Search

Marine-derived actinobacteria isolated from sponge Cliona varians and soft coral Eunicea fusca were screened for antibacterial activity against acne-related bacteria, specifically Staphylococcus epidermidis ATCC 14990, methicillin-resistant Staphylococcus aureus ATCC BAA44, and Cutibacterium acnes ATCC 6919. Cytotoxicity assays were performed on human dermal fibroblast (HDFa) and keratinocyte (HaCaT) cell lines to assess the safety profile of the extracts. Chemical characterization was conducted using high-performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS). Among the extracts, six derived from Kocuria sp., Rhodococcus sp., Nocardia sp., Micrococcus sp., and Streptomyces sp. demonstrated significant antibacterial activity. Notably, extract Z9.216 from Kocuria sp. exhibited the highest efficacy, inhibiting S. epidermidis by 68%, S. aureus by 93%, and C. acnes by 98.7% at a concentration of 0.003 mg/mL, which was comparable to the standard antibiotics erythromycin and vancomycin, while maintaining over 90% cell viability in both HDFa and HaCaT cell lines. Untargeted metabolomic analysis suggested that antibacterial activity might be associated with compounds from the chemical families of alkaloids, terpenoids, and fatty acids, among others. These findings highlight the therapeutic potential of marine actinobacteria in underexplored environments as a promising strategy for treating acne vulgaris, a chronic inflammatory skin condition., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2025 De La Hoz-Romo, Díaz, Gómez-León, Quintero and Villamil.)

Probiotics face harsh conditions during their transit through the gastrointestinal tract (GIT) of fish because of low-pH environments and intestine fluid. Therefore, the evaluation of probiotic viability under simulated gastrointestinal conditions is an important step to consider for probiotic supplementation in fish feed prior to in vivo trials. Therefore, this study aimed to evaluate the effect of stomach and intestinal simulated conditions on the viability of encapsulated Lactococcus lactis A12 using an in vitro digestion model for tilapia. A Box Behnken design was used to evaluate the potential effect of three factors, namely stomach pH, residence time in the stomach, and enzyme quantity, on the viability of encapsulated Lactococcus lactis A12. As the main results, low pH (4.00), long residence time (4 h), and enzyme quantity (2.68 U of total protease activity) led to lower final cell counts after the phases of the stomach and intestine. Encapsulated probiotic bacteria showed higher viability ( p < 0.05) and antibacterial activity ( p < 0.05) against the pathogen Streptococcus agalactiae than non-encapsulated bacteria. The results suggest that L. lactis A12 survives in GIT conditions and that the proposed in vitro model could be used to explore the viability of probiotic bacteria intended for fish feed supplementation.

UV filters in current sunscreen formulations can have negative effects on human health, such as endocrine disruption and allergic reactions, as well as on the environment, including bioaccumulation and coral health toxicity. As a result, there is a need to find alternative compounds that serve as safer and more ecofriendly active ingredients. This study successfully isolated actinomycetes from the octocoral Eunicea fusca and assessed their potential as producers of photoprotective compounds. The use of bio-based chemical agents, particularly natural products, has been a highly effective strategy for discovering bioactive compounds, especially in marine invertebrates and their associated microbiota. Eighteen bacterial isolates were obtained and subsequently employed to prepare raw methanolic extracts from seven-day submerged cultures in Zobell marine broth. The resulting extracts were screened for 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging capacity and characterized by total phenolic and flavonoid content measurements. After screening, the Gordonia hongkongensis EUFUS-Z928-derived raw extract exhibited the best antioxidant profile, i.e. DPPH and ABTS radical scavenging of 4.93 and 6.00 µmol Trolox per gram of extract, respectively, and selected for further photoprotection-related analysis. Thus, this extract demonstrated a UV-absorbing capacity of 46.33% of the in vitro sun protection factor calculated for 30 µg/mL oxybenzone but did not exhibit any cytotoxicity on human dermal fibroblasts (HDFa cell line) at concentrations up to 500 µg/mL. The liquid chromatography-mass spectrometry chemical characterization of this extract showed compounds with structural features associated with free radical scavenging and UV absorption (i.e. photoprotection-related activities). These findings highlighted the potential of the microbiota associated with E. fusca and confirmed the feasibility of exploiting its metabolites for photoprotection-related purposes., Competing Interests: Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

