Effect of temporal thermal stress on Penaeus vannamei: Growth performance and physiological plasticity.

The present study evaluated the effect of temporal periods of hypothermia and hyperthermia, followed by an optimal temperature recovery phase on the growth, survival, and physiological response of Penaeus vannamei. Post-larvae were exposed to stress periods for 7 and 14 days at 22 °C and 32 °C each, followed by a recovery phase at 28 °C to complete seven experimental weeks, and were compared with a control group maintained at 28 °C. Weight gain, specific growth rate, feed intake, feed conversion ratio, and survival were weekly determined. Muscle, hepatopancreas, and hemolymph were sampled on the 14th day of the recovery phase for biochemical composition, and antioxidant and digestive enzyme activities determination. The shrimp presented a higher growth rate during short-term hyperthermia in contrast to shrimp under hypothermia that presented compensatory growth after thermal stress when the temperature was restored at 28 °C. Hyperthermia increased 12-13% the feed intake while this was diminished 21-29% by the hypothermia periods. Shrimp undergo metabolic adjustments following thermal stress, with short hypothermia increasing the lipase activity and lipid storage in the hepatopancreas, while short hyperthermia also enhances chymotrypsin activity and leads to higher protein and lipid accumulation. Conversely, prolonged hyperthermia induces greater energy consumption, depleting lipid and glycogen stores, while hypothermia causes scarce mobilization of energy reserves during recovery phase. Antioxidant enzyme activities were not affected by short-thermal stress (7d), while prolonged thermal stress (14d) significantly affected SOD, CAT, and GPx activities. The present study provides important insights into the physiological plasticity of P. vannamei during recovery from thermal stress.

Transcription factor NAC1 activates expression of peptidase-encoding AtCEPs in roots to limit root hair growth.

Plant genomes encode a unique group of papain-type Cysteine EndoPeptidases (CysEPs) containing a KDEL endoplasmic reticulum (ER) retention signal (KDEL-CysEPs or CEPs). CEPs process the cell-wall scaffolding EXTENSIN (EXT) proteins that regulate de novo cell-wall formation and cell expansion. Since CEPs cleave EXTs and EXT-related proteins, acting as cell-wall-weakening agents, they may play a role in cell elongation. The Arabidopsis (Arabidopsis thaliana) genome encodes 3 CEPs (AtCPE1-AtCEP3). Here, we report that the genes encoding these 3 Arabidopsis CEPs are highly expressed in root-hair (RH) cell files. Single mutants have no evident abnormal RH phenotype, but atcep1-3 atcep3-2 and atcep1-3 atcep2-2 double mutants have longer RHs than wild-type (Wt) plants, suggesting that expression of AtCEPs in root trichoblasts restrains polar elongation of the RH. We provide evidence that the transcription factor NAC1 (petunia NAM and Arabidopsis ATAF1, ATAF2, and CUC2) activates AtCEPs expression in roots to limit RH growth. Chromatin immunoprecipitation indicates that NAC1 binds to the promoter of AtCEP1, AtCEP2, and, to a lower extent, AtCEP3 and may directly regulate their expression. Inducible NAC1 overexpression increases AtCEP1 and AtCEP2 transcript levels in roots and leads to reduced RH growth while the loss of function nac1-2 mutation reduces AtCEP1-AtCEP3 gene expression and enhances RH growth. Likewise, expression of a dominant chimeric NAC1-SRDX repressor construct leads to increased RH length. Finally, we show that RH cell walls in the atcep1-3 atcep3-2 double mutant have reduced levels of EXT deposition, suggesting that the defects in RH elongation are linked to alterations in EXT processing and accumulation. Our results support the involvement of AtCEPs in controlling RH polar growth through EXT processing and insolubilization at the cell wall.

Conversion of 2-methyl-4-styrylquinolines into 2,4-distyrylquinolines: synthesis, and spectroscopic and structural characterization of five examples.

