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Gram stains of Brucella spp. growing in blood culture broth may be mistaken for common Gram-positive organisms. Gram stains from three clinical laboratories of blood culture bottles that ultimately yielded Brucella spp. The slow-growing Gram-positive bacilli (A), Gram-positive rods and Gram-positive cocci in chains (B), and Gram-positive cocci in pairs and chains (C) were mistaken initially for streptococci or diphtheroids. Following streaking of blood culture broth on agar plates, Gram stains of colonies yielded small, Gram-negative organisms more typical of Brucella (Table 2).
Source publication
During 2015-2017, 11 confirmed brucellosis cases were reported in New York City, leading to 10 Brucella exposure risk events (“ Brucella events”) in 7 clinical laboratories (CLs). Most patients traveled to endemic countries and presented with histories and findings consistent with brucellosis. CLs were not notified that specimens might yield a haza...
Citations
... Manipulating unknown Brucella isolates on an open bench rather than in a biosafety cabinet (BSC) exposes many workers through aerosolization and increases the risk of LAI. In a recent assessment of the risk of exposure to brucellosis in laboratory workers in New York from 2015 to 2017, Brucella exposure incidents occurred in 10 of 11 confirmed brucellosis cases [26]. In the present case, brucellosis was clinically suspected, and the worker wore a mask and conducted all work in a Class II BSC, so there was no exposure. ...
... The worker was monitored for fever but did not develop symptoms. The use of MALDI-TOF MS is increasing, and safe work practices, including working with slow-growing organisms in a BSC and not using MALDI-TOF MS unless a biothreat agent is excluded, are recommended [26]. ...
Background
It is challenging to diagnose brucellosis in nonendemic regions because it is a nonspecific febrile disease. The accurate identification of Brucella spp. in clinical microbiology laboratories (CMLs) continues to pose difficulties. Most reports of misidentification are for B. melitensis, and we report a rare case of misidentified B. abortus.
Case presentation
A 67-year-old man visited an outpatient clinic complaining of fatigue, fever, and weight loss. The patient had a history of slaughtering cows with brucellosis one year prior, and his Brucella antibody tests were negative twice. After blood culture, the administration of doxycycline and rifampin was initiated. The patient was hospitalized due to a positive blood culture. Gram-negative coccobacilli were detected in aerobic blood culture bottles, but the CML's lack of experience with Brucella prevented appropriate further testing. Inaccurate identification results were obtained for a GN ID card of VITEK 2 (bioMérieux, USA) and matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI–TOF MS) using a MALDI Biotyper (Bruker, Germany). The strain showed 100.0% identity with Brucella spp. according to 16S rRNA sequencing. MALDI–TOF MS peaks were reanalyzed using the CDC MicrobeNet database to determine Brucella spp. (score value: 2.023). The patient was discharged after nine days of hospitalization and improved after maintaining only doxycycline for six weeks. The isolate was also identified as Brucella abortus by genomic evidence.
Conclusion
Automated identification instruments and MALDI–TOF MS are widely used to identify bacteria in CMLs, but there are limitations in accurately identifying Brucella spp. It is important for CMLs to be aware of the possibility of brucellosis through communication with clinicians. Performing an analysis with an additional well-curated MALDI–TOF MS database such as Bruker security-relevant (SR) database or CDC MicrobeNet database is helpful for quickly identifying the genus Brucella.
... Laboratory-acquired infections are important sources of transmission when specimens or cultures containing the organism are not handled using the proper biosafety precautions. 42,43 ...
The process of bacterial nomenclature change has evolved in complexity over time and continues to be an iterative process that is not without challenges. The importance and feasibility of such changes varies between basic researchers, clinical microbiologists, and clinicians. In recent years, clinically-relevant changes have been made across gram-positive and gram-negative organism groups, as well as the mycobacteria. Updated clinical laboratory accreditation requirements state that clinical laboratories must update their reporting practices in the case of clinically-relevant nomenclature changes. These updates may significantly impact various sectors of healthcare including antimicrobial stewardship, laboratory protocols, and infection prevention procedures and policies. While regularly updating bacterial nomenclature aims to improve the accuracy and consistency of our microbial language, the potential impact of such changes must be considered.
