Collection and characterization of semen from Chilean rose tarantulas (Grammostola rosea).

OBJECTIVE: To establish a nonterminal semen collection method for use in captive Chilean rose tarantulas (Grammostola rosea) and to evaluate tools for investigating morphology and viability of spermatozoa. ANIMALS: 7 mature male Chilean rose tarantulas. PROCEDURES: Each tarantula was anesthetized in a 500-mL induction chamber containing a cotton ball infused with 2 mL of isoflurane. Semen collection was performed by applying direct pressure to the palpal bulbs (sperm storage organs) located on the distal segment of the palpal limbs. Morphology of spermatozoa was examined by light microscopy and transmission and scanning electron microscopy. Propidium iodide and a fluorescent membrane-permeant nucleic acid dye were used to evaluate cell viability. RESULTS: Semen was collected successfully from all 7 tarantulas. Microscopic examination of semen samples revealed coenospermia (spherical capsules [mean ± SD diameter, 10.3 ± 1.6 µm] containing many nonmotile sperm cells [mean number of sperm cells/capsule, 18.5 ± 3.8]). Individual spermatozoa were characterized by a spiral-shaped cell body (mean length, 16.7 ± 1.4 µm; mean anterior diameter, 1.5 ± 0.14 µm). Each spermatozoon had no apparent flagellar structure. The fluorescent stains identified some viable sperm cells in the semen samples. CONCLUSIONS AND CLINICAL RELEVANCE: The described technique allowed simple and repeatable collection of semen from Chilean rose tarantulas. Semen from this species was characterized by numerous spherical capsules containing many nonmotile spermatozoa in an apparently quiescent state. Fluorescent staining to distinguish live from dead spermatozoa appeared to be a useful tool for semen evaluation in this species.

Dapagliflozin monotherapy in drug-naïve patients with diabetes: a randomized-controlled trial of low-dose range.

AIMS: Many patients with type 2 diabetes are suboptimally managed with currently available therapies. Dapagliflozin, a sodium-glucose co-transporter-2 inhibitor, has shown efficacy in reducing diabetic hyperglycaemia. This study assessed efficacy of three lower doses in recently diagnosed patients. METHODS: This phase 3, randomized, double-blind, placebo-controlled study assigned treatment-naïve patients to placebo or dapagliflozin monotherapy (1, 2.5 or 5 mg) daily for 24 weeks. Patients were antidiabetic drug-naïve with inadequate glycaemic control [haemoglobin A1c (HbA1c) ≥7.0 and ≤10.0%]. The primary efficacy endpoint was change in HbA1c from baseline. Secondary endpoints included changes in body weight and fasting plasma glucose (FPG), and proportions achieving HbA1c <7%. RESULTS: A total of 282 patients with type 2 diabetes were randomly assigned to one of four treatment groups. Baseline characteristics were similar across groups. At week 24, mean HbA1c reduction was significantly greater with dapagliflozin: -0.68% for 1 mg, -0.72% for 2.5 mg, -0.82% for 5 mg, versus 0.02% for placebo (p < 0.0001); compared to mean baseline values of 7.8-8.1%. Mean FPG reduction was significantly greater for all dapagliflozin groups versus placebo (p < 0.02), as was mean weight reduction (p < 0.003). During the treatment period, 19.1% of placebo-treated patients received rescue medication or discontinued because of poor glycaemic control versus 6.9, 4.1 and 5.9% for dapagliflozin 1, 2.5 and 5 mg, respectively. Percentages of patients experiencing ≥1 adverse event were similar across groups. CONCLUSION: Dapagliflozin at doses of 1, 2.5 and 5 mg/day is effective in reducing glycaemic levels and body weight in treatment-naïve patients with type 2 diabetes. Dapagliflozin was generally well tolerated. This insulin-independent mechanism suggests a new treatment for type 2 diabetes.

Soybean, palm kernel, and animal-vegetable oils and vitamin E supplementation effect on lipid oxidation stability of sous vide chicken meat.

