Luteal vascularity and embryo dynamics in mares during early gestation: Effect of age and endometrial degeneration

The present study characterized the luteal status and the dynamic of the conceptus during the first 20 days of gestation in mares with different ages and degrees of endometrial degeneration. Total area of the corpus luteum (CL), luteal vascularity, CL area with blood signals, progesterone concentrations (P4), embryonic vesicle diameter, number of embryonic location changes, embryonic fixation position and uterine contractility were evaluated. In Experiment 1, mares ≤6 years of age (Young group, 5.6 ± 0.2 years, n = 7 mares) and mares ≥15 years of age (Old group, 17.2 ± 0.9 years, n = 6 mares) were used to investigate the effect of age. In Experiment 2, the luteal and embryonic parameters were compared between mares with minimal (Mild group, endometrial category I, n = 9 mares) and severe (Severe group, endometrial category III, n = 7 mares) endometrial degeneration. The Old and Severe groups had greater (p ≤ 0.04) total CL area and reduced luteal vascularity (p ≤ 0.04) than the Young and Mild groups, respectively. However, P4 levels and CL area with blood signals were similar (p ≥ 0.8) between the groups. A negative effect of age (p < 0.01), but not of endometrial degeneration (p = 0.6), was found for the embryonic vesicle diameter. The conceptus mobility was high (p > 0.1) until day 14 of gestation in the Severe group, while a reduced number of changes of the embryo location was detected earlier (p < 0.05) in the Old group. In conclusion, the newly formed CL of aged mares and mares with severe endometrial degeneration suffered a structural remodelling to safeguard the local blood supply and the continuous P4 output during early gestation. Moreover, an earlier reduction of the embryonic mobility and a delayed development of the conceptus were associated with advanced age, regardless of the degree of endometrial degeneration.     

IRMS as a tool to obtain the carbon turnover (δ13C) in organs of weaned piglets fed glutamic acid and nucleotides

Early weaning of piglets causes stress characterized by a decrease in feed intake followed by a decline in growth rates; thus, a fast recovery represents an essential step for proper growth of these animals. Considering that IRMS is a potential tool for non‐destructive sampling and the fact that it provides time‐integrated estimate of assimilated and not just ingested nutrients turned possible its application to evaluate the effects of dietary nucleotides and glutamate on carbon turnover (δ¹³C) in organs of weanling piglets. At day 0, three piglets were slaughtered (prior to diet switch), the remaining eighty‐four piglets weaned at 21‐day‐old were randomly assigned in a complete block design with a 2 × 2 factorial arrangement of treatments (two Nu levels: 0 and 0.1% and two Glu levels: 0 and 1%), being three piglets per treatment slaughtered on trial days 3, 6, 9, 14, 21, 35 and 49. The samples were analysed by IRMS and adjusted to first‐order equation by a non‐linear regression analysis using NLIN of SAS, in order to establish exponential graphics. After that, the turnover data were submitted to analysis of variance using GLM of SAS. The turnover value (t_95%) verified for spleen was faster (p < 0.05) when glutamate was supplemented in diets. For pancreas and liver, the turnover rates were faster (p < 0.05) for the mixture of additives. However, for renal tissue, the turnover rate (t_95%) was greater (p < 0.05) for the free additive diet. The results obtained suggest that the mixture of additives was more efficient to develop the digestive tract at post‐weaning phase, taking into account the functional importance of pancreas and liver for nutrients’ digestion and processing.     

