A dose-finding,long-term study on the use of calcium chloride in saline solution as a method of nonsurgical sterilization in dogs: evaluation of the most effective concentration with the lowest risk

Background

Canine overpopulation is a global issue with serious health and welfare implications. Nonsurgical methods of sterilization could yield positive impacts on this problem, but no long-term data on such methods are available. The objective of the current investigation was to determine the effects of intratesticular injections of calcium chloride dihydrate (CaCl₂) in saline in dogs over a one year period. Five concentrations (0%, 10%, 20%, 30%, 60%) of CaCl₂ in saline were administered via intratesticular injection to groups of 10 dogs each. Total sperm count and motility, blood levels of testosterone, and side effects were examined at 0, 2, 6, and 12 months post-injection (PI). Testicular size and semen volume were examined at 0 and 12 months PI.

Results

Total sperm count, semen volume and testosterone showed significant dose-dependent decreases upon treatment with 10%-60% CaCl₂ compared with either the control group (0% CaCl₂) or baseline for each treatment group. Azoospermia was achieved for at least 12 months PI in 60% and 80% of treated dogs after administration of a 10% and 20% CaCl₂, respectively. Treatment with 30% or 60% CaCl₂ resulted in azoospermia in 100% of dogs, but more side effects were observed, while no side effects were noticed at lower doses. For each treatment group, testosterone levels had decreased an average of 35%-70% at 6 months following treatment. However, testosterone levels rebounded by the 12-month time point in all groups except the highest dosage group (60% CaCl₂), which remained at the low end of physiological range throughout the study. Sperm motility dropped to zero or near zero in all dogs treated with CaCl₂. Testicular size was significantly smaller at 12 months PI for all groups when compared to baseline.

Conclusions

This first long-term study confirms reports of the efficacy of CaCl₂ sterilization. However, at dosages free of adverse events, calcium chloride in saline may not provide permanent sterilization as previously believed. Future work should explore optimized solvents to increase the permanence of the well-tolerated 20% formulation.     

Dynamics of slow-moving landslides from permanent scatterer analysis

High-resolution interferometric synthetic aperture radar (InSAR) permanent scatterer data allow us to resolve the rates and variations in the rates of slow-moving landslides. Satellite-to-ground distances (range changes) on landslides increase at rates of 5 to 7 millimeters per year, indicating average downslope sliding velocities from 27 to 38 millimeters per year. Time-series analysis shows that displacement occurs mainly during the high-precipitation season; during the 1997-1998 El Ni?o event, rates of range change increased to as much as 11 millimeters per year. The observed nonlinear relationship of creep and precipitation rates suggests that increased pore fluid pressures within the shallow subsurface may initiate and accelerate these features. Changes in the slope of a hill resulting from increases in the pore pressure and lithostatic stress gradients may then lead to landslides.     

Effects of a diet enriched with eicosapentaenoic,docosahexaenoic and glutamine on cytokines as immunological markers for systemic inflammation in bitches before and after ovariohysterectomy

The post-operative period can generate immunological stress and can be modulated through supplementation with the omega-3 series of polyunsaturated fatty acids. This study aimed to evaluate the effects of diets enriched with high doses of eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids and glutamine on inflammatory mediators in dogs before and after ovariohysterectomy (OVH). Twelve female dogs were divided into two groups: group A was fed a commercial diet without the addition of EPA and DHA, and group B was fed an experimental diet enriched with EPA and DHA (0.2 g/100 kcal). Experimental diet intake initiated 21 days before surgery and continued until 30 days after OVH. Parameters measured were serum cytokines (TNF-α, IL-6 and IL-10), C-reactive protein (CRP), IGF-1, lymphoproliferation and body composition before and after surgery. Statistical analyses were performed with SAS software considering the effects of age and diet and their interactions, and means were compared by the Tukey test. There was no difference between groups in body weight (p = .682), lean mass (p = .101) and body fat (p = .103). There were no group differences in serum concentrations of TNF-α, IL-6, IL-10, IGF-1, CRP and the percentage of lymphocyte proliferation. However, a time effect for TNF-α was observed (p < .001), in which T_0P (10 days after the surgical procedure) presented lower values of this cytokine when compared to the other evaluation time points; and interaction effects between group and time were observed for serum concentrations of IL-6 (p < .001) and IL-10 (p = .002). OVH procedure was not considered invasive enough to increase inflammatory cytokines after 30 days of surgery, as well as the dosage of the EPA and DHA used before and after the surgery did not modulate the inflammatory markers.     

Proteomic profile of pre-implantational ovine embryos produced in vivo

The present study was conducted to decipher the proteome of in vivo-produced pre-implantation ovine embryos. Ten locally adapted Morana Nova ewes received hormonal treatment and were inseminated 12 hr after ovulation. Six days later, 54 embryos (morula and blastocyst developmental state) were recovered from eight ewes and pooled to obtain sufficient protein for proteomic analysis. Extracted embryo proteins were analysed by LC-MS/MS, followed by identification based on four database searches (PEAKS, Proteome Discoverer software, SearchGUI software, PepExplorer). Identified proteins were analysed for gene ontology terms, protein clusters and interactions. Genes associated with the ovine embryo proteome were screened for miRNA targets using data sets of TargetScan ( http://www.targetscan.org ) and mIRBase ( http://www.mirbase.org ) servers. There were 667 proteins identified in the ovine embryos. Biological processes of such proteins were mainly related to cellular process and regulation, and molecular functions, to binding and catalytic activity. Analysis of the embryo proteins revealed 49 enriched functional clusters, linked to energy metabolism (TCA cycle, pyruvate and glycolysis metabolism), zona pellucida (ZP), MAPK signalling pathway, tight junction, binding of sperm to ZP, translation, proteasome, cell cycle and calcium/phospholipid binding. Sixteen miRNAs were related to 25 pre-implantation ovine embryo genes, all conserved in human, bovine and ovine species. The interaction network generated by miRNet showed four key miRNAs (hsa-mir-106b-5p; hsa-mir-30-5p; hsa-mir-103a-5p and hsa-mir-106a-5p) with potential interactions with embryo-expressed genes. Functional analysis of the network indicated that miRNAs modulate genes related to cell cycle, regulation of stem cell and embryonic cell differentiation, among others. Retrieved miRNAs also modulate the expression of genes involved in cell signalling pathways, such as MAPK, Wnt, TGF-beta, p53 and Toll-like receptor. The current study describes the first major proteomic profile of 6-day-old ovine embryos produced in vivo, setting a comprehensive foundation for our understanding of embryo physiology in the ovine species.     

