Search Results (17)

Caulerpin, a bis-indole alkaloid isolated from Caulerpa racemosa, has several documented pharmacological activities, including antineoplastic and antiviral properties. This study aimed to evaluate the anti-inflammatory and anti-tubercular potentials of caulerpin and its analogues in RAW 264.7 macrophages infected with Mycobacterium spp. Additionally, we evaluated cytokine production and NLRP3 expression in this infection model. Toxicity tests were performed using Vero E6 and HepG2 cell lines and Artemia salina. Pre-incubation of RAW 264.7 cells with caulerpin and its analogues decreased internalized M. smegmatis and M. tuberculosis H37Ra. Furthermore, treatment of M. smegmatis-infected macrophages with caulerpin and its analogues reduced bacterial loads. Caulerpin reduced the CFU count of internalized bacilli in the M. tuberculosis H37Ra infection model. In addition, caulerpin and its diethyl derivative were notably found to modulate IL-1β and TNF-α production in the M. smegmatis infection model after quantifying pro-inflammatory cytokines and NLRP3. Caulerpin and its derivates did not affect the viability of Vero E6 and HepG2 cell lines or nauplii survival in toxicity studies. These findings demonstrate that caulerpin and its analogues exhibit anti-inflammatory activity against Mycobacterium spp. infection in RAW 264.7 macrophages and show promising potential for further efficacy and safety evaluation. Full article

Cardiotonic steroids modulate various aspects of the inflammatory response. The synthetic cardiotonic steroid γ-benzylidene digoxin 15 (BD-15), a digoxin derivative, has emerged as a promising candidate with potential immunomodulatory effects. However, its biological activity remains largely unexplored. This study investigated the anti-mycobacterial and anti-inflammatory effects of BD-15 in an in vitro macrophage infection model with Mycobacterium spp. Unlike digoxin, which showed significant toxicity at higher concentrations, BD-15 exhibited no cytotoxicity in RAW 264.7 cells (a murine macrophage cell line). Both compounds were evaluated in Mycobacterium smegmatis-infected RAW 264.7 cells, reducing bacterial burden without direct bactericidal activity. Additionally, both modulated pro-inflammatory cytokine levels, notably by decreasing tumor necrosis factor alpha (TNF-α) and interleukin-1 beta (IL-1β) levels. BD-15 specifically reduced NOD-, LRR-, and pyrin-domain-containing protein 3 (NLRP3) inflammasome expression and increased interleukin-10 (IL-10) production. Notably, BD-15 reduced colony-forming unit (CFU) counts in Mycobacterium tuberculosis-infected RAW 264.7 cells. Toxicity assays in HepG2 cells (a human liver cancer cell line) showed that BD-15 had minimal hepatotoxicity compared to digoxin, and both demonstrated negligible acute toxicity in an Artemia salina bioassay. These findings revealed the immunomodulatory effects of cardiotonic steroids in a bacterial infection model and highlighted BD-15 as a safer alternative to digoxin for therapeutic applications. Full article

Determining molecular markers for parasites provides a useful tool for their identification, particularly for larval stages with few distinguishable diagnostic characters. Avian cestodes play a key role in the food webs and biodiversity of hypersaline wetlands, yet they remain understudied. Using naturally infected Artemia, we identified cestode larvae (cysticercoids), assessed their genetic diversity, and explored phylogenetic relationships in relation to larval morphology and waterbird final hosts. We obtained partial 18S rDNA sequences for 60 cysticercoids of the family Hymenolepidae infecting Artemia spp. from seven localities and three countries (Spain, the USA, and Chile). We present the first DNA sequences for six taxa: Confluaria podicipina, Fimbriarioides sp., Flamingolepis liguloides, Flamingolepis sp. 1, Flamingolepis sp. 2, and Hymenolepis californicus. Intraspecific sequence variation (0.00–0.19% diversity) was lower than intergroup genetic distance (0.7–14.75%). Phylogenetic analysis revealed three main clades: 1—Flamingolepis, 2—Fimbriarioides, 3—Confluaria and Hymenolepis, all of which separated from hymenolepidids from mammals and terrestrial birds. This clear separation among taxa is congruent with previous morphological identification, validating the 18S gene as a useful marker to discriminate at generic/species level. Working with intermediate hosts allows the expansion of knowledge of taxonomic and genetic diversity of cestodes in wildlife, as well as elucidation of their life cycles. Full article

