Design of Lactococcus lactis Strains Producing Garvicin A and/or Garvicin Q, Either Alone or Together with Nisin A or Nisin Z and High Antimicrobial Activity against Lactococcus garvieae

Lactococcus garvieae is a main ichthyopathogen in rainbow trout (Oncorhynchus mykiss, Walbaum) farming, although bacteriocinogenic L. garvieae with antimicrobial activity against virulent strains of this species have also been identified. Some of the bacteriocins characterized, such as garvicin A (GarA) and garvicin Q (GarQ), may show potential for the control of the virulent L. garvieae in food, feed and other biotechnological applications. In this study, we report on the design of Lactococcus lactis strains that produce the bacteriocins GarA and/or GarQ, either alone or together with nisin A (NisA) or nisin Z (NisZ). Synthetic genes encoding the signal peptide of the lactococcal protein Usp45 (SPusp45), fused to mature GarA (lgnA) and/or mature GarQ (garQ) and their associated immunity genes (lgnI and garI, respectively), were cloned into the protein expression vectors pMG36c, which contains the P32 constitutive promoter, and pNZ8048c, which contains the inducible PnisA promoter. The transformation of recombinant vectors into lactococcal cells allowed for the production of GarA and/or GarQ by L. lactis subsp. cremoris NZ9000 and their co-production with NisA by Lactococcus lactis subsp. lactis DPC5598 and L. lactis subsp. lactis BB24. The strains L. lactis subsp. cremoris WA2-67 (pJFQI), a producer of GarQ and NisZ, and L. lactis subsp. cremoris WA2-67 (pJFQIAI), a producer of GarA, GarQ and NisZ, demonstrated the highest antimicrobial activity (5.1- to 10.7-fold and 17.3- to 68.2-fold, respectively) against virulent L. garvieae strains.Sección Dptal. de Nutrición y Ciencia de los Alimentos (Veterinaria)Fac. de VeterinariaTRUEMinisterio de Ciencia, Innovación y Universidades (MCIU)Universidad Complutense de Madrid and Banco de SantanderUniversidad Complutense de Madridpu

Clonal Diversity and Antimicrobial Resistance of Methicillin-Resistant Staphylococcus pseudintermedius Isolated from Canine Pyoderma

[EN] The emergence of methicillin-resistant Staphylococcus pseudintermedius (MRSP) antimicrobial resistance and epidemic genetic lineages is posing a challenge in veterinary medicine due to the limited therapeutical options. MRSP has been identified as an important canine pyoderma pathogen. Thus, we aimed to characterize the antimicrobial resistance and clonal lineages of MRSP isolated from canine cutaneous pyoderma. Thirty-one MRSP isolates recovered from pyoderma were further characterized. The antimicrobial susceptibility testing of the isolates was performed by the KirbyBauer disc diffusion method against 14 antimicrobial agents. The presence of antimicrobial and virulence genes was carried out by PCR. Multilocus sequence typing was performed in all isolates. All strains had a multidrug-resistant profile showing resistance mainly to penicillin, macrolides and lincosamides, aminoglycosides, tetracycline and trimethoprim-sulfamethoxazole, which was encoded by the blaZ, ermB, msr(A/B), aac(6′ )-Ie-aph(2′′ )-Ia, aph(3′ )-IIIa, ant(4′ )-Ia, tetM, tetK and dfrG genes. All isolates harbored the lukS-I/lukF-I virulence factors. Isolates were ascribed to nine previously described sequence types (STs): ST123, ST339, ST727, ST71, ST537, ST45, ST1029, ST118 and ST1468; and to five STs first described in this study: ST2024, ST2025, ST2026, ST2027 and ST2028. In this study, most isolates belonged to ST123 (n = 16), which belongs to CC71 and is the most common clone in Europe. All isolates were multidrug-resistant, which may impose a serious threat to animal healthSIThis research was funded by the Ministerio de Ciencia, Innovación y Universidades (Spain, Project RTI2018-098267-R-C33) and the Junta de Castilla y León (Consejería de Educación, Spain, Project LE018P20). This work was funded by the R&D Project CAREBIO2: Comparative assessment of antimicrobial resistance in environmental biofilms through proteomics—towards innovative thera- nostic biomarkers, with reference NORTE-01-0145-FEDER-030101 and PTDC/SAU-INF/30101/2017, financed by the European Regional Development Fund (ERDF) through the Northern Regional Operational Program (NORTE 2020) and the Foundation for Science and Technology (FCT). This work was supported by the Associate Laboratory for Green Chemistry-LAQV, which is financed by national funds from FCT/MCTES (UIDB/50006/2020 and UIDP/50006/2020). Vanessa Silva is grateful to FCT (Fundação para a Ciência e a Tecnologia) for financial support through the PhD grant SFRH/BD/137947/201

