The Mississippi Sandhill Crane (Grus canadensis pulla), the most endangered North American crane, is considered critically endangered and is protected by Federal and State law. Substantial funding has established the Mississippi Sandhill Crane National Wildlife Refuge in Southern Mississippi and an artificial insemination / breeding facility at the Audubon Center for Research of Endangered Species in New Orleans, Louisiana to promote species recovery. In spite of extensive time, labor, and money invested in captive propagation, juvenile Mississippi Sandhill Cranes suffer substantial mortality due to predation by bobcats (Lynx rufus), coyotes (Canis latrans) and red tailed hawks (Buteo jamaicensis) upon reintroduction to the refuge. Studies have shown decreased avian mortality in reintroduction programs incorporating antipredator conditioning. Appropriate antipredator behaviors are likely innate in cranes, however the object at which to direct these behaviors may require social learning in lieu of the normally long period of parental care known in this species. An antipredator conditioning program was conducted prior to release for 2 years in juvenile Mississippi Sandhill Cranes using live tame predators and conspecific presence to teach predator recognition and appropriate responses. Death of juvenile cranes upon reintroduction to the refuge due to predation has not occurred since the inception of the program. However, factors such as an increase in predator control or differing weather conditions may have contributed to these results. Behavioral results strongly suggest that the presence of adult cranes during antipredator conditioning of subadult cranes is of benefit. With the presence of an adult pair of cranes (models), subadult cranes show significantly more vigilance in the form of the tall alert behavior. Contact call and guard call occurrence were associated with age, however appropriate vocal response to predator presence occurred regardless of whether a model was present. No cranes were harmed during antipredator conditioning procedures, and time and money expenditures were minimal. Antipredator conditioning programs for cranes can be relatively simple and inexpensive with minimal risk to participants. We strongly recommend similar procedures be incorporated into other avian endangered species reintroduction programs.

Hearing loss in dogs and cats following dental or ear procedures performed under anesthesia has recently been reported. The most likely causes for this acute-onset deafness were considered to be mechanical or vascular. Jaw manipulation as a possible cause of acute-onset deafness in the dog was investigated in the current study. Structures adjacent to the temporomandibular joint were of interest because changes in jaw orientation could disrupt vessels and nerves in this area. Current descriptions of the anatomy of the vasculature supplying the canine ear are either incomplete or inconsistent. Another considered cause was a jaw-ear connection via a ligamentous remnant of Meckels cartilage. A ligament from the jaw to the ear, while recognized in humans, has not been described in the dog. The goal of this research was to provide more accurate anatomical descriptions of relevant canine juxta-articular structures to support future physiological studies. The occurrence of hearing loss in dogs and cats following procedures performed under anesthesia was documented. Occurrence was low, with only 62 identified cases between 2002 and 2009, but the true occurrence may be greater. No relationship was observed between hearing impairment following these procedures and breed, gender, size of dogs, or anesthetic drug used; however, older animals may be more vulnerable. New anatomical variations of three arteries are described in this study. (1) The rostral tympanic artery is a branch of the mandibular ramus and is accommodated by a small foramen located within a depression medial to the temporomandibular joint. (2) A rostral auditory tube branch of the caudal deep temporal artery was identified. (3) The origin of the caudal auricular artery occurred opposite the lingual artery in 25% of dissected specimens, contrary to published descriptions. The rostral tympanic artery and the rostral auditory tube branch may be susceptible to damage during jaw manipulation due to their locations. Variability of the caudal auricular artery can create problems during surgical procedures. The tympanomandibular ligament, a remnant of Meckels cartilage and equivalent to the human sphenomandibular ligament, was established as a jaw-ear connection in the dog. Tension on this ligament did not produce malleus movement.

