Identification of the first bacteriocin isolated in Lebanon extracted via a modified adsorption-desorption method and its potential food application

Introduction: The raw goat milk is considered as a good source of novel bacteriocinogenic lactic acid bacteria (LAB) strains. Method: The bacteriocin, which named enterocin CMUL20-2 was secreted by Enterococcus faecium CMUL20-2. This bacterial strain was originally isolated from raw goat’s milk, was extracted by using a modified adsorption-desorption method and purified via RP-HPLC. antimicrobial activity was tested against several pathogenic and spoilage microbes. Results: The enterocin CMUL20-2 showed a strong adsorption on cell wall of producer strain even in acidic environment which facilitate its extraction in only two simple steps. The recovered purified enterocin has decreased procedure time and diminished the number of undesirable molecules present in Rogosa and Sharpe (MRS) broth. The recovered enterocin showed antimicrobial activity against several foodborne pathogenic and spoilage microbes. Conclusion:  The recovered enterocin was able to tolerate a variety of food chain conditions such as high temperature, pH and storage stability, and it can be a good candidate to protect food from spoilage microbes &nbsp

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Background: Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. // Methods: We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung's disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. // Findings: We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung's disease) from 264 hospitals (89 in high-income countries, 166 in middle-income countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in low-income countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. // Interpretation: Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between low-income, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

Global, regional, and national burden of disorders affecting the nervous system, 1990–2021: a systematic analysis for the Global Burden of Disease Study 2021

BackgroundDisorders affecting the nervous system are diverse and include neurodevelopmental disorders, late-life neurodegeneration, and newly emergent conditions, such as cognitive impairment following COVID-19. Previous publications from the Global Burden of Disease, Injuries, and Risk Factor Study estimated the burden of 15 neurological conditions in 2015 and 2016, but these analyses did not include neurodevelopmental disorders, as defined by the International Classification of Diseases (ICD)-11, or a subset of cases of congenital, neonatal, and infectious conditions that cause neurological damage. Here, we estimate nervous system health loss caused by 37 unique conditions and their associated risk factors globally, regionally, and nationally from 1990 to 2021.MethodsWe estimated mortality, prevalence, years lived with disability (YLDs), years of life lost (YLLs), and disability-adjusted life-years (DALYs), with corresponding 95% uncertainty intervals (UIs), by age and sex in 204 countries and territories, from 1990 to 2021. We included morbidity and deaths due to neurological conditions, for which health loss is directly due to damage to the CNS or peripheral nervous system. We also isolated neurological health loss from conditions for which nervous system morbidity is a consequence, but not the primary feature, including a subset of congenital conditions (ie, chromosomal anomalies and congenital birth defects), neonatal conditions (ie, jaundice, preterm birth, and sepsis), infectious diseases (ie, COVID-19, cystic echinococcosis, malaria, syphilis, and Zika virus disease), and diabetic neuropathy. By conducting a sequela-level analysis of the health outcomes for these conditions, only cases where nervous system damage occurred were included, and YLDs were recalculated to isolate the non-fatal burden directly attributable to nervous system health loss. A comorbidity correction was used to calculate total prevalence of all conditions that affect the nervous system combined.FindingsGlobally, the 37 conditions affecting the nervous system were collectively ranked as the leading group cause of DALYs in 2021 (443 million, 95% UI 378–521), affecting 3·40 billion (3·20–3·62) individuals (43·1%, 40·5–45·9 of the global population); global DALY counts attributed to these conditions increased by 18·2% (8·7–26·7) between 1990 and 2021. Age-standardised rates of deaths per 100 000 people attributed to these conditions decreased from 1990 to 2021 by 33·6% (27·6–38·8), and age-standardised rates of DALYs attributed to these conditions decreased by 27·0% (21·5–32·4). Age-standardised prevalence was almost stable, with a change of 1·5% (0·7–2·4). The ten conditions with the highest age-standardised DALYs in 2021 were stroke, neonatal encephalopathy, migraine, Alzheimer's disease and other dementias, diabetic neuropathy, meningitis, epilepsy, neurological complications due to preterm birth, autism spectrum disorder, and nervous system cancer.InterpretationAs the leading cause of overall disease burden in the world, with increasing global DALY counts, effective prevention, treatment, and rehabilitation strategies for disorders affecting the nervous system are needed