Evaluación farmacodinámica y análisis físico-químico de dos formulaciones de propofol usadas en infusión objeto-controlada

BACKGROUND AND OBJECTIVES: There are several formulations of propofol available to the anesthesiologist for clinical use. The aim of this study was to analyze the physicochemical properties, pharmacodynamic effect, and pharmaceutical and clinical equivalence of the reference drug propofol as well as a similar formulation. METHOD: Sixteen volunteers were enrolled in this randomized, double-blind, and paired study of Diprivan® and Propovan® formulations. Formulations were given as target-controlled infusion with target concentration of 3.0 μg.mL-1 for 15 minutes. Variables studied were the area under the curve (AUC) of the bispectral index (BIS) graph regarding time, minimum BIS reached and time to reach it, and recovery time. The two formulations were sent to analysis of particle size of lipid emulsion, surface potential, and active principle quantification. RESULTS: There was no difference between the formulations when comparing AUC, minimum BIS reached and time to reach it. The similar formulation recovery time was lower compared to the reference formulation (eight and 10 min, respectively, p = 0.014). Mean particle size of lipid emulsion, surface potential, and active ingredient quantification were similar for both formulations. CONCLUSION: There was no clinically significant difference between the use of propofol, reference Diprivan®, and the similar Propovan® during infusion. However, the recovery time was longer with the reference drug. Although analysis of both formulations studied show similar results regarding its physicochemical characterization, further studies should be conducted to justify this difference.JUSTIFICATIVA Y OBJETIVOS: Existen varias formulaciones de propofol para el uso clínico que están disponibles para el anestesiólogo. El objetivo de este estudio, fue analizar las propiedades físico-químicas, el efecto farmacodinámico y la equivalencia farmacéutica y clínica del fármaco referencia de propofol y una formulación similar. MÉTODO: Dieciséis voluntarios participaron en este estudio aleatorio, doble ciego y pareado entre las formulaciones Diprivan® y Propovan®. Las formulaciones fueron administradas en un régimen de infusión objeto-controlada con una concentración objetivo de 3,0 µg.mL-1 durante 15 minutos. Las variables estudiadas fueron el área bajo la curva (ASC) del gráfico del índice bispectral (BIS) con relación al tiempo, el BIS mínimo alcanzado y el tiempo para tal, y el tiempo de recuperación. Las dos formulaciones se sometieron a los análisis de tamaño de partículas de la emulsión lipídica, potencial de superficie y cuantificación del principio activo. RESULTADOS: No hubo diferencia entre las formulaciones cuando se comparó la ASC, el BIS mínimo alcanzado y el tiempo transcurrido para tal. El tiempo de recuperación con la formulación similar fue menor con relación a la referencia (8 y 10 min, respectivamente, p = 0,014). El tamaño promedio de partículas de la emulsión lipídica, potencial de superficie y la cuantificación del principio activo, fueron similares en las dos formulaciones. CONCLUSIONES: No hubo diferencia clínica significativa entre el uso de propofol referencia Diprivan® y su similar Propovan® durante la infusión. Sin embargo, el tiempo de recuperación se extendió más con el fármaco de referencia. Aunque los análisis de las formulaciones estudiadas muestren resultados similares en cuanto a su caracterización físico-química, otros estudios deben ser realizados para justificar tal diferencia.JUSTIFICATIVA E OBJETIVOS: Existem várias formulações de propofol para uso clínico à disposição do anestesiologista. O objetivo desse estudo foi analisar as propriedades físico-químicas, o efeito farmacodinâmico e a equivalência farmacêutica e clínica do fármaco referência de propofol e uma formulação similar. MÉTODOS: Dezesseis voluntários participaram desse estudo aleatório, duplamente encoberto e pareado entre as formulações Diprivan® e Propovan®. As formulações foram administradas em regime de infusão alvo-controlada com concentração-alvo de 3,0 µg.mL-1 por 15 minutos. As variáveis estudadas foram a área sob a curva (ASC) do gráfico do índice bispectral (BIS) em relação ao tempo, o BIS mínimo atingido e o tempo para tal e o tempo de recuperação. As duas formulações foram submetidas às análises de tamanho de partículas da emulsão lipídica, potencial de superfície e quantificação de princípio ativo. RESULTADOS: Não houve diferença entre as formulações quando se comparou a ASC, BIS mínimo atingido e o tempo decorrido para tal. O tempo de recuperação com a formulação similar foi menor em relação à referência (oito e 10 min, respectivamente, p = 0,014). O tamanho médio de partículas da emulsão lipídica, potencial de superfície e a quantificação de princípio ativo foram semelhantes nas duas formulações. CONCLUSÃO: Não houve diferença clínica significativa entre o uso de propofol referência Diprivan® e seu similar Propovan® durante a infusão. Entretanto, o tempo de recuperação foi mais prolongado com o fármaco referência. Embora as análises com as duas formulações estudadas mostrarem resultados semelhantes quanto a sua caracterização físico-química, outros estudos devem ser realizados para justificar tal diferença.Instituto Penido Burnier Sociedade Brasileira de Anestesiologia Centro de Ensino e TreinamentoCentro Médico de CampinasHospital Santa SofiaUniversidade de São Paulo Faculdade de Ciências FarmacêuticasUniversidade Federal de São Paulo (UNIFESP)UNIFESPSciEL

