Expression study on the porcine PTHLH gene and its relationship with sow teat number

Teat number is an important trait in sows that should accompany the increase in litter size that has been achieved in the last decades through selection. We have previously identified a genome-wide significant QTL for teat number in porcine chromosome SSC5 by means of an experimental Meishan by Iberian F(2) intercross population. In the present report, we have studied the porcine parathyroid hormone-like hormone (PTHLH) gene, which maps to SSC5, as a candidate gene for this trait, as PTHLH is involved in nipple formation during embryogenesis and nipple development during pregnancy and lactation. We have found that porcine PTHLH gene is transcribed into three mRNA species differing in the 5'UTR region. Two of these variants are reported in pigs here for the first time: one was similar to variant 1 described in humans while the other, which was generated by the retention of two small introns, has not been identified before in any other species. In addition, mRNA expression profile for two of the mRNA variants was assessed in 19 pig tissues. Porcine PTHLH showed a widespread expression as it was present in all tested tissues and relative expression of each variant was tissue dependent. Finally, we have performed an association study between a non-synonymous mutation in the coding region of this gene and sow teat number. The PTHLH polymorphism was segregating in our Meishan by Iberian F(2) population at intermediate allelic frequencies. We compared here six different statistical models to choose the one with a better fit and a lower degree of complexity. However, despite the potential negative effect of the PTHLH mutation in the signal peptide of this protein, we did not detect any association between the PTHLH genotype and the sow teat number phenotype, concluding that the causal mutation of the observed QTL is very likely not related to this gene.     

Seroprevalence and isolation of Toxoplasma gondii from free-range chickens from Espírito Santo state, southeastern Brazil

Prevalence of Toxoplasma gondii infection in 510 free-range (FR) chickens (380 from 33 small farms, and 130 from a slaughter house for FR chickens) from Espírito Santo state, southeastern Brazil, was investigated. Antibodies to T. gondii were sought using commercial indirect haemagglutination (IHAT, Imuno-HAI Toxo(?), Wama Diagnóstica, S?o Paulo, Brazil, cut-off 1:16) and the modified agglutination test (MAT, cut-off 1:25) tests. Attempts were made to isolate viable T. gondii from seropositive chickens by bioassay in mice. Pooled samples of brain, heart and quadriceps muscle of one thigh (total 40 g) from 64 chickens with IHAT titers of ≥ 1:16 were minced, digested in pepsin and bioassayed in mice. Antibodies to T. gondii were found in 40.4% (206/510) FR chickens by IHAT (titer ≥ 1:16) and 38.8% (198/510) by MAT (titer ≥ 1:25); concordance between IHAT and MAT was 81.6% (kappa index=0.614). Viable T. gondii was isolated (designated TgCkBr234-281) from 48 of 64 (75%) seropositive (IHAT titers ≥ 1:32) FR chickens. Most isolates of T. gondii were virulent for mice; 100% of mice inoculated with 44 of 48 isolates died of toxoplasmosis within 30 days post inoculation (p.i). An epidemiological investigation revealed that people living in rural areas have little knowledge about the parasite and about the risk of acquiring it from raw meat. Results indicated that the locally available IHAT was useful for screening of chicken sera for T. gondii antibodies.     

Phenology,photosynthesis, and phosphorus in European beech (Fagus sylvatica L.) in two forest soils with contrasting P contents

Phosphorus (P) is often a limiting macronutrient in temperate forests, but knowledge on the phenological and physiological responses of beech (Fagus sylvatica L.) to P deficiency is scarce. In this study, young beech trees were excavated with intact soil cores from two German forests, Unterlüss (LUE) with low soil P and Bad Brückenau (BBR) with high soil P concentrations. The trees were transferred to identical climatic conditions. In the subsequent growth phase phenological stages during bud burst and leaf unfolding were recorded; biomass production and total P concentrations in different tissues were measured. Seasonal fluctuations in photosynthesis and of soluble P in wood and bark exudates were determined. BBR beeches grew faster and produced more and larger leaves than the LUE beeches. Leaf extension and unfolding were delayed in LUE compared with BBR beeches, but not the time point of bud break. All plant tissues of BBR trees contained higher total P concentrations than those of LUE trees. Strong seasonal fluctuations for P in exudates of beech transport tissues, wood and bark, indicated higher P supply in BBR than in LUE plants, especially at the beginning of the growth phase until leaf maturity. Photosynthetic activity of LUE beeches was lower than that of BBR beeches due to stomatal limitations as the result of anatomically smaller stomatal pore widths, but not as the result of acute biochemical limitation of photosynthesis. Our results suggest that developmental retardation and lower photosynthesis under low P availability may be adaptation mechanisms that adjust the acquisition and recycling of P resources to seasonal growth demand.     