There is an increasing demand in precooked chicken meat products for restaurants and catering services. Because cooked chicken meat develops lipid oxidation relatively fast, sous vide chicken meat was studied to assess its shelf-life. Six hundred Cobb x Ross broilers were fed for 6 wk with a basal corn-soybean meal diet including soybean, palm kernel, or animal-vegetable oil, each supplemented with 33 or 200 mg/kg of dl-alpha-tocopheryl acetate. Broilers were randomly assigned into 6 treatments and 4 repetitions with 25 birds each. Boneless breast or thigh muscle pieces were dissected into 5 x 5 x 5 cm cubes, vacuum-packed, cooked in water bath (until 74 degrees C internal temperature), chilled, and stored at 4 degrees C for 1, 5, 10, 25, and 40 d. For each storage day, each pouch contained 3 pieces of meat, either breast or thigh. Thiobarbituric acid reactive substances analysis, to quantify malonaldehyde (MDA) values, was conducted to estimate the lipid oxidation development. Nonheme iron values of cooked meat were analyzed. Fatty acid methyl esters analysis was performed in chicken muscle to determine its fatty acid composition. There was no interaction between dietary fat and vitamin E level in all of the variables studied except in nonheme iron. Dietary fat significantly influenced the fatty acid composition of the muscle (P < 0.01), but it did not affect the MDA values, regardless of differences in the muscle fatty acid composition between treatments. Supplementation of the high level of vitamin E significantly reduced the MDA values in both breast and thigh meat (P < 0.01). The maximum MDA values were observed at d 40 of storage in thigh and breast meat in animal-vegetable and soybean oil treatments with the low levels of vitamin E, 0.91 and 0.70 mg/kg, respectively. Nonheme iron values in thigh meat differed between treatments at 1 or 25 d of storage but not in breast meat. In conclusion, refrigerated sous vide chicken meat has a prolonged shelf-life, which is enhanced by dietary supranutritional supplementation of vitamin E.

Growth characteristics, reproductive performance, and evaluation of their associative relationships in Brangus cattle managed in a Chihuahuan Desert production system1.

Balancing growth and reproductive performance in beef cattle managed in desert environments is challenging. Our objectives were to 1) evaluate trends in growth and reproductive traits, and 2) assess associative relationships between growth characteristics and reproductive performance in a Brangus herd managed in a Chihuahuan Desert production system from 1972 to 2006. Data were from bull (n = 597) and heifer calves (n = 585; 1988 to 2006) and cows (n = 525; repeated records of cows, n = 2,611; 1972 to 2006). Variables describing the growth curve of each cow were estimated using a nonlinear logistic function (each cow needed 6 yr of data). Mixed-effect models and logistic regression were used to analyze trends across years in growth and reproductive traits (both continuous and categorical). For continuous traits of calves, a slight cubic response (P < 0.01) described the dynamics of birth weight, 205-d BW, and 365-d BW across years. For categorical traits of females, positive linear trends (P < 0.05) across years were observed in percent pregnant as yearlings, calved at 2 yr of age, and first-calf heifer rebreeding (slopes ranged from 0.007 to 0.014%/yr). Autumn cow BW increased gradually until 1997 (509 kg +/- 8.8) and then decreased gradually by 0.6 kg/yr, whereas pregnancy percentage decreased gradually until 1995 (78.4% +/- 1.0) and then increased slightly by 0.2%/yr. A quadratic effect best described the dynamics of these 2 variables across years (P < 0.01) as well as estimates describing the growth curve of each cow. Specifically, asymptotic BW and age increased (P < 0.05) from 1972 to 1983 and 1990, respectively. Asymptotic age then decreased by 27% from 1983 to 1996 (P < 0.05). The maturing rate index was negatively correlated with age at first calving and calving interval (r = -0.42 and -0.18, P < 0.01), which suggested that early-maturing cows had enhanced fertility in this environment and production system. In summary, minimal changes were observed in measures of growth in bulls and heifers in a Brangus herd managed in the Chihuahuan Desert. Opposing relationships were observed among measures of cow size and fertility; as growth curves shifted toward earlier maturity, measures of reproductive performance suggested that fertility improved.

Population genetics of the understory fishtail palm Chamaedorea ernesti-augusti in Belize: high genetic connectivity with local differentiation.