The current emphasis within the cosmetic market on sustainable ingredients has heightened the exploration of new sources for natural, active components. Actinomycetota , recognized for producing pigments with bioactive potential, offer promising functional cosmetic ingredients. This study aimed to optimize pigment and antioxidant metabolite production from the Gordonia hongkongensis strain EUFUS-Z928 by implementing the Plackett-Burman experimental design and response surface methodology. Extracts derived from this strain exhibited no cytotoxic activity against human primary dermal fibroblast (HDFa, ATCC® PCS-201-012™, Primary Dermal Fibroblast; Normal, Human, Adult). Eight variables, including inoculum concentration, carbon and nitrogen source concentration, NaCl concentration, pH, incubation time, temperature, and stirring speed, were analyzed using the Plackett-Burman experimental design. Subsequently, factors significantly influencing pigment and antioxidant metabolite production, such as temperature, inoculum concentration, and agitation speed, were further optimized using response surface methodology and Box-Behnken design. The results demonstrated a substantial increase in absorbance (from 0.091 to 0.32), DPPH radical scavenging capacity (from 27.60% to 84.61%), and ABTS radical scavenging capacity (from 17.39% to 79.77%) compared to responses obtained in the isolation medium. The validation of the mathematical model accuracy exceeded 90% for all cases. Furthermore, liquid chromatography coupled with mass spectrometry (LC-MS) facilitated the identification of compounds potentially responsible for enhanced pigment production and antioxidant capacity in extracts derived from G. hongkongensis . Specifically, six carotenoids, red-orange pigments with inherent antioxidant capacity, were identified as the main enhanced compounds. This comprehensive approach effectively optimized the culture conditions and medium of a G. hongkongensis strain, resulting in enhanced carotenoid production and antioxidant capacity. Beyond identifying bioactive compounds and their potential cosmetic applications, this study offers insights into the broader industrial applicability of these extracts. It underscores the potential of G. hongkongensis and hints at the future utilization of other untapped sources of rare actinomycetes within the industry., Competing Interests: Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Background: Probiotics are viable microorganisms that when administered in adequate amounts confer health benefits to the host. In fish, probiotic administration has improved growth, and immunological parameters. For this reason, it is necessary production of probiotic bacteria, however, commercial culture mediums used for probiotic growth are expensive, so the design of a "low" cost culture medium is necessary. Therefore, this research aimed to produce a potential multistrain probiotic preparation composed of L. lactis A12 and Priestia species isolated from Nile tilapia (Oreochromis niloticus) gut using an agro-industrial by-products-based culture medium., Results: A Box-Behnken design with three factors (whey, molasses, and yeast extract concentration) was used. As the main results, a high concentration of three components enhanced the viability of L. lactis A12, however, viable cell counts of Priestia species were achieved at low molasses concentrations. The Optimal conditions were 1.00% w/v whey, 0.50% w/v molasses, and 1.50% w/v yeast extract. L. lactis A12 and Priestia species viable counts were 9.43 and 6.89 Log 10 CFU/mL, respectively. L. lactis A12 concentration was higher (p < 0.05) in the proposed medium compared to commercial broth., Conclusions: It was possible to produce L. lactis A12 and Priestia species in co-culture conditions. Whey and molasses were suitable components to produce the multistrain preparation. The cost of the proposed culture medium was 77.54% cheaper than the commercial medium. The proposed culture medium could be an alternative to commercial mediums for the production of this multistrain probiotic., (© 2023. The Author(s).)

Objective: Data on the frequency of cardiovascular disease in people living with human immunodeficiency virus from low- and middle-income countries is scarce., Methods: We performed an observational study based on data from a historical cohort of people living with human immunodeficiency virus in Colombia during a 10-year follow-up to describe the prevalence of cardiovascular risk factors and their behavior according to CD4 count., Results: One thousand patients were initially included, out of which 390 had a 10-year follow-up. The mean age was 34 (standard deviation 10) years, and 90% were male. We observed an increase in the prevalence of dyslipidemia (29%-52%, p  < 0.001) and obesity (1.1%-3.5%, p  < 0.001). Major cardiovascular events occurred in less than 1% of patients. Patients with a CD4 count <200 cells/mm 3 had a higher frequency of acute myocardial infarction and obesity., Conclusion: Over time, people living with human immunodeficiency virus present with an increasing prevalence of cardiovascular risk factors, particularly those with a lower CD4 count., Competing Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2023.)

Members of the phylum Actinomycetota (formerly Actinobacteria ) have historically been the most prolific providers of small bioactive molecules. Although the genus Streptomyces is the best-known member for this issue, other genera, such as Gordonia , have shown interesting potential in their specialized metabolism. Thus, we combined herein the result of a comprehensive literature survey on metabolites derived from Gordonia strains with a comparative genomic analysis to examine the potential of the specialized metabolism of the genus Gordonia . Thirty Gordonia -derived compounds of different classes were gathered (i.e., alkaloids, amides, phenylpropanoids, and terpenoids), exhibiting antimicrobial and cytotoxic activities, and several were also isolated from Streptomyces (e.g., actinomycin, nocardamin, diolmycin A1). With the genome data, we estimated an open pan-genome of 57,901 genes, most of them being part of the cloud genome. Regarding the BGCs content, 531 clusters were found, including Terpenes, RiPP-like, and NRPS clusters as the most frequent clusters. Our findings demonstrated that Gordonia is a poorly studied genus in terms of its specialized metabolism production and potential applications. Nevertheless, given their BGCs content, Gordonia spp. are a valuable biological resource that could expand the chemical spectrum of the phylum Actinomycetota , involving novel BGCs for inspiring innovative outlines for synthetic biology and further use in biotechnological initiatives. Therefore, further studies and more efforts should be made to explore different environments and evaluate other bioactivities.

Aims: This work aims to characterize the microbial diversity of the encrusting sponge Cliona varians, a pore-forming and coral reef bioeroding marine sponge of emerging spread related to ocean acidification., Methods and Results: We analysed the microbiome composition by 16S V4 amplicon next-generation sequencing in a community of the bioeroding coral reef encrusting/excavating marine sponge Cliona varians thriving at the Southern Caribbean Sea. About 87.21% and 6.76% of the sequences retrieved were assigned to the domain Bacteria and Archaea. The most predominant operational taxonomic units were classified as members of the order Rhizobiales and family Nitrosopumilaceae, representing members of not yet characterized genera. Features found strictly conserved in the strain/genomic representatives reported in those microbial taxa are nitrogen fixation and transformation., Conclusion: Our results suggest, in accordance with recent results, that these microbiome members and associated functions could be contributing to the biological fitness of the sponge to be able to colonize and bioerode in environments with low access and scarce availability of nitrogen sources., Significance and Impact of Study: Coral reefs bioresources such as sponge holobionts are intriguing and complex ecosystem units. This study contributes to the knowledge of how C. varians microbiota is composed or shaped, which is crucial to understand its ecological functions., (© 2022 Society for Applied Microbiology.)