The effect of heat treatment of whole cottonseed on site and extent of protein digestion in dairy cows

Six mature Holstein cows in late lactation and fitted with permanent ruminal, duodenal and ideal cannulae were used in a double 3 X 3 Latin-square design to study the effect of roasting or extruding of cottonseed on protein degradation in the rumen and availability of amino acids in the small intestine. Raw whole cottonseed (CS), extruded whole cottonseed (ECS), and roasted whole cottonseed (RCS) were fed in diets containing 17% crude protein and composed of 42% whole cottonseed, 26% corn grain and 29% hay supplemented with dry molasses, vitamins and minerals. The diets were isocaloric and contained 1.86 Mcal NEL/kg. Acid detergent lignin was used as a digesta marker, and diaminopimelic acid as a bacterial marker. Ruminal ammonia concentration was higher (P less than .05) for CS than for ECS and RCS (15.7, 12.7 and 10.2 mM). Concentrations of isobutyrate (1.3 vs 0.8 and 0.8 mM) and isovalerate (1.8 vs 1.1 and 1.1 mM) were higher (P less than .05) for CS compared with ECS and RCS. Non-ammonia nitrogen flow to the duodenum (g/d) was 248, 300 and 350 for CS, ECS, and RCS, with RCS higher (P less than .05) that in CS. Apparent absorption of total amino acids in the small intestine was .80, 1.01, and 1.24 kg/d, for CS, ECS, and RCS, respectively, with RCS higher (P less than .05) than CS. Apparent absorption of essential and non-essential amino acids was 376, 425; 458, 548;610 and 628 g/d for CS, ECS and RCS respectively, with RCS higher (P less than .05) than CS.     

Unmanned aerial vehicle to evaluate frost damage in coffee plants

Damage caused by frost on coffee plants can impact significantly in the reduction of crop quality and productivity. Remote sensing can be used to evaluate the damage caused by frost, providing precise and timely agricultural information to producers, assisting in decision making, and consequently minimizing production losses. In this context, this study aimed to evaluate the potential use of multispectral images obtained by unmanned aerial vehicle (UAV) to analyze and identify damage caused by frost in coffee plants in different climatic favorability zones. Visual evaluations of frost damage and chlorophyll content quantification were carried out in a commercial coffee plantation in Southern Minas Gerais, Brazil. The images were obtained from a multispectral camera coupled to a UAV with rotating wings. The results obtained demonstrated that the vegetation indices had a strong relationship and high accuracy with the frost damage. Among the indices studied the normalized difference vegetation index (NDVI) was the one that had better performances (r?=?? 0.89, R²?=?0.79, MAE?=?10.87 e RMSE?=?14.35). In a simple way, this study demonstrated that multispectral images, obtained from UAV, can provide a fast, continuous, and accessible method to identify and evaluate frost damage in coffee plants. This information is essential for the coffee producer for decision-making and adequate crop management.

     

Effects of Melatonin Implants During Non‐Breeding Season on Sperm Motility and Reproductive Parameters in Rasa Aragonesa Rams

The effect of melatonin implants administered during non‐breeding season in Rasa Aragonesa rams on sperm motility parameters and other reproductive traits was assessed. In a first experiment, two Rasa Aragonesa rams were implanted (with melatonin group M), remaining other two males as control group (C). Semen of each group was collected from 1 May to 23 June, twice or three times a week, and motility parameters were assessed using a computer‐assisted sperm analysis system. Melatonin increased the percentage of progressive motile spermatozoa, particularly during 46–75 days after melatonin implantation (p < 0.01). In experiment 2, M and C in vitro fertilization ability had been determined by zona‐pellucida binding assays, using spermatozoa from experiment 1, obtained 60–70 days after melatonin was implanted. A significantly higher number of spermatozoa attached per oocyte was observed in frozen‐thawed immature ovine oocytes incubated with sperm from M animals than in those incubated with sperm from the C group (p < 0.01). Finally, a field assay (experiment 3) was performed. In this case, five Rasa Aragonesa rams were implanted with melatonin and three remained as control group. Sperm doses from those animals were used for artificial insemination of 2608 Rasa Aragonesa ewes from 39 different farms at non‐breeding season. Fertility, litter size and fecundity were studied. Semen from melatonin implanted rams seemed to increase both fertility and fecundity in ewes inseminated with spermatozoa obtained 46–60 days after implantation (p < 0.1). Thus, melatonin treatment in rams during non‐breeding season modifies sperm motility parameters and seems to improve the fertilization parameters obtained.     