Four new 2,4-distyrylquinolines and one 2-styryl-4-[2-(thiophen-2-yl)vinyl]quinoline have been synthesized using indium trichloride condensation reactions between aromatic aldehydes and the corresponding 2-methylquinolines, which were themselves prepared using Friedländer annulation reactions between mono- or diketones and (2-aminophenyl)chalcones: the products have all been fully characterized by spectroscopic and crystallographic methods. 2,4-Bis[(E)-styryl]quinoline, C25H19N, (IIa), and its dichloro analogue, 2-[(E)-2,4-dichlorostyryl]-4-[(E)-styryl]quinoline, C25H17Cl2N, (IIb), exhibit different orientations of the 2-styryl unit relative to the quinoline nucleus. In each of the 3-benzoyl analogues {2-[(E)-4-bromostyryl]-4-[(E)-styryl]quinolin-3-yl}(phenyl)methanone, C32H22BrNO, (IIc), {2-[(E)-4-bromostyryl]-4-[(E)-4-chlorostyryl]quinolin-3-yl}(phenyl)methanone, C32H21BrClNO, (IId), and {2-[(E)-4-bromostyryl]-4-[(E)-2-(thiophen-2-yl)vinyl]quinolin-3-yl}(phenyl)methanone, C30H20BrNOS, (IIe), the orientation of the 2-styryl unit is similar to that in (IIa), but the orientation of the 4-arylvinyl units show considerable variation. The thiophene unit in (IIe) is disordered over two sets of atomic sites having occupancies of 0.926 (3) and 0.074 (3). There are no hydrogen bonds of any kind in the structure of (IIa), but in (IId), a single C-H...O hydrogen bond links the molecules into cyclic centrosymmetric R22(20) dimers. A combination of C-H...N and C-H...π hydrogen bonds links the molecules of (IIb) into a three-dimensional framework structure. A combination of three C-H...π hydrogen bonds links the molecules of (IIc) into sheets, and a combination of C-H...O and C-H...π hydrogen bonds forms sheets in (IIe). Comparisons are made with the structures of some related compounds.

A three-step pathway from (2-aminophenyl)chalcones to novel styrylquinoline-chalcone hybrids: synthesis and spectroscopic and structural characterization of three examples.

Three new styrylquinoline-chalcone hybrids have been synthesized using a three-step pathway starting with Friedländer cyclocondensation between (2-aminophenyl)chalcones and acetone to give 2-methyl-4-styrylquinolines, followed by selective oxidation to the 2-formyl analogues, and finally Claisen-Schmidt condensation between the formyl intermediates and 1-acetylnaphthalene. All intermediates and the final products have been fully characterized by IR and 1H/13C NMR spectroscopy, and by high-resolution mass spectrometry, and the three products have been characterized by single-crystal X-ray diffraction. The molecular conformations of (E)-3-{4-[(E)-2-phenylethenyl]quinolin-2-yl}-1-(naphthalen-1-yl)prop-2-en-1-one, C30H21NO, (IVa), and (E)-3-{4-[(E)-2-(4-fluorophenyl)ethenyl]quinolin-2-yl}-1-(naphthalen-1-yl)prop-2-en-1-one, C30H20FNO, (IVb), are very similar. In each compound, the molecules are linked into a three-dimensional array by hydrogen bonds, of the C-H...O and C-H...N types in (IVa), and of the C-H...O and C-H...π types in (IVb), and by two independent π-π stacking interactions. By contrast, the conformation of the chalcone unit in (E)-3-{4-[(E)-2-(2-chlorophenyl)ethenyl]quinolin-2-yl}-1-(naphthalen-1-yl)prop-2-en-1-one, C30H20ClNO, (IVc), differs from those in (IVa) and (IVb). There are only weak hydrogen bonds in the structure of (IVc), but a single rather weak π-π stacking interaction links the molecules into chains. Comparisons are made with some related structures.

A Broad-Host-Range Phage Cocktail Selectively and Effectively Eliminates Vibrio Species from Shrimp Aquaculture Environment.