Immunomodulatory effects of bacteriocinogenic and non-bacteriocinogenic Lactococcus cremoris of aquatic origin on rainbow trout (Oncorhynchus mykiss, Walbaum)

11 Pág.Lactic Acid Bacteria (LAB) are a group of bacteria frequently proposed as probiotics in aquaculture, as their administration has shown to confer positive effects on the growth, survival rate to pathogens and immunological status of the fish. In this respect, the production of antimicrobial peptides (referred to as bacteriocins) by LAB is a common trait thoroughly documented, being regarded as a key probiotic antimicrobial strategy. Although some studies have pointed to the direct immunomodulatory effects of these bacteriocins in mammals, this has been largely unexplored in fish. To this aim, in the current study, we have investigated the immunomodulatory effects of bacteriocins, by comparing the effects of a wild type nisin Z-expressing Lactococcus cremoris strain of aquatic origin to those exerted by a non-bacteriocinogenic isogenic mutant and a recombinant nisin Z, garvicin A and Q-producer multi-bacteriocinogenic strain. The transcriptional response elicited by the different strains in the rainbow trout intestinal epithelial cell line (RTgutGC) and in splenic leukocytes showed significant differences. Yet the adherence capacity to RTgutGC was similar for all strains. In splenocyte cultures, we also determined the effects of the different strains on the proliferation and survival of IgM+ B cells. Finally, while the different LAB elicited respiratory burst activity similarly, the bacteriocinogenic strains showed an increased ability to induce the production of nitric oxide (NO). The results obtained reveal a superior capacity of the bacteriocinogenic strains to modulate different immune functions, pointing to a direct immunomodulatory role of the bacteriocins, mainly nisin Z.This work was supported by the Ministerio de Ciencia e Innovación (MICINN, Madrid, Spain) (projects RTI2018-094907-B-I00, and PID2020-113268RB-I00), and the Universidad Complutense de Madrid (UCM) (project FEI16/54). DC was supported by a contract from the project RTI2018-094907-B-I00. JF was supported by a FEI16/54 contract and held a predoctoral contract from UCM. LD-F was supported by a contract from the “Programa Investigo” (Ministerio de Trabajo y Economía Social, Madrid, Spain), funded by the EU (NextGenerationEU). JB was supported by the Atracción de Talento, program of the Comunidad de Madrid, Spain (2018-T1/BIO-10158).Peer reviewe

