Conception rate,uterine infection and embryo quality after artificial insemination and natural breeding with a stallion carrier of Pseudomonas aeruginosa: a case report

Background

Pseudomonas aeruginosa may cause venereal disease and infertility in horses. A Pseudomonas aeruginosa - carrier stallion, often unresponsive to artificial vagina collection, was used to naturally breed mares. Semen collected from the same stallion was also used to perform artificial inseminations. Pregnancy rates, embryo quality and incidence of uterine infection were compared between inseminated or naturally-bred mares.

Methods

P. aeruginosa was isolated from swabbing of the penis, prepuce and distal urethra of the stallion. Before being bred or inseminated, clitoral/vestibular samples were collected from all mares, and cultured for isolation of P. aeruginosa. At the first observed estrus, endometrial swabs were also collected. All mares subjected to natural mating (NS) were re-evaluated for P.aeruginosa by culture of clitoral and endometrial swabs. Artificial inseminations (AI) were performed either with fresh-extended semen (11 AI/7 mares) or frozen semen (10 AI/7 mares). The stallion was also used to breed 3 mares (4 services). For embryo collection, 2 mares were inseminated with fresh-extended semen (1 AI/mare), and 2 additional mares were inseminated with frozen semen (2 AI/mare). Two mares were naturally-bred with a total of 9 services, for embryo collection. All mares were examined after AI or natural service (NS), for uterine pathologies. Embryo recoveries were attempted passing a catheter with inflatable cuff connected to a sterile flexible 2-way flushing catheter, through the cervix. Flushed media was recovered into an Em-Con filter, and embryos searched using a stereoscope. Embryos were graded from 1 (excellent) to 4 (degenerated/dead).

Results

Pregnancy rates obtained after NS was 50% per cycle. However, more than half of the NS resulted in uterine disease, while uterine pathology was seen only in 22% of the time following AI. Half of the mares bred by NS got positive to P. aeruginosa. Percentage of embryo recovery rates was identical after AI or NS (66.7%). The 4 embryos recovered after AI were classified as Grade 1, while after NS only 2 out of the 6 recovered embryos were Grade 1.

Conclusion

a) there was no evidence of reduced fertilization after AI or NS, b) a numerically higher incidence of uterine disease was noticed after NS, c) venereal transmission of P. aeruginosa after NS was confirmed, d) a lower percentage of G1 embryos may be obtained after NS. Overall, the data supports the indication for P. aeruginosa-carrier stallions to be bred by AI rather than by NS, and raises the possibility that P. aeruginosa may affect embryo quality.     

Antioxidant Compounds Recovery from Juçara Residue by Thermal Assisted Extraction

This study aimed to recover bioactive compounds by solid-liquid extraction from the agro-industrial residue obtained during juçara fruits processing into pulp. A preliminary study using different solvents (methanol, ethanol and water) indicated ethanol in aqueous solution as the best solvent for antioxidants recovery. Then, a Box-Behnken design was applied considering as independent variables the solvent composition (30–70% ethanol in water), temperature (30–70 °C) and time (30–60 min), in order to evaluate the effects of these factors on antioxidant activity in juçara extract. Results showed that the extracts with higher antioxidant activity were obtained using 30% ethanol at 70 °C for 60 min; measurements included ABTS and DPPH assays, determination of total phenolic content and total monomeric anthocyanins. Furthermore, the effect of pH in antioxidants recovery was evaluated. For this purpose, the 30% ethanol solution was acidified to pH 1 and 2 with HCl. Principal component analysis showed the formation of three distinct groups: one characterized by high bioactive compounds content (pH 1.0), another with superior antioxidant activity (pH 5.75, non-acidified), and finally the group at pH 2 presenting the worst concentrations in the evaluated responses. HPLC analysis showed the presence of cyanidin-3-O-rutinoside and cyanidin-3-O-glucoside in the extracts. Therefore, the conventional solid-liquid extraction using renewable solvent can be successfully applied to recover bioactive compounds from juçara residue, which can be used by different food industries.     