Microbiome components and bacterial isolates related to healthy and epitheliocystis states in aquaculture cycles of cobia fish were studied. We detected well-defined 16S rRNA amplicon gene sequence variants showing differential abundance in healthy or diseased cycles. Isolation trials were performed, and experimental tests were used to determine probiotic potential of the bacterial strains obtained from water, tissues or live food used in this aquaculture model. The taxonomic affiliation of these strains was cross-compared against microbiome components, finding that some of them had close or identical affiliation to the abundant types found in healthy cycles. Strains belonging to the groups already identified as predominant by culture-independent means were screened as potential probiotics based on desirable activities such as antagonism and antibiosis against marine pathogenic bacteria, quorum quenching, bile acid resistance, antibiotic sensitivity and enzymatic activities for improved nutrient digestion. We have also found that in the tracking of microbiome composition across different developmental stages of cobia, healthy cycles exhibited a consistent high relative abundance of a Mesobacillus sp., while in the diseased cycle the emergence of a Vibrio sp. was observed. Our study suggests that epithelocystis in cobia is associated with a displacement of a symbiotic microbiome community linked to the increase frequency of Vibrio species., Competing Interests: Authors declare no conflict of interest., (© 2022 The Authors.)

Acne vulgaris is a multifactorial disease that remains under-explored; up to date it is known that the bacterium Cutibacterium acnes is involved in the disease occurrence, also associated with a microbial dysbiosis. Antibiotics have become a mainstay treatment generating the emergence of antibiotic-resistant bacteria. In addition, there are some reported side effects of alternative treatments, which indicate the need to investigate a different therapeutic approach. Natural products continue to be an excellent option, especially those extracted from actinobacteria, which represent a prominent source of metabolites with a wide range of biological activities, particularly the marine actinobacteria, which have been less studied than their terrestrial counterparts. Therefore, this systematic review aimed to identify and evaluate the potential anti-infective activity of metabolites isolated from marine actinobacteria strains against bacteria related to the development of acne vulgaris disease. It was found that there is a variety of compounds with anti-infective activity against Staphylococcus aureus and Staphylococcus epidermidis , bacteria closely related to acne vulgaris development; nevertheless, there is no report of a compound with antibacterial activity or quorum-sensing inhibition toward C. acnes , which is a surprising result. Since two of the most widely used antibiotics for the treatment of acne targeting C. acnes were obtained from actinobacteria of the genus Streptomyces , this demonstrates a great opportunity to pursue further studies in this field, considering the potential of marine actinobacteria to produce new anti-infective compounds.

Metallothionein-like protein concentrations (MT) and three functionally defined fractions of cholinesterase activity (ChE) were quantified in gill and digestive gland homogenates of tropical cup oysters from 5 nearshore locations in the Colombian Caribbean and correlated with sediment and tissue metal (9 metals) and pesticide (22 organophosphates, OPs, and 20 organochlorines-OCPs), as well as water physical-chemical parameters (salinity, pH, temperature, and dissolved oxygen). Tissue and sediment pesticide concentrations were below detection limits in all samples, whereas sediment and tissue metal concentrations exceeded environmental thresholds at several locations. Tissue MT and ChE biomarkers varied by a factor of 5-6 between locations. Inhibition of cholinesterase activity was negligible for all 5 sites, despite spatial-temporal variation in ChE activity, consistent with below-detection OP concentrations. Tissue MT and ChE biomarkers correlated with tissue and metal sediment concentrations, yet, statistically significant covariance between biomarkers and water chemistry parameters was also observed, indicating that both, metal concentrations and physical-chemical variables, are likely to be responsible for generating the observed spatial-temporal variations in biomarker patterns., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Octocorals are among the most prolific sources of biologically active compounds. A significant part of their specialized metabolites richness is linked to the abundance of their associated microbiota. Consequently, research on the bioprospecting potential of microorganisms associated with these marine invertebrates has gained much interest. Here, we describe the draft genome of Gordonia hongkongensis strain EUFUS-Z928 isolated from the octocoral Eunicea fusca . The genome was assembled de novo from short-read whole-genome sequencing data. Additionally, functional annotation of predicted genes was performed using the RAST tool kit, including genome mining for specialized metabolite biosynthetic gene clusters using the antiSMASH v6.0 tool. The genome sequence data of G. hongkongensis EUFUS-Z928 can provide information for further analysis of the potential biotechnological use of this microorganism and guide the characterization of other related actinobacterial isolates. Likewise, this information increases the analytical capacity for studying the genus Gordonia ., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Author(s).)

Marine invertebrates are a significant source of biologically active compounds. Recent studies have highlighted the role of microbiota associated with marine invertebrates in the production of bioactive compounds. Corals and sponges are the main marine invertebrates producing bioactive substances, and Symbiodiniaceae dinoflagellates are well-recognized endosymbionts with corals and sponges playing vital functions. The biological properties of Symbiodiniaceae-derived compounds have garnered attention in the past decades owing to their ecological implications and potentiality for bioprospecting initiatives. This study aims to systematically review studies on bioactivities and potential biotechnological applications of Symbiodiniaceae-derived compounds. The PRISMA guidelines were followed. Our study showed that anti-inflammatory and vasoconstrictive activities of Symbiodiniaceae-derived compounds have been the most investigated. However, very few studies have been published, with in vitro culturing of Symbiodiniaceae being the most significant challenge. Therefore, we surveyed for the metabolites reported so far, analyzed their chemodiversity, and discussed approaches to overcome culturing-related limitations., Competing Interests: The authors declare that there are no conflicts of interest regarding the publication of this paper., (Copyright © 2021 Jeysson Sánchez-Suárez et al.)