Draft Genome Sequence of Lactococcus lactis Subsp. cremoris WA2-67: A Promising Nisin-Producing Probiotic Strain Isolated from the Rearing Environment of a Spanish Rainbow Trout (Oncorhynchus mykiss, Walbaum) Farm

Probiotics are a viable alternative to traditional chemotherapy agents to control infectious diseases in aquaculture. In this regard, Lactococcus lactis subsp. cremoris WA2-67 has previously demonstrated several probiotic features, such as a strong antimicrobial activity against ichthyopathogens, survival in freshwater, resistance to fish bile and low pH, and hydrophobicity. The aim of this manuscript is an in silico analysis of the whole-genome sequence (WGS) of this strain to gain deeper insights into its probiotic properties and their genetic basis. Genomic DNA was purified, and libraries prepared for Illumina sequencing. After trimming and assembly, resulting contigs were subjected to bioinformatic analyses. The draft genome of L. cremoris WA2-67 consists of 30 contigs (2,573,139 bp), and a total number of 2493 coding DNA sequences (CDSs). Via in silico analysis, the bacteriocinogenic genetic clusters encoding the lantibiotic nisin Z (NisZ) and two new bacteriocins were identified, in addition to several probiotic traits, such as the production of vitamins, amino acids, adhesion/aggregation, and stress resistance factors, as well as the absence of transferable antibiotic resistance determinants and genes encoding detrimental enzymatic activities and virulence factors. These results unveil diverse beneficial properties that support the use of L. cremoris WA2-67 as a probiotic for aquaculture

Genomic Sequence of Streptococcus salivarius MDI13 and Latilactobacillus sakei MEI5: Two Promising Probiotic Strains Isolated from European Hakes (Merluccius merluccius, L.)

FEI16/54 Universidad Complutense de Madrid 2018-T1/BIO-10158 Programa Atracción del Talento de la Comunidad de MadridSimple Summary: In fish farming, diseases have commonly been fought with the abusive use of antibiotics, which have caused antibiotic (multi)resistances in bacteria. Consequently, it is necessary to explore safe and environmentally friendly alternative approaches to improve the fish health and avoid the treatment of bacterial diseases with antibiotics such as the use of probiotics. This study focuses on bioinformatic and functional analyses of the genome sequences of Streptococcus salivarius MDI13 and Latilactobacillus sakei MEI5, two Lactic Acid Bacteria (LAB) isolated from the gut of European hakes (Merluccius merluccius, L.), a highly valued marine fish for Spanish gastronomy. The potential probiotic characteristics of both bacteria, and the lack of antibiotic resistance genes and virulence were confirmed. In addition, genes encoding three antimicrobial peptides (known as bacteriocins) were identified in the genome of S. salivarius MDI13. One of these in vitro-synthesized bacteriocins (BlpK) showed antimicrobial activity against two fish pathogens (namely Lactococcus garvieae and Streptococcus parauberis). Altogether, our results suggest that S. salivarius MDI13 and L. sakei MEI5 have a strong potential as probiotics to prevent bacterial diseases in fish farming.Abstract: Frequently, diseases in aquaculture have been fought indiscriminately with the use of antibiotics, which has led to the development and dissemination of (multiple) antibiotic resistances in bacteria. Consequently, it is necessary to look for alternative and complementary approaches to chemotheraphy that are safe for humans, animals, and the environment, such as the use of probiotics in fish farming. The objective of this work was the Whole-Genome Sequencing (WGS) and bioinformatic and functional analyses of S. salivarius MDI13 and L. sakei MEI5, two LAB strains isolated from the gut of commercial European hakes (M. merluccius, L.) caught in the Northeast Atlantic Ocean. The WGS and bioinformatic and functional analyses confirmed the lack of transferable antibiotic resistance genes, the lack of virulence and pathogenicity issues, and their potentially probiotic characteristics. Specifically, genes involved in adhesion and aggregation, vitamin biosynthesis, and amino acid metabolism were detected in both strains. In addition, genes related to lactic acid production, active metabolism, and/or adaptation to stress and adverse conditions in the host gastrointestinal tract were detected in L. sakei MEI5. Moreover, a gene cluster encoding three bacteriocins (SlvV, BlpK, and BlpE) was identified in the genome of S. salivarius MDI13. The in vitro-synthesized bacteriocin BlpK showed antimicrobial activity against the ichthyopathogens Lc. garvieae and S. parauberis. Altogether, our results suggest that S. salivarius MDI13 and L. sakei MEI5 have a strong potential as probiotics to prevent fish diseases in aquaculture as an appropriate alternative/complementary strategy to the use of antibiotics.Ministerio de Ciencia, Innovación y Universidades (España)Ministerio de Trabajo y Economía Social (España)European CommissionUniversidad Complutense de MadridComunidad de MadridSección Dptal. de Nutrición y Ciencia de los Alimentos (Veterinaria)Fac. de VeterinariaTRUEpu

