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Summary Mixed lymphocyte culture and serological typing of NIH and Sinclair miniature swine indicate that the two herds share a common SLA haplotype. NIH haplotype a (International Haplotype H10) appears identical to Sinclair haplotype B, which has significant effects on the penetrance of Sinclair swine cutaneous malignant melanoma (SSCM). Offspring of crosses between melanoma-bearing Sinclair swine homozygous for the B haplotype and non-melanoma NIH aa swine have tumour incidence identical to Sinclair melanoma BB× Sinclair non-melanoma BB offspring. Our results provide further support for the involvement of the swine leucocyte antigen (SLA) complex in the inheritance of SSCM, and identify a new source of non-tumour animals that have all of the genes for SSCM except those at the tumour-initiator locus.

Adrenocortical responsiveness to turning stress was examined in wild, reproductively-active olive ridley sea turtles (Lepidochelys olivacea) in relation to their mass nesting (arribada) behavior. We hypothesized that the high sensitivity threshold (HST) observed in ovipositing sea turtles is associated with a diminished sensitivity of the hypothalamo-pituitary-adrenal (HPA) axis to stressful stimuli in arribada females. We tested this hypothesis by determining whether arribada females exhibited an increased activation threshold of the HPA axis to an imposed stressor (turning stress). Mean basal corticosterone (B) and glucose levels were below 1.0 ng/ml and 60 mg/dl, respectively. Basal B remained unchanged throughout a 24-hr period in basking females. Most animals responded to turning stress with elevated mean B levels (up to 6.5 ng/ml after 6 hr) and no increase in circulating glucose. Nearly 50% of females (and none of the males) were refractory to the stimulation. Males exhibited the most rapid response, with B levels significantly elevated by 20 min over basal levels. Among females, arribada and solitary nesters exhibited a slower rate of response than basking, non-nesting animals. These results demonstrate that olive ridleys exhibit stress-induced changes in circulating B which are slower than those observed in most reptilian and in mammalian, avian, and piscine species. Furthermore, the presence of refractory females and the relatively slower increase in B in arribada and solitary nesters indicate a hyporesponsiveness of the HPA axis to turning stress in nesting olive ridleys. The hyporesponsiveness may be part of a mechanism to facilitate arribada nesting. J. Exp. Zool. 284:652-662, 1999.

The seasonal reproductive cycle of the Kemp's ridley sea turtle (Lepidochelys kempi) was studied under seminatural conditions at the Cayman Turtle Farm, Grand Cayman, British West Indies, from June 1987 to July 1988. Male L. kempi displayed a prenuptial rise in serum testosterone 4 to 5 months prior to the mating period (March). Male testosterone then declined sharply during the mating period. Female L. kempi also displayed a prenuptial rise in serum testosterone, estradiol, and total calcium 4 to 6 months prior to the mating period (March). Female testosterone and estradiol declined during the nesting period (April to July) immediately following the mating period (March). Elevated levels in female estradiol and total calcium corresponded with the period of vitellogenesis as determined from gel electrophoresis and ultrasonography. Serum thyroxine also fluctuated seasonally with elevated levels observed in females associated with the period of vitellogenesis. L. kempi displayed a distinct seasonal reproductive cycle in captivity. Nesting in the captive study group corresponded with nesting in the wild population at Rancho Nuevo, Mexico (April to July). Female endocrine cycles during the nesting period were similar to those observed in the wild population.

Selectively-bred Sinclair miniature swine exhibit a high incidence of congenital malignant melanoma which either proves fatal (10-15% of tumor-bearing piglets) or spontaneously regresses with a biphasic immunological phenomenon (85-90%) and no recurrence of malignancy. Mononuclear leukocytes were isolated from cutaneous melanomas and peripheral blood specimens collected from melanomatous (tumor-bearing) Sinclair swine during second-phase regression, and from peripheral blood specimens collected from non-melanomatous (tumor-free) Sinclair swine and control Hanford swine. Leukocyte identities were determined with single- and dual-parameter indirect immunofluorescence assays via flow cytometry. Assays for the specific surface antigens CD45, CD2, CD4, CD8, CD1, MHC class II, and N1 were employed to develop immunophenotypic profiles within the gated lymphocyte clusters from each TIL and PBL suspension. Significantly more CD8+ T-lymphocytes were identified in TIL suspensions than in peripheral blood leukocyte (PBL) suspensions (P < and = 0.05), regardless of breed or tumor status. Conversely, PBL suspensions contained significantly higher percentages of CD4+ T-lymphocytes than the levels found in TIL suspensions (P < and = 0.05). Virtually all TIL were MHC class II+, whereas the percentages of PBL expressing this antigen were markedly lower (P < and = 0.05). The percentages of T-lymphocytes co-expressing CD4 and CD8, a normal subset unique to swine, were generally consistent in all TIL and PBL suspensions examined. The results of this study have firmly established the immunophenotypic identities of cells associated with the second-phase regression phenomenon of this melanoma and have identified specific variations in the leukocyte profiles of the respective TIL and PBL suspensions.