Testicular shape,scrotal skin thickness and testicular artery blood flow changes in bulls of different ages

The aim of this study was to compare the biometric testicular characteristics, skin thickness and haemodynamics of the testicular artery of 12- and 24-month-old bulls using Doppler ultrasonography, the study was conducted using 48 indicus-taurus animals. The scrotal circumference (SC) and biometry characteristics of the bulls were measured to calculate the testicular volume. Doppler ultrasonography was used to obtain the haemodynamic values of the testicular artery. The skin thickness and volume were lower (p<.01) in the younger bulls (12 months:4.68 ± 0.68 mm; 168.76 ± 47.96 cm³) versus 24 months (5.05 ± 0.89; 499.73 ± 129.24 cm³) animals (p<.01). During diastole, mean velocity was lower in the 12 months (7.98 ± 3.83) than in the 24 months (11.37 ± 4.15) animals (p <.05). The 12-month-old animals had higher pulsatility and resistivity indices (0.49 ± 0.02; 0.51 ± 0.20) compared to the 24-month-old animals (0.32 ± 0.16; 0.40 ± 0.15) (p < .05). The final testicular end velocity was lower in animals with long/moderate-shaped (L/M) (7.31 ± 2.91) than in those moderate/oval-shaped (M/O) (11.48 ± 3.88) testicles (p < .05). Animals with L/M testes presented higher pulsatility values and resistivity indices (0.51 ± 0.05; 0.55 ± 0.04) compared to animals with M/O shape (0.29 ± 0.20; 0.36 ± 0.15). We showed that the blood flow of the supra testicular artery between the two evaluated ages differed, and that 24-month-old bulls presented better thermoregulation capacity. Animals with a long/moderate testicular format presented a greater vascular resistance, which was imposed on the blood flow due to the anatomical differences in the testicular artery, resulting in lower velocity, and indicating better heat dissipation in this format.     

Variation within improved cultivars and landraces of lucerne in Central Italy

Summary A variety must be distinguishable, uniform, stable and of sufficient productivity to be officially registered. In Italy landraces of lucerne are still widely used because they are characterized by a good persistence and productivity in their area of origin and adaptation.The landrace from Casalina (20 km south of Perugia, Central Italy) has higher dry matter production than many other commercial cultivars. The objective of this paper was to measure the variation of the landrace Casalina and compare it with that of the commonest cultivars available on the market and with some lines selected from the landrace Casalina.The experiment consisted of 11 entries: Casalina, two selections from Casalina (one for high seed yield and one for frequent cuttings), the registered landrace Italia Centrale and 7 registered cultivars. Each entry was represented by 80 genotypes transplanted in the field 60×40 cm apart in a randomized block design with 4 replications. In 1991 the following characters were recorded on a single plant basis: time of first flowering, height, number of shoots and dry matter yield at the first harvest, time of second flowering, leafiness, height, number of shoots and dry matter yield at the second harvest, height, number of shoots and dry matter yield at the third and fourth harvest. Height, number of shoots and dry matter yield at the first harvest, dry matter at the second and third harvest were recorded during the 1992 season.Casalina was as variable as the other entries in 77% of the comparisons; it was more variable in 8% of the comparisons and less variable in 15% of the comparisons. In conclusion, the variation of landrace Casalina is equal to or less than that of registered varieties so that it could be directly registered at the National Registry of Varieties.     

The role of rock fragments in crack and soil structure development: a laboratory experiment with a Vertisol

A large number of factors that influence soil surface cracking behaviour in swell‐shrink soils have been studied, but there are few studies on the influence of rock fragments on crack development. Effects of rock fragments have been investigated with reference to the soil pore system but they have mainly been studied indirectly through measurements of water flow and/or bulk density changes. A laboratory experiment was carried out with repacked soil samples that were prepared by adding rock fragments of three different sizes (2–4, 4–8 and 12–16 mm) to a Vertisol at two different rates (10 and 25% by volume). Soil image analysis procedures were applied in order to describe quantitatively the network of surface cracks and the pore system to a depth of 8 cm below the surface, which developed after wetting‐drying cycles. Mean values and standard deviations of surface crack or pore widths decreased with the increase in rock fragment content and the decrease in rock fragment size, while the density of the ‘skeleton’ of the crack and pore networks increased. Rock fragments also induced vertical homogenization of the soil structure. The number of rock fragments was a key factor in determining some crack network characteristics, inducing inverse variations in mean width and skeleton density of the crack or pore network. Overall results suggested that rock fragments acted as triggering points for crack and pore development in Vertisols. The findings of this work provide a contribution to the understanding of the mechanisms of crack network and soil structure development that are induced by stones in swell‐shrink soils.