Terminalia petiolaris A. Cunn. Ex Benth. (genus: Terminalia, family: Combretaceae) is native to Australia. Terminalia spp. have traditionally been used to treat various ailments, including bacterial infections. Solvents of varying polarity were used to extract compounds from leaves of this species, and the extracts were tested against a panel of bacteria, including antibiotic-resistant strains. The methanolic and water extracts showed substantial inhibitory activity against several bacteria, including antibiotic-resistant strains in both disc diffusion and liquid dilution assays. Combining these extracts with selected conventional antibiotics enhanced the inhibition of bacterial growth for some combinations, while others showed no significant interaction. In total, two synergistic, twenty-five additive, twenty-three non-interactive and one antagonistic interaction were observed. The methanolic and ethyl acetate plant extracts were found to be non-toxic in Artemia franciscana nauplii toxicity assays. A liquid chromatography–mass spectrometry metabolomics analysis identified several flavonoid compounds, including miquelianin, trifolin and orientin, which might contribute to the observed activities. The potential modes of these active extracts are further discussed in this study. Full article

Bacterial communities associated with fish larvae are highly influenced by the microbiota of live prey used as feed (rotifers or Artemia), generally dominated by bacterial strains with a low degree of specialization and high growth rates, (e.g., Vibrionaceae), which can be detrimental to larvae. Co-cultivation of microalgae used in the enrichment of Artemia (e.g., Phaeodactylum tricornutum, or Chlorella minutissima) with Vibrio-antagonistic probiotics belonging to the Roseobacter clade bacteria (e.g., Phaeobacter spp. or Ruegeria spp.) was studied. The introduction of the probiotics did not affect microalgae growth or significantly modify the composition of bacterial communities associated with both microalgae, as revealed by DGGE analysis. The inoculation of P. tricornutum with Ruegeria ALR6 allowed the maintenance of the probiotic in the scale-up of the microalgae cultures, both in axenic and non-axenic conditions. Using Ruegeria-inoculated P. tricornutum cultures in the enrichment of Artemia reduced the total Vibrionaceae count in Artemia by 2 Log units, therefore preventing the introduction of opportunistic or pathogenic bacteria to fish larvae fed with them. Full article

Solar salterns and salt marshes are unique ecosystems with special physicochemical features and characteristic biota. Currently, there are very few studies focused on the impacts of pollution on these economic and ecological systems. Unfortunately, diversified pollution (metals, Polycyclic Aromatic Hydrocarbons, etc.) has been detected in these complex ecosystems. These hypersaline environments are under increasing threat due to anthropogenic pressures. Despite this, they represent a valuable source of microbial diversity, with taxa displaying special features in terms of environmental remediation capacities as well as economical species such as Artemia spp. (Branchiopoda) and Dunaliella salina (Chlorophyta). In this review, we discuss the impacts of pollution on these semi-artificial systems. Therefore, we have indicated the sentinel species identified in plankton communities, which can be used in ecotoxicological investigations in solar salterns. In future, researchers should increase their interest in pollution assessment in solar salterns and salt marshes. Full article

Hypersaline waters are unique polyextreme habitats, where the salinity limits species richness. There are main patterns of a relationship between salinity and the species richness of free-living aquatic animals, but for parasitic organisms, general regularities have not yet been established. There are quite numerous data on parasites in hypersaline waters worldwide; however, they have not been summarized before. This review tries to fill this gap by summarizing the available data. All parasites, 85 species and forms, found in hypersaline waters belong to five phyla: Platyhelminthes, Nematoda, Acanthocephala, Cnidaria, and Arthropoda. Platyhelminthes are the most diverse phylum with the highest species richness in class Cestoda. Most species were noted in hypersaline waters with a salinity of no more than 100 g·L⁻¹. The total number of parasitic species decreases exponentially with an increase in salinity. The number of free-living animal species inhabiting waters with a salinity from 35 to 210 g·L⁻¹ is approximately 12 times higher than that of parasitic ones in all intervals of this salinity range. Salinity influences parasite richness and composition in two ways—directly and through the availability of hosts. Free-living crustaceans were hosts of most parasite species in hypersaline waters. Artemia spp., the most halotolerant animals, are an intermediate host for 22 species and unidentified forms of parasites. Full article

