Effect of different egg yolk-based extenders on the quality of ovine cauda epididymal spermatozoa during storage at 4°C

Cauda epididymal spermatozoa were obtained from testicles collected from abattoir(s). The pooled sperm samples were divided into four aliquots. Each aliquot was washed separately with the buffer of respective extender and finally extended with the four extenders viz. egg yolk–citrate (EYC), egg yolk–citrate–fructose (EYCF), Tris–citric acid–egg yolk–fructose (TCEYF) and egg yolk–Mcillvaine glucose (EYMG) and preserved at 4°C. The per cent sperm motility for EYC, EYCF, TCEYF and EYMG at 0 h was 50.83%, 56.67%, 75.00% and 31.67%, respectively, and at 72 h was 24.17% (EYC), 30.83% (EYCF), 51.67% (TCEYF) and 7.50% (EYMG). The corresponding figures for live sperm count at 0 h was 83.17%, 86.33%, 90.42% and 81.75% and at 72 h was 64.75%, 73.92%, 76.00% and 57.67%. The corresponding figures for mean per cent intact acrosome at 0 h was 95.33%, 95.50%, 90.92% and 97.25% and at 72 h was 86.17%, 83.92%, 77.58% and 86.33%. The sperm motility was significantly (p < 0.05) higher for TCEYF at different h of preservation from 0 h through 72 h. The sperm motility, live sperm count and per cent intact acrosome declined significantly (p < 0.05) with the advancement of storage time in all the four extenders. Our study concluded that TCEYF was best out of the extenders studied for preservation of cauda epididymal spermatozoa after double centrifugation and extension at 4°C up to 72 h of preservation. However, EYCF also has better potential for the preservation of cauda epididymal spermatozoa as viability was in close proximity and acrosomal integrity was higher compared with TCEYF extender.     

Extending the viability of emu spermatozoa during in vitro storage by manipulation of temperature and diluent potassium concentration

1. Survival of emu spermatozoa during in vitro storage is not affected by increasing the extracellular [K⁺] to the point where it does not adversely affect spermatozoa function.

2. In three experiments, the effects were studied of [K⁺] in a diluent in the range 12·5–80?mM/l on emu spermatozoa survival for up to 48?h at 5, 10 or 20°C.

3. At the end of the storage period, spermatozoa viability, motility, fertilising ability and morphology were measured.

4. In Experiment 1, spermatozoa viability and morphology were adversely affected after storage (P?M/l [K⁺] whereas spermatozoa motility decreased as [K⁺] increased from 12·5 to 80?mM/l.

5. In Experiment 2, during storage at 5°C, the spermatozoa viability was comparable among any of the diluents (standard or modified) but morphology was better (P? 6. In Experiment 3, after 48?h of storage in a diluent containing 40?mM/l of [K⁺], the spermatozoa functions were better preserved at 10°C than at 5 or 20°C.

7. It is concluded that a higher than physiological level of potassium can be used in a diluent without detrimental effect on emu spermatozoa survival during 48?h storage and that the best outcome was with storage at 10°C rather than 5 or 20°C.     

The shelf‐life of uneviscerated and eviscerated chicken carcasses stored at 10°C and 4°C

1. Uneviscerated and eviscerated chicken carcasses were processed together and stored in groups of 10 at 10°C and 4°C; a further 10 eviscerated carcasses were wrapped in “polythene” bags and stored at 4°C.

2. The bacteriological condition of the uneviscerated and eviscerated carcasses prior to storage was very similar.

3. At 10 °C the eviscerated carcasses developed a slight “off” odour in 3 to 4 d (average 3.5 d) whilst the first signs of greening in the uneviscerated carcasses occurred in 4 to 6 d (average 5 d).

4. At 4 °C wrapped eviscerated carcasses developed slight “off” odour in 5 to 6 d (average 5.6 d) whilst the unwrapped eviscerated carcasses varied considerably in their shelf‐life from 5 to 11 d (average 7.9 d). After 18 d the uneviscerated carcasses were still quite acceptable and no bacteria were found in the breast muscle (i.e. < 150/g).

5. Bacteriological examinations made of the skin of the three groups of chickens stored at 4 °C confirmed the differences obtained in shelf‐life; Pseudomonas spp. were found to be the predominant spoilage organisms in each case.     

Stallion spermatozoa selected by single layer centrifugation are capable of fertilization after storage for up to 96?h at 6°C prior to artificial insemination

Background

One of the challenges faced by equine breeders is ensuring delivery of good quality semen doses for artificial insemination when the mare is due to ovulate. Single Layer Centrifugation (SLC) has been shown to select morphologically normal spermatozoa with intact chromatin and good progressive motility from the rest of the ejaculate, and to prolong the life of these selected spermatozoa in vitro. The objective of the present study was a proof of concept, to determine whether fertilizing ability was retained in SLC-selected spermatozoa during prolonged storage.

Findings

Sixteen mares were inseminated with SLC-selected sperm doses that had been cooled and stored at 6°C for 48 h, 72 h or 96 h. Embryos were identified in 11 mares by ultrasound examination 16–18 days after presumed ovulation.

Conclusion

SLC-selected stallion spermatozoa stored for up to 96 h are capable of fertilization.     

