Short-term consumption of the mycotoxin zearalenone by pubertal gilts causes persistent changes in the histoarchitecture of reproductive tissues.

Consumption of zearalenone (ZEN) detrimentally affects tissues and systems throughout the body, and these deleterious effects are especially pronounced in swine. The objectives of this project were to determine the effects of short-term consumption of ZEN (at concentrations that could be found on-farm) on growth, carcass weight, liver weight, and reproductive tissues of pubertal gilts, and to determine if the effects are transient or persistent. Cross-bred gilts (107.25 ± 2.69 kg) were randomly assigned to one of three feed treatments: 1) solvent only for 21 d (CON; n = 10), 2) ZEN for 7 d followed by 14 d of solvent (ZEN-7; 6 mg/d; n = 10), and 3) ZEN for 21 d (ZEN-21; 6 mg/d; n = 10). Body weights were collected at the beginning and end of the experiment (189.1 ± 0.8 and 211.1 ± 0.8 d of age, respectively). Carcass weights and tissues were collected at harvest. There were no treatment-based differences in growth, carcass, liver, or reproductive tissue weights. Histological analyses revealed differences based on treatment and the interaction between treatment and luteal status. The thickness of the ampullary muscularis declined with ZEN exposure (P < 0.05), while the isthmic epithelial cell height (P < 0.01) and uterine endometrial thickness (P < 0.02) increased. Interestingly, the thickness of the isthmic muscularis, uterine myometrium, and epithelial cell height only differed in the presence of a corpus luteum. Uterine epithelial cell height in the luteal phase was lowest in ZEN-7 pigs (P < 0.01). The isthmic muscularis in the luteal phase was thinner in pigs from both ZEN treatments (P < 0.01). Conversely, the luteal-stage myometrium was thicker in pigs from both ZEN treatments (P < 0.01). The discovery of these tissue-based differences during the luteal phase is particularly concerning since this corresponds with the time when embryos would be affected by the functional competency of the oviduct and uterus. The results of this work demonstrate that short-term consumption of ZEN produces microscopic, but not macroscopic alterations in reproductive organs which are likely to have negative effects on their subsequent function and that these differences persist even after ZEN consumption ceases. Taken together, these results indicate that it is insufficient to rely solely on outwardly visible symptoms as indicators of zearalenone exposure, as detrimental effects on reproductive tissues were found in the absence of phenotypic and morphologic changes.

The mycotoxin zearalenone is a common contaminant of livestock feed. The consumption of zearalenone is particularly problematic for pigs as they are very sensitive to its effects. This study evaluated the effects of zearalenone on growth, carcass weight, liver weight, and reproductive tissues in young female pigs. Thirty pigs were split across three treatment groups. The control group was given standard feed (no zearalenone added) for 21 d, the second group received zearalenone-treated feed for 7 d followed by 14 d of standard feed, and the third group received zearalenone-treated feed for the full 21 d. Pigs receiving the treated feed exhibited no visible symptoms associated with zearalenone consumption. There were also no treatment-related differences in growth, carcass weight, liver weight, or reproductive tract weight. Histological analyses of both the oviduct and uterus revealed changes in tissue thickness that could indicate potential impairments in reproductive organ function. Changes in tissue layer thickness were especially prominent in the luteal phase. This interaction between the treatment and the presence of a corpus luteum is noteworthy because tract function during the luteal phase is imperative for fertilization and early embryonic development.

Circannual changes in progesterone secretion in intact ewes, luteinizing hormone secretion in ovariectomized estradiol-implanted ewes, and prolactin secretion in three sheep breeds anticipated to differ in seasonality of reproduction.

Changes in progesterone secretion in intact ewes (7 or 9 per breed) and luteinizing hormone secretion in ovariectomized, estradiol-implanted ewes (9 or 10 per breed) were monitored for 12 mo in Suffolk, tropically adapted St. Croix, and OOS ewes. The OOS line is a composite population of 50% Dorset, 25% Rambouillet, and 25% Finnish Landrace breeding that was selected for 10 yr for ability to lamb in October and early November. Ewes were isolated from rams, and blood samples were collected twice weekly. Circulating prolactin concentrations were also determined from blood samples collected near the summer and winter solstice and vernal and autumnal equinox. Intact OOS ewes entered anestrus later, began the subsequent breeding season sooner, and had a shorter seasonal anestrus than Suffolk and St. Croix ewes (P ≤ 0.005). St. Croix ewes did not differ from Suffolk ewes in date of onset or cessation of breeding or duration of anestrus (P ≥ 0.06). Breed differences in duration of luteinizing hormone inhibition in ovariectomized ewes were essentially identical to those observed for duration of anestrous. Prolactin concentrations varied during the year: annual changes were larger in relatively seasonal Suffolk ewes than in tropically-derived St. Croix ewes (P<0.01), and OOS ewes were intermediate to, and tended to differ from (P<0.10), the other two breeds. We conclude that OOS ewes developed by selection for fertility in spring matings had an abbreviated seasonal anestrus that is one of the shortest ever reported for temperate breeds, and that tropical St. Croix sheep did not have a shorter seasonal anestrus than Suffolk sheep under temperate conditions and ram isolation.

Exogenous gamma-glutamyl cycle compounds supplemented to in vitro maturation medium influence in vitro fertilization, culture, and viability parameters of porcine oocytes and embryos.