The role of genetic factors involved in the determination of risk of cutaneous malignant melanoma (CMM) in humans remains unclear owing to genetic heterogeneity and reliance on simplistic models of inheritance. Here, we report a statistical genetic analysis of cutaneous malignant melanoma in Sinclair swine (SSCM), a unique animal model for human CMM. Using complex segregation analysis a two-locus model involving an unknown major locus and a second locus that lies within or close to the swine leukocyte antigen (SLA) complex jointly determine risk of SSCM in pedigreed animals. These loci also influence severity of affection, accounting for approximately 20% of the phenotypic variation in quantitative tumour burden.

Summary This paper reports the quantitative analysis of the historical database of a herd of Sinclair swine affected by cutaneous malignant melanoma. The herd was under partial and non-systematic selection for melanoma susceptibility (animals having at least one tumour during the first 6 weeks of life). Weighted selection differentials for the number of tumours at birth and the number of tumours at 6 weeks were generally positive and between )0.43 and 4.76 tumours for the number of tumours at 6 weeks. Estimates of the heritability for number of tumours at birth and at 6 weeks using 1934 animals were 0.27 (� 0.03) and 0.25 (� 0.03), respectively. The estimate of the genetic correlation between these two traits was 0.95 (� 0.03). Genetic trends were positive for the number of tumours at birth and at 6 weeks. In spite of positive selection differentials and a moderate heritability, there was a negative phenotypic trend in the number of tumours. Natural selection might be acting in a direction opposite to artificial selection in the Sinclair herd. The slopes of the regression of the number of tumours at birth, at 6 weeks, and melanoma susceptibility on individual inbreeding coefficients were non-significant, indicating no evidence of dominance. The number of live-born pigs was lower in litters from parents susceptible to the disease (p < 0.01).

The swine leukocyte antigen (SLA) haplotype B is associated with increased penetrance of the tumor traits in Sinclair swine cutaneous melanoma (SSCM). We established a series of Sinclair × Hanford swine crosses to facilitate genetic mapping of the tumor-associated loci. In this study, the SLA diversity in the founding animals was characterized for effective selection of maximum tumor penetrance in the pedigrees. Using the sequence-based typing (SBT) method we identified a total of 29 alleles at five polymorphic SLA loci ( SLA-1 , SLA-3 , SLA-2 , DRB1 and DQB1 ) representing six class I and five class II haplotypes. We subsequently developed a rapid PCR-based typing assay using sequence-specific primers (PCR-SSP) to efficiently follow the SLA types of the crossbred progeny. In a total of 469 animals we identified three crossovers within the class I region and three between the class I and class II regions, which corresponded to recombination frequencies of 0.39% and 0.56%, respectively. We also confirmed the presence of two expressed SLA-1 loci in three of the class I haplotypes and were able to determine the relative chromosomal arrangement of the duplicated loci in two haplotypes. This study furthers our understanding of the allelic architecture and polymorphism of the SLA system and will facilitate the mapping of loci associated with the expression of SSCM.

The sex ratio of immature loggerhead sea turtles, Caretta caretta, inhabiting the Atlantic coastal waters of Florida was investigated. Blood samples were obtained from 223 turtles that were captured in the intake channel of a power plant on Hutchinson Island. A serum androgen sexing technique was utilized to sex individual turtles. The sex ratio of the turtles (2.1 female: 1.0 male) differed significantly from 1:1 and thus appears to differ from predictions of sex allocation theory. These observations are consistent with those of a previous study, and collectively the results suggest that the sex ratio of immature C. caretta inhabiting the Atlantic coastal waters of the United States is significantly female biased: approximately two females per male.