... The event represents a tragic and tangible reminder of the high transmissibility of members of the genus and the potential role of brucellae as bioterrorism agents [2]. Although the Lanzhou outbreak stands out because of its enormous size, it should be pointed out that brucellosis is one of the most common organisms transmitted in the laboratory setting, and smaller LAB clusters have repeatedly occurred worldwide [3,4]. ...
... Even in areas endemic to zoonosis, the diagnosis of brucellosis is not initially considered in a substantial fraction of patients [17]. Under these circumstances, the physicians may fail to alert the CML that the patients' specimens might contain a hazardous pathogen and should be handled with appropriate safety precautions [3]. The vague and unspecific manifestations of the disease may also result in a delay in recognition of outbreaks of LAB and failure to implement corrective measures and prevent additional cases [4]. ...
... Each of the individual links of the identification chain is prone to error, misidentifying the isolate and causing LAB. Furthermore, because of the effective veterinarian control of the zoonosis, the disease has become uncommon in industrialized countries, and personnel working at CMLs have become unfamiliar with the phenotypic characteristics of the genus [3]. Gram stain plays an early and key role in correctly identifying Brucella species. ...
Brucellosis is one of the most common etiologies of laboratory-acquired infections worldwide, and handling of living brucellae should be performed in a Class II biological safety cabinet. The low infecting dose, multiple portals of entry to the body, the wide variety of potentially contaminated specimens, and the unspecific clinical manifestations of human infections facilitate the unintentional transmission of brucellae to laboratory personnel. Work accidents such as spillage of culture media cause only a small minority of exposures, whereas >80% of events result from unfamiliarity with the phenotypic features of the genus, misidentification of isolates, and unsafe laboratory practices such as working on an open bench without protective goggles or gloves or the aerosolization of bacteria. The bacteriological diagnosis of brucellae by traditional methods is simple and straightforward but requires extensive manipulation of the isolates, and, nowadays, many laboratory technicians are not familiar with the genotypic features of the genus, resulting in inadvertent exposure and contagion. Detection of brucellar infections by culture-independent molecular methods is safe, but the identification of the organism using MALDI-TOF technology is not hazard-free, requiring an initial bacterial inactivation step to avoid transmission. Unfortunately, these novel and safer methods are costly and frequently unavailable in resource-limited endemic countries.
... Brucellosis is the most common bacterial laboratoryacquired infection worldwide [23]. Clinical laboratory workers have failed to recognize suspicious isolates and have manipulated unknown isolates on open benches, using procedures that aerosolized Brucella, increasing their exposure risk to biological hazards [24]. In this study, nine laboratory-acquired infection events were found in the southern region, a brucellosis-emerging area [25]. ...
... In New York City, more than 200 occupational exposures occurred because of the generation of infectious Brucella spp. aerosols [24]. In our study, we observed a high risk of developing laboratory-acquired brucellosis in microbiological laboratory workers. ...
Brucellosis is a severe public health problem in China. However, analysis on related infection events is lacking. We performed a systematic analysis of brucellosis laboratory infection and vaccine infection events from 2006 to 2019 in China based on the published literatures. Our analysis showed that most laboratoryBrucellainfections in hospitals were found in Southern China. The identification and handling of suspected samples ofBrucellainfection without following the recommended biosafety protection was the main risk factor. It is important to strengthen the preventive awareness of clinical laboratory staff and physicians, while highlighting the compulsory handling and identification of suspectedBrucella-infected samples in biosafety facilities and following biosafety practices. However, a severe Brucella infection accident at the Northeast Agricultural University, with 28 positive cases, showed that strengthening the management in teaching experiments of students in the veterinary-related profession is essential. However, cluster S2 vaccine strain infection events caused by vaccination and production were mainly observed in Northern China. Strengthening vaccination skills, personal protection, and improving the biosafety management of vaccine production and implementing regular risk surveillance is mandatory. Our analysis provides helpful clues for control of public health events involving brucellosis, as well as implementing intervention strategies is urgent.