Effect of β‐Carotene Supply During Close‐up Dry Period on the Onset of First Postpartum Luteal Activity in Dairy Cows

The aim of this study was to examine the effect of β‐carotene supply during the close‐up dry period on the onset of first postpartum luteal activity in dairy cows. Twelve cows were supplied with 2000 mg of β‐carotene (20 g Rovimix^®β‐Carotene containing 10%β‐carotene; DSM Nutrition Japan K.K., Tokyo, Japan) by oral administration daily from day 21 before expected calving date to parturition. Fourteen cows (control) did not receive β‐carotene supplementation. Blood samples were obtained on days 21, 14 and 7 before expected calving date and on days 1, 7, 14, 21 postpartum. When the plasma progesterone concentration exceeded 1 ng/ml by day 21 postpartum, luteal activity was assumed to have been initiated. The result showed that serum β‐carotene concentrations in the β‐carotene cows were higher than in the control cows during the experimental period (p < 0.01). The number of cows with the onset of luteal activity by day 21 postpartum was 9/12 in the β‐carotene cows and 4/14 in the control cows (p < 0.05). Retinol, certain metabolic parameters and metabolic hormones concentrations did not differ between β‐carotene and control cows. In addition, serum retinol concentration in β‐carotene cows without luteal activity was lower than in β‐carotene cows with luteal activity (p < 0.05), and serum gamma‐glutamyl transpeptidase concentration in β‐carotene cows with luteal activity (p < 0.05) and control cows without luteal activity (p < 0.01) was higher than in control cows with luteal activity. In conclusion, β‐carotene supply during the close‐up dry period may support the onset of luteal activity during early lactation in dairy cows.     

Evaluation of Potential Thrombin Inhibitors from the White Mangrove (Laguncularia racemosa (L.) C.F. Gaertn.)

The aim of this work was to verify the effects of methanol (MeOH) and hydroalcoholic (HA) extracts and their respective partition phases obtained from white mangrove (Laguncularia racemosa (L.) C.F. Gaertn.) leaves on human thrombin activity. Among the extracts and phases tested, only the ethyl acetate and butanolic partitions significantly inhibited human thrombin activity and the coagulation of plasma in the presence of this enzyme. Chromatographic analyses of the thrombin samples incubated with these phases revealed that different compounds were able to interact with thrombin. The butanolic phase of the MeOH extract had the most potent inhibitory effects, reducing enzymatic activity and thrombin-induced plasma coagulation. Two glycosylated flavonoids in this partition were identified as the most potent inhibitors of human thrombin activity, namely quercetin-3-O-arabinoside (QAra) and quercetin-3-O-rhamnoside (Qn). Chromatographic analyses of thrombin samples incubated with these flavonoids demonstrated the chemical modification of this enzyme, suggesting that the MeOH extract contained other compounds that both induced structural changes in thrombin and diminished its activity. In this article, we show that despite the near absence of the medical use of mangrove compounds, this plant contains natural compounds with potential therapeutic applications.     

Testing the relationship between instantaneous peak flow and mean daily flow in a Mediterranean Area Southeast Spain

Extreme hydrologic events have great importance in Southeastern Spain regarding the personal and economic damage they imply. The commonly-used design parameter for hydraulic structures is the maximum annual instantaneous stream flow recorded in conventional gauging stations. However, the majority of available data in Southeastern Spain is mean daily stream flows. This paper explores possible linear relationships between annual instantaneous peak discharge (IPF) and the corresponding (MDF) mean daily stream flow. This relationship was previously explored by other authors such as Fuller [Fuller, W.E., 1914. Flood flows. Trans. Am. Soc. Civ. Eng., 77: 564–617]. Non-linear responses of IPF–-MDF were observed in several study basins. The use of Principal Components Analysis (PCA) allowed characterizing the most important topographic and hydrological attributes of the basins and provided important information about variables that should be included in IPF–MDF regional equations. The key factor to justify the different IPF–MDF relationships in a relatively small area is the nature of the extreme events and their effects on semi-arid soil conditions. In addition, a regional equation to estimate IPF from MDF was developed. This equation was applied to a series of flow of nine stations of the Southeast Basin of Spain, and a significant improvement was achieved when applying this formula in comparison to the traditional method of Fuller. This study indicates possible restrictions to take into account when traditional hydrological model are applied in semi-arid areas.