Histological alterations in gills of shrimp Litopenaeus vannamei in low‐salinity waters under different stocking densities: Potential relationship with nitrogen compounds

Two experimental modules with different stocking densities (M1 = 70 and M2 = 120 shrimp /m²) were examined weekly over a culture cycle in tanks with low‐salinity water (1.9 g/L) and zero water exchange. Results showed survival rates of 87.7 and 11.9% in M1 and M2, respectively. Water temperature, pH, dissolved oxygen, electrical conductivity and chlorophyll a were not significantly (p > .05) different between modules. In contrast, the concentrations of nitrogen compounds were significantly (p < .05) different between modules, except nitrite‐N (M2 were 2.31 ± 1.38 mg/L N‐TAN, 0.18 ± 0.49 mg/L N‐NO₂^? and 6.83 ± 6.52 mg/L N‐NO₃^?; in M1: 0.97 ± 0.73 mg/L N‐TAN, 0.05 ± 0.21 mg/L N‐NO₂^? and 0.63 ± 0.70 mg/L N‐NO₃^?). When waters of both modules reached higher levels of ammonia and nitrite, histological alterations were observed in gills. The histological alterations index (HAI) was higher in M2 (5‐112) than in M1 (2‐22).     

Poor maternal nutrition inhibits muscle development in ovine offspring

Background

Maternal over and restricted nutrition has negative consequences on the muscle of offspring by reducing muscle fiber number and altering regulators of muscle growth. To determine if over and restricted maternal nutrition affected muscle growth and gene and protein expression in offspring, 36 pregnant ewes were fed 60%, 100% or 140% of National Research Council requirements from d 31 ± 1.3 of gestation until parturition. Lambs from control-fed (CON), restricted-fed (RES) or over-fed (OVER) ewes were necropsied within 1 d of birth (n = 18) or maintained on a control diet for 3 mo (n = 15). Semitendinosus muscle was collected for immunohistochemistry, and protein and gene expression analysis.

Results

Compared with CON, muscle fiber cross-sectional area (CSA) increased in RES (58%) and OVER (47%) lambs at 1 d of age (P < 0.01); however at 3 mo, CSA decreased 15% and 17% compared with CON, respectively (P < 0.01). Compared with CON, muscle lipid content was increased in OVER (212.4%) and RES (92.5%) at d 1 (P < 0.0001). Muscle lipid content was increased 36.1% in OVER and decreased 23.6% in RES compared with CON at 3 mo (P < 0.0001). At d 1, myostatin mRNA abundance in whole muscle tended to be greater in OVER (P = 0.07) than CON. Follistatin mRNA abundance increased in OVER (P = 0.04) and tended to increase in RES (P = 0.06) compared with CON at d 1. However, there was no difference in myostatin or follistatin protein expression (P > 0.3). Phosphorylated Akt (ser473) was increased in RES at 3 mo compared with CON (P = 0.006).

Conclusions

In conclusion, maternal over and restricted nutrient intake alters muscle lipid content and growth of offspring, possibly through altered gene and protein expression.     

Insights into an undescribed high-elevation lake (6,170 m a.s.l.) on Volcán Llullaillaco: A physical and microbiological view

1. High elevation lakes are extreme ecosystems and serve as sentinels of various global changes.
2. An expedition to Volcán Llullaillaco in 1996 discovered an unstudied high-elevation lake (6,170 m a.s.l.) that probably was formed as a result of the past eruptive events or climatic processes such as glacial retreat in the lake basin.
3. This article describes an initial physical characterization of the lake and its microbial communities derived from two sampling expeditions in 2013 and 2016.
4. The microbial community in the lake, with an area between 1.2 and 1.4 ha and a depth of 6.8 m, was dominated by Proteobacteria, Actinobacteria and Haloarchaea. In addition, 26 bacterial isolates were identified within the genera Subtercola, Xylophilus, Rhodanobacter, Mesorhizobium and Pseudomonas.
5. Lago Llullaillaco is one of the highest recorded lakes in the world, and this study highlights the unique microbial diversity of this aquatic ecosystem and the importance of its preservation to understand the complex biological processes under polyextreme conditions.