... These properties have been evaluated in the past in the context of biological weapons' study facilities [6]: Brucella species have traditionally been included in biodefense research lists, while Brucella suis was the first pathogen to be weaponized in 1952. The same properties are also responsible for its significant burden on laboratory-acquired infections [7,8], numerous of which have been reported from China, even since 1936 [9,10]. In endemic areas, clinicians are aware that microbiology laboratory personnel should be preemptively informed about the possibility of brucellosis when dealing with blood cultures of patients, and ideally these should be handled at Biosafety Level 3 (BSL-3), or, when not applicable, with extreme caution and appropriate personal protective equipment (PPE), at BSL-2. ...
An inadequacy in sanitizing processes in a biopharmaceutical plant in Lanzhou, China, during July and August 2019, led to the aerosolization of Brucella that was subsequently spread through wind in nearby settlements and academic institutes, resulting in more than 10,000 human brucellosis cases, as of November 2020. The leak, possibly the largest laboratory accident in the history of infectious diseases, underlines the particular characteristics of Brucella that have made the pathogen a historical entity in biodefense research and a major cause of laboratory-associated infections. It further underlines the need for enhanced vigilance and strict regulatory interventions in similar facilities.
... Furthermore, it is now well established that Brucella spp. is an intracellular bacterium that escapes destruction by macrophages and causes severe mitochondrial fragmentation after 48 hours of bacterial entry into different cell types [37,38]. Therefore, antibiotics for the treatment of brucellosis must have the ability to kill the bacterium by entering macrophages [39]. In our study, a lower sensitivity pattern was observed only for chloramphenicol (33 and 38%). ...
Introduction: Foodborne resistant bacteria have become a challenge to food security. Milk and milk products are easy vectors of transmission of foodborne pathogens, these being the main sources of human infection by antimicrobial resistant pathogens. The present study aimed at making a comparative approach of the antibiotic sensitivity/resistance of 3 bacterial strains (Escherichia coli, Salmonella spp. and Brucella spp.) isolated from milk, drinking water and green fodder consumed by cows in the West Cameroon region (Central Africa). Methodology: A total of 48 raw milk samples, 48 water samples and 48 green fodder samples were collected during the year 2020 and subjected to culture and identification of Escherichia coli, Salmonella spp. and Brucella spp. Antibiotic susceptibility testing was performed using the antibiotic disc diffusion method. Results: Escherichia coli isolates showed high resistance (56-100%) to ampicillin, amoxicillin/clavulanic acid, cefotaxime, ceftazidime and ceftriaxone in all three samples. Salmonella spp. isolates showed resistance to ampicillin only (62, 67 and 67%). Brucella spp. strains isolated from raw milk and drinking water showed high sensitivity (78-100%) to azithromycin, doxycycline, ciprofloxacin, levofloxacin, gentamicin, rifampicin and trimethoprim/sulfamethoxazole, streptomycin and tetracycline. Antibiotic sensitivity/resistance to Escherichia coli and Salmonella spp. strains largely did not differ between samples (P>0.05). No difference in sensitivity/resistance (P>0.05) of Brucella spp. strains isolated from milk and water was observed with respect to the 10 antibiotics tested. Conclusions: The emergence of resistance to various antibiotics commonly used in medical and veterinary practices has important implications for public health. It seems necessary to strengthen of the regulations covering the sale and prescription of antibiotics.
... Adherence of MLT to infection control guidelines is of the utmost importance to reduce their risk of infection with SARS-CoV-2 [20,21]. The adherence to these guidelines can be influenced by multiple factors including educational level, training on laboratory safety, and years of experience [22]. Therefore, it is essential to assess the adherence of MLT to infection control practices during the pandemic to ensure the safety of medical teams and patients [23,24]. ...
Background:
The adherence of medical laboratory technicians (MLT) to infection control guidelines is essential for reducing the risk of exposure to infectious agents. This study explored the adherence of MLT towards infection control practices during the COVID-19 pandemic.
Method:
The study population consisted of MLT (n = 444) who worked in private and government health sectors in Jordan. A self-reported survey was used to collect data from participants.