Global burden of 288 causes of death and life expectancy decomposition in 204 countries and territories and 811 subnational locations, 1990–2021: a systematic analysis for the Global Burden of Disease Study 2021

BACKGROUND Regular, detailed reporting on population health by underlying cause of death is fundamental for public health decision making. Cause-specific estimates of mortality and the subsequent effects on life expectancy worldwide are valuable metrics to gauge progress in reducing mortality rates. These estimates are particularly important following large-scale mortality spikes, such as the COVID-19 pandemic. When systematically analysed, mortality rates and life expectancy allow comparisons of the consequences of causes of death globally and over time, providing a nuanced understanding of the effect of these causes on global populations. METHODS The Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2021 cause-of-death analysis estimated mortality and years of life lost (YLLs) from 288 causes of death by age-sex-location-year in 204 countries and territories and 811 subnational locations for each year from 1990 until 2021. The analysis used 56 604 data sources, including data from vital registration and verbal autopsy as well as surveys, censuses, surveillance systems, and cancer registries, among others. As with previous GBD rounds, cause-specific death rates for most causes were estimated using the Cause of Death Ensemble model-a modelling tool developed for GBD to assess the out-of-sample predictive validity of different statistical models and covariate permutations and combine those results to produce cause-specific mortality estimates-with alternative strategies adapted to model causes with insufficient data, substantial changes in reporting over the study period, or unusual epidemiology. YLLs were computed as the product of the number of deaths for each cause-age-sex-location-year and the standard life expectancy at each age. As part of the modelling process, uncertainty intervals (UIs) were generated using the 2·5th and 97·5th percentiles from a 1000-draw distribution for each metric. We decomposed life expectancy by cause of death, location, and year to show cause-specific effects on life expectancy from 1990 to 2021. We also used the coefficient of variation and the fraction of population affected by 90% of deaths to highlight concentrations of mortality. Findings are reported in counts and age-standardised rates. Methodological improvements for cause-of-death estimates in GBD 2021 include the expansion of under-5-years age group to include four new age groups, enhanced methods to account for stochastic variation of sparse data, and the inclusion of COVID-19 and other pandemic-related mortality-which includes excess mortality associated with the pandemic, excluding COVID-19, lower respiratory infections, measles, malaria, and pertussis. For this analysis, 199 new country-years of vital registration cause-of-death data, 5 country-years of surveillance data, 21 country-years of verbal autopsy data, and 94 country-years of other data types were added to those used in previous GBD rounds. FINDINGS The leading causes of age-standardised deaths globally were the same in 2019 as they were in 1990; in descending order, these were, ischaemic heart disease, stroke, chronic obstructive pulmonary disease, and lower respiratory infections. In 2021, however, COVID-19 replaced stroke as the second-leading age-standardised cause of death, with 94·0 deaths (95% UI 89·2-100·0) per 100 000 population. The COVID-19 pandemic shifted the rankings of the leading five causes, lowering stroke to the third-leading and chronic obstructive pulmonary disease to the fourth-leading position. In 2021, the highest age-standardised death rates from COVID-19 occurred in sub-Saharan Africa (271·0 deaths [250·1-290·7] per 100 000 population) and Latin America and the Caribbean (195·4 deaths [182·1-211·4] per 100 000 population). The lowest age-standardised death rates from COVID-19 were in the high-income super-region (48·1 deaths [47·4-48·8] per 100 000 population) and southeast Asia, east Asia, and Oceania (23·2 deaths [16·3-37·2] per 100 000 population). Globally, life expectancy steadily improved between 1990 and 2019 for 18 of the 22 investigated causes. Decomposition of global and regional life expectancy showed the positive effect that reductions in deaths from enteric infections, lower respiratory infections, stroke, and neonatal deaths, among others have contributed to improved survival over the study period. However, a net reduction of 1·6 years occurred in global life expectancy between 2019 and 2021, primarily due to increased death rates from COVID-19 and other pandemic-related mortality. Life expectancy was highly variable between super-regions over the study period, with southeast Asia, east Asia, and Oceania gaining 8·3 years (6·7-9·9) overall, while having the smallest reduction in life expectancy due to COVID-19 (0·4 years). The largest reduction in life expectancy due to COVID-19 occurred in Latin America and the Caribbean (3·6 years). Additionally, 53 of the 288 causes of death were highly concentrated in locations with less than 50% of the global population as of 2021, and these causes of death became progressively more concentrated since 1990, when only 44 causes showed this pattern. The concentration phenomenon is discussed heuristically with respect to enteric and lower respiratory infections, malaria, HIV/AIDS, neonatal disorders, tuberculosis, and measles. INTERPRETATION Long-standing gains in life expectancy and reductions in many of the leading causes of death have been disrupted by the COVID-19 pandemic, the adverse effects of which were spread unevenly among populations. Despite the pandemic, there has been continued progress in combatting several notable causes of death, leading to improved global life expectancy over the study period. Each of the seven GBD super-regions showed an overall improvement from 1990 and 2021, obscuring the negative effect in the years of the pandemic. Additionally, our findings regarding regional variation in causes of death driving increases in life expectancy hold clear policy utility. Analyses of shifting mortality trends reveal that several causes, once widespread globally, are now increasingly concentrated geographically. These changes in mortality concentration, alongside further investigation of changing risks, interventions, and relevant policy, present an important opportunity to deepen our understanding of mortality-reduction strategies. Examining patterns in mortality concentration might reveal areas where successful public health interventions have been implemented. Translating these successes to locations where certain causes of death remain entrenched can inform policies that work to improve life expectancy for people everywhere. FUNDING Bill & Melinda Gates Foundation

