Cytokine mRNA expression and pathological findings in pigs inoculated with African swine fever virus (E-70) deleted on A238L

African swine fever virus (ASFV) induces a variety of immune responses and clinical forms in domestic pigs. As it is the only member of the Asfarviridae family, ASFV encodes many novel genes not encoded by other virus families. Among these genes, A238L may regulate the synthesis of pro-inflammatory cytokines, controlled mainly by NFkappaB and NFAT pathways. In this study, we inoculated two groups of pigs, one with the ASFV highly virulent E-70 isolate, deleted on A238L gene, and the other group with the parental E-70 isolate. No significant differences were observed in the clinical signs or pathology between both groups. However, the TNF-alpha mRNA expression was strongly enhanced in the PBMC from pigs inoculated with the virus deleted in A238L, reinforcing the role of the A238L gene in the inhibition of the NFkappaB pathway of expression of cytokines. No up-regulation of pro-inflammatory cytokines was observed in the PBMC of animals inoculated with the E-70 isolate, even though apoptosis and haemorrhages were evident and might be related to the presence of bystander monocyte-macrophages expressing these cytokines. Other studies using ASFV deleted in other genes inoculated in the natural hosts should be performed to gain further insight into the role of these genes in the pathogenesis of ASF.     

Replacement of fishmeal by vegetable meal mix in the diets of Litopenaeus vannamei reared in low‐salinity biofloc system: effect on digestive enzymatic activity

Biofloc (consortium of diverse microorganisms associated to suspending substrates) was developed from waste of shrimp Litopenaeus vannamei postlarvae culture under low salinity (5 g L^?1) to provide an additional nutritious biomass and reduce fishmeal inclusion in feeds in a 28‐day indoor shrimp nursery trial conducted in 15 experimental containers (250 L stocked at 600 org m^?3). Four experimental diets (isoproteic and isocaloric) containing different percentage of fishmeal: 0%, 10%, 20% and 30% substituted by vegetable meal mix (corn, sorghum and wheat) were formulated and elaborated. A control treatment consisted of a commercial feed. The main water quality parameters were monitored, and no significant differences were found among treatments. The growth and survival were similar among treatments. In general, digestive enzymatic activities showed differences being greater in the biofloc system compared with clear water. It was concluded that low‐salinity shrimp nursery could be successfully developed with minimum inclusion of fishmeal in feeds, without significant effect on production response. The adjustment of C : N ratio allowed the increase of microbial biomass in the bioflocs, which contributed to maintain good water quality, provide live food and enhance digestive enzymatic activity of cultured organisms.     

Effect of human tubal fluid medium and hyperactivation inducers on stallion sperm capacitation and hyperactivation

Conventional in vitro fertilization has not yet been implemented in the equine species. One of the main reasons has been the inability to develop a culture medium and incubation conditions supporting high levels of stallion sperm capacitation and hyperactivation in vitro. Although different culture media have been used for this purpose, human tubal fluid (HTF) medium, widely used in the manipulation of human and mice gametes, has not been reported so far in stallion sperm culture. The first part of this study aimed to compare HTF and Whitten's media on different stallion sperm quality and capacitation variables. Additionally, the effect of procaine, aminopyridine and caffeine in both media was evaluated on sperm motility parameters at different incubation times. Integrity and destabilization of the plasma membrane were evaluated by merocyanine 540/SYTOX Green (MC540), mitochondrial membrane potential (?Ψm) using tetramethylrhodamine methyl ester perchlorate (TMRM), acrosome membrane integrity by PNA/FITC and tyrosine phosphorylation by P‐tyrosine mouse mAb conjugated to Alexa Fluor® by flow cytometry. Motility parameters were evaluated using the integrated semen analysis system (ISAS®). We found no differences between Whitten's and HTF media and incubation time in terms of sperm viability, uninduced acrosome membrane damage or mitochondrial membrane potential at 30‐ and 120‐min incubation. Membrane fluidity (MC540) increased in both media at 30‐ and 120‐min incubation compared to noncapacitating conditions. Similarly, tyrosine phosphorylation increased in both media in capacitating conditions at 2‐ and 4‐hr incubation compared to noncapacitating conditions. Although procaine showed the best result in terms of sperm hyperactivated motility in both media, aminopyridine also showed parameters consistent with the hyperactivation including an increase in curvilinear velocity and decrease in straightness. In conclusion, HTF medium and aminopyridine equally support capacitation‐related parameters in stallion sperm.     