Summary Epidermal growth factor (EGF) receptors were detected in plasma membrane preparations of equine hoof wall laminar tissue at concentrations comparable to that of equine liver. Scatchard analysis of the equilibrium binding data suggested the presence of two classes of EGF binding sites in most of the controls (plasma membranes from clinically normal horses); a high-affinity class and a more numerous low-affinity class. The dissociation constant of the low-affinity class of EGF-specific receptors (KD=1 × 10−9M) is in reasonable agreement with other values established for the EGF receptor. The variability between individual estimates for the KD of the high-affinity receptor class precluded an accurate estimate for those sites. A possible explanation is discussed. The high-affinity binding sites were uniformly absent in plasma membranes prepared from horses affected by chronic laminitis. Autoradiographic analysis localised the EGF receptors primarily to the secondary epidermal laminae, with an apparent greater density over the proliferative basal keratinocytes. Little label was associated with the dermal or the keratinised primary epidermal laminae. Tissue from horses with chronic laminitis had EGF receptors located uniformly over the hyperplastic epidermal keratinocytes. These data suggest that an EGF-mediated response may be involved in the hyperproliferative response that is characteristic of chronic laminitis.

A heterologous radioreceptor binding assay (RRA) has been developed capable of detecting nanogram amounts of epidermal growth factor (EGF) receptor-binding activity in equine urine. The binding parameters of [125I]mEGF (murine EGF) to EGF receptors on equine plasma membranes are in good agreement with values from other EGF-RRA systems. The dissociation constant estimated from equilibrium methods (KD = 4 X 10(-10) M) is in reasonable agreement with that determined from the rate constants (KD = 6 X 10(-10) M) and is in good agreement with values determined in other species. The assay is specific for equine EGF (eEGF) receptor-binding activity and capable of detecting less than 0.34 nM eEGF receptor-binding activity in urine. Equine EGF receptor-binding activity in equine urine form adult horses varied widely between samples (8.5 +/- 6.5 nM). This variability was somewhat reduced when values were adjusted for dilutional effects using urine creatinine as an indicator (3.6 +/- 2.0 nanomoles/g creatinine). No significant differences were demonstrated between the means of EGF binding activity concentrations in clinically normal horses and horses affected by chronic laminitis.

Raynaud's phenomenon (RP) and equine laminitis in the horse are medical enigmas. Clinical and scientific data were compared to evaluate the degree of similarity that exists between these two peripheral vascular diseases. Data indicate that certain pathologic and pharmacologic aspects seem to have common features. Some of the correlations maybe due simply to both diseases having ischemia of the distal digits as a pathologic component. The exact etiology of the ischemia is not known for either disease. The results of this study suggest the hypothesis that RP and laminitis are the same disease in different species. This hypothesis can be tested more efficiently when the pathophysiology of both conditions is better documented. It is possible that comparative studies will promote advances in the understanding of both RP and laminitis. The fact that equine laminitis can be experimentally induced is of potential value in such future studies.

Changes in serum concentrations of gonadotropins and gonadal steroids during the periovulatory period were monitored in green, Chelonia mydas , and loggerhead, Caretta caretta , sea turtles. Turtles were from natural populations that nest on a coral island on the Great Barrier Reef. After nesting, each turtle was transferred to a holding tank and held for a maximum of 8 days. A time series of blood samples was obtained from each of five sea turtles (three C. mydas and two C. caretta ) starting immediately after nesting and then at approximately 12-hr intervals until the time of release. Prior to release back into the ocean, each turtle was examined by laparoscopy to verify that ovulation had occurred. Serum concentrations of follicle-stimulating hormone (FSH), luteinizing hormone (LH), progesterone (PRO), and testosterone (T) in both species exhibited significant changes during this period. Surges of FSH, LH, and PRO were evident within approximately 20 to 50 hr after each turtle had nested. The significant change in FSH concentration during the periovulatory period is the first such report for a reptile. Coincident with maximal concentrations of FSH, LH, and PRO was a decline in T concentrations in both species. Estradiol-17β concentrations were near or below assay sensitivity in the C. mydas , whereas those in the C. caretta were detectable but exhibited no significant changes. The dynamic changes in FSH, LH, PRO, and T concentrations are consistent with the hypothesis that these hormones facilitate specific physiological events during ovulation and egg production.

