Assessment of Different Functional Parameters of Frozen–Thawed Buffalo Spermatozoa by Using Cytofluorimetric Determinations

Flow cytometry is a useful tool that provides an accurate, objective and rapid evaluation of semen quality. The use of this technique could significantly improve the quality of buffalo semen samples used in artificial insemination. This study was carried out to evaluate, by flow cytometry, frozen–thawed buffalo spermatozoa quality parameters such as sperm viability by SYBR‐14/propidium iodide staining; mitochondrial function by JC‐1 potentiometric probe; sperm chromatin stability (SCSA) by acridine orange; and acrosome reaction (AR) by FITC‐PNA staining. Semen samples from five Italian Mediterranean buffalo bulls were used. Sperm viability was not different between bulls and ranged from 33.4% to 43.6%. A consistent rate (55.1 ± 10.8%) of sperm cells showed high mitochondrial membrane potential (Δψ^high), with no significant differences between subjects. Sperm chromatin structure assay differed significantly between the five buffalo bulls; moreover, data showed high stability within each buffalo. DNA fragmentation indexes (DFI), such as %‐DFI, ‐DFI, SD‐DFI, were 11.2 ± 8.6, 153.3 ± 24.6 and 81.6 ± 21.2, respectively. Regarding AR, the percentage of acrosome‐reacted live (ARL) and acrosome‐reacted dead (ARD) spermatozoa was 0.3 ± 0.2 and 15.3 ± 5.5, respectively. This functional parameter differed significantly between buffalo bulls and showed high stability. Following to Ca²⁺ ionophore A23187 for 3 h, AR significantly differed between subjects and was characterized by an increase in both ARL (10.8%) and ARD population (22.0%). This study indicates that flow cytometry could be a useful tool for a quick multiparametric evaluation of sperm quality in buffalo. In particular, SCSA and AR resulted in sperm functional parameters sensitive enough for the diagnosis of frozen‐thawed semen fertilizing potential.     

Evaluation of calving seasons and marketing strategies in Northern Great Plains beef enterprises: I. Cow-calf systems

A bioeconomic computer model was used to evaluate alternate calving seasons in a cow-calf enterprise under range conditions representative of the Northern Great Plains. The simulated ranch utilized a rotational breeding system based on Hereford and Angus and had a fixed forage base (4,500 animal unit months of native range, 520 t of grass hay, and 183 t of alfalfa hay). Calving seasons studied were spring (SP, beginning March 15), summer (SU, beginning May 15), and fall (FA, beginning August 15). Weaning dates were October 31, December 15, and February 1, for SP, SU, and FA. The SP system was also simulated with a 5% increase in calf mortality (SP-IM), and SU with early weaning on October 31 (SU-EW). Herd size for the fixed resource was 509, 523, 519, 560, and 609 cows exposed per year for SP, SP-IM, SU, SU-EW, and FA, respectively. Corresponding values for weight weaned per cow exposed were 206, 186, 193, 153, and 145 kg. Steer calves, nonreplacement heifer calves, and cull cows were sold at the time of weaning. Quarterly cattle and feed prices used were representative of the peak, descending, valley, and ascending phases of the 1990s cattle cycle adjusted for inflation. Estimates of ranch gross margin (gross returns minus variable costs) were greatest for SP, followed by SP-IM, SU, SU-EW, and FA, and the ranks were consistent across phases of the cattle cycle. Differences between ranch gross margin for SP-IM and SU were small. In beef enterprises representative of the Northern Great Plains, with a restricted grazing season, limited access to low-cost, high-quality grazeable forage, and with calves sold at weaning, switching from early spring to a summer or fall calving date is not expected to improve profitability. If delaying calving improves calf survival, then calving in early summer may be a competitive choice.     

Long‐term Effects of Pyrethrin and Cyfluthrin,a Type II Synthetic Pyrethroid,Insecticide Applications on Bull Reproductive Parameters

The objectives of this study were to determine effects of cyfluthrin and pyrethrin spray products, used in combination with cyfluthrin topical and ear tag applications, on bull reproductive parameters over 18 weeks. Angus or Angus x Simmental bulls were randomly assigned to one of three treatment groups: (i) no exposure to pyrethrins/cyfluthrin (CONT; n = 10), (ii) cyfluthrin ear tag and topical applications (ET; n = 10), or (iii) cyfluthrin ear tag, topical, premise spray and pyrethrin fog spray applications (ET+S; n = 8). Bull body weight was measured every 3 week, and body condition score and scrotal circumference were recorded on weeks 0, 9 and 18. Semen and serum were collected every 3 weeks for sperm evaluation and testosterone measurement, respectively. There was a treatment × week interaction (p < 0.01) for sperm with primary defects; bulls in CONT group had a greater (p = 0.01) percentage of sperm with primary defects than bulls treated with insecticides at week 18. Overall and progressive sperm motility, normal sperm morphology, secondary sperm defects and serum testosterone concentrations changed (p < 0.01) over time in all bulls; however, treatment did not affect (p ≥ 0.13) any of these parameters. There were also no treatment effects (p ≥ 0.08) on bull body weight, body condition score or scrotal circumference. The use of pyrethrin‐ and cyfluthrin‐based insecticides, regardless of application, did not negatively affect reproductive parameters in beef bulls when administered over 18 weeks.     

Manganese‐iron interactions in the plant‐soil system

Dicotyledonous plants had significantly higher Mn and Fe intake rates on a near neutral soil, had a significantly higher Mn intake rate on a slightly calcareous soil, but had lower Mn and Fe intake rates on a calcareous soil, than monocotyledonous plants. This dependency on soil reaction suggests that dicots utilize primarily a chemical reduction mechanism, whereas monocots utilize some less pH‐dependent mechanism (possibly Mn(III)‐, Fe(III)‐organic complexation) to mobilize soil Mn and Fe. Soluble soil Mn and Fe fractions in the rhizosphere were consistently positively correlated with each other, as were Mn and Fe intake rates. These results suggest that for soil‐grown plants, Mn and Fe uptake was positively interrelated because both Mn and Fe were mobilized by similar root processes.     