Genomic and Functional Evaluation of Two Lacticaseibacillus paracasei and Two Lactiplantibacillus plantarum Strains, Isolated from a Rearing Tank of Rotifers (Brachionus plicatilis), as Probiotics for Aquaculture

Author Contributions: Conceptualization, L.M.C. and P.P.; methodology, D.C., L.D.-F., J.F. and E.M.-A.; software, D.C. and L.D.-F.; validation, J.F. and J.B.; formal analysis, D.C., J.F. and P.E.H.; investigation, D.C., L.D.-F., J.F. and E.M.-A.; resources, J.B., P.P. and L.M.C.; data curation, E.M.- A.; writing—original draft preparation, D.C.; writing—review and editing, L.M.C., P.P. and P.E.H.; visualization, D.C., J.F., P.E.H. and E.M.-A.; supervision, L.M.C. and P.P.; project administration, L.M.C. and J.B.; funding acquisition, L.M.C. and J.B. All authors have read and agreed to the published version of the manuscript.Aquaculture plays a crucial role in meeting the increasing global demand for food and protein sources. However, its expansion is followed by increasing challenges, such as infectious disease outbreaks and antibiotic misuse. The present study focuses on the genetic and functional analyses of two Lacticaseibacillus paracasei (BF3 and RT4) and two Lactiplantibacillus plantarum (BF12 and WT12) strains isolated from a rotifer cultivation tank used for turbot larviculture. Whole-genome sequencing (WGS) and bioinformatics analyses confirmed their probiotic potential, the absence of transferable antibiotic resistance genes, and the absence of virulence and pathogenicity factors. Bacteriocin mining identified a gene cluster encoding six plantaricins, suggesting their role in the antimicrobial activity exerted by these strains. In vitro cell-free protein synthesis (IV-CFPS) analyses was used to evaluate the expression of the plantaricin genes. The in vitro-synthesized class IIb (two-peptide bacteriocins) plantaricin E/F (PlnE/F) exerted antimicrobial activity against three indicator microorganisms, including the well-known ichthyopathogen Lactococcus garvieae. Furthermore, MALDI-TOF MS on colonies detected the presence of a major peptide that matches the dimeric form of plantaricins E (PlnE) and F (PlnF). This study emphasizes the importance of genome sequencing and bioinformatic analysis for evaluating aquaculture probiotic candidates. Moreover, it provides valuable insights into their genetic features and antimicrobial mechanisms, paving the way for their application as probiotics in larviculture, which is a major bottleneck in aquaculture.Ministerio de Ciencia, Innovación y Universidades (España)Sección Dptal. de Nutrición y Ciencia de los Alimentos (Veterinaria)Fac. de VeterinariaTRUEpu

Avaliação farmacodinâmica e análise físico-química de duas formulações de propofol usadas em infusão alvo-controlada Evaluación farmacodinámica y análisis físico-químico de dos formulaciones de propofol usadas en infusión objeto-controlada Pharmacodynamic evaluation and physical/chemical analysis of two formulations of propofol used in target-controlled infusion