Findings:
More than 87% of the participants reported adherence to hand-washing guidelines and using personal protective equipment (PPE) when interacting with patients (74.5%), and handling clinical samples (70.0%). Besides, 88.1%, 48.2%, and 7.7% reported wearing of lab coats, face masks, and goggles, at all times, respectively. The majority reported increased adherence to infection control practices during the COVID-19 pandemic. This includes increased PPE use at the workplace (94.2%), increased frequency of disinfection of laboratory surfaces (92.4%) and laboratory equipment (86.7%), and increased frequency of handwashing/use of antiseptics (94.6%). Having a graduate degree was significantly associated with increased adherence of participants to the daily use of goggles/eye protection (p = 0.002), and the use of PPE while handling clinical samples (p = 0.011). Having work experience of >10 years was associated with increased adherence to the use of PPE while handling clinical samples (p = 0.001).
Conclusion:
MLT reported very good adherence with most assessed infection control practices. In addition, they reported increased conformity with infection control guidelines during the COVID-19 pandemic.
... Brucellosis is a widespread zoonotic disease that is caused by bacteria and is categorized as a bacterial human disease (1). The World Organization for Animal Health (OIE) lists brucellosis as a multi-animal comorbidity (2), and brucellosis is a second-category animal infectious disease in China (3). ...
Brucellosis is a common zoonosis in China, resulting in abortion in animals. Outbreaks of abortion in blue foxes caused by Brucella infection have rarely been reported. In the present study, 3–5 mL blood samples collected from the femoral veins of 10 abortuses of blue foxes were assessed by RBPT (Rose Bengal plate test) and SAT (serum tube agglutination test) to preliminarily investigate the source of infection for the clustering of abortion events at a blue fox farm in Heilongjiang Province. Screening experiments showed that all 10 blood samples were positive in the RBPT, while only eight blood samples out of the 10 were positive in the SAT. Subsequently, 10 tissue samples (spleen, lungs, stomach contents, and afterbirth) from the same 10 foxes were assessed using AMOS (acronym for B. abortus, melitensis, ovis, and suis)-PCR (polymerase chain reaction), and sequencing analysis was performed on amplification products to verify the results of the serology survey. Results showed a spectral band of ~731 bp in these samples. BLAST showed sequences of AMOS-PCR products in this study to be 100% similar (E = 0.0) to sequences in B. melitensis strain from GenBank. These data preliminarily indicated that the blue fox's outbreak of abortion events was caused by brucellosis via the B. melitensis strain. Then 726 serum samples were tested by RBPT and SAT to determine the prevalence of brucellosis on the farm. A comprehensive epidemiological and reproductive status survey of the infected blue fox population was performed. The seropositive rate was found to be 67.90% (493/726) by RBPT and 41.32% (300/726) by SAT. The technicians had stopped feeding the foxes with chicken carcasses and instead fed them raw ground sheep organs (lungs, tracheae, placentae, and dead sheep fetuses) infected by B. meliteneis strains, and that this change in diet caused the outbreak of abortion events. The high abortion rate (55%) and low cub survival rate (65%) were the most distinctive features of the outbreak; these factors led to severe economic losses. Feeding cooked sheep/goat offal and strict breeding management is necessary for disease prevention.
... On the other hand, in vitro antimicrobial susceptibility testing of Brucella can be performed by several other techniques, including agar dilution, broth microdilution, and the E-test (34). However, antimicrobial susceptibility testing for highly zoonotic bacteria such as Brucella needs a laboratory with level-3 biosafety, which may not be accessible in many regions (35). Moreover, traditional antibiotic susceptibility tests require several passaging of Brucella, which is a high-risk and time-consuming activity (22). ...
