The intact perivitelline layer (PL) at ovulation was composed of a meshwork of anastomosing fibers uniformly enveloping the hen's ovum.

Because elevated ambient temperatures decrease fertility, this study was designed to segregate the male and female contribution to heat stress infertility in broiler breeders. Eighty hens and 16 roosters at 21 wk of age were divided equally among two heat stress (S) and two control (C) temperature chambers. For a 10-wk pretreatment period, all birds were maintained at an ambient temperature of 21.1 C and 40% relative humidity. Following the pretreatment period, birds in the S chambers were acclimated for 1 wk at a constant temperature of 29.4 C after which the temperature in the S chambers was increased to 32.2 C for 8 wk. The temperature in the two C chambers was maintained at 21.1 C. Hens in each chamber were artificially inseminated on a weekly basis with 5 x 10(7) sperm per 50 microL from either C or S males. Egg production, semen volume, spermatocrit, and percentage dead sperm were similar during the acclimation period, even though body temperature was significantly elevated in S birds (41.8 vs 41.3 C). Sperm penetration of the perivitelline layer overlying the germinal disc (GD) was decreased in eggs from hens inseminated with semen from S males compared to eggs from hens inseminated with semen from C males (9.5 vs 23.4 sperm per GD). Following the acclimation period, body temperature remained elevated in the S birds compared to the C birds (42.2 vs 41.3 C). Also, egg production was depressed in the S vs C hens (55.8 vs 82.9%). Semen volume, spermatocrit, and percentage dead sperm were not affected by S treatment. However, when hens were inseminated with semen from S males, sperm penetration of the perivitelline layer overlying the GD and egg fertility were decreased compared to hens inseminated with semen from C males (5.4 vs 14.9 sperm per GD, 45.5 vs 73.8% fertility). In conclusion, the male bird appears to contribute more to heat stress infertility than the female.

A reexamination of the fertilizing ability of cock spermatozoa from the testis, epididymis and vas deferens was accomplished through the use of intramagnal insemination. Intramagnal insemination of spermatozoa taken from the testes, epididymides and vas deferentia resulted in fertility levels of 85-90% during the first week and levels of 67-90% the second week after a single insemination. In contrast, vaginal insemination of testicular spermatozoa resulted in a total absence of fertile eggs. Vaginal and intramagnal insemination of ejaculated spermatozoa resulted in fertility levels of 70 and 71%, respectively, during the first week and levels of 42 and 53%, respectively, the second week after a single insemination. There were no significant differences in hatchability of fertile eggs from hens inseminated with semen from different regions of the male tract or by different routes of insemination.

In vitro incubation of cock spermatozoa with perivitelline layer (PL) from recently ovulated ova of the hen resulted in binding of spermatozoa to the PL and activation of the acrosome reaction. A simple quantitative technique was developed for assessing these events. Following incubation of the PL (0.5 cm2 sections) with spermatozoa, the PL section was rinsed and stained with Schiff's reagent. Microscopic examination revealed holes in the PL that were assumed to be sites of spermatozoa penetration. Utilizing this technique, a correlation was demonstrated between sperm concentration and the number of spermatozoa attaching to the PL and undergoing an acrosome reaction. Pre-treatment of spermatozoa with solubilized PL inhibited spermatozoa binding to pieces of intact PL. The PL overlying the germinal disc and a similarly sized section of PL from another area of the ovum were removed and incubated separately with spermatozoa (1 x 10(5) sperm/100 microliters). Spermatozoa showed preferential attachment and digestion of the PL from the germinal disc area (809 sperm/mm2) as compared to PL from other areas of the ovum (608 sperm/mm2). Spermatozoa attached to the PL in a circular, doughnut-shaped fashion in the area directly over the germinal disc.

The purpose of the present study was to define the role of the male broiler breeder in heat-induced infertility. Seventy-two Arbor Acres roosters were individually caged at 21 wk of age and divided equally among three heated (H) and three control (C) temperature chambers. Control temperature chambers were held at 21 C. After an 8-wk pretreatment period (20 C), an 8-wk treatment period was conducted in which the temperature in all three of the H chambers was varied from week to week according to the following schedule: Week 1, 27 C; Week 2 through Week 4, 32 C; and Week 5 through Week 8, 21 C. On a weekly basis, semen was pooled by room and inseminated into 12 groups of 10 hens each (2 groups per room). During the 1st wk when males were maintained at 27 C for 12 h, in vivo sperm-egg penetration was reduced by 48% as compared to data obtained when males were maintained at 21 C. Fertility, in vivo sperm-egg penetration, and uterovaginal sperm storage were decreased when semen from males exposed to 32 C was used to inseminate hens as compared to insemination with semen from C males. However, during this same period, the ability of sperm to bind and penetrate the egg, as determined by in vitro sperm-egg penetration, was similar between sperm from C and H males. After lowering the temperature in the H chambers back to 21 C, in vivo sperm-egg penetration as a result to insemination with semen from H males was analogous to results obtained when C males were used for insemination. Immediately after decreasing the temperature in the H chambers, fertilization of eggs by sperm from H males increased to a level similar to that obtained when eggs were fertilized by sperm from C males but then declined again during the later weeks.

A technique was developed to assess the number of cock spermatozoa penetrating the perivitelline layer (PL) in oviposited eggs in vivo. Two trials were conducted to test this technique and to establish correlation values between fertility and sperm penetration (SP). First, three Athens Canadian Randombred males, previously tested as having high fertility (100%), were each housed with seven hens. Sperm penetration was determined from eggs laid over a 3-d period (n = 41) with the mean number of spermatozoa penetrating the PL overlying the germinal disc (GD; 1.35 mm2 area) and nongerminal disc (NGD) areas being 162.8 and 8.4, respectively. Following removal of the males, SP was monitored to establish its duration with an average of 4.6 eggs analyzed per male per day. Mean sperm penetration during this period declined from 167.0 to .2 and from 9.2 to 0 for the GD and NGD regions, respectively. The mean duration of SP was 15.7 and 11.3 d for the GD and NGD PL, respectively. The duration of fertility was also established to be 14.0 d. There was a positive correlation between sperm penetration of the GD PL and fertility from eggs laid by naturally mated hens (r = .89, P < .001). In the second trial, three groups (1, 2, or 3) of 16 hens (35 wk of age) each were artificially inseminated weekly for 4 consecutive wk with either 100, 50, or 25 million sperm/50 microL, respectively. Inseminations were repeated weekly for 12 consecutive wk. Mean values were obtained from each of three 4-wk periods and used as replicates. Mean SP values from the GD PL for Groups 1, 2, and 3 were 402, 19.5, and 14.1, with fertility values of 95.8, 92.4, and 83.3%, respectively. Each replicate mean was obtained from approximately 24 eggs per group per day postinsemination. A significant correlation between SP of the GD PL and fertility (r = .90, P < .001) was established using artificial insemination of hens.

The cortical organization of the hen's ovum at ovulation was examined. A fibrous reticulum,

In-vitro culture of 1-cell fertilized rabbit ova for 6 hr at a temperature corresponding to elevated body temperature (40\ s=deg\ C) resulted in increased post-implantation embryonic mortality following the transfer of such eggs to synchronous pseudopregnant females. This increased mortality was not observed when culture temperatures corresponded to normal body temperature (38\ s=deg\ C) or in cultures that were begun after completion of the first cleavage.It is concluded that the early rabbit embryo is directly affected by the increased maternal body temperature that accompanies thermal stress of the female. Such effects may not become apparent until the late stages of embryonic development.