JUSTIFICATIVA E OBJETIVOS: Existem várias formulações de propofol para uso clínico à disposição do anestesiologista. O objetivo desse estudo foi analisar as propriedades físico-químicas, o efeito farmacodinâmico e a equivalência farmacêutica e clínica do fármaco referência de propofol e uma formulação similar. MÉTODOS: Dezesseis voluntários participaram desse estudo aleatório, duplamente encoberto e pareado entre as formulações Diprivan® e Propovan®. As formulações foram administradas em regime de infusão alvo-controlada com concentração-alvo de 3,0 µg.mL-1 por 15 minutos. As variáveis estudadas foram a área sob a curva (ASC) do gráfico do índice bispectral (BIS) em relação ao tempo, o BIS mínimo atingido e o tempo para tal e o tempo de recuperação. As duas formulações foram submetidas às análises de tamanho de partículas da emulsão lipídica, potencial de superfície e quantificação de princípio ativo. RESULTADOS: Não houve diferença entre as formulações quando se comparou a ASC, BIS mínimo atingido e o tempo decorrido para tal. O tempo de recuperação com a formulação similar foi menor em relação à referência (oito e 10 min, respectivamente, p = 0,014). O tamanho médio de partículas da emulsão lipídica, potencial de superfície e a quantificação de princípio ativo foram semelhantes nas duas formulações. CONCLUSÃO: Não houve diferença clínica significativa entre o uso de propofol referência Diprivan® e seu similar Propovan® durante a infusão. Entretanto, o tempo de recuperação foi mais prolongado com o fármaco referência. Embora as análises com as duas formulações estudadas mostrarem resultados semelhantes quanto a sua caracterização físico-química, outros estudos devem ser realizados para justificar tal diferença.JUSTIFICATIVA Y OBJETIVOS: Existen varias formulaciones de propofol para el uso clínico que están disponibles para el anestesiólogo. El objetivo de este estudio, fue analizar las propiedades físico-químicas, el efecto farmacodinámico y la equivalencia farmacéutica y clínica del fármaco referencia de propofol y una formulación similar. MÉTODO: Dieciséis voluntarios participaron en este estudio aleatorio, doble ciego y pareado entre las formulaciones Diprivan® y Propovan®. Las formulaciones fueron administradas en un régimen de infusión objeto-controlada con una concentración objetivo de 3,0 µg.mL-1 durante 15 minutos. Las variables estudiadas fueron el área bajo la curva (ASC) del gráfico del índice bispectral (BIS) con relación al tiempo, el BIS mínimo alcanzado y el tiempo para tal, y el tiempo de recuperación. Las dos formulaciones se sometieron a los análisis de tamaño de partículas de la emulsión lipídica, potencial de superficie y cuantificación del principio activo. RESULTADOS: No hubo diferencia entre las formulaciones cuando se comparó la ASC, el BIS mínimo alcanzado y el tiempo transcurrido para tal. El tiempo de recuperación con la formulación similar fue menor con relación a la referencia (8 y 10 min, respectivamente, p = 0,014). El tamaño promedio de partículas de la emulsión lipídica, potencial de superficie y la cuantificación del principio activo, fueron similares en las dos formulaciones. CONCLUSIONES: No hubo diferencia clínica significativa entre el uso de propofol referencia Diprivan® y su similar Propovan® durante la infusión. Sin embargo, el tiempo de recuperación se extendió más con el fármaco de referencia. Aunque los análisis de las formulaciones estudiadas muestren resultados similares en cuanto a su caracterización físico-química, otros estudios deben ser realizados para justificar tal diferencia.BACKGROUND AND OBJECTIVES: There are several formulations of propofol available to the anesthesiologist for clinical use. The aim of this study was to analyze the physicochemical properties, pharmacodynamic effect, and pharmaceutical and clinical equivalence of the reference drug propofol as well as a similar formulation. METHOD: Sixteen volunteers were enrolled in this randomized, double-blind, and paired study of Diprivan® and Propovan® formulations. Formulations were given as target-controlled infusion with target concentration of 3.0 μg.mL-1 for 15 minutes. Variables studied were the area under the curve (AUC) of the bispectral index (BIS) graph regarding time, minimum BIS reached and time to reach it, and recovery time. The two formulations were sent to analysis of particle size of lipid emulsion, surface potential, and active principle quantification. RESULTS: There was no difference between the formulations when comparing AUC, minimum BIS reached and time to reach it. The similar formulation recovery time was lower compared to the reference formulation (eight and 10 min, respectively, p = 0.014). Mean particle size of lipid emulsion, surface potential, and active ingredient quantification were similar for both formulations. CONCLUSION: There was no clinically significant difference between the use of propofol, reference Diprivan®, and the similar Propovan® during infusion. However, the recovery time was longer with the reference drug. Although analysis of both formulations studied show similar results regarding its physicochemical characterization, further studies should be conducted to justify this difference