The protective effects of a phage cocktail composed of vB_Vc_SrVc2 and vB_Vc_SrVc9 were tested in Pacific white shrimp (Litopenaeus vannamei) postlarvae, which were originally isolated from diseased shrimps and selected due to their broad-host-range properties against several pathogenic Vibsrio species. We used culture-dependent and culture-independent approaches to explore its effect on bacterial communities associated with shrimp postlarvae. Both methods revealed that the levels of Vibrio species were significantly reduced after phage cocktail administration. Phage-treated shrimp also exhibisuppted lesser damage and higher lipid accumulation in B cells of the hepatopancreas, as revealed by histopathological examination. Taken together, this study provides clear evidence that phage therapy can selectively and effectively reduce Vibrio species, thereby providing an environmentally safe alternative to the prophylactic use of antibiotics in shrimp aquaculture.

Effect of Sample Sources on Heavy Metal Concentration Measured in Beta Vulgaris Organs.

Aim: Heavy metal concentration [mg/dL, MP] in soil and the transfer to vegetable organs may have a sampling effect. We compared the [MP] in soil and organ samples of Beta vulgaris collected in sites with socioeconomic differences potentially inducing phytotoxicity. Materials and Methods: Samples of Beta vulgaris and soils (n = 4 per sample of soil and plant material) were randomly collected from two distant geographic areas (Mosquera and Sibaté, Cundinamarca, Colombia). We determined the [MP] using acid digestion of HCl : HNO3 [1 : 1]; the [MP] was obtained by atomic absorption in Varian AA-140 and Shimadzu AA-7000 equipment. A two-way ANOVA estimated the effect (partial η2) of the sampling site and metal type on the [MP] and transfer to the vegetable. Results: In Sibaté, the means (SD) of As_1.44 (0.18), Co_1.09 (0.51), Cr_6.21 (0.33), Ni_0.22 (0.02), and Pb_4.17 (0.87) were higher than in Mosquera (As_1.06 (0.21), Co_0.81 (0.19), Cr_3.72 (0.51), Ni_0.13 (0.04), and Pb_1.69 (0.40)) (p value <0.05). The effect of the interaction between the metal type and Beta vulgaris organs on the [MP] (0.801) in Sibaté was more meaningful than in Mosquera (0.430). Additionally, there was a strong correlation (Spearman's ρ > 0.8, p value <0.001) between [MP_soil] and [MP_plants] and between the transfer of metals to the plant and to the leaves. Discussion. The sampling location has a differential effect on the [MP] in soil and the transfer to Beta vulgaris. Given the differential effect described, the monitoring and phytoremediation strategies must be adjusted to scenarios with potentially phytotoxic conditions.

Class III Peroxidases PRX01, PRX44, and PRX73 Control Root Hair Growth in Arabidopsis thaliana.

Root hair cells are important sensors of soil conditions. They grow towards and absorb water-soluble nutrients. This fast and oscillatory growth is mediated by continuous remodeling of the cell wall. Root hair cell walls contain polysaccharides and hydroxyproline-rich glycoproteins, including extensins (EXTs). Class-III peroxidases (PRXs) are secreted into the apoplastic space and are thought to trigger either cell wall loosening or polymerization of cell wall components, such as Tyr-mediated assembly of EXT networks (EXT-PRXs). The precise role of these EXT-PRXs is unknown. Using genetic, biochemical, and modeling approaches, we identified and characterized three root-hair-specific putative EXT-PRXs, PRX01, PRX44, and PRX73. prx01,44,73 triple mutation and PRX44 and PRX73 overexpression had opposite effects on root hair growth, peroxidase activity, and ROS production, with a clear impact on cell wall thickness. We use an EXT fluorescent reporter with contrasting levels of cell wall insolubilization in prx01,44,73 and PRX44-overexpressing background plants. In this study, we propose that PRX01, PRX44, and PRX73 control EXT-mediated cell wall properties during polar expansion of root hair cells.

Composite of methyl polysiloxane and avocado biochar as adsorbent for removal of ciprofloxacin from waters.