Cutaneous malignant melanoma of Sinclair Swine (SSCM) is a heritable, congenital neoplasm which either proves fatal to the neonatal animal or undergoes spontaneous regression. Four SSCM cell lines, UISO-SSCM-433, UISO-SSCM-438, UISO-SSCM-5052, and UISO-SSCM-8093, were derived from biopsy specimens of primary tumors removed from swine at 26, 8, and 8 weeks of age, and 15 weeks gestation, respectively. Morphologic features, DOPA oxidase staining, and abnormal karyotype were suggestive of malignant melanoma. Each cell line was morphologically heterogeneous in culture with dendritic, spindle- and cuboidal-shaped cells. Pigmented melanosomes and DOPA oxidase activity were present in all cell lines at passages 20-22. UISO-SSCM-433 and UISO-SSCM-5052 contained hypodiploid and hypotetraploid sublines whereas UISO-SSCM-438 and UISO-SSCM-8093 were hypodiploid and hypotetraploid, respectively. At later passages, all cell lines presented evolutionary, karyotypic changes; the same chromosomes were involved in the alterations, however. Chromosomes 2, 6, 13, and 14 were the most affected, exhibiting numerical and structural alterations in all four cell lines. Despite the presence of multiple chromosomal anomalies in all cell lines, each with a unique set of chromosomal markers, clonal growth was not detected in soft agar, nor were any of the lines tumorigenic following s.c. inoculation in athymic mice. This suggests that the loss of malignant potential in SSCM may be inherent.

Adult male loggerhead sea turtles, Caretta caretta, exhibited a "prenuptial" spermatogenic cycle that was coincident with increased concentrations of serum testosterone (T). Serum T was high during the months when migration and mating have been recorded for males. In contrast to females, males appear to be annual breeders. Nine reproductively active female C. caretta (as verified through laparoscopy) were tagged with sonic transmitters and were repeatedly bled prior to migration. Four months prior to the nesting season, the ovaries of reproductively active females had hundreds of vitellogenic follicles of approximately 1.5 cm in diameter (i.e., half the size of ovulatory follicles). Approximately 4-6 weeks prior to migration from feeding grounds to mating and nesting areas, serum estradiol-17 beta (E2) concentrations increased significantly and remained high for approximately 4 weeks, suggesting a period of increased vitellogenesis. During a 1- to 2-week period prior to migration, serum E2 decreased significantly, while serum T concentrations increased (at least) until the time of migration. Serum T, E2, and progesterone (PRO) were elevated during nesting if a turtle was going to nest again during that nesting season. During the last nesting of a season, turtles had low serum concentrations of T, E2, and Pro. The prenuptial pattern of gonadal recrudescence and gonadal steroid production in both male and female C. caretta contrasts with those of many temperate freshwater turtles, and this type of reproductive pattern may have been facilitated by adaptation to a tropical marine environment.

1986. Kinematics of swallowing in the yellow rat snake, Elaphe obsoleta quadrivittata: a reappraisal. Japan. J. Herpetol. 11:96-109. , AND J. E. HAVERLY. 1993. Drinking by the common boa, Boa constrictor. Copeia 1993:808-818. MCDOWELL, S. B. 1972. The species of Stegonotus (Serpentes, Colubridae) in Papua New Guinea. Zool. Meded. Rijksmuseum Nat. Hist., Leiden 47:6-26. MINTON, S. A., JR., AND J. A. ANDERSON. 1963. Feeding habits of the Kukri snake, Oligodon taeniolatus. Herpetologica 19:147. , AND H. B. BECHTEL. 1958. Another Indiana record of Cemophora coccinea and a note on eggeating. Copeia 1958:47. 80. Evolutionary patterns in adv nced MORI, A., N. NARUMI, AND K. V. KARDONG. 1992. Unusual defensive behavior in Oligodon formosanus (Serpentes: Colubridae): head-slashing and tailstriking. J. Herpetol. 26:213-216. SAVITZKY, A. H. 1981. Hinged teeth in snakes: an adaptation for swallowing hard-bodied prey. Science 212:346-349.

Arginine vasotocin (AVT) and neurophysin (NP) levels were measured by radioimmunoassay in two species of sea turtle, the olive ridley, Lepidochelys olivacea, and the loggerhead, Caretta caretta, during the brief period of nesting and oviposition. In both species, AVT was low in animals which were not reproductively active. AVT was also low at the time animals emerged from the surf to nest, but increased significantly during oviposition and then declined as the animals returned to the water. NP increased in concert with AVT, also reaching highest levels during oviposition. In both species, however, NP levels remained elevated over prenesting levels at the time of return to the water. These findings are consistent with the hypothesis that an AVT-neurophysin complex is released from the neurohypophysis during nesting, and that AVT is a physiological regulator of oviducal contractions in sea turtles.