     

Differential expression of defence-related proteins in Vigna unguiculata (L. Walp.) seedlings after infection with Fusarium oxysporum

This work aimed to study the induction of defence proteins in cowpea seedlings during the first days after infection with the fungi Fusarium oxysporum f. sp. cubenses and F. oxysporum f. sp. phaseoli. Cowpea seeds, after disinfection, were transferred to Petri dishes containing 0.5% agar and, after germination, were infected with a drop of a suspension containing 0.5 × 10⁴ spores ml⁻¹. Seedlings were collected at 24, 48, 72 and 96 h after infection and were dissected into leaves, hypocotyls, roots, cotyledons and teguments, which were measured and weighed for morphometric analysis. The agar medium was also analyzed. Enzymatic assays of proteic extracts yielded antimicrobial peptides detected by Western blotting. The cowpea seedlings showed a complex pattern of induction and repression of defence proteins in response to infection by both pathogens. Furthermore, morphometric analysis showed differences between infected and control seedlings. Infected samples did not at any time exhibit chitinase activity, but did exhibit different β-1,3-glucanase and peroxidase activities. Western blotting for lipid transfer protein (LTP) demonstrated its presence in all parts of the infected seedlings. Exuded proteins, also obtained from cowpea seeds in the germination medium, were separated by SDS-PAGE and tricine gel electrophoresis. The analysis showed that some proteins were exuded from moistened cowpea seeds, particularly after F. oxysporum infection.     

Enrichment of Adult Artemia Biomass and Squid Mantle Muscle,Dosidicus gigas,with Different Ascorbic Acid (L‐Ascorbyl‐2‐Monophosphate‐Na/Ca) Concentrations

L‐ascorbyl‐2‐monophosphate‐Na/Ca (AMP‐Na/Ca) was used as a vitamin C source to investigate its ascorbic acid (L‐AA) enrichment and retention in boosted Artemia biomass (AB) and squid mantle muscle (SM). Different doses of AMP‐Na/Ca (500, 1000, and 1500 AMP‐Na/Ca mg/kg) were gradually dissolved into the culture tanks at time 0 (T₀) and at each hour until Hour 6 (T₆). Samples of AB and SM were taken for AMP‐Na/Ca and L‐AA analysis at T₀, T₁, T₂, T₃, T_4, T₅, T₆, T₁₂, and T₂₄. There were no significant differences (P > 0.05) among the AB groups at T₁. The T₆ enrichment analysis for AB resulted in significant differences (P < 0.05) in the AMP‐Na/Ca content for the 1500 mg/kg treatment, in which the initial concentration (0.001 ± 0.002 mg/kg) increased by more than 16‐fold. For all AB enrichment treatments, the AMP‐Na/Ca content demonstrated a decrease (32–11%) for the T₆, T₁₂ and T₂₄ analysis. The T₁ analysis for SM at the higher AMP‐Na/Ca enrichment concentration registered 30 mg/kg of L‐AA and decreased (27.6%) at T₆. This study demonstrated that AB and SM can be boosted with AMP‐Na/Ca.     

Effects of Inorganic and Organic Fertilization on Physicochemical Parameters,Bacterial Concentrations,and Shrimp Growth in Litopenaeus vannamei Cultures with Zero Water Exchange

To identify ways to improve water quality and shrimp production in closed systems, two parallel experiments (one in tanks and one in ponds) were conducted using Pacific white shrimp, Litopenaeus vannamei, cultures. In both experiments, the effects of inorganic (Nutrilake®) and organic (molasses) fertilization on physicochemical parameters, bacterial concentrations, and shrimp performance under zero water exchange were evaluated. Fertilization with both molasses and Nutrilake enhanced the feed conversion rate, as well as shrimp survival and production. In tanks, the shrimp survival and production rates were highest in the molasses treatment, but this effect was not observed in ponds. In ponds, fertilization with Nutrilake increased nitrogen and phosphorus concentrations more than did the controls and molasses treatments toward the end of the experiment. In tanks, fertilization with molasses reduced ammonia concentrations toward the end of the experiment, but the same effect was not observed in ponds. In ponds, fertilization reduced the proportion of Vibrio spp. bacteria, which most likely reduced the incidence of disease from these potentially pathogenic organisms. In both culture systems, fertilization increased the proportion of Bacillus spp., which most likely enhanced food availability.     

