ABSTRACT
The objective of the current study was to investigate the echotextural and hemodynamic changes of ewes entering the dry period with or without subclinical mastitis. B-mode and color Doppler ultrasonography were applied to 12 Chios ewes (6 with healthy udders (group A) and 6 with subclinical mastitis (group B)) before the dry period, during the dry period (the involution phase, steady state, and transition phase), and postpartum. The color Doppler of the mammary arteries was used to evaluate them according to the pulsatility index (PI), resistive index (RI), end-diastolic velocity (EDV), time-averaged maximum velocity (TAMV), blood flow volume (BFV), and artery diameter (D). Udder parenchyma images, analyzed by Echovet v2.0, were used to evaluate the mean value (MV), standard deviation (SD), gradient mean value (GMV), gradient variance (GV), contrast (Con), entropy (Ent), gray value distribution (GVD), run length distribution (RunLD), and long run emphasis (LRunEm). In the involution phase, the PI was higher in group B compared to group A (p ≤ 0.05). The PI and RI were higher postpartum, whereas the EDV, TAVM, and D were higher in the transition phase (p ≤ 0.05). Neither the period nor the ewe group affected the MV, SD, GMV, GV, Con, and GVD values (p ≤ 0.05). In the steady state, the LRunEm was higher in group B, but postpartum, it was higher in group A (p ≤ 0.05). In conclusion, B-mode and Doppler can reveal differences (i) between healthy ewes and ewes with subclinical mastitis and (ii) among the different periods studied. Further research is needed on the blood flow and echotexture indices of the udders of ewes with unilateral subclinical mastitis.
ABSTRACT
The aim of the study was to investigate the effect of iron oxide (Fe) and silver (Ag) nanoparticles (NPs) on ram semen. A skim milk extender without antibiotics was used as a diluent of 21 ejaculates (8 rams; 2-3 ejaculates/ram). The groups of control (C; semen without NPs), Fe NPs (3.072 mg Fe3O4/mL semen), and Ag NPs (2.048 mg Ag-Fe/mL semen) were incubated (15 °C; 30 min), and then a magnetic field was used for NPs' removal. Standard microbiological procedures were performed for all groups. Post-treated samples were stored (15 °C) for 24 h, and sperm variables (kinetics by computer assisted sperm analysis (CASA); viability; morphology; HOST; DNA integrity) were evaluated at 6 and 24 h. Semen data were analyzed by a mixed model for repeated measures and microbiological data with Student's t-test for paired samples. At 6 h of storage, VCL and rapid movement-spermatozoa, and at 24 h, total/progressive motility and amplitude of lateral head displacement (ALH) were significantly decreased in group Ag compared to control. In group Fe, progressive/rapid movement-spermatozoa were significantly lower compared to control after 24 h of storage. Only in group Ag was a significant reduction of total bacterial count revealed. In conclusion, the examined Fe NPs demonstrated slight antibacterial effect, while the examined Ag NPs provided higher antibacterial properties accompanied by cytotoxicity.
ABSTRACT
The aim of this study was to investigate the effect of experimental Toxoplasma gondii infection on ram sperm quality. Five months old, pre-pubertal, rams were divided into four groups (n = 8 per group). Group A was the control group; the remaining animals received per os (p.o.) 5000 oocysts per ram. Group B did not receive treatment post-infection (p.i.). Group C received sulphadimidine (intermuscular injection (i.m.) 33 mg/kg for eight days; every 48 hrs) two months p.i. and Group D received the same drug twice (24 hours p.i. and two months later). Blood samples were collected every 15 days to detect serum immunoglobulin G (IgG). Epididymal sperm samples were analyzed for concentration, kinetics, morphology/viability, functional membrane integrity, DNA integrity, and the presence of parasite DNA. Histopathological examination was performed on the testes. The IgG titres in infected groups raised two weeks p.i. and remained high for four months. Higher values were noticed in viability and functional membrane integrity in positive spermatozoa in the control group compared to other groups, level of significance p < 0.05. Abnormal sperm was higher in groups C and D vs. A and C vs. B (p < 0.05). T. gondii DNA was detected in three sperm samples of the infected rams (12.5%). Histopathology revealed similar findings with little variation among all infected groups, characterized mostly by increased interstitial connective tissue, non-purulent inflammation, and presence of seminiferous tubules with spermatogenic cell depletion, which increased gradually from D to C and B groups. In conclusion Toxoplasmosis in pre-pubertal age negatively affected mature ram sperm quality, while sulphadimidine administration failed to alter this.
