Ph Saratsis

Effect of antioxidant supplementation in semen extenders on semen quality and reactive oxygen species of chilled canine spermatozoa.

The objective of this study was to evaluate quality of chilled dog semen processed with extenders containing various antioxidants. Single ejaculates from five dogs were always pooled and evaluated for concentration, sperm motility, progressive motility (RSF-movement), viability, acrosomal integrity and by the hypo-osmotic swelling (HOS)-test. Also, superoxide (O(2)(-)) production, hydroxyl radicals (OH) and total reactive oxygen species (tROS) were determined. Pooled semen was divided in seven aliquots (for control and test conditions), which were diluted to a final concentration of 67x10(6)spermatozoa/ml with TRIS-glucose-egg yolk extender with or without the following supplements: control (without antioxidants), vitamin C (0.5mM), N-acetyl-l-cysteine (NAC; 0.5mM), taurine (0.2mM), catalase (100u/ml), vitamin E (0.1mM) and 5-(4-dimethylamino-phenyl)-2-phenyl-penta-2,4-dienoic acid (B16; 0.1mM). The semen aliquots were chilled and preserved at 4 degrees C. Portions of chilled semen were removed at 24 and 72h, and semen quality was evaluated after rewarming. At 24h the mean (+/-S.E.M.) sperm motility was higher (p<0.001) when vitamin E, taurine and B16 were added in the extender, whereas more spermatozoa with RSF-movement were observed (p<0.001) in the vitamin E, catalase, B16 and taurine groups. Sperm viability was higher (p=0.040) in B16 and vitamin E groups and the percentage of swollen spermatozoa was higher (p=0.002) only in the B16 group. Acrosomal integrity and OH were not significantly influenced by any of the antioxidants tested. Superoxide production was significantly lower when vitamin C, B16 and vitamin E were added in semen extenders compared with the control (p=0.017). All antioxidant groups, except vitamin C and NAC, contained less tROS compared to the control group, but only the B16 group value differed significantly (p=0.05). At 72h sperm motility was higher (p<0.001) when vitamin E, catalase, B16, taurine and NAC were added in the extender. More spermatozoa with RSF-movement were observed (p<0.001) in the vitamin E, catalase, B16, taurine and NAC treatment groups. Sperm viability was higher (p=0.001) when vitamin E, B16, taurine and vitamin C were added in semen extenders. HOS-test percentages were higher (p=0.016) in the B16, vitamin E, catalase and NAC groups. Acrosomal integrity was not influenced in any case. Production of O(2)(-) was significantly higher using catalase compared to all the other groups (p=0.006), while OH was not significantly influenced by any of the antioxidants tested. The addition of vitamin E, catalase and B16 in semen extenders resulted in significantly lower tROS values compared with the controls (p<0.0005). The results suggest that vitamin E and B16 had the most pronounced effect in preserving semen quality of chilled dog spermatozoa.

Ultrasonographic appearance of clinically healthy testicles and epididymides of rams

The clinically healthy testicles and epididymides of 31 rams were imaged inside and outside the breeding period, by using a real time ultrasound scanner. A scanning technique based on multiple imaging planes from the caudal and the lateral surface of the genitalia was employed. Optimum imaging was achieved by using a 6.0 MHz frequency sector transducer. The testicular parenchyma appeared homogeneous with a coarse medium echo-pattern. The mediastinum testis was present in 87% of rams and 77% of testicles; its median echogenicity score was 2 (range: 0-3) among rams aged 13 months or older and 1 among rams aged less than 13 months (P = 0.001). The tail of the epididymis was always clearly visible; it appeared less echoic than the testicular parenchyma and with a heterogeneous structure. The epididymal body was not visible, whilst the epididymal head was consistently partially imaged. The pampiniform plexus was clearly imaged as a dome-shaped structure masking the upper part of the head of the epididymis. The scrotal septum was seen in lateral sonograms as a highly echogenic line between the testicles. The scrotal skin formed a thick hyper-echoic peripheral structure.

Incidence risk and aetiology of mammary abnormalities in dry ewes in 10 flocks in Southern Greece

In a field investigation of 10 flocks in Southern Greece, 3367 dairy ewes were examined twice, in order to estimate the incidence risk and the aetiology of mammary abnormalities during the dry-period. Abnormal secretion, lumps, nodules, diffuse hardness, abscesses and cysts were the abnormalities detected. The cumulative incidence of mammary abnormalities during the dry-period was 5.1% (95% confidence interval: 4.4-5.8%); 47% of the cases detected developed during the first three weeks after cessation of lactation. Despite variation in the flock size, there was no between-flock variation in the risk of a ewe developing mammary abnormalities. Staphylococci (Staphylococcus aureus and coagulase-negative isolates) were the most frequently isolated bacteria from mammary samples; Actinomyces pyogenes, Clostridium perfringens, streptococci and Escherichia coli were also isolated. Resistance was encountered among the staphylococcal isolates.