Snapshots Analyses for Turbidity Measurements in Water

Turbidity has been used as an effective indicator of water quality and it is regulated by national control agencies as a water potability parameter. This work proposes a simple, low-cost, and easily reproducible method based on digitally processed snapshots of electromagnetic radiation beam through a scattering medium in order to measure turbidity. The formazin polymer was used as the standard reference in the experiments to prepare samples from 0.1 to 100 NTU for high turbidity range and from 0.02 to 10 FNU for low turbidity range. The device design is comprised of a webcam and an LED/laser as a light sensor and a radiation source, respectively. The captured scattered light snapshot can be decomposed digitally in color components values and correlated with the turbidity parameter. It is possible to obtain differently device performances changing the light sensor device configuration. The linear regression analyses have shown a distinct relationship among red, green, blue, and grayscale components and the turbidity. At high range, the green component present the best LOD and LOQ values 0.64 and 2.10, respectively, operating with an ordinary webcam and white LED. Nevertheless, the best device performances were obtained with dedicated Raspberry Pi camera modules and white LED for low range turbidity reaching LOD=0.027 and LOQ=0.087 FNU. The figures of merit show recovery between (97.50?101.95%), repeatability (2.11%), within-laboratory reproducibility (2.25%), and limits of quantification (0.087 FNU). The achieved performance device shows the effectiveness of digitally processed snapshots obtained with digital cameras for turbidity measurements.     

Ciprofloxacin Adsorption on ZnO Supported on SBA-15

Most drugs are synthesized by human medicine both for the treatment of men and animals and are also produced to maintain their physical and chemical properties for a time sufficient to serve a therapeutic purpose in treatments of some kind of illness. Ciprofloxacin is an antibiotic synthetically obtained in 1987 and belongs to the family of fluoroquinolones and is currently prescribed in certain treatments. This work was developed with the objective of evaluating the adsorption of the ciprofloxacin antibiotic in solution on zinc oxide (ZnO) supported on SBA-15-type mesoporous silica. The results showed that the post-synthesis method is effective in impregnating zinc oxide in SBA-15 and its structure has not been damaged and has not lost its organization in the hexagonal 2D planes. The ZnO-SBA-15 (10%) sample adsorbed 69.10% of ciprofloxacin (25 mg/L) in 180 min. Freundlich adsorption model was observed with the correlation factor of R²?=?0.9999, for the adsorbent ZnO-SBA-15 (10%), which showed the best sample. The kinetics was classified as pseudo-second order, as well as the thermodynamic parameters were determined, showing that the process has a spontaneous nature and a value of ΔH°?=?4.677 kJ/mol, evidencing that the process has the nature of physiosorption.     

Phenotypic,molecular and pathological characterization of motile aeromonads isolated from diseased fishes cultured in Uruguay

Information about motile aeromonads from aquaculture systems of the Neotropical region is scarce. The aim of this study was to characterize motile Aeromonas isolated from ornamental and consumable fishes cultured in Uruguay. Biochemical and molecular methods were used for species identification. Antimicrobial susceptibility and the presence of virulence genes were evaluated. Genetic diversity was analysed by rep‐PCR, and virulence of the most representative isolates was determined by calculating the fifty lethal dose in experimentally challenged fish (Australoheros facetus). Aeromonas hydrophila and A. veronii were the most prevalent identified species (38.2% and 32.4%, respectively), whereas A. allosacharophila, A. bestiarium, A. caviae and A. punctata were less prevalent. This study constitutes the first report of these last four species in Uruguay. All isolates were resistant to at least three antimicrobials, and 82.3% of them showed multidrug resistance. Virulence genotypes were correlated with the Aeromonas species and haemolytic activity. The genotype act+/alt+/ast+/ela+/lip+ was the most prevalent (26.5%). A correlation between virulence genotypes and Aeromonas species was found. A. punctata showed a clonal structure according to rep‐PCR analysis, whereas other species showed high genetic diversity. The number of virulence genes of the isolates was related with virulence according to the experimental challenge assays.     

Moderate salinities enhance growth performance of Nile tilapia (Oreochromis niloticus) fingerlings in the biofloc system

The biofloc technology (BFT) has recently gained attention as an economic and environmentally sustainable system for aquaculture. The use of BFT with salinized water could be used to minimize the negative effects of nitrogenous waste and improve the growth performance of Nile tilapia. This work evaluated the growth performance, survival, gill lesions, and fillet composition of tilapia fingerlings (Oreochromis niloticus) reared during 70 days with different salinities (0, 4, 8, 12, and 16 g/L) in a biofloc previously developed. The results indicated that the use of mature bioflocs alone was sufficient to avoid fingerling mortality due to nitrite peaks. Moreover, the salinized water, especially between 4 and 8 g/L (maximum points around 6 g/L), can be recommended in BFT to improve the growth performance of tilapia in the initial culture phase. The salinity level that was evaluated did not affect fillet composition nor the occurrence of gill lesions. The total ammonia nitrogen concentration was lower in freshwater than in salinized water (p < 0.05). Nitrite peaks in salinities of 12 and 16 g/L appeared later and were higher in comparison to other treatments. Considering minimal water exchange in BFT, the use of salinized water in this system may be environmentally viable.     