Sunscreen and sunblock are crucial skincare products to prevent photoaging and photocarcinogenesis through the addition of chemical filters to absorb or block ultraviolet (UV) radiation. However, several sunscreen and sunblock ingredients, mostly UV filters, have been associated with human and environmental safety concerns. Therefore, the exploration and discovery of promising novel sources of efficient and safer compounds with photoprotection-related activities are currently required. Marine invertebrates, particularly their associated microbiota, are promising providers of specialized metabolites with valuable biotechnological applications. Nevertheless, despite Actinobacteria members being a well-known source of bioactive metabolites, their photoprotective potential has been poorly explored so far. Hence, a set of methanolic extracts obtained from Cliona varians -derived actinomycetes was screened regarding their antioxidant and UV-absorbing capacities (i.e., photoprotection-related activities). The active extract-producing strains were identified and classified within genera Streptomyces, Micrococcus, Gordonia, and Promicromonospora. This is the first report of the isolation of these microorganisms from C. varians (an ecologically important Caribbean coral reef-boring sponge). The in vitro cytotoxicity on dermal fibroblasts of oxybenzone and the selected active extracts revealed that oxybenzone exerted a cytotoxic effect, whereas no cytotoxic effect of test extracts was observed. Accordingly, the most active (SPFi > 5, radical scavenging > 50%) and nontoxic (cell viability > 75%) extracts were obtained from Streptomyces strains. Finally, LC-MS-based characterization suggested a broad chemical space within the test strains and agreed with the reported streptomycetes' chemodiversity. The respective metabolite profiling exposed a strain-specific metabolite occurrence, leading to the recognition of potential hits. These findings suggest that marine Streptomyces produce photoprotectants ought to be further explored in skincare applications.

Sun overexposure is associated with the development of diseases that primarily affect the skin, which can lead to skin cancer. Among the main measures of photoprotection is the use of sunscreens. However, there is currently concern about the reported harmful effects to both humans and the environment due to several of the sunscreen ingredients available on the market. For this reason, the search for and development of new agents with photoprotective properties is required. In searching for these metabolites, researchers have turned their attention to microbial sources, especially the microbiota in unusual hostile environments. Among the diverse microorganisms available in nature, Actinobacteria and specifically Streptomyces , have been shown to be a source of metabolites with various biological activities of interest, such as antimicrobial, antitumor and immunomodulator activities. Herein, we present the results of a systematic review of the literature in which Streptomyces isolates were studied as a source of compounds with photoprotective properties. A meta-analysis of the structure-property and structure-activity relationships of those metabolites identified in the qualitative analysis phase was also carried out. These findings indicate that Streptomyces are a source of metabolites with potential applications in the development of new, safe and more eco-friendly sunscreens.

In the present study, the sensitivity and concentration dependence of three functionally-defined components of cholinesterase activity (total: T-ChE; eserine-sensitive: Es-ChE; and eserine-resistant: Er-ChE) were quantified in the gill, digestive gland and adductor muscle of the tropical cup oyster Saccostrea sp., following acute (96h) aqueous exposure to commercial formulations of the organophosphate (OP) insecticide chlorpyrifos and the neonicotinoid (NN) imidacloprid (concentration range: 0.1-100mg/L), as well as to dissolved cadmium and copper (concentration range: 1-1000μg/L). Oysters (1.5-5.0cm shell length), field-collected from a boating marina in Santa Marta, Colombia (Caribbean Sea) were exposed in the laboratory to each substance at five concentrations. T-ChE, Es-ChE, and Er-ChE activity were quantified in the three tissues in pools of 5 individuals (3 replicates per concentration), before and after inhibition with the total cholinesterase inhibitor eserine (physostigmine, 100µM). Oysters exposed to chlorpyrifos, imidacloprid and Cd showed reduced T-ChE and Es-ChE activity in gills at highest exposure concentrations, with Es-ChE activity being inhibited proportionally more so than T-ChE, whereas Er-ChE activity showed no significant concentration-response. Digestive gland also showed diminished T-ChE, Es-ChE and Er-ChE activity for highest chlorpyrifos and Cd concentrations relative to controls, but an increase of T-ChE and Er-ChE activity at the highest imidacloprid concentration (100mg/L). For Cu, T-ChE, Es-ChE and Er-ChE activities in gills and digestive gland were elevated relative to controls in oysters exposed to Cu concentrations > 100µg/L. In adductor muscle, T-ChE, Es-ChE and Er-ChE activity showed no apparent pattern for any of the four xenobiotics and concentration levels tested. Although this study confirms acute (96h) concentration-dependent reduction of tissue T-ChE and Es-ChE activity in gills and digestive glands of Saccostrea sp. exposed to high concentrations of chlorpyrifos (100mg/L), significant changes in T-ChE, Es-ChE and Er-ChE were also caused by exposure to Cd and Cu at concentrations > 100µg/L and by exposure to imidacloprid (100mg/L), indicating that cholinesterase activity is not a specific biomarker of organophosphate exposure in this species, but, rather, a biomarker of diverse xenobiotic exposure., (Copyright © 2018 Elsevier Inc. All rights reserved.)