Study of the antimicrobial and antioxidant activity of essential oil extracted from Lebanese plants

La résistance aux antibiotiques est devenue une véritable préoccupation. Ainsi, la recherche de composés antimicrobiens est plus que jamais d’actualité. Les plantes constituent une source potentielle de ces composés. La thèse présente une étude détaillée de la composition chimique et décrit l’activité antimicrobienne d’huiles essentielles issues de trois plantes : Micromeria barbata, Eucalyptus spp. et Juniperus excelsa. L’influence de plusieurs paramètres tels que le type d’organe, l’altitude de croissance de la plante, la durée de l’extraction, la méthode de séchage sur la qualité et l’activité de l’huile est abordée. Les résultats montrent, dans le cas de M. barbata non fleurie, que le pourcentage maximal (99.54 %) de l’huile est obtenu à la plus haute altitude (1700 m). Cependant, le pourcentage du Pulgeone, le composé principal de l’huile essentielle, diminue graduellement lorsque l’altitude augmente. Par ailleurs, l’activité des huiles essentielles extraites des plantes aux trois différentes altitudes est équivalente pour les différentes souches testées.L’activité antibactérienne et antifongique de l’huile essentielle de M. barbata est également mise en évidence contre différentes espèces bactériennes, y compris celles qui résistent aux carbapénèmes, pour les champignons opportunistes et les dermatophytes L’activité antimicrobienne de ces trois huiles essentielles est évaluée vis-à-vis de deux souches de Mycobacterium tuberculosis (dont une XDR), deux espèces atypiques : Mycobacterium kansasii et Mycobacterium gordonae ainsi que sur Paenibacillus larvae. Les résultats montrent que les trois huiles possèdent une activité notoire.Antibiotic resistance has become a real concern. Thus, the research of antimicrobial compounds is more relevant than ever. The aromatic plants are a potential source of these compounds. The thesis presents an advanced study of the chemical composition and describes the antimicrobial activity of essential oils from three plants: Micromeria barbata (M.barbata), Eucalyptus spp. and Juniperus excelsa (J.excelca). The influence of several parameters such as organ type, elevation of plant growth,duration of extraction, and drying method has been discussed .The results reveal that non – flowering of M..barbata has the maximum percentage (99.54%) of the oil is collected at the altitude (1700 m). However, the percentage of Pulgeone, the main component of the essential oil gradually decreases with increasing altitude. Furthermore, the activity of essential oils extracted from plants in three different altitudes is similar for the different microbial strains tested.The essential antibacterial and antifungal oil M. barbata is also highlighted against different bacterial species, including those resistant to carbapenems, for opportunistic fungi and dermatophytesThe antimicrobial activity of three essential oils is evaluated against two strains of Mycobacterium tuberculosis (including XDR), two atypical species: Mycobacterium kansasii and Mycobacterium gordonae well as Paenibacillus larvae ,the causative agent of American foulbrood disease that infect the honey bees larvae . Results show that three oils have a significant anti-mycobacterial activity

