A novel framework for analyzing conservation impacts: evaluation, theory, and marine protected areas.

Environmental conservation initiatives, including marine protected areas (MPAs), have proliferated in recent decades. Designed to conserve marine biodiversity, many MPAs also seek to foster sustainable development. As is the case for many other environmental policies and programs, the impacts of MPAs are poorly understood. Social-ecological systems, impact evaluation, and common-pool resource governance are three complementary scientific frameworks for documenting and explaining the ecological and social impacts of conservation interventions. We review key components of these three frameworks and their implications for the study of conservation policy, program, and project outcomes. Using MPAs as an illustrative example, we then draw upon these three frameworks to describe an integrated approach for rigorous empirical documentation and causal explanation of conservation impacts. This integrated three-framework approach for impact evaluation of governance in social-ecological systems (3FIGS) accounts for alternative explanations, builds upon and advances social theory, and provides novel policy insights in ways that no single approach affords. Despite the inherent complexity of social-ecological systems and the difficulty of causal inference, the 3FIGS approach can dramatically advance our understanding of, and the evidentiary basis for, effective MPAs and other conservation initiatives.

Characterization and cryopreservation of semen from endangered markhor goats (Capra falconeri heptneri) with evaluation of reproductive seasonality.

The aims of this study were to determine the reproductive seasonality of four captive markhor goats (Capra falconeri heptneri), to characterize semen collected by electroejaculation, and to compare extenders and processing techniques for semen cryopreservation. Over the course of 1 yr, mean monthly scrotal circumference, serum testosterone, and fecal testosterone were measured and found to be inversely associated with day length. Maximum scrotal circumference (25.2 +/- 0.9 cm), serum testosterone (521.0 +/- 103.4 ng/dl), and fecal testosterone (382.5 +/- 90.3 ng/g) occurred in November, when day length was short (9.7 +/- 0.1 hr). Once a month for 3 mo (December, January, and February), bucks were anesthetized for electroejaculation and semen evaluation. Semen samples were divided into six aliquots for extension and cryopreservation in soy-based Bioxcell or Tris-based extender with 5 or 15% egg yolk, with and without centrifugation. Samples were then thawed for repeat evaluation 1-3 mo later. Postthaw evaluation revealed no significant differences between centrifuged and noncentrifuged samples. Sperm in Tris 5% and 15% egg yolk displayed higher total motility at 0, 3, and 6 hr postthaw and higher progressive motility postthaw compared with sperm in Bioxcell (P < 0.05). Sperm in Bioxcell displayed higher viability than sperm in both Tris-egg yolk extenders (P < 0.01), more intact acrosomes than sperm in Tris-15% egg yolk (P < 0.05), and a tendency for more intact acrosomes than sperm in Tris-5% egg yolk (P < 0.10). Sperm in Tris-5% egg yolk tended to have a higher percentage of morphologically normal sperm compared with Bioxcell (P < 0.10). This study provides evidence that markhor goats exhibit seasonality in scrotal circumference and testosterone levels and that centrifugation may be eliminated from the processing of markhor semen.

Spermatogenesis in testis xenografts grafted from pre-pubertal Holstein bulls is re-established by stem cell or early spermatogonia.

Xenografting of testis explants into recipient mice has resulted in successful restoration of spermatogenesis in several species. Most studies have utilized neonatal donor tissue, although a few have used prepubertal testes. In Holstein bulls, prepubertal development of the testis occurs between 16 and 32 weeks of age. The purpose of the present study was to determine the optimal age during prepubertal development of Holstein bulls for testis grafting. Explants of testis tissue from Holstein bulls between 12 and 32 weeks of age (2 bulls/age; 6 ages) were subcutaneously grafted into castrated or intact immunocompromised mice (n=8/age), then recovered after 75 and 173 days (n=4 mice/grafting period) and evaluated histologically for spermatogenic progression. Seminiferous tubules were assigned a score based on the most advanced type of germ cell present within the tubule and the average for all tubules scored (n=25) within an explant was calculated. Scores for all explants per mouse (n=6) were averaged to give a single spermatogenic progression score per mouse. No difference in spermatogenic progression of grafts between intact and castrated recipients was observed. Spermatocytes were observed in testis grafts from bulls of all ages 75 days post-grafting. At 173 days, the spermatogenic progression score for explants derived from 20 weeks bulls was greater than all ages except 12 weeks donors (p<0.05), with 8% of tubules containing spermatids. Donor material from bulls older than 20 weeks had lesser spermatogenic progression scores largely attributed to the greater number of atrophic tubules in grafts from older donors. Grafts from 28 and 32 weeks donors showed signs of degeneration by 75 days post-grafting, with 30 and 55% atrophic tubules, respectively, and lesser spermatogenic efficiency scores. By 173 days post-grafting, 72% of tubules in explants from 32 weeks donors were atrophic. The results of the present study suggest that the early stages of prepubertal development are optimal for testis grafting while advanced spermatogenesis in the donor tissue prior to grafting had a negative effect on graft development. Spermatogenesis within the grafts apparently needs to be re-established by spermatogonial stem cells or early spermatogonia.