This investigation was aimed at contributing to develop a suitable multi-biomarker approach for pollution monitoring in mangrove-lined Caribbean coastal systems using as sentinel species, the mangrove cupped oyster, Crassostrea rhizophorae. A pilot field study was carried out in 8 localities (3 in Nicaragua; 5 in Colombia), characterized by different environmental conditions and subjected to different levels and types of pollution. Samples were collected in the rainy and dry seasons of 2012-2013. The biological effects at different levels of biological complexity (Stress-on-Stress response, reproduction, condition index, tissue-level biomarkers and histopathology) were determined as indicators of health disturbance, integrated as IBR/n index, and compared with tissue burdens of contaminants in order to achieve an integrative biomonitoring approach. Though modulated by natural variables and confounding factors, different indicators of oyster health, alone and in combination, were related to the presence of different profiles and levels of contaminants present at low-to-moderate levels. Different mixtures of persistent (As, Cd, PAHs) and emerging chemical pollutants (musk fragrances), in combination with different levels of organic and particulate matter resulting from seasonal oceanographic variability and sewage discharges, and environmental factors (salinity, temperature) elicited a different degree of disturbance in ecosystem health condition, as reflected in sentinel C. rhizophorae. As a result, IBR/n was correlated with pollution indices, even though the levels of biological indicators of health disturbance and pollutants were low-to-moderate, and seasonality and the incidence of confounding factors were remarkable. Our study supports the use of simple methodological approaches to diagnose anomalies in the health status of oysters from different localities and to identify potential causing agents and reflect disturbances in ecosystem health. Consequently, the easy methodological approach used herein is useful for the assessment of health disturbance in a variety of mangrove-lined Caribbean coastal systems using mangrove cupped oysters as sentinel species., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Metallothioneins and vitellogenins are low molecular weight proteins that have been used widely in environmental monitoring as biomarkers of exposure and damage to metals and estrogenic compounds, respectively. In the present study, the responses of metallothionein and vitellogenin tissue concentrations were measured following acute (96h) aqueous exposures to cadmium in Saccostrea sp., a tropical cup oyster native to the Western Pacific Ocean that has recently established itself in the Caribbean Sea. Adult oysters (1.5-5.0cm shell length) collected from the municipal marina of Santa Marta, Colombia (Caribbean Sea) and acclimated for 5days in the laboratory, were exposed to Cd at five concentrations (0, 1, 10, 100 and 1000μg/L) and their tissues (gills, digestive gland and adductor muscle) were analyzed in pools of 5 individuals (3 replicates per concentration). Metallothioneins in digestive glands of oysters exposed to Cd concentrations≥100μg/L showed a significant increase, from 8.0 to 14.8μg MT/mg total protein, whereas metallothionein concentrations in gills increased to lesser extent, and no differences were observed in adductor muscle. Metallothionein concentrations in digestive gland and gills correlated directly with whole soft tissue Cd concentrations (ranging from 2 to 297μg/g dw Cd). Vitellogenin in homogenates of oyster gonad tissue, after 96h of exposure to 1000μg/L Cd, were significantly lower (0.04mg P/g gonad) compared to control oysters (0.68mg P/g gonad), suggestive of an anti-estrogenic effect of Cd at high concentrations, whereas no significant changes in vitellogenin concentrations were observed at intermediate Cd exposure concentrations. This study confirms acute responses of metallothionein and vitellogenin concentrations in tissues of Saccostrea sp. exposed to high concentrations of cadmium (Cd≥100μg/L, 96h). The present results are first step towards validating the use of these two proteins as biomarkers of metal exposure in this species., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background: Multiple sclerosis (MS) is a disease whose physiopathogenesis shows a complex interaction of genetic and environmental factors. Given that those factors have not been documented in our country, we describe the clinical and demographic characteristics from a sample of patients with MS., Methods: We carried out an observational, descriptive, cross-sectional, and retrolective study in a Center for Demyelinating Diseases. We took the information from the clinical records of a sample of patients with multiple sclerosis, who arrived to the center from April 2014 to July 2015., Results: We obtained data from 313 patients, out of which 65.5 % were women. Mean age was 41 years (SD 11.22). Minimum age of diagnosis was 12 years and maximum, 66 years; mean age of diagnosis was 32 years (SD 9.72). With regards to clinical variables, 3.4 % presented radiologically isolated syndrome (RIS), 82 % relapsing-remitting MS (RRMS), 13.9 % secondary-progressive MS (SPMS), and 0.8 % primary-progressive MS (PPMS). Of all the patients, 10 % had first or second degree relatives with diagnosis of this disease; 16 % had foreign ancestors; 27 % were smokers. Treatment consisted of glatiramer acetate, 28 %; intramuscular interferon beta 1a, 18 %; subcutaneous interferon beta 1a, 16 %; subcutaneous interferon beta 1b, 30 %; fingolimod, 3 %; and others, 5 %., Conclusions: Clinical and demographic characteristics are similar to those reported in international literature. More studies would be needed to typify Mexican population with MS.