Nonsurgical artificial insemination (AI) procedures were performed using frozen, thawed semen in two adult female addax. Pregnant mares' serum gonadotropin (PMSG) (300 IU) and prostaglandin (PG) (125 μg) was administered intramuscularly to female 1 on days −3 and −1, respectively, where day 1 was the first day of AI. Only one injection of PG was used, as a low serum progesterone (PROG) concentration (0.5 ng/ml) and a low pregnanediol glucuronide (PdG) concentration (0.36 ng PdG/mg creatinine) had been recorded on day −13. A total of 219 × 106 motile sperm was deposited in the os cervix or anterior vagina on days 1–4. High serum PROG levels (9.4–15.8 ng/ml) and high urinary PdG levels (1.35–1.63 ng PdG/mg creatinine) were observed on days 23, 31, and 39, and the fetal heart was seen beating during an ultrasound exam on day 45. A male calf was born on day 266, 8.7 mo after insemination. This is the first live birth resulting from AI of an antelope with frozen, thawed semen. The Synchro-mate-B system of estrous control was used in female 2. The implant was inserted, and the injection was given on day −11 per the manufacturer's instructions. The implant was removed on day −1, and a total of 220 × 106 motile sperm was deposited in the os cervix or anterior vagina on days 1 and 2. High serum PROG levels (17.5–24.5 ng/ml) were observed on days 13, 21, and 29. Although the fetal heartbeat was observed during an ultrasound examinataion on day 42, the fetus was not carried to term. Details of the radioimmunoassay used to determine the luteinizing hormone immunoreactivity of urine samples are presented.

Naturally elevated serum testosterone levels do occur in mares and may be of adrenal or ovarian origin, due to lesions in these organs themselves or to a lesion in the hypothalamo—pituitary axes. These elevated levels work on sexually dimorphic areas of the body to result in certain masculine characteristics. Of the mares exhibiting these characteristics, some are normal with respect to androgen levels, some have tumors of either the adrenal cortex or the ovary, and the others may have impairments in enzymatic transformation which result in the elevated serum testosterone levels.

Blood samples and testicular measurements were obtained from 4-8 captive adult collared peccaries monthly for 18 months and from wild adult males during summer (N = 16) and winter (N = 22) seasons. Serum concentrations of testosterone were determined by radioimmunoassay. Semen samples were collected monthly by electroejaculation from captive males for 1 year. Serum testosterone concentrations and testicular measurements varied in a low-amplitude circannual pattern, with maximum mean testosterone concentrations in fall and winter (1150-1400 pg/ml) and minimum values in summer (500-700 pg/ml). Circannual rhythms appeared to be related to dominance. Serum testosterone levels in wild males generally were lower than in captive males, although this difference was not significant (P greater than 0.05). Semen characteristics did not exhibit a circannual rhythm. These results suggest that the male peccary remains reproductively fertile throughout the year, yet may undergo a facultative summer quiescence influenced by ambient temperature and social factors.

The primary structure of ovine hypothalamic hypophysiotropic luteinizing hormone-releasing factor, LRF, has been established as pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH 2 by hydrolysis of the peptide with chymotrypsin or pyrrolidone-carboxylylpeptidase and by analysis of the products by an Edman-dansylation sequencing technique, as well as by mass spectrometry of the derived phenylthiohydantoins. A decapeptide with the proposed primary structure, prepared by total synthesis, gave the same result on sequencing. The synthetic decapeptide possesses the same biological activities as the native ovine LRF. The amino-acid sequence of ovine LRF is identical to that already published for porcine LRF.

Epidermal growth factor (EGF) receptor binding kinetics and EGF-mediated stimulation of DNA synthesis and cellular proliferation were studied in cultured vascular smooth muscle cells (VSMC) from the equine thoracic aorta. Binding studies, using murine 125I-labeled EGF, indicate the presence of a single class of high-affinity binding sites (apparent KD = 2.8 X 10(-11) M), with an estimated maximal binding capacity of 5,800 sites/cell. EGF stimulated [3H]thymidine uptake in confluent quiescent monolayers in a dose-dependent fashion, half-maximal stimulation occurring at 7.5 X 10(-11) M. Likewise, EGF-mediated cellular proliferation was dose dependent (50% effective dose = 5 X 10(-11) M) under reduced serum concentrations. Equine VSMC contain specific receptors for EGF, and EGF can stimulate DNA synthesis and proliferation in these cultured cells, which suggests that EGF may participate in the proliferative changes observed in equine distal digital peripheral vascular disease.

