluid and urine samples. The bacterial culture results demonstrated that the major species that were detected in either the prostatic fluid or urine samples were Staphylococcus spp., Escherichia coli, Pseudomonas spp., Streptococcus spp., Proteus mirabilis and Klebsiella pneumoniae. Staphylococcus spp. (26.5 %, 43/162) and Escherichia coli (26.1 %, 12/46) were the most prevalent species from the prostatic fluid and urine samples, respectively. Statistical tests revealed that there were no significantly different prevalence levels among the isolated bacteria between the prostatic fluid and urine samples.Imipenem and gentamicin were the most potent antimicrobial drugs tested against the bacterial isolates in the present study. However, the administration of imipenem to treat prostatitis and cystitis in dogs was of concern. Interestingly, there were no significant differences in the antimicrobial drug susceptibility trends between the prostatic fluid and urine samples. Based on these results, a urine sample might be considered as an optional sample for bacterial cultures and antimicrobial drug susceptibility testing when it is not possible to collect a prostatic fluid sample.

Testing physiologically relevant conditions in minimal inhibitory concentration assays.

The minimal inhibitory concentration (MIC) assay uses agar or broth dilution methods to measure, under defined test conditions, the lowest effective concentration of an antimicrobial agent that inhibits visible growth of a bacterium of interest. This assay is used to test the susceptibilities of bacterial isolates and of novel antimicrobial drugs, and is typically done in nutrient-rich laboratory media that have little relevance to in vivo conditions. As an extension to our original protocol on MIC assays (also published in Nature Protocols), here we describe the application of the MIC broth microdilution assay to test antimicrobial susceptibility in conditions that are more physiologically relevant to infections observed in the clinic. Specifically, we describe a platform that can be applied to the preparation of medium that mimics lung and wound exudate or blood conditions for the growth and susceptibility testing of bacteria, including ESKAPE pathogens. This protocol can also be applied to most physiologically relevant liquid medium and aerobic pathogens, and takes 3-4 d to complete.

A novel concentration gradient microfluidic chip for high-throughput antibiotic susceptibility testing of bacteria.

Antibiotic resistance has become a serious threat to food safety and public health globally. Therefore, the development of a sensitive, quick, and simple method for antibiotic susceptibility testing is an urgent and crucial need. A novel concentration gradient microfluidic chip was designed in this work to generate antibiotic concentration gradient, culture bacteria, and produce fluorescence emission. An in-house-assembled fluorescence detection platform was constructed, and experiments were conducted to verify the linearity of the generated concentration gradient, explore the appropriate incubation time and flow rate for the microfluidic chip, and study the effect of long-term acid-based food processing on antibiotic susceptibility testing. Experimental results show that the concentration gradient generated by the microfluidic chip exhibited good linearity, stability, and controllability. The appropriate flow rate and incubation time for the microfluidic chip were 2μL/min and 5 h, respectively. The use of this microfluidic chip for testing antibiotic resistance of Salmonella to ofloxacin and ampicillin generated results that were completely consistent with test results obtained using the gold-standard method. Furthermore, Salmonella showed greater sensitivity to antibiotics under strong acid conditions, confirming the potential influence of acid-based food processing on antibiotic susceptibility testing of real samples. The designed microfluidic chip provides a high-throughput, sensitive, and rapid antibiotic susceptibility testing method that combines the microfluidic chip and the fluorescence detection platform. The application of this method would facilitate determination of antibiotic-resistant bacterial strains for clinicians and researchers, and enable monitoring of changes in bacterial resistance during food processing.

Precision Medicine in the Diagnosis and Management of Orthopedic Biofilm Infections.

Orthopedic biofilm infections are difficult to treat and require a multidisciplinary approach to diagnostics and management. Recent advances in the field include methods to disrupt biofilm, sequencing tools, and antibiotic susceptibility tests for bacteria residing in biofilm. The observation of interclonal differences in biofilm properties of the causative microorganisms, together with considerations of comorbidities and polypharmacy in a growing aging population, calls for a personalized approach to treat these infections. In this article, we highlight aspects of precision medicine that may open new perspectives in the diagnosis and management of orthopedic biofilm infections.

MAPt: A Rapid Antibiotic Susceptibility Testing for Bacteria in Environmental Samples as a Means for Bioterror Preparedness

Antibiotic resistance of bio-threat agents holds major concerns especially in light of advances in methods for engineering pathogens with antibiotic resistance. Preparedness means for rapid identification and prompt proper medical treatment are of need to contain the event and prevent morbidity and spreading of the disease by properly treating exposed individuals before symptoms appearance. Herein, we describe a novel, rapid, simple, specific, and sensitive method named Micro-Agar-PCR-test (MAPt), which determines antibiotic susceptibility of bio-terror pathogens, directly from environmental samples, with no need for any prior isolation, quantification, or enrichment steps. As proof of concept, we have used this approach to obtain correct therapeutic antibiotic minimal inhibitory concentration (MIC) values for the Tier-1 select agents, Bacillus anthracis, Yersinia pestis, and Francisella tularensis, spiked in various environmental samples recapitulating potential bioterror scenarios. The method demonstrated efficiency for a broad dynamic range of bacterial concentrations, both for fast-growing as well as slow-growing bacteria and most importantly significantly shortening the time for accurate results from days to a few hours. The MAPt allows us to address bioterror agents-contaminated environmental samples, offering rational targeted prophylactic treatment, before the onset of morbidity in exposed individuals. Hence, MAPt is expected to provide data for decision-making personal for treatment regimens before the onset of symptoms in infected individuals.