Effect of N‐acetyl‐L‐cysteine Supplementation in Semen Extenders on Semen Quality and Reactive Oxygen Species of Chilled Canine Spermatozoa

The objective of this study was to evaluate the quality of chilled dog semen processed with extenders containing various concentrations of N-acetyl-L-cysteine (NAC). Ejaculates from five dogs were collected, pooled and evaluated for concentration, motility, rapid steady forward movement (RSF-movement), viability, acrosomal integrity and by the hypo-osmotic swelling test (HOST). In addition, superoxide anion (O(2)(-*)) production, hydroxyl radicals (OH(*)) and total reactive oxygen species (tROS) were determined. The pool was divided into five aliquots, which were diluted to a final concentration of 66.66 x 10(6) spermatozoa/ml with Tris-glucose-egg yolk extender containing one of the following concentrations of NAC (0, 0.5, 1, 2.5 or 5 mm). The semen aliquots were chilled and preserved at 4 degrees C. Semen quality was evaluated after rewarming at 72 h. Sperm motility was significantly higher with the 0.5 mm concentration compared with the control group (p = 0.001). Rapid steady forward movement was higher with the 0.5 and 1 mm concentrations compared with the control and 5 mm group (p < 0.001). Viability and HOST percentages were not significantly altered. Compared with the control, the 5 mm concentration showed significantly reduced percentages of spermatozoa with normal acrosomes (p = 0.049). None of the ROS values at 72 h were significantly affected by the presence of NAC in semen extenders, although all NAC concentrations showed lower O(2)(-*) and OH(*) values compared with the control. Only the concentrations of 1 and 5 mm inhibited the significant increase of tROS values after 72 h, compared with the fresh semen value. In conclusion, NAC supplementation of semen extenders is beneficial to semen motility of canine spermatozoa during chilling with the 0.5 mm concentration being the most effective, although no significant ROS inhibition was observed at 72 h.

The effect of experimentally induced subclinical mastitis on the milk yield of dairy ewes

Abstract In each of the two experiments, subclinical mastitis was induced by inoculating both mammary glands of Karagouniko and of Chios-cross ewes with a Staphylococcus epidermidis. After inoculation, the somatic cell counts of the milk of the inoculated ewes increased (p

Clinical and epidemiological findings during ram examination in 47 flocks in southern Greece.

In a field investigation of 47 flocks in south Greece, a clinical examination for fertility evaluation was done on 293 rams. In each ram, a general-health examination, examination of the genital organs and libido evaluation were carried out. In 143 (49%) of the 293 rams examined, at least one abnormality was recorded. The median within-flock prevalence risk (PR) of at least one abnormality was 58.0%, that of general-health abnormalities was 33%, that of genital abnormalities was 20% and that of libido abnormalities 14%. The principal abnormalities observed during the general-health examination were interdigital dermatitis, injuries of the sole, blowfly myiasis and brisket sores. The principal abnormalities observed during the examination of the genitalia were testicular hypoplasia, nodules into the testicular parenchyma, ulcerative lesions characteristic of orf in the prepuce and blowfly myiasis on the preputial orifice. The principal abnormalities observed during the evaluation of libido were incomplete erection and inability to mount the ewe. A statistically significant (P<0.05) positive association was recorded between: ram:ewe ratio and general-health abnormalities PR; general-health abnormalities PR and at least one abnormality PR; genital abnormalities PR and libido abnormalities PR; genital abnormalities PR and at least one abnormality PR; lambing percentage and total lambs born per ewe. A statistically significant (P<0.05) negative association was recorded between: ram age and general-health abnormalities PR; ram age and any abnormality PR; general-health abnormalities PR and lambing percentage.

Efficiency of methods used for estrous synchronization in indigenous Greek goats

The efficiency of estrus synchronization and reproductive performance was evaluated in 120 indigenous goats (Capra prisca) in the breeding season. Treatments were fluorogestone acetate (FGA)-impregnated sponges, FGA-impregnated sponges plus intramuscular injection of 400 IU pregnant mare serum gonadotrophin (PMSG), Double intramuscular injection of prostaglandin F2α (PGF2α), FGA-impregnated sponges plus intramuscular injection of PGF2α, and FGA-impregnated sponges plus intramuscular injection of PGF2α and 400 IU of PMSG. Estrous response to treatment was ≥95% in all treatments. Onset of estrus and estrous duration varied significantly among treatments, whereas fertility rate, gestation length, prolificacy and birth type did not differ (P>0.05). The use of FGA-impregnated sponges, and FGA-impregnated sponges plus intramuscular injection of 400 IU PMSG was effective in the attainment of early and compact synchronization of estrus in goats.