Formation of an Antral Follicle–like Structure of Bovine Cumulus–Oocyte Complexes Embedded Individually or in Groups in Collagen Gels

Culture techniques of antral follicle–like structure (AFLS) derived from cumulus–oocyte complexes (COCs) might provide important insights into follicular development and oocyte maturation. This study was undertaken to investigate the effects of embedding bovine COCs individually (one COC) or in groups (4–5 COCs) in collagen gels on the formation of AFLS and the meiotic status of oocytes. The observations of AFLS formation were performed every second day for 14 days. The AFLS was formed at Day 2 or 4 after the start of culture (Day = 0), irrespective of the culture methods. The mean diameters of AFLS during Days 4–14 using the individual culture method were significantly higher (p < 0.05) than those using the group culture method. However, the AFLS formation rate in the individual culture method was significantly lower compared to that in the group culture method (26.1% vs 62.7%, p < 0.01). Almost all oocytes had undergone the germinal vesicle breakdown stage, irrespective of the culture method or AFLS formation. In conclusion, comparison with the individual culture method revealed that the mean diameters of AFLS in the group culture method were smaller, but more COCs formed AFLS. The group culture method might be useful for evaluating the various hypotheses of follicular formation and interfollicular communication. However, improvement of the group culture system is necessary to prevent the meiotic resumption of oocytes, because the AFLS formation is dependent on the cumulus/granulosa cells surrounding oocytes.     

Evaluation of calving seasons and marketing strategies in Northern Great Plains beef enterprises. II. Retained ownership systems

Two bioeconomic computer models were used to evaluate calving seasons in combination with calf marketing strategies for a range-based cow-calf enterprise in the Northern Great Plains. Calving seasons studied were spring (SP, calving beginning March 15 and weaning October 31), spring with calf mortality increased by 5% (SP-IM), summer (SU, calving beginning May 15 and weaning December 31), summer with early weaning (SU-EW, calving beginning May 15 and weaning October 31), and fall (FA, calving beginning August 15 and weaning February 1). Marketing scenarios for steer calves and nonreplacement heifer calves were as follows: sold after weaning (WS), backgrounded in Montana and sold as feeder cattle (WBS), backgrounded in Montana and then fed to slaughter BW in Nebraska (WBFS), and shipped to Nebraska at weaning and fed to slaughter BW (WFS). Quarterly inflation-adjusted cattle and feedstuff prices were representative of the 1990s cattle cycle. Cumulative gross margin (CGM), the sum of ranch gross margin and net return from retained ownership was used to compare systems. At the peak of the cattle cycle, all forms of retained ownership (WBS, WBFS, WFS) were profitable for all calving seasons, but during the descending phase, only WBS increased CGM markedly over WS for SU-EW. At the cycle valley, retained ownership was not profitable for SP and SP-IM, whereas WBFS and WFS were profitable for SU and SU-EW, and all forms of retained ownership were profitable for FA. During the ascending phase, retained ownership was profitable for all calving season-marketing combinations. Systems with the greatest CGM at each phase of the cattle cycle were FA-WFS, SP-WBS, FA-WFS, and FA-WFS at the peak, descending, valley, and ascending phases, respectively. In beef enterprises representative of the Northern Great Plains, with a restricted grazing season and limited access to low-cost, good-quality grazeable forage, no single calving season and no single combination of calving season and calf marketing is expected to be superior throughout the cattle cycle. Fall calving systems most often benefit from retained ownership through slaughter.     

Oestrogen and Progesterone Receptors and COX‐2 Expression in Endometrial Biopsy Samples During Maternal Recognition of Pregnancy in Llamas (Lama glama)

Endometrial expression of oestrogen receptor‐α (ERα), progesterone receptor (PR) and cyclooxigenase‐2 (COX‐2) was evaluated in non‐pregnant and pregnant llamas during the period when luteolysis/maternal recognition of pregnancy is expected to occur. Females (n = 28) were divided into two groups: non‐pregnant llamas were induced to ovulate with a Buserelin injection, and endometrial biopsies were obtained on day 8 (n = 5) or 12 (n = 5) post‐induction of ovulation. Animals of the pregnant group (n = 18) were mated with a fertile male. Pregnancy was confirmed by the visualization of the embryo collected by transcervical flushing in 5 of 9 animals on day 8 post‐mating and by progesterone profile on day 12 post‐mating in 4 of 9 animals, when endometrial biopsies were obtained. An immunohistochemical technique was used to evaluate receptors population and COX‐2 expression. Pregnant llamas showed a higher percentage of positive cells and stronger intensity for ERα than for non‐pregnant llamas in stroma on day 8 and in the luminal epithelium on day 12 post‐induction of ovulation, while a deep decrease in endometrial PR population was reported in pregnant llamas on that day in luminal and glandular epithelia and stroma. In the luminal epithelium, COX‐2 expression was lower in pregnant than in non‐pregnant animals. Briefly, the increase of ERα in pregnant llamas gives further support to the hypothesis that oestrogens are involved in the mechanism of maternal recognition of pregnancy. Endometrial PR decrease in pregnant llamas might be a necessary event to allow the expression of proteins involved in conceptus attachment, a mechanism widely accepted in other species. Moreover, embryo seems to attenuate maternal PGF_2α secretion during early pregnancy by decreasing the endometrial expression of COX‐2 in the luminal epithelium of pregnant llamas.