Ayahuasca denotes an Amazonian psychotropic plant tea obtained from Banisteriopsis caapi, which contains beta-carboline (b-carboline) alkaloids, chiefly harmine, harmaline and tetrahydroharmine. The tea usually incorporates the leaves of Psychotria viridis, which are rich in N,N-dimethyltryptamine (DMT), a psychoactive 5-HT2A agonist. The beta-carbolines reversibly inhibit monoamine-oxidase (MAO), effectively preventing oxidative deamination of the orally inactive DMT and allowing its absorption and access to the central nervous system. Despite increased use of the tea worldwide, easy to perform and validated methods for its characterization do not exist and the metabolism and excretion of DMT and the b-carbolines has not been studied systematically in humans following ayahuasca consumption. Thus, we developed a liquid chromatographyelectrospray ionization-tandem mass spectrometry procedure for the simultaneous quantification of the major alkaloid components of ayahuasca, including several known and potential metabolites. The assay was applied to a variety of ayahuasca samples and modified to be applicable to human blood and urine samples before and after consumption of ayahuasca. The major components present in ayahuasca samples were tetrahydroharmine and harmine, followed by DMT and harmaline. The major metabolite of DMT was the corresponding N-oxide, DMT-N-oxide which was found in both blood plasma and urine, although not detectable in ayahuasca samples. Less than 1% of the administered DMT dose was detected in urine or blood plasma, despite the inhibition of monoamine oxidase afforded by the presence of the harmala alkaloids in ayahuasca. The major harmala alkaloid excreted was tetrahydroharmine. The methods developed would be suitable for the study of ayahuasca in human and ethnobotanical research, as well as in forensic examinations of ayahuasca preparations. The characteristics of the methods suggest that their sensitivity, selectivity and reproducibility are adequate for use in further toxicological and clinical research on ayahuasca as well as functioning as an assay to screen biological samples for endogenous hallucinogens. Based on the results of these studies we also present a critical review of 69 published studies reporting the detection in human body fluids of three indole alkaloids that possess differing degrees of psychedelic activity. Suggestions for the future directions of ayahuasca and endogenous psychedelics research are offered.

Achondroplasia and hypochondroplasia are two forms of skeletal dysplasias caused predominantly by single base mutations in the fibroblast growth factor receptor 3 gene (FGFR-3). The mutation for achondroplasia is a G1138A/C substitution and the mutation for hypochondroplasia (occurring about 50% of the time) is a C1620A/G substitution. Recent genetic studies have shown that spontaneous mutations for achondroplasia and hypochondroplasia occur exclusively on the paternally derived chromosome, suggesting that these mutations occur preferentially during spermatogenesis. For unknown reasons, the mutation rates at these FGFR-3 nucleotides appear to occur at a much higher frequency than nucleotide specific mutation rates observed in other human genetic diseases. The purpose of this study was to develop an assay that could detect the frequencies of achondroplasia and hypochondroplasia causing mutations in human sperm. A Needle-in-a-Haystack PCR/RE/LCR selection technique has been developed that measures single base changes, commonly single base substitution mutations, at sensitivities of one mutant allele in one cell in up to 10⁷ wild-type cells. This technique was modified and designed for the achondroplasia and hypochondroplasia base sites 1138 and 1620 of the FGFR-3 gene. With the development of this technique, future studies could focus on determining the frequencies of the mutations in the sperm of fathers of affected children and the frequencies of the mutations in the sperm of the normal population. These studies will help elucidate the paternal age effect, have important implications in genetic counseling and provide a novel method by which to study genetic disease in humans.

Much debate surrounds the topic of women choosing to deliver their infants at home with the services of a midwife. The outcomes of women beginning labor at home, ultimately delivering at home, and the infants born at home were studied in order to determine the safety of home birth among expectant women choosing to be attended by a midwife licensed to practice by the state of Louisiana. Trends associated with those choosing home birth and the frequency of home birth were also analyzed. The study was conducted using retrospective records of clients accepted during the study years 1986-2000. During the study years, the midwives began care on 284 women. Of those women, 225 delivered at home, 31 were transported to the hospital at some point, 7 voluntarily left the midwifes care, and 20 were referred out for various reasons. The only cause of infant mortality in the home births was congenital anomalies. There were no maternal deaths or significant maternal morbidity. The years with the largest number of home births were 1997, 1998, and 1999 with 32, 35, and 32 births respectively. During these three years there were three actively practicing midwives, with a fourth serving only six women total. The same three midwives were also practicing in 2000, but the number of home births dropped sharply to 20 due to difficulties attaining physician back-up. Most women who chose home birth had already had a child. Only 20.8% were primiparous. Complications and obstetric procedures were infrequent, outcomes were good, and the breastfeeding rate was 100% with most babies being nursed within one hour of birth. The cesarean section rate for all women who began labor with the intention of delivering at home was 6.2%, much lower than the national cesarean rate which averages 22%. Comparisons were also made to vital statistics data available for East Baton Rouge Parish and national records Home birth attended by a licensed midwife is safe and has outcomes as good as or better than that of hospital birth. Home birth with a trained midwife should be a option available to low-risk women.