Secretion of multi-protein migratory complex induced by Toxoplasma gondii infection in macrophages involves the uPA/uPAR activation system

Toxoplasmosis is a world wide spread zoonosis caused by Toxoplasma gondii, an obligate intracellular parasite that is able to disseminate into deep tissues and cross biological barriers, reaching immunoprivileged sites such as the brain and retina. The parasite is able to infect macrophages and dendritic cells for dispersal throughout the body. However, the molecular mechanisms or outcomes of the subversion of the host cell are largely unknown. Recently our group established that metalloproteinases are involved in migration of infected macrophages. Herein, we evaluated the recruitment of host invasive machinery components in T. gondii infected murine macrophages. We showed by immunoprecipitation assays that MMP-9, CD44 TIMP-1 and uPAR were secreted as a multi-protein complex by infected macrophages. Zymographic analysis revealed that MMP-9 was present in its pro- and active form. Moreover, inhibition of uPA/uPAR pathway by PAI-1 decreased secretion of MMP-9 active forms, as well those associated to uPAR and TIMP-1, but not to CD44. Data presented here suggest that MMP-9 is secreted as a multiprotein complex by T. gondii infected macrophages, similar to that observed in metastatic cells. We further speculate that uPA/uPAR system is involved in the expression/secretion of complexes containing active MMP-9 forms.     

Poor maternal nutrition inhibits muscle development in ovine offspring

Background:Maternal over and restricted nutrition has negative consequences on the muscle of offspring by reducing muscle fiber number and altering regulators of muscle growth.To determine if over and restricted maternal nutrition affected muscle growth and gene and protein expression in offspring,36 pregnant ewes were fed 60%,100%or 140%of National Research Council requirements from d 31 ± 1.3 of gestation until parturition.Lambs from control-fed(CON),restricted-fed(RES) or over-fed(OVER) ewes were necropsied within 1 d of birth(n = 18) or maintained on a control diet for 3 mo(n = 15).Semitendinosus muscle was collected for immunohistochemistry,and protein and gene expression analysis.Results:Compared with CON,muscle fiber cross-sectional area(CSA) increased in RES(58%) and OVER(47%)lambs at 1 d of age(P 0.01);however at 3 mo,CSA decreased 15%and 17%compared with CON,respectively(P 0.01).Compared with CON,muscle lipid content was increased in OVER(212.4%) and RES(92.5%) at d 1(P 0.0001).Muscle lipid content was increased 36.1%in OVER and decreased 23.6%in RES compared with CON at 3mo(P 0.0001).At d 1,myostatin mRNA abundance in whole muscle tended to be greater in OVER(P = 0.07) than CON.Follistatin mRNA abundance increased in OVER(P = 0.04) and tended to increase in RES(P = 0.06) compared with CON at d 1.However,there was no difference in myostatin or follistatin protein expression(P 0.3).Phosphorylated Akt(ser473) was increased in RES at 3 mo compared with CON(P = 0.006).Conclusions:In conclusion,maternal over and restricted nutrient intake alters muscle lipid content and growth of offspring,possibly through altered gene and protein expression.     

Effect of light limitation on the water quality,bacterial counts and performance of Litopenaeus vannamei postlarvae reared with biofloc at low salinity

The aim of this study was to evaluate the effect of light limitation on the water quality, bacterial counts and performance of Litopenaeus vannamei postlarvae reared with biofloc at low salinity (≈9 g L^?1). Two treatments were designed: T₁ = culture with natural sunlight and T₂ = culture in darkness. After 28 days, in both treatments, the final weight of shrimp was over 0.6 g with a specific growth rate over 7.4% d^?1, and a survival rate over 70%. In both treatments, Vibrio sp. concentration presented low values (culture with natural sunlight = 0.1 to 9.9 × 10² CFU mL^?1, culture in darkness = 0.4 to 11.7 × 10² CFU mL^?1) and Bacillus sp. had high values (culture with natural sunlight = 0.7 to 66.0 × 10⁴ CFU mL^?1, culture in darkness = 0.7 to 65.8 × 10⁴ CFU mL^?1). All water quality parameters remained within the ranges suitable for shrimp culture, except for alkalinity during the first stage of the study. Although in some sampling periods some significant differences were found in bacterial counts and water quality parameters, shrimp productive performance under culture with biofloc at low salinity was not affected significantly by light limitation.     

The wheat AMT2 (AMmonium Transporter) family,possible functions in ammonium uptake and pathogenic/symbiotic interactions

Ammonium uptake into wheat roots relies primarily on two AMmonium Transporters of subfamily one, while the wheat genome comprises 4 to 6 AMT2 type transporters. Plant AMT2s generally show functions in root-to-shoot translocation or pathogenic and symbiotic plant–microorganism interactions. We addressed the activity of TaAMT2s in ammonium transport. Nitrogen-dependent expression implicated a physiological function in ammonium uptake for TaAMT2;1 and in ammonium distribution for TaAMT2;2-6.