Comparison of the efficacy of two insecticides for the management of Dactylopius opuntiae on prickly pear cactus in Lebanon and monitoring of the insecticides residues dissipation rates in fruits and cladodes

International audienceEnvironmental conditions and availability of new cultivated areas of prickly pear cactus in Lebanon appeared lately to be favourable for the survival and development of D. opuntiae which has become highly noxious. To thwart use of random chemicals and broad spectrum pesticides by local farmers, efficacy of sulfoxaflor (Closer™ 240 SC, rate 0.179 kg a.i/ha) and diflubenzuron (Dimilin® 25 W, rate 0.329 kg a.i/ha) was compared. Individual insecticidal solutions of each applied alone or in mixing with mineral oil (spray oil 7E, rate 1.010 kg a.i/ha) on potted cladodes in laboratory conditions (T = 23 ± 4 °C, RH = 58–63%) showed a sharp decrease in the number of individuals within colonies 48 h after treatment especially among first- and second-instar larvae. Similarly, the same insecticidal solutions applied on separate plots in field condi-tions (T = 20–35 °C, RH = 55–75%) according to the Good Agricultural Practices showed efficacious control of the pest development. D. opuntiae mortality mean percentage reached 80% and 72%, respectively, for sulfoxaflor and diflubenzu-ron 96 h after treatment. The added mineral oil acted as a synergist and enhanced efficacy of both insecticides. Mortality mean percentages increased to 92% and 88%, respectively. Insecticides residues content over time in fruits and cladodes were afterwards monitored. Samples were harvested at 3 days interval and residues extracted by the QuEChERS method and quantified by LC–MS. Dissipation rates after 30 days of treatment attained 70.8% and 80.6% in fruits and 98.5% and 74.0% in cladodes, respectively, for sulfoxaflor and diflubenzuron. Half-life time varied accordingly and was higher for fruits treated with sulfoxaflor (14.20 days) than for those treated with diflubenzuron (11.18 days). On contrary, for clad-odes, half-life time of the former (4.18 days) was lower than that of the latter (15.47 days). The addition of mineral oil into the spray solution of the insecticides increased their persistency in the crop. Consequently, half-life times increased and dissipation rates decreased. Findings of this study should enlarge the spectrum of chemical means for D. opuntiae management and promote resilience of the crop. Analytical method applied for residues analysis should be useful for registration of the insecticides on the crop and MRLs setting