A Morphological Study of Cultured Endodermal Cells of Chick Embryo: Characteristics of Adhesion, Spreading and Locomotion

A study is made of the morphological characteristics of the endodermic cells of the stage 5 chick embryo by means of in vitro cell culture techniques. The scanning electron microscope revealed that the endodermic cells in cultures were rounded, tended to be smooth and had few blebs and microvilli. Regarding cell projections typical of culture cells, such as filopodia, lamellipodia and pseudopodia, there was a noteworthy scarcity after 12 h growth, although greater cellular activity was observed at 24 h, characterized by the presence of filopodia and an ability of the cells to form clusters on the substratum. These facts show the morphological and adhesion movements of the endodermal cells studied to be related mainly with the presence of filopodia as the most abundant cell projections.     

Epidemiological pattern and risk factors associated with bovine viral-diarrhoea virus (BVDV) infection in a non-vaccinated dairy-cattle population from the Asturias region of Spain

A survey of bovine viral-diarrhoea virus (BVDV) infection was carried out in a non-vaccinated cattle population from the Asturias region of Spain in 1997 to assess seroprevalence and identify risk factors associated with infection. Twenty-eight herds were included; 529 cows were bled. Information regarding the herd and each animal sampled were recorded through a personal interview with the farmer. The true prevalence was estimated to be 21%. According to the antibody-age profiles and the herd-management characteristics, no persistently infected animals were suspected at that time within the herds sampled. Random-effects logistic regression found two major factors associated with seropositivity: age and cow origin. Results suggested that BVDV infection could be controlled in that area by livestock-trade control (without vaccines). In addition, an increasing risk of abortion was not observed when cows were seropositive to both BVDV and Neospora caninum infections.     

Correlation of lycopene measured by HPLC with the L, a, b color readings of a hydroponic tomato and the relationship of maturity with color and lycopene content

Tomatoes (Lycopersicon esculentum cv. Laura) were separated, according to the ripening stage, by a sensory panel into seven groups, and color was measured on the tomato surface with a Minolta Chroma meter. The L, a, b, hue, chroma, and lycopene content were plotted against the maturity stages of the tomatoes, and several good correlations were found. The a/b ratio and the lycopene content were the parameters that allowed six of seven maturity groups in the tomato to be statistically distinguished. The lycopene content, measured by HPLC, was also correlated with the color measurements, and the a, a/b, and (a/b)(2) color factors produced the best regressions. An estimation of the lycopene content in tomatoes can be achieved by using a portable chroma meter, with a possible field usage application. Equations to calculate the lycopene content of tomatoes based on the color readings are reported.     

Estimation of phosphate adsorption by Acrisols of western Spain from electrical conductivity of potassium dihydrogen phosphate extracts of soil

Estimation of the phosphate adsorption capacity in highly weathered acid soils is crucial for adopting appropriate phosphate fertilization practices. In this work, the salt sorption effect was used to develop a method for estimating phosphate adsorption capacity from the decrease in electrical conductivity of a potassium dihydrogen phosphate solution brought in contact with acidic soil samples from surface and subsurface horizons of Acrisols. The method is fast and simple and requires only the use of a conductimeter.     

Hg, Cu, Pb, Cd, and Zn Accumulation in Macrophytes Growing in Tropical Wetlands

The concentrations of Hg, Cu, Pb, Cd, and Zn accumulated by regional macrophytes were investigated in three tropical wetlands in Colombia. The studied wetlands presented different degrees of metal contamination. Cu and Zn presented the highest concentrations in sediment. Metal accumulation by plants differed among species, sites, and tissues. Metals accumulated in macrophytes were mostly accumulated in root tissues, suggesting an exclusion strategy for metal tolerance. An exception was Hg, which was accumulated mainly in leaves. The ranges of mean metal concentrations were 0.035?C0.953 mg g^?1 Hg, 6.5?C250.3 mg g^?1 Cu, 0.059?C0.245 mg g^?1 Pb, 0.004?C0.066 mg g^?1 Cd, and 31.8?363.1 mg g^?1 Zn in roots and 0.033?C0.888 mg g^?1 Hg, 2.2?C70.7 mg g^?1 Cu, 0.005?C0.086 mg g^?1 Pb, 0.001?C0.03 mg g^?1 Cd, and 12.6?C140.4 mg g^?1 Zn in leaves. The scarce correlations registered between metal concentration in sediment and plant tissues indicate that metal concentrations in plants depend on several factors rather than on sediment concentration only. However, when Cu and Zn sediment concentrations increased, these metal concentrations in tissues also increased in Eichhornia crassipes, Ludwigia helminthorriza, and Polygonum punctatum. These species could be proposed as Cu and Zn phytoremediators. Even though macrophytes are important metal accumulators in wetlands, sediment is the main metal compartment due to the fact that its total mass is greater than the corresponding plant biomass in a given area.