Several diseases have a significant impact on American oyster populations in the Atlantic coasts of North America. Knowledge about the responses of oysters to pathogenic challenge could help in identifying potential markers of disease resistance and biomarkers of the health status of an oyster population. A previous analysis of the transcriptome of resistant and susceptible American oysters in response to challenge with the bacterial pathogen Roseovarius crassostreae, as well as sequencing of suppression subtractive hybridization libraries from oysters challenged with the protozoan parasite Perkinsus marinus, provided a list of genes potentially involved in disease resistance or susceptibility. We investigated the patterns of inducible gene expression of several of these genes in response to experimental challenge with the oyster pathogens R. crassostreae, Vibrio tubiashii, and P. marinus. Oysters showing differential susceptibility to R. crassostreae demonstrated differential patterns of expression of genes coding for immune (serine protease inhibitor-1, SPI1) and stress-related (heat shock protein 70, HSP70; arginine kinase) proteins 30 days after challenge with this bacterial pathogen. Differential patterns of expression of immune (spi1, galectin and a matrix metalloproteinase) and stress-related (hsp70, histone H4, and arginine kinase) genes was observed in hemocytes from adult oysters challenged with P. marinus, but not with V. tubiashii. While levels of spi1 expression in hemocytes collected 8 and 21 days after P. marinus challenge were negatively correlated with parasite load in oysters tissues at the end of the challenge (62 days), levels of expression of hsp70 in hemocytes collected 1-day after challenge were positively correlated with oyster parasite load at 62 days. Our results confirm previous research on the role of serine protease inhibitor-1 in immunity and disease resistance in oysters. They also suggest that HSP70 and histone H4 could be used as a markers of health status or disease susceptibility in oysters., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Several histones and histone-derived peptides have been shown to have antimicrobial activity and a potential role in innate immune defenses. A histone H4 sequence was identified in a subtractive suppression library containing genes upregulated in American cupped oysters, Crassostrea virginica, in response to challenge with the protozoan parasite Perkinsus marinus. Oyster histone H4 protein levels significantly increased in hemocyte lysates and cell free hemolymph of oysters experimentally challenged with P. marinus. The complete histone H4 coding sequence of C. virginica was cloned into a Saccharomyces cerevisiae yeast expression system and recombinant expression was confirmed using SDS-PAGE analysis and western blot. Delivery of yeast cells expressing recombinant oyster histone H4 into the gut of brine shrimp, Artemia salinas, challenged with a streptomycin resistant strain of Vibrio anguillarum resulted in a significant and dose-dependent decrease in the load of V. anguillarum. Purified recombinant histone H4 showed antimicrobial activity against V. anguillarum and Escherichia coli at micromolar concentrations, but did not affect the viability of P. marinus in culture. These results support the role of histone H4 in the defense of oysters against bacterial infection and validate the use of a novel oyster antimicrobial H4 in a yeast feed-based delivery system for the treatment of bacterial infections in aquaculture applications., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Traumatic brain injury (TBI) produces several cellular changes, such as gliosis, axonal and dendritic plasticity, and inhibition-excitation imbalance, as well as cell death, which can initiate epileptogenesis. It has been demonstrated that dysfunction of the inhibitory components of the cerebral cortex after injury may cause status epilepticus in experimental models; we proposed to analyze the response of cortical interneurons and astrocytes after TBI in humans. Twelve contusion samples were evaluated, identifying the expression of glial fibrillary acidic protein (GFAP) and calcium-binding proteins (CaBPs). The study was made in sectors with and without preserved cytoarchitecture evaluated with NeuN immunoreactivity (IR). In sectors with total loss of NeuN-IR the results showed a remarkable loss of CaBP-IR both in neuropil and somata. In sectors with conserved cytoarchitecture less drastic changes in CaBP-IR were detected. These changes include a decrease in the amount of parvalbumin (PV-IR) neurons in layer II, an increase of calbindin (CB-IR) neurons in layers III and V, and an increase in calretinin (CR-IR) neurons in layer II. We also observed glial fibrillary acidic protein immunoreactivity (GFAP-IR) in the white matter, in the gray-white matter transition, and around the sectors with NeuN-IR total loss. These findings may reflect dynamic activity as a consequence of the lesion that is associated with changes in the excitatory circuits of neighboring hyperactivated glutamatergic neurons, possibly due to the primary impact, or secondary events such as hypoxia-ischemia. Temporal evolution of these changes may be the substrate linking severe cortical contusion and the resulting epileptogenic activity observed in some patients.

Species of Perkinsus are responsible for high mortalities of bivalve molluscs world-wide. Techniques to accurately estimate parasites in tissues are required to improve understanding of perkinsosis. This study quantifies the number and tissue distribution of Perkinsus marinus in Crassostrea virginica by modern stereology and immunohistochemistry. Mean total number of trophozoites were (mean +/- SE) 11.80 +/- 3.91 million and 11.55 +/- 3.88 million for the optical disector and optical fractionator methods, respectively. The mean empirical error between both stereological approaches was 3.8 +/- 1.0%. Trophozoites were detected intracellularly in the following tissues: intestine (30.1%), Leydig tissue (21.3%), hemocytes (14.9%), digestive gland (11.4%), gills (6.1%), connective tissues (5.7%), gonads (4.1%), palps (2.2%), muscle (1.9%), mantle connective (0.8%), pericardium (0.7%), mantle epithelium (0.1%), and heart (0.1%). The remaining 0.6% were found extracellularly. Percentages of trophozoite stages were (mean +/- SE): large, log-phase trophonts, i.e., signet rings, 97.0 +/- 1.2%; meronts, 2.0 +/- 0.9%; clusters of small, log-phase trophonts, i.e., merozoites, 1.0 +/- 0.5%. Levels of infection in hemocytes and Leydig tissue were representative of total parasite intensity. These techniques are a powerful tool to follow parasite distribution and invasion, and to further explore mechanisms of Perkinsus spp. pathogenesis in bivalves.