PRÉVALENCE DE LISTERIA MONOCYTOGENES DANS LE LAIT CRU DE VACHE AU LIBAN NORD

Al Kassaa Imad, Khaled el Omari, Marwa Saati, Bachar Ismail and Monzer Hamze. 2016. Prevalence of Listeria monocytogenes in raw cow milk in north Lebanon. Lebanese Science Journal, 17(1): 39-45. Listeriosis, although a zoonosis, is an invasive disease that can affect newborns, pregnant women and immunocompromised adults. Clinical manifestations can be expressed by febrile gastroenteritis, invasive forms including severe sepsis, meningitis, rhombencephalitis, prenatal infections and abortions. Species of Listeria bacteria are ubiquitous and adaptable to the environment in animal and plant foods. This study aimed to determine the prevalence of Listeria monocytogenes in 100 samples of fresh cow milk collected from different areas of North Lebanon. Listeria monocytogenes was detected by using the Grand VIDAS technique (Biomérieux France). The results obtained revealed the absence of Listeria monocytogenes in all analyzed samples

In-vitro evaluation of the antibacterial activity of the essential oils of Micromeria barbata, Eucalyptus globulus and Juniperus excelsa against strains of Mycobacterium tuberculosis (including MDR), Mycobacterium kansasii and Mycobacterium gordonae

Background: Mycobacterium spp. are responsible for several diseases, particularly in immunocompromised populations. The spreading of the resistance to antimycobacterial drugs is a significant problem to the public health and requires to find out a new and innovative alternative for the treatment of drug resistant mycobacterial strains. In this study, the antimycobacterial activity of Micromeria barbata, Eucalyptus globulus and Juniperus excelsa essential oils extracted from Lebanese plants was investigated against selected Mycobacterium spp. strains. Methods: Several dilutions of the three aforementioned essential oils were studied for antimycobacterial activity against four Mycobacterium spp. strains: Mycobacterium tuberculosis subsp. tuberculosis (ATCC (R) 27294 (TM)), multidrug-resistant M. tuberculosis (CMUL 157), Mycobacterium kansasii Hauduroy (ATCC (R) 12478 (TM)) and Mycobacterium gordonae Bojalil et al. (ATCC (R) 14470 (TM)). Results: Even with high dilutions, all tested essential oils showed a high antimycobacterial activity against targeted strains. Our data showed that M. barbata, E. globulus and J. excelsa essential oils totally inhibit the mycobacterial growth whatever the tested strains for the dilution 1/250, 1/100 and 1/250, respectively. Conclusion: To our knowledge, this is the first study regarding the antimycobacterial activity of essential oils in Lebanon. Our data show promising results, and encourage to investigate more on these medicinal plants, especially M. barbata

Pepsin and Trypsin Treatment Combined with Carvacrol: An Efficient Strategy to Fight <i>Pseudomonas aeruginosa</i> and <i>Enterococcus faecalis</i> Biofilms

Biofilms consist of microbial communities enclosed in a self-produced extracellular matrix which is mainly responsible of biofilm virulence. Targeting this matrix could be an effective strategy to control biofilms. In this work, we examined the efficacy of two proteolytic enzymes, pepsin and trypsin, to degrade P. aeruginosa and E. faecalis biofilms and their synergistic effect when combined with carvacrol. The minimum dispersive concentrations (MDCs) and the contact times of enzymes, as well as the minimal inhibitory concentrations (MICs) and contact times of carvacrol, were determined against biofilms grown on polystyrene surfaces. For biofilms grown on stainless steel surfaces, the combined pepsin or trypsin with carvacrol treatment showed more significant reduction of both biofilms compared with carvacrol treatment alone. This reduction was more substantial after sequential treatment of both enzymes, followed by carvacrol with the greatest reduction of 4.7 log CFU mL−1 (p P. aeruginosa biofilm and 3.3 log CFU mL−1 (p E. faecalis biofilm. Such improved efficiency was also obvious in the epifluorescence microscopy analysis. These findings demonstrate that the combined effect of the protease-dispersing activity and the carvacrol antimicrobial activity could be a prospective approach for controlling P. aeruginosa and E. faecalis biofilms