Mexico is a center of diversification for the Cactaceae family, with 69% of the species recorded as endemic. Certain members of the Cactaceae family have been chemically analyzed to relate their medicinal use with their phytochemistry. Here, the phytochemistry and bioactivity of ethanol extracts of Ferocactus echidne, F. latispinus, and Mammillaria geminispina were evaluated. A preliminary phytochemical analysis was performed, detecting the presence of saponins, tannins, cardiotonic glycosides, and sesquiterpene lactones. The presence of nicotinic acid in F. echidne and F. latispinus was identified by GC-MS. Other compounds found in the extracts of these three species were gentisic acid, diosmetin, chlorogenic acid, N-methyltyramide, and hordenine. The antioxidant activity was estimated with the DPPH free radical scavenging test. To determine the toxicity of the extracts, the in vivo model of Artemia spp. was used. In addition, the cytotoxicity of the extract was tested on C6, HaCaT, THP-1, and U937 cell lines, while the inflammatory activity was tested by measuring the secretion of cytokines using macrophage cells. The three species showed different bioactivities, including antioxidant, antimicrobial, cytotoxic, and anti-inflammatory activities. To the best of our knowledge, the results presented here are the first described for these species. Full article

Many works have been devoted to the study of the molecular genetic diversity of Artemia in different regions; however, there are regions such as Crimea, the largest peninsula in the Black Sea, which has seen few studies. Artemia specimens from several Crimean hypersaline lakes were analyzed using the mitochondrial marker cytochrome oxidase C (COI). The analyzed individuals from bisexual populations formed clades with the species A. salina, A. urmiana, A. sinica, and A. monica (=A. franciscana). A. sinica and A. monica had not been recorded in Crimea previously. In Lake Adzhigol, the three species A. urmiana, A. sinica, and A. monica were found at the same time, which has not been noted anywhere before. In the Crimean lakes, a total of 10 haplotypes were found, six of them for the first time: Once for A. monica, once for A. sinica, and four for A. salina. Those haplotypes may be regarded as endemic to Crimea. In the 1990s, experiments were carried out in Lake Yanyshskoe using mainly purchased cysts of Artemia, so A. monica and A. sinica were introduced into Crimea and could then have easily been spread by birds to other Crimean lakes. Full article

Seven medicinal plants from Chiapas, Mexico, used by Native Americans were analyzed, aiming to improve the understanding of their medicinal properties through the evaluation of various biological activities, i.e., bactericidal, antioxidant, α-glucosidase inhibition, and toxicity, to provide a scientific basis for the management of infectious and hyperglycemic diseases in the Mexican southeast. Plant extracts were obtained from Cordia dodecandra, Gaultheria odorata, Heliotropium angiospermum, Justicia spicigera, Leucaena collinsii spp. collinsii, Tagetes nelsonii, and Talisia oliviformis through maceration techniques using methanol and chloroform (1:1). Minimum Inhibitory Concentration (MIC) was employed to determine the antibacterial activity against Staphylococcus aureus, Enterobacter faecalis, Escherichia coli, Enterobacter aerogenes, Enterobacter cloacae, Klebsiella pneumoniae, and Pseudomonas aeuroginosa. The antiradical/antioxidant activity was determined by the 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays and antihemolytic activity using the 2,2’-Azobis(2-amidinopropane) dihydrochloride radical (APPH). The anti-α-glucosidase activity was evaluated in vitro through the chromogenic PNPG assay. The toxicity was assessed using the brine shrimp lethality assay. The highest antimicrobial activity was displayed by T. nelsonii, mainly against E. faecalis and P. aeuroginosa. The extracts of L. collinsii, J. spicigera, and T. nelsonii possess antioxidant properties with EC₅₀ < 50 μg/mL. J. spicigera and T. nelsonii extracts showed the highest antihemolytic activity with IC₅₀ < 14 μg/mL. T. nelsonii exhibited a remarkable inhibitor effect on the α-glucosidase enzyme and the greatest toxic effect on Artemia salina with IC₅₀ = 193 ± 20 μg/mL and LD₅₀ = 14 ± 1 μg/mL, respectively. According to our results, G. odorata, J. spicigera, T. nelsonii, and T. oliviformis extracts contained active antimicrobial compounds. At the same time, T. nelsonii stands to be a possible source of effective antineoplastic and antihyperglycemic compounds. Full article