Identification, localization, and sequencing of fetal bovine VASA homolog.

The vasa gene, first described in Drosophila, is purported to be important in germ cell development. Vasa is present across several invertebrate and vertebrate taxa, including frogs, fish, chickens, and humans. Vasa, a DEAD (asparagine-glutamine-alanine-asparagine) box protein shown to function as an RNA helicase in vitro, has not been investigated previously in fetal stage cattle. Total RNA was extracted from bovine fetal gonads obtained at 35-55 days, 55-80 days, and 80-120 days of gestation to amplify a 296 bp reverse transcription polymerase chain reaction (RT-PCR) product using primers for human vasa. The complete coding sequence of bovine vasa was cloned with 5' and 3' random amplification of cDNA ends polymerase chain reaction (RACE-PCR) and subsequently identified as bovine vasa homolog (BVH). Northern blot analysis revealed that among the tissues examined (gonad, liver, heart, brain, and femur), the vasa gene was expressed in the gonad. This localization, the conserved pattern of gene expression, and the gene sequence suggests that BVH plays a role in bovine germ cell development as proposed for other mammalian species.

Heat shock reduces developmental competence and alters spindle configuration of bovine oocytes.

Heat shock may enhance the thermotolerance of, or cause detrimental effects on, a variety of cell types or organisms, depending on the duration and intensity of the thermal challenge. Experiments were designed to investigate the effect of heat shock on the developmental competence and cytoskeletal structures of bovine oocytes following IVF. In Experiment 1, bovine cumulus-oocyte complexes (COCs) were subjected to standard IVM culture conditions for 20 h and were then randomly allocated to groups for heat shock at 42 degrees C for 0 (control), 1, 2, or 4h. The oocytes were fertilized after heat shock and followed by culture in KSOM for 8d. There were no significant differences in cleavage rates, but blastocyst formation (27% versus 44%) and total cell number per blastocyst (82+/-21 versus 108+/-36; mean+/-S.D.) were lower in the 4-h heat shock group compared to the control (P<0.05). Trophectoderm, but not ICM, cell numbers were decreased (P<0.05) in the 4-h heat shock group compared to the control. Alterations in the meiotic spindle of IVM oocytes (n=120-126) were examined after 1 to 4-h of heat shock in Experiments 2 and 3. The metaphase spindle became elongated or aberrant and smaller following heat shock, compared to the non-heat shock oocytes (P<0.05). The basis for changes in spindle configuration and the differential decrease in trophectoderm cell numbers after heat shock are not clear, but may lead to reduced embryonic development and perhaps the low pregnancy rate of domestic animals during hot seasons.

Effect of preparing and loading multiple insemination guns on conception rate in two large commercial dairy herds.

In large herds, inseminators frequently thaw multiple straws of semen and prepare several insemination guns at once. The aim of this study was to measure the effect of breeding order, the order that thawed straws are used, on conception rate in commercial dairy herds. A single professional inseminator, utilizing semen from five suppliers, performed 2629 inseminations over 30 months. Up to four straws were thawed at one time with the restriction that straws were used within 20 min of thawing. Straws were thawed per supplier's recommendations, with 66.4% of the straws pocket thawed and the remainder warm-water thawed. Conception was determined by a pregnancy check at 42 days. Data were modeled by multiple logistic regression analysis, which included herd, breeding order, lactation number, times bred, month bred and year bred. Breeding order had no significant effect on the probability of conception: Cows 1-4 achieved adjusted conception rates of 0.36, 0.41, 0.37, and 0.39, respectively. Odds ratios (and 95% confidence intervals) were 1.00 (1.00-1.00), 1.22 (0.99-1.49), 1.04 (0.82-1.32), and 1.12 (0.86-1.45), respectively. Associated laboratory studies, which evaluated the effect of post-thaw holding time on percentage of sperm with progressive motility, found mean values at 20 min holding time to be largely unchanged from mean values at 5 min. Thawing temperatures were 6, and 35 degrees C and holding temperatures were 6, 22, and 35 degrees C. The length of the trial and the wide use of semen from multiple suppliers with differing thawing methods suggests that under similar conditions, a careful and experienced inseminator can thaw multiple straws of semen and prepare insemination guns sufficient to breed up to four cows within 20 min, without an adverse effect on conception.