Gastroenteritis caused by the nematode parasite Haemonchus contortus is a serious concern for small ruminants in the tropics and subtropics. Control is traditionally by anthelmintics and pasture management, however this specific nematode parasite has become resistant to many anthelmintics and research is now focused on novel control methods including biological control using the nematode-trapping fungus, Duddingtonia flagrans. The objective of this study was to investigate the effectiveness of feeding chlamydospores of the fungus at a dosage of 5 x 105 spores/kg of BW to ewe/lamb pairs (Trial 1) and weaned lambs (Trial 2) for a total of 17 and 8 weeks, respectively. For Trial 1, 34 pregnant ewes were randomly allocated into Treated and Control groups of 17 each and each group grazed on separate pastures for 21 weeks. For Trial 2, 30 lambs from Trial 1 were randomly allocated into Treated and Control groups of 15 each and each group grazed the same pastures as in Trial 1. In both Trials, results from fecal cultures, pasture larval recovery and tracer animals showed that the Treated group pasture had lower infectivity. This reduced pasture infectivity did not translate into reduced infection levels in the ewe/lamb pairs in Trial 1, but infection levels were reduced in Trial 2. There was no effect on weight gain in either Trial. It can be concluded from this study that Duddingtonia flagrans can be used as a biological control agent to reduce pasture infectivity, however, infection levels and productivity may not be affected in sheep during one grazing season.

Acute laminitis is a severely debilitating disease of the laminae of the equine digit; however, the mechanism(s) of pathogenesis have yet to be fully elucidated. In physiologic states, the endothelium synthesizes substances, such as nitric oxide (NO; vasodilator) and endothelin-1 (ET-1; profound vasoconstrictor), which play a crucial role in vasomotor regulation. The overall hypothesis is that the initiating factor in the onset of acute laminitis is a disruption in the balance between NO and ET-1, which leads to digital vasoconstriction and subsequent laminar ischemic necrosis. In vitro studies with digital vessels from healthy horses and horses with naturally-acquired laminitis determined that ET-1 caused concentration-dependent, sustained contraction of arteries and more profound contraction of veins, and incubation with the nonselective ET receptor antagonist (PD145065) at a 10-5 M concentration abolished these contractile effects. ET-1 was then administered into the digit of healthy conscious horses, which resulted in reduced blood flow and the ET antagonist, especially in combination with a NO donor, reversed these reductions. Naturally-acquired laminitic horses had a trend for increased jugular and cephalic venous plasma ET-like immunoreactivity, and horses during the development of black walnut extract (BWE)-induced laminitis developed increased digital venous plasma ET-like immunoreactivity. After validation for equine tissues, ET-1 immunohistochemical staining was conducted on digital vascular and laminar tissues, but no notable differences were found between healthy and naturally-acquired or experimentally-induced laminitic horses. During the developmental stages of BWE-induced laminitis, digital blood flow initially decreased followed by hyperemia, corresponding with demonstration of clinical signs of laminitis. Administration of the ET antagonist, and the antagonist combined with a NO donor, improved Starling force alterations by improving digital vascular resistances and blood flow. Utilizing digital vessel rings from BWE-treated horses, ET-1 caused a concentration-dependent contraction in vitro that was abolished by the ET antagonist. Endothelium-dependent vasodilation was decreased in these vessels, demonstrating possible altered endothelial function due to BWE administration. Based on the results of these studies, ET-1 appears to play a role in the pathophysiology of acute laminitis in horses and continued investigations evaluating ET antagonists as preventative and therapeutic agents for this devastating disease are warranted.

Gastrointestinal tract disease is the leading natural cause of death in horses and horses with ischemic intestinal disease have the greatest mortality. We hypothesized there is basal synthesis of endothelin-1 (ET-1) in the intestinal tract of healthy horses that is likely involved in regulating vasomotor tone, secretion and motility and that ET-1 synthesis increases with gastrointestinal tract disease, which may be involved in the pathophysiology of these disorders. Plasma ET-like immunoreactivity was increased in horses with naturally-acquired gastrointestinal disease, compared with normal horses; values were greatest in horses with large intestinal strangulation obstruction, enterocolitis and peritonitis. There was an association between ET-1 levels and survival, PCV and duration of signs of pain. Immunohistochemical staining for ET-1 was present in surface epithelium, villi, muscularis and serosa of numerous intestinal segments in healthy horses. Staining was also present in submucosal vessels with veins staining more intense than arteries. Staining appeared more diffuse and intense in samples from horses with intestinal strangulation obstruction. Polymerase chain reaction analysis revealed the presence of ET-1 gene expression in numerous intestinal segments of normal horses. These findings suggest ET-1 is involved in physiologic functions such as regulation of secretion, vasomotor tone and motility, and that increased ET-1 with strangulation obstruction may be involved in the pathophysiology of these disorders. ET-1 caused sustained, concentration-dependent increases in cecal longitudinal smooth muscle tone in vitro, but the magnitude of contraction was less than that induced by carbachol. Pre-incubation of tissues with ETA (BQ-123) and ETB (IRL-1038) receptor antagonists alone did not inhibit ET-1 induced contraction. However, contractile responses were inhibited when tissues were incubated with both antagonists (10-5 M) together, suggesting both ETA and ETB receptors mediate the contraction. Electric field stimulation did not change the contractile response. These studies indicate a physiologic role of ET-1 in the equine gastrointestinal tract and that increased synthesis and release occurs with gastrointestinal tract disease, especially ischemic conditions, and may contribute to the pathophysiology of these disorders. Further studies involving ET-1 and ET antagonists appear warranted.