Two carbon composite materials were prepared by mixing avocado biochar and methyl polysiloxane (MK). Firstly, MK was dissolved in ethanol, and then the biochar was added at different times. In sample 1 (R1), the time of adding biochar was immediately after dissolving MK in ethanol, and in sample 2 (R2), after 48 h of MK dissolved in ethanol. The samples were characterized by nitrogen adsorption/desorption measurements obtaining specific surface areas (SBET) of 115 m2 g-1 (R1) and 580 m2 g-1 (R2). The adsorbents were further characterized using scanning electron microscopy, FTIR and Raman spectroscopy, adsorption of vapors of n-heptane and water, thermal analysis, Bohem titration, pHpzc, and C H N elemental analysis. R1 and R2 adsorbents were employed as adsorbents to remove the antibiotic ciprofloxacin from the waters. The t1/2 and t0.95 based on the interpolation of Avrami fractional-order were 20.52 and 246.4 min (R1) and 14.00 and 157.6 min (R2), respectively. Maximum adsorption capacities (Qmax) based on the Liu isotherm were 10.77 (R1) and 63.80 mg g-1 (R2) for ciprofloxacin. The thermodynamic studies showed a spontaneous and exothermic process for both samples, and the value of ΔH° is compatible with physical adsorption.

Longitudinal Study after Sputnik V Vaccination Shows Durable SARS-CoV-2 Neutralizing Antibodies and Reduced Viral Variant Escape to Neutralization over Time.

Recent studies have shown a temporal increase in the neutralizing antibody potency and breadth to SARS-CoV-2 variants in coronavirus disease 2019 (COVID-19) convalescent individuals. Here, we examined longitudinal antibody responses and viral neutralizing capacity to the B.1 lineage virus (Wuhan related), to variants of concern (VOC; Alpha, Beta, Gamma, and Delta), and to a local variant of interest (VOI; Lambda) in volunteers receiving the Sputnik V vaccine in Argentina. Longitudinal serum samples (N = 536) collected from 118 volunteers obtained between January and October 2021 were used. The analysis indicates that while anti-spike IgG levels significantly wane over time, the neutralizing capacity for the Wuhan-related lineages of SARS-CoV-2 and VOC is maintained within 6 months of vaccination. In addition, an improved antibody cross-neutralizing ability for circulating variants of concern (Beta and Gamma) was observed over time postvaccination. The viral variants that displayed higher escape to neutralizing antibodies with respect to the original virus (Beta and Gamma variants) were the ones showing the largest increase in susceptibility to neutralization over time after vaccination. Our observations indicate that serum neutralizing antibodies are maintained for at least 6 months and show a reduction of VOC escape to neutralizing antibodies over time after vaccination. IMPORTANCE Vaccines have been produced in record time for SARS-CoV-2, offering the possibility of halting the global pandemic. However, inequalities in vaccine accessibility in different regions of the world create a need to increase international cooperation. Sputnik V is a recombinant adenovirus-based vaccine that has been widely used in Argentina and other developing countries, but limited information is available about its elicited immune responses. Here, we examined longitudinal antibody levels and viral neutralizing capacity elicited by Sputnik V vaccination. Using a cohort of 118 volunteers, we found that while anti-spike antibodies wane over time, the neutralizing capacity to viral variants of concern and local variants of interest is maintained within 4 months of vaccination. In addition, we observed an increased cross-neutralization activity over time for the Beta and Gamma variants. This study provides valuable information about the immune response generated by a vaccine platform used in many parts of the world.

Combining machine learning and texture analysis to differentiate mediastinal lymph nodes in lung cancer patients.

Evaluate whether texture analysis associated with machine learning approaches could differentiate between malignant and benign lymph nodes. A total 18 patients with lung cancer were selected, with 39 lymph nodes, being 15 malignant and 24 benign. Retrospective computed tomography scans were utilized both with and without contrast medium. The great differential of this work was the use of 15 textures from mediastinal lymph nodes, with five different physicians as operators. First and second order statistical textures such as gray level run length and co-occurrence matrix were extracted and applied to three different machine learning classifiers. The best machine learning classifier demonstrated a variability of less than 5% among operators. The support vector machine (SVM) classifier presented 95% of the area under the ROC curve (AUC) and 89% of sensitivity for sequences without contrast medium. SVM classifier presented 93% of AUC and 86% of sensitivity for sequences with contrast medium. Texture analysis and machine learning may be helpful in the differentiation between malign and benign lymph nodes. This study can aid the physician in diagnosis and staging of lymph nodes and potentially reduce the number of invasive analysis to histopathological confirmation.