Physical properties of bovine cervical mucus during normal and induced (progesterone and/or PGF2α) estrus

Ninety two Friesian cows were used to determine physical properties of cervical mucus collected during normal estrus and estrus induced. Estrus was induced using either progesterone (P4) releasing intravaginal devices (PRID) and/or prostaglandin F2alpha (PGF2alpha). The animals were assigned to 4 groups (no treatment, a PRID for 12 days plus an injection of 1000 IU PMSG at the removal of the PRID, a double injection of 3 mL PGF2alpha 11 days apart, and a PRID for 7 days plus an injection of PGF2alpha 24 h before the removal of PRID). A number of cows with normal estrus exhibited three consecutive estrus cycles after calving. Cows that had not shown estrus for three months after calving had their reproductive system palpated twice at 10-day intervals, to determine their ovarian activity. Then PRID and/or PGF2alpha was administered to cows that were found to have a palpable corpus luteum in one of two palpations (cycling cows). The cows of the three induced estrous groups were artificially inseminated (AI) twice, while those with normal estrus received only a single AI. Cervical mucus samples were collected from all cows 5 to 30 min before the first AI. Additionally, samples of cervical mucus were collected from 20 cows at their first estrus after the induced estrus. The results are summarized as follows: 1) The physical properties of cervical mucus were similar in the first three normal consecutive estrus cycles after calving. 2) The physical properties of cervical mucus in normal estrus after calving were similar to those in the first estrus after an induced estrus. 3) The pH values for normal estrus were similar to those for induced estrus. 4) Viscosity of cervical mucus in the normal estrous group was significantly lower than that in the induced estrus. Furthermore, significant differences were noticed among the three induced estrous groups. 5) Spinnbarkeit, crystallization and receptivity of cervical mucus (penetration test) were significantly higher in the normal estrous group than in the induced estrous groups, while no difference was detected among induced estrus groups. 6) Pregnancy rates in the normal estrus group were the same as in the induced estrus groups. 7) The percentages of cows in the induced estrous groups that produced cervical mucus with similar viscosity, spinnbarkeit and receptivity (penetration test) characteristics as the normal estrus group, was very low.

Quality and reactive oxygen species of extended canine semen after vitamin C supplementation

The objective of this study was to evaluate the quality of extended dog semen processed with diluents containing various concentrations of vitamin C. Ejaculates from five dogs were collected, pooled and evaluated for concentration, sperm motility, rapid steady forward movement (RSF-movement), viability, acrosomal integrity and by the hypo-osmotic swelling test. Also, superoxide (O(2)(-)*) production, hydroxyl radicals (OH*) and total reactive oxygen species (tROS) were determined. The pool was divided in five aliquots, which were diluted to a final concentration of 66 x 10(6) spermatozoa/ml with a Tris-glucose-egg yolk extender containing one of the following concentrations of vitamin C (0, 0.1, 0.5, 1 or 2.5 mM). The semen aliquots were chilled and preserved at 4 degrees C. Portions of chilled semen were removed at 24 and 72 h, and semen quality was evaluated after rewarming. This process was repeated 10 times in pooled semen of the same origin and data were analysed by one-way analysis of variance. At both times, none of the semen quality parameters were positively influenced (p>0.05) by vitamin C supplementation. At 24 h, none of the reactive oxygen species (O(2)(-)*, OH*, tROS) were significantly altered. At 72 h, significant reductions of O(2)(-)* production were observed by the concentrations of 0.1, 0.5 and 2.5 mM compared with the 0 mM concentration (p=0.049). Also, at 72 h, the 2.5 mM concentration showed significantly lower OH* values in comparison with the control group (p=0.048). In conclusion, addition of vitamin C to semen extenders does not benefit the quality of canine extended spermatozoa.

The use of cytological examination of vaginal smears (CEVS) in the selection of rabbits for superovulation.

In the present study, we examined two factors associated with the reproduction of rabbit does, cytology of vaginal smears and color of vulva, as potential predictors of the success of superovulation treatment. Vulval color and vaginal smear cytology were assessed in 55 young New Zealand does. Superovulation was then induced by a single administration of eCG (20 IU/kg BW i.m.). Does were artificially inseminated 3 days later, followed by intravenous administration of hCG (120 IU per animal). Does were classified with regard to vulval color (white, rose, red, purple), and the predominant cell type in the vaginal smear (i.e. parabasal, intermediate, superficial, or anuclear). Furthermore, we categorized cells into two groups because we usually observed parabasal and intermediate cells (Group A), and superficial and anuclear cell (Group B) in the same smear. Does were humanely killed 19 h after administration of hCG and the total numbers of corpora lutea (CLs), oocytes, and zygotes (i.e. 1-cell embryos) were determined. The zygotes were assessed by morphological appearance and classified as normal or abnormal. The color of the vulva at the time of eCG treatment did not predict the success of superovulation in terms of the number of CLs, oocytes or zygotes. Does with predominantly superficial cells in vaginal smears yielded significantly fewer CLs and oocytes-zygotes (OZ) compared to does with predominantly parabasal, intermediate, or anuclear cells (P<0.05). Does with predominantly superficial cells in vaginal smears tended to yield fewer normal zygotes (nZ), but this reached significance only when compared to does with predominantly parabasal cells (P<0.05). Does in Group A yielded significantly more (P<0.05) CLs, OZ and nZ compared to does in Group B. Does with predominantly parabasal and intermediate cells in vaginal smears and rose color vulva tended to yield more OZ and nZ (P<0.05). These results suggest that the cytology of vaginal smears may help identify does with a significantly higher likelihood of yielding low numbers of CLs, oocytes, or nZ.