Sixteen pregnant collared peccaries were assigned to four experimental diets representing two levels of crude protein and two levels of digestible energy. Serum levels of progesterone, estradiol-17β(E 2 ), and glucocorticoids were measured by radioimmunoassay. There was no significant dietary effect ( P > 0.05) associated with any hormone during gestation. Progesterone did not differ between days 11 and 140 of gestation ( X ± SE = 36.48 ± 1.11 ng / ml , N = 72, began to decline during the last week prepartum, and continued to decline within 24 hr of birth. Glucocorticoid concentrations remained level throughout gestation ( X ± SE = 6.57 ± 0.45 μ g / dl , N = 74 ). E 2 levels were low during the first 90 days of gestation, rose significantly ( P 10.11 ± 1.73 pg / ml ( X ± SE , N = 8) at Days 81–90 gestation to 49.07 ± 12.87 pg/ml ( N = 3) at 2–4 days prepartum (Days 141–147 of gestation and declined rapidly to baseline levels within 24 hr of farrowing. Litter size had no effect on progesterone or E 2 concentrations. There was no significant dietary effect on litter size or gestation length. During lactation, glucocorticoids and E 2 remained stable and did not differ by diet, while progesterone concentrations appeard to be affected by diet. Individuals on the high energy-high protein diet returned to normal ovarian function before animals on the other three diets. Results indicated that the pregnant collared peccary can maintain a reproductive environment that allows for fetal development in the face of moderate caloric or protein restriction and that the lactating peccary can undergo a postpartum ovulation in the presence of good nutrition.

Serial physiological responses were examined for 150 min from captive collared peccaries during immobilization with ketamine hydrochloride. Rectal temperatures decreased significantly (P less than 0.01) during anesthesia. Serum concentrations of total proteins, albumin, cholesterol, alanine aminotransferase, and calcium declined significantly (P less than 0.05) during the first 45 min post-immobilization before stabilizing. Concentrations of lactate dehydrogenase and alkaline phosphatase in sera showed similar but nonsignificant (P greater than 0.05) trends. Inorganic phosphorus and aspartate aminotransferase concentrations increased significantly (P less than 0.05) throughout the trial. Concentrations of serum glucose and glucocorticoid during the immobilization period were highly variable between individuals. Serum electrolytes, urea nitrogen, creatinine, gammaglutamyl transferase and progesterone were not significantly (P greater than 0.05) affected by immobilization. Elevations in serum testosterone were noted. Results indicated appropriate sampling times relative to immobilization for assay of particular serum biochemicals and steroid hormones during investigations of the physiology of the collared peccary.

Pigmented tumors resembling cutaneous melanoma were first reported in Sinclair miniature swine in 1967. Since that time, carefully planned breeding has established that this is an inherited malignancy the natural history of which mimics human cutaneous melanoma in a number of ways. Because of these characteristics, miniature swine melanoma appears to be an effective model with which to investigate the mechanisms influencing initiation, growth, and progression of human melanoma. This investigation characterized histologically the cutaneous melanoma in miniature swine and compared the findings with human neoplasm. Primary cutaneous melanoma in swine has been reclassified and standardized according to the classifications currently in vogue in human melanoma. Our results suggest that the condition in miniature swine is histologically similar to that in humans. These observations will provide a basis for interpretation of the results derived in the biologic studies performed in this model.