ABSTRACT
This study examined the effect of Fe3O4 nanoparticles on boar semen. Beltsville thawing solution without antibiotics was used to extend ejaculates from 5 boars (4 ejaculates/boar). Semen samples of control group (C) and group with Fe3O4 (Fe; 0.192 mg/mL semen) were incubated under routine boar semen storage temperature (17 °C) for 0.5 h and nanoparticles were removed by a magnetic field. Before and after treatment, aliquots of all groups were cultured using standard microbiological methods. The samples after treatment were stored (17 °C) for 48 h and sperm parameters (computer-assisted sperm analyzer (CASA) variables; morphology; viability; hypo-osmotic swelling test (HOST); DNA integrity) were evaluated at storage times 0, 24, 48 h. Semen data were analyzed by a repeated measures mixed model and microbial data with Student's t-test for paired samples. Regarding CASA parameters, Fe group did not differ from C at any time point. In group C, total motility after 24 h and progressive motility after 48 h of storage decreased significantly compared to 0 h. In group Fe, linearity (LIN) after 48 h and head abnormalities after 24 h of storage increased significantly compared to 0 h. The microbiological results revealed a significant reduction of the bacterial load in group Fe compared to control at both 24 and 48 h. In conclusion, the use of Fe3O4 nanoparticles during semen processing provided a slight anti-microbiological effect with no adverse effects on sperm characteristics.
ABSTRACT
This study investigated correlations among ram semen quality parameters, testicular vascularity and echotexture. Measurements were performed biweekly for a year, before (BE) and after ejaculation (AE), on seven Chios breed rams [three rams (group A) and four rams (group B) of 2-6 and 9-13 years old, respectively], considering breeding (1) and non-breeding period (2). Hemodynamics [Pulsatility Index (PI), Resistive Index (RI)] were evaluated in each testis. Images of testicular parenchyma were analysed to evaluate echogenicity [Mean Value (MV), Contrast (Con), Gray Value Distribution (GVD), Run Length Distribution (RunLD), Long Run Emphasis (LRunEm)]. Semen volume (V), concentration (C), kinetics (CASA), morphology (Sperm Blue), viability (Eosin-Nigrosin) and DNA fragmentation (Acridine Orange) were assessed. In group A/2, semen V was correlated to left testis PI and right testis RI, BE (r = 0.4, p = 0.013/r = 0.6, p < 0.001, respectively), and to right testis PI and RI, AE (r = 0.5, p < 0.001). In A/1/AE, semen C was correlated to left testis PI (r = -0.5, p = 0.003). In group A/1/BE, RI of both testes was correlated to total and tail abnormalities (left: r = -0.5, p = 0.008/ p = 0.004, respectively - right: r = -0.4/r = -0.5, p = 0.01/ p = 0.008, respectively). In group B/1/BE, sperm tail and total abnormalities were correlated to left testis Con (r = 0.5, p = 0.02/ p < 0.001, respectively), whereas in A/2/BE, they were correlated to left testis LRunEm (r = -0.8/r = -0.7, p < 0.001, respectively). In group A/1/BE, DNA damage was correlated to right testis LRunEm (r = -0.6, p < 0.001) and tail abnormalities to left testis PI (r = -0.5, p = 0.03). In group B/1/AE, right testis LRunEm, GVD, RunLD, were correlated to sperm DNA integrity (r = -0.9, p < 0.001). Immotile spermatozoa and viability were correlated to left testis Con in group B/2/AE (r = -0.6/r = 0.6, p < 0.001) and to right testis MV in group A/1/AE (r = -0.6/r = 0.6, p < 0.001). In conclusion, alterations in both ram testes blood flow and echotexture seem to be moderately correlated with sperm assessed parameters, depending on season and ram age.