We investigated the role of nitric oxide (NO) in the responses of the Eastern oyster, Crassostrea virginica, to the protozoan parasite Perkinsus marinus, causative agent of Dermo disease. P. marinus induced a slight but significant increase in NO production by oyster hemocytes in vitro, comparable to the increase induced by the immune stimulants phorbol myristrate acetate (PMA) and lipopolysaccharide (LPS). P. marinus also activated the NO response in oysters in vivo, as shown by induction of a protein reacting with a universal NO synthase (NOS) antibody in hemocytes and the presence of high levels of nitrite in plasma. Treatment of experimentally infected oysters with the NOS inhibitor, Nomega-nitro-L-arginine methyl ester (L-NAME) resulted in a transient decrease in NO levels in oyster plasma and a significant increase in the number of parasites at early time points after infection. The NO donor, S-nitroso-N-acetyl-penicillamine (SNAP) caused a significant inhibition in the proliferation of P. marinus cultured cells after 24 h of incubation. These results indicate that NO has a role in decreasing parasite loads at early time points after infection.

Two episodes of mortality of cultured carpet shell clams (Ruditapes decussatus) associated with bacterial infections were recorded during 2001 and 2002 in a commercial hatchery located in Spain. Vibrio alginolyticus was isolated as the primary organism from moribund clam larvae that were obtained during the two separate events. Vibrio splendidus biovar II, in addition to V. alginolyticus, was isolated as a result of a mixed Vibrio infection from moribund clam larvae obtained from the second mortality event. The larval mortality rates for these events were 62 and 73%, respectively. Mortality was also detected in spat. To our knowledge, this is the fist time that these bacterial species have been associated with larval and juvenile carpet shell clam mortality. The bacterial strains were identified by morphological and biochemical techniques and also by PCR and sequencing of a conserved region of the 16S rRNA gene. In both cases bacteria isolated in pure culture were inoculated into spat of carpet shell clams by intravalvar injection and by immersion. The mortality was attributed to the inoculated strains, since the bacteria were obtained in pure culture from the soft tissues of experimentally infected clams. V. alginolyticus TA15 and V. splendidus biovar II strain TA2 caused similar histological lesions that affected mainly the mantle, the velum, and the connective tissue of infected organisms. The general enzymatic activity of both live cells and extracellular products (ECPs), as evaluated by the API ZYM system, revealed that whole bacterial cells showed greater enzymatic activity than ECPs and that the activity of most enzymes ceased after heat treatment (100 degrees C for 10 min). Both strain TA15 and strain TA2 produced hydroxamate siderophores, although the activity was greater in strain TA15. ECPs from both bacterial species at high concentrations, as well as viable bacteria, caused significant reductions in hemocyte survival after 4 h of incubation, whereas no significant differences in viability were observed during incubation with heat-killed bacteria.

The effect of a pathogenic Vibrio pelagius, isolated during a mass mortality of turbot larvae, on the non-specific immune response of turbot, Scophthalmus maximus (L.), macrophages was studied both in vitro and in vivo. The in vitro treatment of head kidney (HK) macrophages with viable V. pelagius caused a significant inhibition of the chemiluminescence (CL) response in comparison with untreated macrophages, while incubation with heat-killed bacteria did not affect this response. In vivo, the intraperitoneal injection of V. pelagius resulted in a significant inhibition of the CL response in infected fish at days 1 and 4 post-infection compared with the control fish response. The HK macrophage nitric oxide (NO) production was enhanced by in vitro incubation with intermediate doses of viable V. pelagius (5 x 10(3) and 5 x 10(4) bacteria mL(-1)) and higher doses of the heat-killed bacteria (5 x 10(4)-5 x 10(6) bacteria mL(-1)). In both cases, the NO inhibitor N-omega -nitro-L-arginine was capable of down-regulating the specific NO induction caused by incubation with the bacterial treatments. In contrast, incubation with ECPs at higher doses caused a reduction in NO production. In vivo, a significant enhancement in NO production was also observed in macrophage supernatants at day 10 post-infection. Lysozyme concentration in the serum was also significantly increased in the experimentally infected fish at days 4 and 10 post-injection. In addition, viable V. pelagius and its ECPs significantly reduced HK macrophage viability in vitro, whereas no significant differences in viability were observed during the incubation with heat-killed bacteria. As NO production was enhanced in the experimentally infected fish, the inhibitory effect of the NO donor, S-nitroso-acetyl-penicillamine (SNAP), was tested in vitro in a cell-free assay. The results showed that growth of V. pelagius was significantly inhibited using SNAP at a high concentration (1 mM).

A bacterial strain, characterized as Vibrio pelagius (Hq 222), was isolated from a turbot, Scophthalmus maximus (L.), larvae mass mortality in a commercial fish farm in Spain. Turbot larvae, post-larvae (0.2 g) and juveniles (5 and 15 g) were experimentally infected. The bacterium appeared to be very virulent for larvae and post-larvae, LD50 being < 5 bacteria mL(-1) for larvae 1 week post-infection and 3.9 x 10(5) bacteria mL(-1) in post-larvae at day 12 post-infection. The bacterial strain was recovered in pure culture from the internal organs of infected fish. Histological lesions in post-larvae exhibited swelling and necrosis of gill secondary lamellae, sloughing of intestinal mucosa and necrosis of haematopoietic tissue in the kidney. Vibrio pelagius (Hq 222) was able to grow in sterile sea water when incubated at room temperature or at 15 degrees C. Vibrio pelagius (Hq 222) was more adherent to the turbot cell lines TV-1 and TF than Escherichia coli. In both cell lines, the number of adhered bacteria increased with incubation time.