The administration of probiotics via live feeds, such as Artemia and rotifers, has gained significant attention. Moreover, indiscriminate use of antibiotics in conventional aquaculture practices in order to prevent or control disease outbreaks has resulted in the occurrence of residues and antimicrobial resistance. Thus, the application of eco-friendly feed additives, such as probiotics, as a safer alternative has received increasing attention in recent years. However, only minimal information on the administration of probiotics via freshwater cladoceran Moina micrura is available despite being commonly used for larval and post-larval feeding of freshwater crustaceans and fish. Thus, this study aimed to evaluate the application of Bacillus pocheonensis strain S2 administered via M. micrura to red hybrid tilapia (Oreochromis spp.) larvae. Bacillus pocheonensis that has been previously isolated from Spirulina sp. was subjected to preliminary in vitro evaluation of antagonistic properties. The agar well-diffusion assay revealed that this probiont could inhibit the growth of Streptococcus agalactiae and Aeromonas hydrophila. The size of inhibition zones ranged from 8.8 ± 0.2 to 18.2 ± 0.4 mm. Moina micrura was later used as a biological model in preliminary in vivo bacterial challenge assays to evaluate the efficacy of B. pocheonensis in protecting the host from diseases. Moina micrura was pre-enriched with B. pocheonensis at ${10}^4$ and ${10}^6 \mathrm{CFU} {\mathrm{mL}}^{-1}$ before S. agalactiae and A. hydrophila were introduced into the culture. The study revealed that B. pocheonensis at ${10}^4 \mathrm{CFU} {\mathrm{mL}}^{-1}$ was able to significantly enhance the survival of M. micrura after being challenged with both pathogens (63 ± 3%) in comparison to the control group. The relative percentage survival (RPS) of M. micrura was highest (p < 0.05) when treated with B. pocheonensis at both concentrations ${10}^4$ and ${10}^6 \mathrm{CFU} {\mathrm{mL}}^{-1}$ (38.33) after being challenged against S. agalactiae. To assess the efficacy of B. pocheonensis in protecting red hybrid tilapia against streptococcosis, the larvae were fed with either unenriched (control) Moina or probiont-enriched Moina daily for 10 days. A significantly (p < 0.05) higher survival rate (77 ± 3%) was observed in larvae fed with probiont-enriched M. micrura compared to other treatments, and the RPS was recorded at 62.90. In addition, the S. agalactiae load was suppressed in larvae fed probiont-enriched M. micrura ($6.84\pm 0.39 \mathrm{CFU} {\mathrm{mL}}^{-1}$) in comparison to the control group ($7.78\pm 0.09 \mathrm{CFU} {\mathrm{mL}}^{-1}$), indicating that the probiont might have contributed to the improvement of tilapia health and survival. This study illustrated that M. micrura was suitable to be used as a vector for probiotics in freshwater fish larvae as an alternative to hazardous antibiotics for disease control. Full article

The high incidence of skeletal anomalies in Senegalese sole (Solea senegalensis) still constitutes a bottleneck constraining its production. There are diverse commercially available products for the enrichment of live preys, but few reports of their influence on skeletogenesis in Senegalese sole. This study evaluated the presence of vertebral anomalies in postlarvae and juvenile Senegalese sole fed with Artemia spp. metanauplii enriched with four commercial products (EA, EB, EC, and ED) in a fish farm. The most frequent alterations consisted of deformations of the neural/haemal arches and spines and fusions and deformations of hypurals, epural, or parhypural. The correspondence analysis ordered fish from each age in separated semiaxis, indicating the presence of different anomaly patterns for the two sampled stages. The results showed only very light changes in the frequency of vertebral abnormalities among tested enrichment products, i.e., individuals from EC and EA lots displayed less vertebral body anomalies and/or vertebral column deviations at 31 and 105 days after hatching, respectively. The existence of a large shared malformation pattern in all the experimental groups leads to impute to the rearing conditions as the main driving factor of the onset of such group of anomalies, probably masking some dietary effect. Full article