Effects of physical and chemical immobilization on hematologic and biochemical variables in captive brown brocket deer (Mazama gouazoubira)

Background: Alterations in cellular and chemical constituents in blood are indicators of physical or chemical restraint in deer. Objectives: The aim of this study was to determine the effects of physical and chemical restraint on hematologic and biochemical analytes in captive brown brocket deer (Mazama gouazoubira). Methods: Six adult deer were physically restrained and randomly anesthetized in a crossover design using 3 protocols (P). In P1, anesthesia was induced directly with isoflurane using a face mask and was maintained with isoflurane delivered by endotracheal tube. In P2, oral premedication with midazolam was followed 1 hour later by induction and maintenance of anesthesia with isoflurane, as in P1. In P3, intravenous anesthesia with a combination of ketamine, xylazine, and atropine was followed by anesthesia with isoflurane, as in P1. Blood was collected immediately after physical restraint (T0) and during anesthesia at 20‐minute intervals (T15, T35, and T55). Hematologic and biochemical constituents were determined using manual methods or automated analyzers, with cortisol measured by enzyme immunoassay. Results: During physical restraint, following food and water restriction, significant hemoconcentration, increased WBC count, and lower cortisol concentration were observed compared with results at T15, T35, and T55. During anesthesia, lymphocyte count was lower in P2 than P1, urea concentration was decreased at T15 using P2, and plasma glucose concentration was increased after T0 (P<.01). Among the anesthetic protocols, there was a trend for decreased cortisol concentration using P2. Conclusions: Physical restraint and anesthesia affect some blood constituents in captive brown brocket deer, and midazolam premedication may modify the intensity of the stress response.     

Characterization of plum procyanidins by thiolytic depolymerization

The phenolic compounds of 'Green Gage' (GG) plums ( Prunus domestica L.), "Rainha Claudia Verde", from a 'protected designation of origin' (PDO), in Portugal, were quantified in both flesh and skin tissues of plums collected in two different orchards (GG-V and GG-C). Analyzes of phenolic compounds were also performed on another GG European plum obtained in France (GG-F) and two other French plums, 'Mirabelle' (M) and 'Golden Japan' (GJ). Thiolysis was used for the first time in the analysis of plum phenolic compounds. This methodology showed that the flesh and skin contain a large proportion of flavan-3-ols, which account, respectively, for 92 and 85% in GJ, 61 and 44% in GG-V, 62 and 48% in GG-C, 54 and 27% in M, and 45 and 37% in GG-F. Terminal units of procyanidins observed in plums are mainly (+)-catechin (54-77% of all terminal units in flesh and 57-81% in skin). The GJ plums showed a phenolic composition different from all of the others, with a lower content of chlorogenic acid isomers and the presence of A-type procyanidins as dimers and terminal residues of polymerized forms. The average degree of polymerization (DPn) of plum procyanidins was higher in the flesh (5-9 units) than in the skin (4-6 units). Procyanidin B7 was observed in the flesh of all GG plums and in the skin of the Portuguese ones. Principal component analysis of the phenolic composition of the flesh and skin of these plums obtained after thiolysis allowed their distinction according to the variety and origin, opening the possibility of the use of phenolic composition for variety/origin identification.     

New qualitative approach in the characterization of antioxidants in white wines by antioxidant free radical scavenging and NMR techniques

The aim of this study was to obtain new information on antioxidant compounds in white wines. For this purpose, white wine degradation was promoted by a forced aged protocol, and six normally aged white wines from different vintages were analyzed. Both normal and forced aged wines were sequentially extracted using hexane and ethyl acetate. Apolar antioxidants were removed using hexane, and polar antioxidants were extracted with ethyl acetate. This last residue was subject to partial re-extraction with hexane and acetone. The antioxidant capacity of the wines and of each fraction was evaluated by two free radical methods, ABTS and DPPH. Normal aging provides a decrease in the total antioxidant capacity of wines. The antioxidant activity of ethyl acetate/acetone extracts was approximately 95% higher than that found for the hexane extracts. Concerning the forced aged wines, results showed that the wine submitted to a temperature of 60 degrees C for 21 days had higher antioxidant activity than that submitted to a temperature of 20 degrees C. With regard to the ethyl acetate/acetone extracts, oxygen and temperature treatment leads to a decrease in their antioxidant activity. NMR analysis was performed in the highest antioxidant capacity organic fractions (ethyl acetate/acetone extracts) and in the aqueous fraction of the control wine (T = 20 degrees C), in order to attempt the characterization of species involved in oxygen protection. Possible structures of antioxidant compounds in white wines were proposed. Two of these are tyrosol-like structures. This molecule is a well-known phenolic compound in wine, and it is reported to have antioxidative effects.