Bacterial susceptibility patterns to cotrimoxazole in urinary tract infections of outpatients and inpatients in Jakarta, Indonesia

BACKGROUND Cotrimoxazole, which has been one of the drugs of choice for urinary tract infections (UTIs) since 1960, must be evaluated to determine whether it is still a relevant drug for this use. This study aimed to assess the susceptibility patterns to cotrimoxazole of the bacteria that cause UTIs from urine samples of female outpatients (community-acquired [CA]-UTI) and inpatients (hospital-acquired [HA]-UTI) in Jakarta.
 METHODS This study was conducted from December 2014 to December 2015. Susceptibility testing of bacteria causing UTIs was conducted on 27 of 311 female outpatient urine samples collected from six clinics in Jakarta, and secondary data susceptibility testing was performed on 27 of 107 urine samples of inpatients from hospitals in Jakarta. These samples were examined in the Clinical Microbiology Laboratory, Faculty of Medicine, Universitas Indonesia, Cipto Mangunkusumo Hospital.
 RESULTS Susceptibility to cotrimoxazole was reported in 83% of the bacteria causing UTIs in CA-UTI and 44% of the bacteria in HA-UTI patients. Klebsiella pneumoniae was the most common cause of CA-UTI, with all isolates susceptible to cotrimoxazole (100%). Conversely, Escherichia coli was the most common cause of HA-UTI but was only susceptible in some isolates (44%). Bacteria from CA-UTI patients were almost twice as susceptible to cotrimoxazole compared with HA-UTI patients (p = 0.003).
 CONCLUSIONS Based on the susceptibility patterns identified, cotrimoxazole can be used as a treatment for CA-UTI but not for HA-UTI patients in Jakarta, Indonesia.

Rapid antibiotic susceptibility testing of bacteria from patients\u2019 blood via assaying bacterial metabolic response with surface-enhanced Raman spectroscopy

Blood stream infection is one of the major public health issues characterized with high cost and high mortality. Timely effective antibiotics usage to control infection is crucial for patients’ survival. The standard microbiological diagnosis of infection however can last days. The delay in accurate antibiotic therapy would lead to not only poor clinical outcomes, but also to a rise in antibiotic resistance due to widespread use of empirical broad-spectrum antibiotics. An important measure to tackle this problem is fast determination of bacterial antibiotic susceptibility to optimize antibiotic treatment. We show that a protocol based on surface-enhanced Raman spectroscopy can obtain consistent antibiotic susceptibility test results from clinical blood-culture samples within four hours. The characteristic spectral signatures of the obtained spectra of Staphylococcus aureus and Escherichia coli—prototypic Gram-positive and Gram-negative bacteria—became prominent after an effective pretreatment procedure removed strong interferences from blood constituents. Using them as the biomarkers of bacterial metabolic responses to antibiotics, the protocol reported the susceptibility profiles of tested drugs against these two bacteria acquired from patients’ blood with high specificity, sensitivity and speed.

Biocide susceptibility testing of bacteria: Development of a broth microdilution method

Biocide susceptibility testing (BST) of bacteria lacks standardised methods. Based on a recently established broth macrodilution BST method, a broth microdilution method for BST was developed. To establish the respective protocol, four reference strains Staphylococcus aureus ATCC® 6538, Enterococcus hirae ATCC® 10541, Escherichia coli ATCC® 10536 and Pseudomonas aeruginosa ATCC® 15442 were investigated for their minimal inhibitory concentrations (MICs) towards quaternary ammonium compounds (benzalkonium chloride), cationic compounds (chlorhexidine), aldehydes (glutardialdehyde) and alcohols (isopropanol) using tryptic soy broth. All combinations of (i) inoculum preparation according to the German Veterinary Medical Society (DVG) or the Clinical and Laboratory Standards Institute (CLSI) with some modifications, (ii) use of 1st subculture (SC) and 2nd SC, (iii) direct colony suspension (DCS) with/without glass beads, and (iv) incubation at 37 °C for 24 h, 48 h, and 72 h were compared using seven independent replications. Overall, the reproducibility was high for all abovementioned strain/biocide/parameter combinations. In total, 86.9 % – 100 % of the results were within ± one dilution step of the mode value. The proposed method for a standardised BST protocol comprises (i) two different inoculum densities, (ii) the use of a fresh overnight culture (1st SC or 2nd SC), (iii) the preparation of the inoculum suspension by either of the two methods using DCS with or without glass beads, and (iv) the incubation at 37 °C for 24 h. This broth microdilution method will help to harmonize BST of bacterial pathogens in routine diagnostics.

Eight rules for improving the quality of papers on the antimicrobial susceptibility of bacteria isolated from aquatic animals.

The methods for antimicrobial susceptibility testing of bacteria, although relatively simple, are not robust. As a consequence, if the data generated in such tests are to be commensurate, all susceptibility tests must be performed using standard protocols. A review of the published literature of antimicrobial susceptibility testing of bacteria from aquatic animals revealed a frequent occurrence of significant errors, particularly with regard to testing methodology, quality control and the use of appropriate interpretive criteria in the performance and reporting of susceptibility tests. This opinion piece provides a set of rules that, if followed, would help authors to avoid these frequently detected shortcomings.

Altitude Effect on the Properties of Honeys from the Region of Jijel (Algeria)

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