Chemical and Genotypic Variations in Aniba rosiodora from the Brazilian Amazon Forest.

Aniba rosiodora has been exploited since the end of the nineteenth century for its essential oil, a valuable ingredient in the perfumery industry. This species occurs mainly in Northern South America, and the morphological similarity among different Aniba species often leads to misidentification, which impacts the consistency of products obtained from these plants. Hence, we compared the profiles of volatile organic compounds (essential oils) and non-volatile organic compounds (methanolic extracts) of two populations of A. rosiodora from the RESEX and FLONA conservation units, which are separated by the Tapajós River in Western Pará State. The phytochemical profile indicated a substantial difference between the two populations: samples from RESEX present α-phellandrene (22.8%) and linalool (39.6%) in their essential oil composition, while samples from FLONA contain mainly linalool (83.7%). The comparison between phytochemical profiles and phylogenetic data indicates a clear difference, implying genetic distinction between these populations.

Single-step pyrolysis for producing magnetic activated carbon from tucumã (Astrocaryum aculeatum) seed and nickel(II) chloride and zinc(II) chloride. Application for removal of nicotinamide and propanolol.

The present research describes the synthesis of new nanomagnetic activated carbon material with high magnetization, and high surface area prepared in a single pyrolysis step that is used for the carbonization, activation, and magnetization of the produced material. The pyrolysis step of tucumã seed was carried out in a conventional tubular oven at 600 °C under N2-flow. It was prepared three magnetic carbons MT-1.5, MT-2.0, MT-2.5, that corresponds to the proportion of biomass: ZnCl2 always 1:1 and varying the proportion of NiCl2 of 1.5, 2.0, and 2.5, respectively. These magnetic nanocomposites were characterized by Vibrating Sample Magnetometer (VSM), X-ray diffraction, scanning electron microscopy, thermogravimetric analysis, hydrophobic/hydrophilic balance, CHN/O elemental analysis, modified Boehm titration, N2 adsorption-desorption isotherms; and pHpzc. All the materials obtained presented Ni particles with an average crystallite size of less than 33 nm. The MT-2.0 was employed for the removal of nicotinamide and propranolol from aqueous solutions. Based on Liu isotherm, the Qmax was 199.3 and 335.4 mg g-1 for nicotinamide and propranolol, respectively. MT-2.0 was used to treat simulated pharmaceutical industry effluents attaining removal of all organic compounds attaining up to 99.1 % of removal.

Ahorcamiento con soga. Presentación de un caso atípico / Hanging with rope. Presentation of an atypical case

RESUMEN Las asfixias mecánicas son aquellas que resultan de un impedimento mecánico a la penetración del aire en las vías respiratorias. Suelen clasificarse atendiendo a la naturaleza del medio mecánico que las origina, y a su modo de actuar. El ahorcamiento no es más que la constricción del cuello, ejercida por un lazo sujeto a un punto fijo, sobre el cual ejerce tracción el propio peso del cuerpo. La muerte, según las circunstancias del caso, puede ocurrir por un mecanismo asfíctico, circulatorio, inhibitorio o por lesión medular (caso raro de presentación). El presente trabajo constituyó un caso atípico de ahorcamiento. El ciudadano que resultó fallecido, al dejarse caer bruscamente del balcón de su vivienda, con el dogal en el cuello (soga) y quedar pendiendo el cuerpo, ocasionó un traumatismo raquimedular severo; lo que trajo como consecuencia que se produjera una fractura de la 3ra vértebra cervical, y una contusión medular que lo llevó a la muerte de forma instantánea por el shock medular ocasionado. Para la discusión del caso se tuvo en cuenta los elementos del lugar del hecho, y el examen del cadáver externa e internamente.