Background: RNA polymerase beta subunit (rpoB) gene analysis in bacterial communities is known as a method for determining rifampin sensitivity and genetic diversity among Brucella spp. Detection of antibiotic resistance among Brucella isolates can be a critical approach to control brucellosis. However, rpoB gene analysis of Brucella melitensis for assessing rifampicin resistance has not yet been performed in Iran, which is considered an endemic area for brucellosis. Objectives: The aim of this study was to analyze the whole sequence of rpoB genes of different B. melitensis isolates from humans to identify the single-nucleotide polymorphisms (SNPs) and mutations related to rifampin resistance and to analyze the genetic diversity of these bacteria in Iran. Methods: Between 2017 and 2019, a total of 156 blood samples along with 12 synovial fluid specimens were collected from brucellosis patients in different Iranian provinces and subjected to bacterial culture in Brucella selective media. Brucella identification was carried out using classical biotyping and molecular examinations. Polymerase chain reaction (PCR)-based amplification of the rpoB gene was performed by specific rpoB primers for whole gene sequencing. The antimicrobial susceptibility of Brucella isolates was assessed using disk diffusion susceptibility tests and minimal inhibitory concentration (MIC) methods. The presence of rifampin-binding sites and SNPs were investigated through rpoB whole gene sequencing. Results: Clinical B. melitensis isolates were obtained from blood (13) and synovial fluid (1) samples of patients from different regions of Iran. The results of MIC and disk diffusion susceptibility tests showed that all the isolates were sensitive to rifampin except for one isolate showing intermediate rifampin resistance based on the standards defined for slow-growing bacteria by the Clinical and Laboratory Standards Institute (CLSI). Gene analysis for identifying the mutations related to rifampin resistance and investigating genetic diversity showed that none of the B. melitensis isolates had missense mutations, confirming the susceptibility of all the studied isolates to rifampin. Conclusions: The present study revealed that rpoB gene analysis could be used for the efficient and precise identifying of the mutations related to rifampin resistance, investigating rifampin binding sites, and genotyping Brucella species. Furthermore, the identification of B. melitensis isolates with intermediate resistance to rifampicin highlighted the importance of periodically carrying out antibiotic susceptibility testing. The molecular detection of rpoB mutations in different Brucella isolates may help to prevent the spread of rifampin-resistant Brucella spp. among humans and livestock.
... However, transmission via blood transfusion and bone marrow transplantation and transplacental transmission from mother to fetus or via breastfeeding have been reported [6,7]. Laboratory-acquired infection is a well-documented infection route in the United States and Asia [8,9]. Clinical signs of brucellosis in humans are highly variable, non-specific, and the disease has similar or very close symptoms to some diseases that cause fever resulting in misdiagnosis and insufficient treatment. ...
Brucellosis is a highly contagious and incapacitating disease of humans, livestock and wildlife species globally. Treatment of brucellosis in animals is not recommended, and in humans, combinations of antibiotics recommended by the World Health Organization are used. However, sporadic antimicrobial-resistant (AMR) isolates and relapse cases have been reported from different endemic regions. In the current study, molecular characterization and antibiotic susceptibility testing using the microdilution method for 35 B. abortus and B. melitensis strains isolated from humans, milk and animal were carried out. Additionally, Next-Generation-Sequence (NGS) technology was applied to confirm Brucella at the species level and investigate AMR and pathogenicity-associated determinants. MALDI-TOF seemed to be a rapid and reliable tool for routine identification of brucellae to the genus level; however, DNA-based identification is indispensable for accurate species identification. Brucella abortus strains were isolated from two human cases and a sheep. Such infections are uncommon in Egypt. Egyptian Brucella strains are still in-vitro susceptible to doxycycline, tetracyclines, gentamicin, ciprofloxacin, levofloxacin, chloramphenicol, streptomycin, trimethoprim/sulfamethoxazole and tigecycline. Probable (no CLSI/EUCAST breakpoints have been defined yet) in-vitro resistance to rifampicin and azithromycin was observed. WGS failed to determine classical AMR genes, and no difference in the distribution of virulence-associated genes in all isolates was found. Isolates of human and non-human origins were still susceptible to the majority of antibiotics used for treatment in humans. The absence of classical AMR genes in genomes of “resistant” Brucella strains may reflect a lack of information in databases, or resistance might not be encoded by single resistance genes. The One Health approach is necessary for tackling brucellosis. Continuous susceptibility testing, updating of breakpoints, assessing mutations that lead to resistance are needed.