Thymus satureioides and Origanum majorana essential oils improve the quality of Beni Arouss buck semen during storage at 4°C

This study aims to investigate the effects of essential oils (EOs), extracted from Thymus satureioides (TS) and Origanum majorana (OM), on Beni Arouss buck semen quality stored in skimmed milk at 4°C. EOs were extracted by hydro-distillation, and the chemical compounds were determined. Ejaculates were collected from six Beni Arouss bucks, once a week for 10 weeks, and they were pooled, divided into five equal aliquots and diluted to 400 × 10⁶ sperm/ml with skimmed milk supplemented with 0.01% of OM EO, 0.01% of TS EO, 0.05% of OM EO and 0.05% of TS EO. Non-supplemented skimmed milk was considered as a control. Semen motility, kinematic parameters, viability, abnormality, membrane integrity and lipid peroxidation were evaluated at 0, 4, 8, 24, 28, 32 and 48 hr of liquid storage at 4°C. The main EO components were carvacrol (31.7%), thymol (28.0%) and borneol (14.4%) for TS, and terpinene-4-ol (31.2%), γ-terpinene (17.4%) and α-terpinene (12.7%) for OM. The results highlighted a dose-dependent effect of TS and OM EOs on all semen quality parameters. 0.01% of both EOs had a beneficial effect on the sperm preservation stored at 4°C compared with control (p < .05) excepted for the straight-line velocity. The 0.05% EO addition had harmful effects during storage particularly for TS EO. In conclusion, 0.01% of TS and OM EOs are recommended to improve the Beni Arouss buck semen preservation at 4°C.     

Effects of anticoagulants and storage (4°C) on packed cell volume (PCV) of Nigerian domestic fowl (Gallus domesticus) and guinea fowl (Numida meleagris)

1. Studies were conducted to determine the effects of anticoagulants and storage (4°C) on the PCV of blood samples from Nigerian domestic fowl (DF) and the guinea fowl (GF).

2. Citrate significantly reduced pre‐storage PCV of the DF in comparison with the effect of ethylenediamine tetra‐acetic acid (EDTA).

3. It further decreased (P<0.05) the PCV of the blood of DF and GF over 3 d of storage; this was similar to the effect of EDTA on the PCV of the GF blood.

4. Citrate and oxalates induced haemolysis of blood of the DF and the GF in storage faster than EDTA, but overall the haemolysis was more pronounced from red cells of the GF than from those of the DF.

5. The mean fall in the PCV of the DF was significant at 3.0 mg EDTA/ml of blood in contrast to the fall in the PCV of the GF blood.     

Effect of gradual acclimation to temperatures up to 44 °C on productive performance of the desert Bedouin fowl,the commercial White Leghorn and the two reciprocal crossbreds

1. The effect of daily exposures to increasing ambient temperatures (for 7 months) on egg production was evaluated in the desert Bedouin fowl of Sinai, a commercial White Leghorn and the two reciprocal cross breds.

2. High ambient temperatures did not adversely affect egg weight, laying rate or output per bird (g egg per day per g body weight) of the acclimated hens.

3. Best productivity was attained during periods of exposure to 38 to 40° C in all breeds.

4. Rates of decrease from maximal productivity to productivity at 42 and 44 °G differed with breed. Productivity of Leghorn and Leghorn x Sinai crossbred decreased curvilinearly above 40 °C, while productivity of Sinai and Sinai ( Leghorn crossbred decreased at 42 °C and then stabilised.

5. When changes in egg weight and laying rate were examined on an individual basis (comparison between successive months), the differences between Sinai and the Leghorn were more pronounced.

6. The results support previous findings that the Sinai breed and its crosses are able to withstand extreme environmental temperatures, reflecting genetic adaptation to desert conditions.     

Stability of hematologic analytes in monkey, rabbit, rat, and mouse blood stored at 4°C in EDTA using the ADVIA 120 hematology analyzer

Background: The time from sampling to analysis can be delayed when blood samples are shipped to distant reference laboratories or when analysis cannot be readily performed. Objective: The objective of this study was to evaluate the stability of hematologic analytes in blood samples from monkeys, rabbits, rats, and mice when samples were stored for up to 72 hours at 4°C. Methods: Blood samples from 30 monkeys, 15 rabbits, 20 rats, and 30 mice were collected into EDTA‐containing tubes and were initially analyzed within 1 hour of collection using the ADVIA 120 analyzer. The samples were then stored at 4°C and reanalyzed at 24, 48, and 72 hours after collection. Results: Significant (P<.0003) changes in hematologic analytes and calculations included increased HCT and MCV and decreased MCHC and cell hemoglobin concentration mean (CHCM) at 72 hours and increased MPV at 24 hours in monkeys; increased MCV at 72 hours and MPV at 48 hours and decreased monocyte count at 24 hours in rabbits; increased MCV and decreased MCHC, CHCM, and monocyte count at 24 hours in rats; increased MCV, red cell distribution width, and MPV and decreased MCHC, CHCM, and monocyte count at 24 hours in mice. Conclusions: Although most of the changes in the hematologic analytes in blood from monkeys, rabbits, rats, and mice when samples were stored at 4°C were analytically acceptable and clinically negligible, the best practice in measuring hematologic analytes in these animals is timely processing of blood samples, preferably within 1 hour after collection.