ABSTRACT Mechanical asphyxias are those resulting from a mechanical interruption of air penetration to the respiratory tract. They are usually classified according to the nature of the mechanical mean originating them and the way they perform. Hanging is no more that the neck constriction, executed by a lace attached to a fixed point, on which the proper weight of the body applies traction. Death, according to the circumstances of the case, may occur by an asphyxiating, circulatory, inhibitory mechanism or by medullar lesion (a rare case of presentation). The current work reports a case of atypical hanging. The person who died, when abruptly jumped down the balcony of his house with the rope around the neck and the body was left pending, suffered a severe spinomedullar trauma, causing a fracture of the 3rd cervical vertebra, and a medullar contusion causing an instantaneous death due to the medullar shock. To discuss the case, the authors took into account elements of the location where it took place, and the external and internal examination of the body.

Evidence for Snake Venom Plasticity in a Long-Term Study with Individual Captive Bothrops atrox.

Variability in snake venom composition has been frequently reported and correlated to the adaptability of snakes to environmental conditions. Previous studies report plasticity for the venom phenotype. However, these observations are not conclusive, as the results were based on pooled venoms, which present high individual variability. Here we tested the hypothesis of plasticity by influence of confinement and single diet type in the venom composition of 13 adult specimens of Bothrops atrox snakes, maintained under captivity for more than three years. Individual variability in venom composition was observed in samples extracted just after the capture of the snakes. However, composition was conserved in venoms periodically extracted from nine specimens, which presented low variability restricted to the less abundant components. In a second group, composed of four snakes, drastic changes were observed in the venom samples extracted at different periods, mostly related to snake venom metalloproteinases (SVMPs), the core function toxins of B. atrox venom, which occurred approximately between 400 and 500 days in captivity. These data show plasticity in the venom phenotype during the lifetime of adult snakes maintained under captive conditions. Causes or functional consequences involved in the phenotype modification require further investigations.

Evaluation of efficiency and selectivity in the sorption process assisted by chemometric approaches: Removal of emerging contaminants from water.

This paper describes, by the first time, a chemometric approach that combines a simple set of the UV-Vis spectra and partial least square regression (PLSR) for measuring the removal of five pharmaceuticals present in simulated hospital effluents by sorption using activated carbon. The use of multivariate calibration allowed the quantification of the remaining concentrations of the studied drugs present in a complex mixture with high accuracy, avoiding the need for the use of sophisticated methodologies based on chromatography. Isothermal sorption studies were performed on single-component solutions containing amoxicillin, paracetamol, propranolol, sodium diclofenac, or tetracycline as well as on a solution containing a mixture of all these 5 compounds. The isotherm data obtained were fitted to the Langmuir, Freundlich and Liu models. It was observed that for each pharmaceutical, the maximum sorption capacity of the activated carbon was higher for the single component than in the mixture. It was observed that the removal of paracetamol, propranolol, and tetracycline, the removal was complete (100%) and for amoxicillin and sodium diclofenac it was at least 92.71 ±â€¯3.15% and 91.82 ±â€¯0.95% respectively, indicating that the avocado seed activated carbon is an adsorbent with high sorption capacity that can remove five pharmaceuticals from simulated hospital effluents.

HER1-based vaccine: Simultaneous activation of humoral and cellular immune response.

The human epidermal growth factor receptor 1 (HER1) is a tumor-associated antigen that has been validated as a clinical target for several passive, non-immune therapies currently approved for the treatment of epithelial tumors. HER1 is an oncogene that not only promotes tumor progression and survival, but also immune escape. Its overexpression in some epithelial malignancies has been correlated with a poor prognosis. We developed an approach to target HER1 by specific active immunotherapy, recognizing the extracellular domain of the receptor, using a combination of VSSP and Montanide ISA 51 as adjuvants. We summarize the results obtained with this vaccine in both the preclinical and clinical settings, emphasizing the importance of the induction of both humoral and cellular responses for the success of cancer vaccines as safe therapeutic alternatives for the treatment of cancer.

Phosphorus, not nitrogen, limits plants and microbial primary producers following glacial retreat.