Total nutrient digestibility and small intestine starch digestion in Nellore and Angus young bulls fed a whole shelled corn diet

Eighteen Nellore and 18 Angus young bulls with BW of 381 ± 12 kg were randomly assigned into two feeding groups (whole shelled corn [WSC] or ground corn with silage [GC]) to evaluate the interaction of breed and diet on total nutrient digestibility, pancreatic α‐amylase, and maltase activity and SLC5A1expression in the small intestine. Experimental diets (DM basis) included (a) a diet containing 30% corn silage and 70% GC and soya bean meal‐based concentrate and (b) a diet containing 85% WSC and 15% of a soya bean meal‐ and mineral‐based pelleted supplement. The treatments were Nellore fed GC diet; Nellore fed WSC diet; Angus fed GC diet; and Angus fed WSC diet. Total faecal collection for the digestibility trial occurred from day 48 until day 50 of the experimental period. Feeding the WSC diet reduced DM and NDF intake (p < 0.01). Angus had greater DM and nutrient intake in kg/day (p < 0.01). However, there was no breed effect on DM and nutrient intakes based on percentage of BW (p > 0.19). Angus had greater starch digestibility (p = 0.03) than Nellore. Cattle fed the WSC diet had greater DM, NDF and starch digestibility (p < 0.01) compared with those fed the GC diet. The activity of pancreatic α‐amylase (U/g of protein) was greater in Nellore (p < 0.01) and was not affected by diet (p = 0.52). In duodenum, maltase activity (U/g of protein) was greater in bulls fed GC diet (p = 0.02). Expression of the gene SLC5A1was not affected by breed or diet (p > 0.05). In conclusion, Nellore had less capacity to digest starch. However, they did not have less pancreatic α‐amylase and duodenal maltase activity compared to Angus. The use of the WSC diet increases DM and total nutrient digestibility.     

Histology and Immunohistochemistry of the Cardiac Ventricular Structure in the Green Turtle (Chelonia mydas)

This study describes the implications of cardiac ventricular microscopy in Chelonia mydas relating to its ability to dive. For this work, 11 specimens of the marine turtle species C. mydas found dead on the coast of Rio Grande do Norte (Northeast Brazil) were used. After necropsy, fragments of the cardiac ventricular wall were fixed in 10% buffered formaldehyde solution for 24 h and then subjected to routine processing for light and scanning electron microscopy (SEM). The ventricle in this species is formed by the epicardium, myocardium and endocardium. The subepicardial layer consists of highly vascularised connective tissue that emits septa to reinforce the myocardium surface. There is an abundant and diffuse subepicardial nerve plexus shown by immunostaining technique. The thickness of the spongy myocardium and the nature of its trabeculae varied between the heart chambers. The endocardium shows no characteristic elements of the heart conduction system. The valves have a hyaline cartilage skeleton, coated by dense irregular connective tissues characterised by elastic fibres. These findings in the green turtle ventricular microscopy are related to hypoxia resistance during diving.     

CLPP and EEA profiles of microbial communities in salt marsh sediments

Objective  Microbial communities are a central component of trophic dynamics and biogeochemical processes on coastal systems, since most of the processes in sediments are mediated by microorganisms and carried out by enzymes. Microorganisms play a key role in decomposition processes in salt marsh sediments, although the significance of microbial dynamics is largely unexplored. A culture-dependent (Ecoplate) and a culture-independent (extracellular enzyme activity [EEA]) approaches were evaluated in their ability to distinguish the catabolic potential among sediments from Tagus estuary salt marshes with different proximities to anthropogenic sources. Methods  Ecoplate was used to analyse the salt marsh community-level physiological profiles (CLPPs). Results were expressed as the net area under the curve for each of the 31 response wells over a 3-day incubation period in two sediment horizons. The catabolic profiles for salt marsh samples were analysed by Principal Component Analysis (PCA) and hierarchic clustering methods. EEA was analysed by fluorescein diacetate (FDA) hydrolysis in two sediment horizons. The FDA is catalysed by extracellular enzymes, i.e. esterases, lipases and partially by proteases. Results were expressed as μg g⁻¹d wt h⁻¹. Results and Discussion  In this study the CLPP and EEA data were not generally correlated. In Corroios salt marsh only in surface sediments higher net areas corresponded to higher extracellular enzymatic activity, and in Alcochete deep sediments lower net areas corresponded to lower enzymatic activity. Although EEA profiles more directly reflect the inherent activity of resident community in each salt marsh sample, the CLPP profiles provide better assessments of diversity. ESS-Submission Editor: Prof. Dr. Peter Schroeder, Institute of Soil Ecology, Department of Rhizosphere Biology, GSF — National Research Center for Environment & Health, Ingolstaedter Landstrasse 1, 85758 Neuherberg, Germany (peter.schroeder@gsf.de)     