The movement of plankton is often dictated by local flow patterns, particularly during storms and in environments with strong flows. Reefs, macrophyte beds, and other immersed structures can provide shelter against washout and drastically alter the distributions of plankton as these structures redirect and slow the flows through them. Advection–diffusion and agent-based models are often used to describe the movement of plankton within marine and fresh water environments and across multiple scales. Experimental validation of such models of plankton movement within complex flow environments is challenging because of the difference in both time and spatial scales. Organisms on the scale of 1 mm or less swim by beating their appendages on the order of 1 Hz and are advected meters to kilometers over days, weeks, and months. One approach to study this challenging multiscale problem is to insert actively moving agents within a background flow field. Open source tools to implement this sort of approach are, however, limited. In this paper, we combine experiments and computational fluid dynamics with a newly developed agent-based modeling platform to quantify plankton movement at the scale of tens of centimeters. We use Artemia spp., or brine shrimp, as a model organism given their availability and ease of culturing. The distribution of brine shrimp over time was recorded in a flow tank with simplified physical models of macrophytes. These simplified macrophyte models were 3D-printed arrays of cylinders of varying heights and densities. Artemia nauplii were injected within these arrays, and their distributions over time were recorded with video. The detailed three-dimensional flow fields were quantified using computational fluid dynamics and validated experimentally with particle image velocimetry. To better quantify plankton distributions, we developed an agent-based modeling framework, Planktos, to simulate the movement of plankton immersed within such flow fields. The spatially and temporally varying Artemia distributions were compared across models of varying heights and densities for both the experiments and the agent-based models. The results show that increasing the density of the macrophyte bed drastically increases the average time it takes the plankton to be swept downstream. The height of the macrophyte bed had less of an effect. These effects were easily observed in both experimental studies and in the agent-based simulations. Full article

The Plectranthus genus is commonly used in traditional medicine due to its potential to treat several illnesses, including bacterial infections and cancer. As such, aiming to screen the antibacterial and cytotoxic activities of extracts, sixteen selected Plectranthus species with medicinal potential were studied. In total, 31 extracts obtained from 16 Plectranthus spp. were tested for their antibacterial and anticancer properties. Well diffusion method was used for preliminary antibacterial screening. The minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) values of the five most active acetonic extracts (P. aliciae, P. japonicus, P. madagascariensis var. “Lynne”, P. stylesii, and P. strigosus) were determined. After preliminary toxicity evaluation on Artemia salina L., their cytotoxic properties were assessed on three human cancer cell lines (HCT116, MCF-7, and H460). These were also selected for mechanism of resistance studies (on NCI-H460/R and DLD1-TxR cells). An identified compound—parvifloron D—was tested in a pair of sensitive and MDR-Multidrug resistance cancer cells (NCI-H460 and NCI-H460/R) and in normal bronchial fibroblasts MRC-5. The chemical composition of the most active extract was studied through high performance liquid chromatography with a diode array detector (HPLC-DAD/UV) and liquid chromatography–mass spectrometry (LC–MS). Overall, P. strigosus acetonic extract showed the strongest antimicrobial and cytotoxic potential that could be explained by the presence of parvifloron D, a highly cytotoxic diterpene. This study provides valuable information on the use of the Plectranthus genus as a source of bioactive compounds, namely P. strigosus with the potential active ingredient the parvifloron D. Full article

Ochratoxin A (OTA) produced by mycotoxigenic fungi (Aspergillus and Penicillium spp.) is an extremely toxic and carcinogenic metabolite. The use of cold plasma to inhibit toxin-producing microorganisms in coffee could be an important alternative to avoid proliferation of mycotoxigenic fungi. Roasted coffee samples were artificially inoculated with A. westerdijikiae, A. steynii, A. versicolor, and A. niger, and incubated at 27 °C over 21 days for OTA production. Samples were cold plasma treated at 30 W input power and 850 V output voltage with helium at 1.5 L/min flow. OTA production in coffee was analyzed by high performance liquid chromatography coupled to a mass spectrometer (HPLC-MS). After 6 min of treatment with cold plasma, fungi were completely inhibited (4 log reduction). Cold plasma reduces 50% of OTA content after 30 min of treatment. Toxicity was estimated for extracts of artificially contaminated roasted coffee samples using the brine shrimp (Artemia salina) lethality assay. Toxicity for untreated roasted coffee was shown to be “toxic”, while toxicity for cold plasma treated coffee was reduced to “slightly toxic”. These results suggested that cold plasma may be considered as an alternative method for the degradation and reduction of toxin production by mycotoxigenic fungi in the processing of foods and feedstuffs. Full article