Current models of ecosystem development hold that low nitrogen availability limits the earliest stages of primary succession, but these models were developed from studies conducted in areas with temperate or wet climates. Global warming is now causing rapid glacial retreat even in inland areas with cold, dry climates, areas where ecological succession has not been adequately studied. We combine field and microcosm studies of both plant and microbial primary producers and show that phosphorus, not nitrogen, is the nutrient most limiting to the earliest stages of primary succession along glacial chronosequences in the Central Andes and central Alaska. We also show that phosphorus addition greatly accelerates the rate of succession for plants and for microbial phototrophs, even at the most extreme deglaciating site at over 5000 meters above sea level in the Andes of arid southern Peru. These results challenge the idea that nitrogen availability and a severe climate limit the rate of plant and microbial succession in cold-arid regions and will inform conservation efforts to mitigate the effects of global change on these fragile and threatened ecosystems.

Administration of Probiotics Improves the Brine Shrimp Production and Prevents Detrimental Effects of Pathogenic Vibrio Species.

In this study, we evaluated a consortium of probiotic bacteria as an environmentally-friendly strategy for controlling pathogenic Vibrio species during the brine shrimp incubation period. Probiotic strains were initially selected on basis of (i) their ability to colonize the cyst surfaces, (ii) their absence of cross-inhibitory effects, and (iii) no detrimental effect on cyst hatching. The cysts and nauplius surfaces were immediately colonized after the application of selected probiotic strains, without detrimental effects on survival. Ten probiotic strains were mixed at similar proportions (probiotic consortium) and evaluated at different concentrations into brine shrimp cultures during incubation and early stages of development. Subsequently, these cultures were challenged with Vibrio parahaemolyticus and Vibrio harveyi. The probiotic consortium was effective to reduce the abundance of pathogenic Vibrio species and to prevent the mortality during Vibrio challenges; however, its effect was concentration-dependent and was successful at a starting concentration of 1.8 × 106 CFU/ml. Our results suggest that this probiotic consortium offers an alternative to antimicrobial agents routinely used to reduce the incidence and prevalence of pathogenic Vibrio species in brine shrimp production.

Molecular mechanisms underlying intraspecific variation in snake venom.

Elucidating the molecular mechanisms underlying snake venom variability provides important clues for understanding how the biological functions of this powerful toxic arsenal evolve. We analyzed in detail individual transcripts and venom protein isoforms produced by five specimens of a venomous snake (Bothrops atrox) from two nearby but genetically distinct populations from the Brazilian Amazon rainforest which show functional similarities in venom properties. Individual variation was observed among the venoms of these specimens, but the overall abundance of each general toxin family was conserved both in transcript and in venom protein levels. However, when expression of independent paralogues was analyzed, remarkable differences were observed within and among each toxin group, both between individuals and between populations. Transcripts for functionally essential venom proteins ("core function" proteins) were highly expressed in all specimens and showed similar transcription/translation rates. In contrast, other paralogues ("adaptive" proteins) showed lower expression levels and the toxins they coded for varied among different individuals. These results provide support for the inferences that (a) expression and translational differences play a greater role in defining adaptive variation in venom phenotypes than does sequence variation in protein coding genes and (b) convergent adaptive venom phenotypes can be generated through different molecular mechanisms. SIGNIFICANCE: Analysis of individual transcripts and venom protein isoforms produced by specimens of a venomous snake (Bothrops atrox), from the Brazilian Amazon rainforest, revealed that transcriptional and translational mechanisms contribute to venom phenotypic variation. Our finding of evidence for high expression of toxin proteins with conserved function supports the hypothesis that the venom phenotype consists of two kinds of proteins: conserved "core function" proteins that provide essential functional activities with broader relevance and less conserved "adaptive" proteins that vary in expression and may permit customization of protein function. These observations allowed us to suggest that genetic mechanisms controlling venom variability are not restricted to selection of gene copies or mutations in structural genes but also to selection of the mechanisms controlling gene expression, contributing to the plasticity of this important phenotype for venomous snakes.