Changes in pigments and plastid ultrastructure during ripening of green-fleshed and yellow-fleshed kiwifruit

Fruit of Actinidia deliciosa have green flesh when ripe, whereas fruit of Actinidia chinensis often have yellow flesh when ripe. The outer and inner pericarps of A. deliciosa fruit retain their chlorophyll during ripening. In those fruit of A. chinensis that become yellow on ripening, the colour change is caused by the disappearance of chlorophylls unmasking the yellow carotenoids already present, rather than by increased synthesis of carotenoids. In fruit of A. deliciosa and those fruit of A. chinensis that remain green, the chloroplasts retain their typical morphology during ripening, whereas in fruit of A. chinensis that become yellow during ripening the chloroplasts of the immature green fruit are transformed into fully developed chromoplasts.     

Contrastant banana accessions for resistance to the burrowing nematode,based on molecular markers RAPD

This work aimed to proceed molecular characterization of seven banana accessions (Borneo, Grand Naine, 1304-06, 4249-05, 0337-02, 0323-03 and 4279-06) resistance to the nematode Radopholus similis. These accessions were selected taking in account the reproduction factor (RF) among 26 banana genotypes from a working collection belonging to Embrapa Mandioca e Fruticultura Tropical. The genomic DNA of the seven accessions was extracted, and 36 decamere primers had been used to obtain RAPD markers. The resulting markers were converted into a matrix of binary data. From that matrix the genetic distance between the accessions were estimated, for further clustering and graphic dispersion analyses. From a total of 521 RAPD markers generated, 420 (81%) were polymorphic, including 140 (27%) potentially promising for application on works related to genetic mapping of the resistance to R. similis. OPE-15, OPH-17, and OPG-09 were the primers that contributed to the highest number of bands promising for genetic mapping of resistance (12, 8, and 8, respectively). The genetic distances between accessions ranged from 0.106 to 0.455, with the longest one observed between cv. Borneo and the genotype 4279-06, considered as highly susceptible and resistant, respectively, to the nematode according to the RF. The graphic dispersion distinguished three groups of accessions, and most of resistant genotypes clustered together in the same group. The most contrastant genotypes for resistance (Borneo and 4249-05) were separated by a genetic distance of 0.374, and possessed a total of 114 polymorphic bands promising for genetic mapping of resistance. In addition, the results of pathogenicity tests were congruent with those obtained by RAPD analyses.     

Development of a real-time PCR method based on duplo target plasmids for determining an unexpected genetically modified soybean intermix with feed components

The occurrence of intermixing, especially that resulting from genetically modified (GM) species, is increasingly becoming a problem in the delicate chain of feed and food quality control. Thus, a strategy is needed for precisely quantifying the presence of intermixing. An analytical assay based on real-time PCR has been developed; it can ascertain the extent of unexpected intermixing of GM soybean with maize meal. Three soybean-maize mix levels, with soybean intermix percentages of, respectively, 0.1, 0.5, and 1%, were prepared to simulate samples containing traces of soybean. As calibrator standards, ad hoc multiple-target pGEM-T plasmids containing soybean and maize reference genes in a 1:1 ratio were constructed. Four different maize endogenous genes, alcohol dehydrogenase 1 (adh1), high-mobility group protein a (hmga), invertase 1 (ivr1), and zein (zein), were assessed, each combined with the soybean endogenous lectin 1 (lect1) gene. Plasmids containing adh1-lect1 and zein-lect1 genes were found to be the most reliable calibration systems for this analysis, providing precise and accurate quantification results. Measuring the percentage of GM soybean intermixing makes it possible to calculate the actual transgenic component of the total sample.