There is anecdotal documentation that intra-articular administration of liquid silicone is used as a lubricant for equine joints. Effects of silicone polymer on equine articular cartilage and synovial fluid have not been evaluated. The objectives were to characterize the effects of intra-articular administration of liquid silicone polymer on clinical, synovial fluid, and microscopic features in normal middle carpal joints of horses, and to determine if silicone migrates to the axillary lymph nodes. Eight adult horses were used in this study. Two mLs of 1,000 centistoke, medical-grade silicone was injected into one randomly selected middle carpal joint. The opposite middle carpal joint was injected with two mLs of 0.9% NaCl. Degree of effusion, range of motion, pain on flexion, joint circumference, and degree of lameness were determined prior to injection, at 2, 8, 12, 24, 48, and 72 hours after injection, and weekly for 6 weeks after injection. Radiographic examination, synovial fluid analysis, and microscopic examination of synovium, articular cartilage and axillary lymph nodes were performed at the conclusion of the study. Joint circumference for silicone-treated carpi was significantly greater (p < 0.05) than saline-treated carpi from eight to 336 hours after injection. There was a significant association between treated limbs and lameness, lameness after flexion, pain elecited upon maximal flexion of carpus and middle carpal effusion in treated limbs. Radiographic examination did not reveal the presence of osseous abnormalities; however, effusion was present in 4/8 silicone-treated carpi and 2/8 saline-treated carpi. Synovial fluid from silicone-treated and saline-treated carpi had normal protein concentrations and total nucleated cell counts. Synovial fluid from 5/8 silicone-treated carpi had refractile cytoplasmic vacuoles within macrophages. All silicone-treated carpi had synovial hypertrophy with 7/8 silicone-treated carpi demonstrating accumulation of vacuolated cells containing silicone. Examination of articular cartilage from saline-treated and silicone-treated carpi did not reveal a significant difference. Silicone was not definitively demonstrated in any axillary lymph nodes. Administration of silicone into the middle carpal joint resulted in synovitis. A single administration of silicone polymer had no apparent effects on articular cartilage within six weeks of administration.

Photobacterium damselae subspecies piscicida, previously known as Pasteurella piscicida, is an important pathogen of hybrid striped bass and many fish species cultured in brackish water in the United States, Japan, Europe, and the Mediterranean. The purpose of this study is to investigate virulence mechanisms that contribute to the pathogenesis of this organism. The ability of P. damselae to survive/replicate within hybrid striped bass macrophages was evaluated with an in vitro killing assay. Results indicated that the numbers of bacteria recovered from macrophages at 3, 6, 12, and 18 hours of incubation increased significantly over time. In contrast, the numbers of Escherichia coli control strain recovered from macrophages declined at the same designated incubation times. Light and electron microscopy confirmed internalization, uptake, and multiplication of bacteria within spacious, clear vacuoles in the macrophages. Using acid phosphatase as a lysosomal marker, it was shown that P. damselae inhibits phagolysosomal fusion. Invasion and replication of P. damselae within epithelioma papillosum carpio (EPC), channel catfish ovary (CCO), and fathead minnow (FHM) cells lines was also evaluated using an in vitro invasion assay. All three cell lines were susceptible to invasion and supported replication of P. damselae. Fathead minnow cells were more susceptible to invasion than the other two cell lines as indicated by greater numbers of infected cells and recovered bacteria at time 0. Using light and electron microscopy, invasion of cells by bacteria was observed as early as 30 minutes after infection, and intracellular bacteria were observed in large, clear, membrane-bound vacuoles. The intracellular location of P. damselae was confirmed using ruthenium red staining to discriminate between the extra- and intra-cellular spaces. Using flow cytometry, results indicated that P. damselae induces apoptosis in phagocytes obtained from hybrid striped bass head kidney and after 12, 18, and 24 hours of incubation, the relative numbers of cells infected with P. damselae showing signs of apoptosis increased over time and were significantly greater than the controls. The relative numbers of apoptotic cells that were infected with the formalin-killed strain increased, but not significantly, above the control after the same designated times of incubation.