In the present work, the effect of nisin on the non-specific immune response of turbot (Scophthalmus maximus L.) leukocytes has been studied both in vitro and in vivo. The head kidney macrophage chemiluminescent (CL) response was significantly increased with intermediate doses of nisin (2.5 and 0.025 micro g ml(-1)) whilst the higher dose (25 micro g ml(-1)) significantly decreased the response after 24h incubation. When the incubation time was extended to 72 h, significant differences between doses were observed and the lower nisin concentration (0.025 micro g ml(-1)) appeared to be the optimum dose for increasing the CL response. The phagocytic activity of HK macrophages was also affected by in vitro nisin treatments. Nisin at 0.25 micro g ml(-1) and 0.025 micro g ml(-1) significantly stimulated the response after 24 and 72 h incubation respectively. Nitric oxide (NO) production by HK macrophages was not influenced by any nisin concentration employed for 24 or 72 h incubationsIn vivo, one week post injection, a slightly but non-significant stimulation of the CL response was observed with the lowest nisin concentration (0.0025 micro g fish(-1)). NO in serum and serum antibacterial index were not significantly affected by nisin treatments. On the other hand, lysozyme concentration in serum was significantly augmented with the lowest nisin dose (0.0025 micro g fish(-1)). The antibacterial effect of nisin against the fish pathogenic bacteria Carnobacterium piscicola (CECT 4020) was also demonstrated in vitro.

In the present work, the effects of several lactic acid bacteria on the immune response of turbot (Scophthalmus maximus) macrophages have been studied both in vitro and in vivo. Out of six lactic acid bacterial strains tested, only heat-killed Lactococcus lactis significantly increased the turbot head kidney macrophage chemiluminescent (CL) response after 24 h of incubation. Nitric oxide (NO) was also significantly enhanced by this bacterium after 72 h of incubation with either viable (10(3) and 10(6) cells/ml) or heat-killed (10(6) cells/ml) bacteria. Viable Leuconostoc mesenteroides (10(6) cells/ml) was also capable of significantly increasing NO production. Since L. lactis proved to be the strain with more effects on the host immune function, further in vivo and in vitro experiments were conducted with this bacterium. The in vitro capacity of L. lactis to adhere to turbot intestinal mucus was positively confirmed. When orally administered, L. lactis significantly increased the macrophage CL response and the serum NO concentration after 7 days of daily administration. The antibacterial effect of the extracellular products from the six LAB strains against the fish-pathogenic bacterium Vibrio anguillarum was also demonstrated in vitro.

Objective: To assess the prevalence of high blood pressure (PHBP) and its association with age, sex, socioeconomic status (SES), and body mass index (BMI = kg/m2)., Material and Methods: A cross-sectional survey was carried out in 1996, in a random sample of 356 adults aged 20 and older, residents of Bucaramanga, Colombia. Weight, height, and blood pressure readings were obtained, plus data on age and gender. Robust linear and log-binomial regression was used to estimate the independent effect of different risk factors on systolic (SBP) and diastolic (DBP) blood pressure and HBP., Results: Mean SBP and DBP were significantly higher in men (122.5 and 88.1 mmHg) than in women (117.2 and 75.4 mmHg). The adjusted SBP increased with age, more in men than in women; DBP did not change with age. For each unit increase in BMI, SBP and DBP increased 0.76 and 0.69 mmHg, respectively. SBP was 6.8 and DBP was 9.8 mmHg higher in low-SES than in high-SES subjects. The adjusted PHBP increased with age and was higher in subjects > or = 50 years than in those 20-30 years old. Sex had no significant effect on PHBP (p = 0.795). For each unit increase in BMI the PHBP increased 3%, and low-SES subjects had a PHBP 1.84 times higher than high-SES subjects., Conclusions: A high PHBP was found. There is a great potential for prevention by weight control. Further studies are needed to confirm the increasing PHBP in low-SES subjects and to identify its causes.

Unlabelled: Intratumor distribution patterns of 99mTc-sestamibi and 14C-2-deoxy-D-glucose were compared in the c-neu OncoMouse, a transgenic mouse that spontaneously develops breast tumors., Methods: Thirty or 60 min after intravenous injection of 5 muCi 14C-2-deoxy-D-glucose and 3 mCi 99mTc-sestamibi into mice (n = 3 per time point) bearing mammary tumors (0.3-1.5 cm), the animals were analyzed for organ and tumor distribution using dual-label, whole-body autoradiography. The retention patterns of the two compounds were related to tumor morphology and viability, based on H&E-stained adjacent sections. For imaging studies, the transgenic mice (n = 9) were anesthetized with pentobarbital, injected intravenously with 5-20 mCi 99mTc-sestamibi and imaged for 60 min using a gamma camera equipped with a 1-mm pinhole collimator., Results: All positively stained tumors retained both agents, with a mean 99mTc-sestamibi tumor retention of 0.38% +/- 0.2% ID/g at 30 min compared to 4.18% +/- 0.62% ID/g for 14C-2-deoxy-D-glucose. Tumor retention of the agents remained the same at 60 min, and neither compound localized within necrotic or cystic regions of the neoplasms. Repeat imaging at 2-8-day intervals indicated a predicted sensitivity to detect a 30% difference in tumor retention of a test versus reference compound in preclinical screening., Conclusion: The c-neu OncoMouse is a useful model for in vivo imaging and provides a spontaneous tumor model for preclinical screening of breast tumor imaging agents.