High concentrations of intracellular glutathione (GSH) enhance in vitro production of porcine embryos. Objectives were: (1) to determine the effects of gamma-glutamyl cycle compound supplements to the IVM medium on IVF and IVC; and (2) to evaluate embryo viability. Porcine oocytes were matured in NCSU 23 medium supplemented with either l-cysteine (3.3 mM), l-cysteamine (150 P < 0.05microM), l-cysteine and l-cystemaine, l-glycine (1, 2.5, or 5 mM), l-glutamate (1, 2.5, or 5 mM), l-alpha-aminobutyrate (3.3mM), beta-mercaptoethanol (BME) (25 microM), l-cysteine and BME, or l-alpha-aminobutyrate and BME. Increases (P < 0.05) in GSH concentrations were observed using l-cysteine, 1.0 mM l-glutamate, l-alpha-aminobutyrate, and l-alpha-aminobutyrate with BME. Oocytes matured with l-alpha-aminobutyrate and BME had a lower (P < 0.05) occurrence of polyspermy during IVF compared to controls and a greater percentage (P < 0.05) of embryos reaching the blastocyst stage compared to other treatment groups. For Objective 2, oocytes were matured in NCSU 23 or NCSU 23 supplemented with l-alpha-aminobutyrate with BME. Embryo cell death was determined using an Annexin V-FITC assay. Supplementation had no effect on the time of cell death. Embryo mortality was increased (P < 0.05) from 24 to 42 h post-IVF, with the greatest occurrence around 36 h. In conclusion, supplementing l-alpha-aminobutyrate and BME into the IVM medium increased intracellular GSH concentrations, decreased the occurrence of polyspermy during IVF, and increased embryo development parameters during IVC, but did not affect cell death during embryo development. The onset of cell death occurred from 24 to 42 h post-IVF, with the greatest occurrence around 36 h post-IVF.

Effects of dietary L-carnitine supplementation on semen characteristics in boars.

In some species, dietary supplementation with L-carnitine has been reported to increase sperm concentration and sperm motility. The objective of these experiments was to test the hypothesis that L-carnitine supplementation improves the semen characteristics of boars. In Experiment 1, boars (258 days of age) were fed daily a control diet (n = 9) or the control diet plus L-carnitine (500mg per day; n = 9 ). Semen was collected weekly from Weeks 0 to 15 and on 4 consecutive days during Week 16. Experiment 2 was similar to Experiment 1 except boars ( n = 10 per treatment) were 504 days of age. For the weekly and intensive collections there were no consistently positive effects of treatment on semen volume, sperm concentration, total spermatozoa, or sperm motility. Spermatozoa from L-carnitine-treated boars did not display an enhanced ability to maintain motility during 7-day liquid storage. In conclusion, indicators of semen quality were not enhanced by dietary supplementation of L-carnitine in boars.

The effect of lutalyse on the training of sexually inexperienced boars for semen collection.

The objective was to determine if i.m. treatments of lutalyse (PGF2alpha; dinoprost tromethamine salt) expedited the training of sexually inexperienced boars for semen collection. Lean-type, terminal-line boars (n = 40; 177.4 +/- 2.4 day of age and 112.8 +/- 2.0 kg body weight) that had not previously experienced natural mating were utilized. Boars were moved individually twice weekly for 6 weeks (total of 12 training sessions) to a semen collection room equipped with an artificial sow. Upon entering the semen collection room, boars received i.m. treatments of either deionized water (4 ml, n = 10) or lutalyse at doses of 5 mg (n = 10), 10 mg (n = 10), or 20 mg (n = 10), and subsequently received a libido score of 1-5 (1 = no interest in the artificial sow; 5 = mounting artificial sow and allowing semen collection). The percentages of boars successfully trained for semen collection during the experimental period were similar (P > 0.05) for controls (20%) and boars receiving 5 mg (30%), 10 mg (20%), or 20 mg (10%) of lutalyse. Average libido score for boars receiving 10 mg lutalyse (2.35 +/- 0.08) was greater (P < 0.05) than for controls (2.14 +/- 0.06). In summary, lutalyse increased libido scores, but did not affect the number of boars trained for semen collection.

Circulating concentrations of luteinizing hormone, testosterone, and growth hormone after naloxone treatment in sexually mature boars.

The effects of naloxone, an antagonist of opioid peptides, on circulating concentrations of luteinizing hormone (LH), testosterone, and growth hormone (GH) were determined in sexually mature boars. Blood samples were collected at 15-min intervals for three hr from five crossbred boars. Two hr after initiation of blood sampling, boars received an i.v. challenge of naloxone (1 mg/kg body weight; n=2) or 0.9% saline (n=3). Twenty-four hr later the experiment was repeated, but boars that previously received naloxone received saline and vice versa. A time by treatment interaction (p=0.09) was detected for concentrations of LH in serum, and levels of LH were greater (p<0.03) after treatment with naloxone compared to saline. Concentrations of testosterone in serum were affected by time (p<0.01), but not treatment (p= 0.59) or treatment by time (p=0.74). A treatment by time interaction (p=0.02) was detected for serum GH concentrations. Levels of GH increased in saline-treated boars (p<0.01), but not in boars receiving naloxone (p>0.1). Our results are consistent with the theory that opioid peptides suppress LH secretion and stimulate GH release in sexually mature boars.