Pharmacodynamic Evaluation and Physical/Chemical Analysis of Two Formulations of Propofol used in Target-Controlled Infusion

Background and objectives: There are several formulations of propofol available to the anesthesiologist for clinical use. The aim of this study was to analyze the physicochemical properties, pharmacodynamic effect, and pharmaceutical and clinical equivalence of the reference drug propofol as well as a similar formulation. Method: Sixteen volunteers were enrolled in this randomized, double-blind, and paired study of Diprivan® and Propovan® formulations. Formulations were given as target-controlled infusion with target concentration of 3.0 μg.mL-1 for 15 minutes. Variables studied were the area under the curve (AUC) of the bispectral index (BIS) graph regarding time, minimum BIS reached and time to reach it, and recovery time. The two formulations were sent to analysis of particle size of lipid emulsion, surface potential, and active principle quantification. Results: There was no difference between the formulations when comparing AUC, minimum BIS reached and time to reach it. The similar formulation recovery time was lower compared to the reference formulation (eight and 10 min, respectively, p = 0.014). Mean particle size of lipid emulsion, surface potential, and active ingredient quantification were similar for both formulations. Conclusion: There was no clinically significant difference between the use of propofol, reference Diprivan®, and the similar Propovan® during infusion. However, the recovery time was longer with the reference drug. Although analysis of both formulations studied show similar results regarding its physicochemical characterization, further studies should be conducted to justify this difference. Keywords: Anesthesia, Intravenous, Consciousness Monitors, Pharmacology, Clinical, Propofo

Antimicrobial Activity, Genetic Relatedness, and Safety Assessment of Potential Probiotic Lactic Acid Bacteria Isolated from a Rearing Tank of Rotifers (Brachionus plicatilis) Used as Live Feed in Fish Larviculture

2024 Descuento MDPIAquaculture is a rapidly expanding agri-food industry that faces substantial economic losses due to infectious disease outbreaks, such as bacterial infections. These outbreaks cause disruptions and high mortalities at various stages of the rearing process, especially in the larval stages. Probiotic bacteria are emerging as promising and sustainable alternative or complementary strategies to vaccination and the use of antibiotics in aquaculture. In this study, potential probiotic candidates for larviculture were isolated from a rotifer-rearing tank used as the first live feed for turbot larvae. Two Lacticaseibacillus paracasei and two Lactiplantibacillus plantarum isolates were selected for further characterization due to their wide and strong antimicrobial activity against several ichthyopathogens, both Gram-positive and Gram-negative. An extensive in vitro safety assessment of these four isolates revealed the absence of harmful traits, such as acquired antimicrobial resistance and other virulence factors (i.e., hemolytic and gelatinase activities, bile salt deconjugation, and mucin degradation, as well as PCR detection of biogenic amine production). Moreover, Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR) analyses unveiled their genetic relatedness, revealing two divergent clusters within each species. To our knowledge, this work reports for the first time the isolation and characterization of Lactic Acid Bacteria (LAB) with potential use as probiotics in aquaculture from rotifer-rearing tanks, which have the potential to optimize turbot larviculture and to introduce novel microbial management approaches for a sustainable aquaculture.Ministerio de Ciencia, Innovación y Universidades (España)Fundação para a Ciência e a Tecnologia (Portugal)Comunidad de MadridEuropean CommissionMinisterio de Trabajo y Economía Social (España)Depto. de Nutrición y Ciencia de los AlimentosFac. de VeterinariaTRUEpubDescuento UC