Metabolic and hormonal responses of eight adult male collared peccaries (Tayassu tajacu) to an ad libitum diet intake, or 25% of an ad libitum intake, were examined. Blood samples for hematological, serum-biochemical and hormonal profiles were collected at three week intervals during the nine week experiment starting 4 August 1983. Males fed on the restricted diet lost an average of 26% of their body weight during the trial, compared to a slight weight gain for those fed ad libitum. Characteristics of the red and white blood cell populations were not influenced by diet intake, with the exception of mean corpuscular volume, which was consistently lower amongst males fed on the restricted diet. Restricted food intake resulted in significantly elevated serum values for urea nitrogen, urea nitrogen:creatinine, urea index, alpha globulin:beta globulin, gamma globulin:albumin, nonesterified fatty acids, alkaline phosphatase and lactate dehydrogenase isozymes (LD1 and LD2). Restricted food intake resulted in significantly lowered serum values for total alpha globulin, alpha-1 globulin, total beta globulin, beta-1 globulin, beta-2 globulin, glucose, triglycerides, calcium, magnesium, sodium, chloride, copper and triiodothyronine. Serum levels of creatinine, total protein, albumin, alpha-2 globulin, uric acid, total bilirubin, cholesterol, aspartate aminotransferase, alanine aminotransferase, gamma glutamyltransferase, lactate dehydrogenase, phosphorus, calcium:phosphorus, potassium, iron, zinc and thyroxine were unaffected by diet intake level. Semen evaluation indicated spermatogenesis was not affected by dietary restriction despite reductions in scrotal circumference and ejaculate gel volume. Serum testosterone levels were significantly lower among males fed on the restricted diet after nine weeks. These data suggest male libido might be depressed during poor range conditions, while maintenance of spermatogenesis might permit them to take immediate advantage of improved range conditions. Blood analysis of metabolic and hormonal function can provide useful information for predicting the adult male's nutritional and reproductive condition.

Publisher Summary The methods described for the extraction of hypothalamic factors from tissues have utilized saline or dilute aqueous acids; however, organic solvents such as glacial acetic acid, ethanol, methanol, butanol, and phenol have been used in extraction of tissues or partitioning of partially purified extracts. Surveys of extraction solvents for thyrotropin releasing factor (TRF) and luteinizing hormone releasing factor (LRF) showed that both of these peptides, as judged by biologic activities, were extracted from crude preparations by aqueous methanol. Less TRF or LRF biologic activity was extracted by absolute methanol and little, if any, by petroleum ether, diethyl ether, chloroform, ethyl acetate, or pyridine. The method described in this chapter employs the extraction of lyophilized powder of hypothalamic tissue by a 90% aqueous ethanol-chloroform mixture. This procedure has been used successfully in programs of isolation and determination of the structures of ovine TRF and LRF2. The yields of hormones extracted by this method are comparable to those obtained using two N acetic acid or glacial acetic acid to extract TRF and LRF.

L'hormone hypothalamique TRF (TSH-releasing factor) est soluble dans l'alcool methylique a 90%, partiellement soluble dans l'alcool methylique absolu ou le chloroforme; TRF n'est pas soluble dans la pyridine. Le chlorure de methylene (dichloromethane) n'extrait pas le TRF d'une solution aqueuse (preparation hautement purifiee, 15000 U/mg). L'hormone hypothalamique LRF (LH-releasing factor) n'est pas soluble dans le chloroforme mais est soluble dans l'alcool methylique absolu ou a 90%.

SummaryPlasma LH levels were measured in male and female rats soon after gonadectomy by means of a solid phase radioimmunoassay method. Male rats responded to gonadectomy by elevating their plasma LH concentrations within 12 hr postcastration; a continuous rise in plasma LH concentration was observed up to day 20 at which date a plateau level was reached. Female rats showed little elevation in plasma LH concentrations up to 10 days after gonadectomy; at this time, the plasma LH level was considerably lower than that found in the male; the stage of estrus during which gonadectomy was performed had no major effect on the animal's ability to elevate plasma LH concentration. At day 30 following gonadectomy, similar levels of plasma LH were observed in both sexes.

With the use of a solid phase radioimmunoassay in which ovine LH can be detected in a concentration of 0.1 ng LH/ml, no immunochemical cross-reaction to a purified preparation of ovine LRF was observed at levels 1 × 105 times greater than the minimum detectable concentration of LH. Biological activity of this preparation of LH-releasing factor (LRF) was studied by rapid intracarotid injection (100 μg) into mature rams (55 and 60 kg body weight) and subsequent assay of plasma LH by radio-immunoassay. Concentrations of plasma LH were found to rise 3 min after injection with a peak at 5 min, after which they returned to preinjection levels within 60 min. Intracarotid injection of purified ovine thyrotropin-releasing factor (TRF) as control produced no detectable changes of plasma LH levels measured by the radioimmunoassay. (Endocrinology 84: 1517, 1969)