BACKGROUND: Developing a greater understanding of population genetic structure in lowland tropical plant species is highly relevant to our knowledge of increasingly fragmented forests and to the conservation of threatened species. Specific studies are particularly needed for taxa whose population dynamics are further impacted by human harvesting practices. One such case is the fishtail or xaté palm (Chamaedorea ernesti-augusti) of Central America, whose wild-collected leaves are becoming progressively more important to the global ornamental industry. We use microsatellite markers to describe the population genetics of this species in Belize and test the effects of climate change and deforestation on its recent and historical effective population size. RESULTS: We found high levels of inbreeding coupled with moderate or high allelic diversity within populations. Overall high gene flow was observed, with a north and south gradient and ongoing differentiation at smaller spatial scales. Immigration rates among populations were more difficult to discern, with minimal evidence for isolation by distance. We infer a tenfold reduction in effective population size ca. 10,000 years ago, but fail to detect changes attributable to Mayan or contemporary deforestation. CONCLUSION: Populations of C. ernesti-augusti are genetically heterogeneous demes at a local spatial scale, but are widely connected at a regional level in Belize. We suggest that the inferred patterns in population genetic structure are the result of the colonization of this species into Belize following expansion of humid forests in combination with demographic and mating patterns. Within populations, we hypothesize that low aggregated population density over large areas, short distance pollen dispersal via thrips, low adult survival, and low fruiting combined with early flowering may contribute towards local inbreeding via genetic drift. Relatively high levels of regional connectivity are likely the result of animal-mediated long-distance seed dispersal. The greatest present threat to the species is the potential onset of inbreeding depression as the result of increased human harvesting activities. Future genetic studies in understory palms should focus on both fine-scale and landscape-level genetic structure.

Serendipitous backyard hybridization and the origin of crops.

Backyard gardens, dump heaps, and kitchen middens are thought to have provided important venues for early crop domestication via generation of hybrids between otherwise isolated plant species. However, this process has rarely been demonstrated empirically. For the majority of polyploid crops, it remains uncertain to what extent hybridization and polyploidization preceded domestication or were precipitated by human activities. Using archaeological, ethnobotanical, geographical, and genetic data, we investigate the extent and significance of predomestication cultivation, backyard sympatry, and spontaneous hybridization for the Mimosoid legume tree Leucaena, which is used as a food crop throughout south-central Mexico. We show that predomestication cultivation was widespread, involved numerous independent transitions from the wild to cultivation, and resulted in extensive artificial sympatry of 2-6 species locally and 13 species in total. Using chloroplast and rapidly evolving nuclear-encoded DNA sequences, we demonstrate that hybridization in Leucaena has been extensive and complex, spawning a diverse set of novel hybrids as a result of juxtaposition of species in cultivation. The scale and complexity of hybridization in Leucaena is significantly greater than that documented for any other Mexican plant domesticates so far. However, there are striking parallels between Leucaena and the other major Mexican perennial domesticates Agave and Opuntia, which show very similar domestication via backyard hybridization pathways. Our results suggest that backyard hybridization has played a central role in Mesoamerican crop domestication and demonstrate that the simple step of bringing species together in cultivation can provide a potent trigger for domestication.

Taxonomy and conservation: A case study from Chamaedorea alternans.

BACKGROUND AND AIMS: The establishment of justified recommendations in conservation biology requires robust taxonomic treatments for the group(s) being considered. Controversial or poorly developed taxonomies can have a negative impact on conservation assessments. One example of a taxonomically difficult and controversial species complex that is important in conservation involves two species of Mexican palms, Chamaedorea tepejilote and C. alternans. The goal of this study was to investigate whether C. alternans and C. tepejilote are genetically distinct within the Los Tuxtlas region of Veracruz. METHODS: Individuals corresponding to the morphology of C. alternans and C. tepejilote were collected from sympatric and allopatric regions within the Los Tuxtlas Biological Station. Eighty-eight samples were genotyped using amplified fragment length polymorphism (AFLP) markers. Cluster and ordination analyses were used to investigate patterns of differentiation. KEY RESULTS: UPGMA and PCO analyses of AFLP profiles recovered two divergent clusters corresponding to morphologically defined C. tepejilote and C. alternans. No intermediate genotypes were observed and five of the 45 loci were fixed for either the presence or absence between the species. The patterns of divergence observed do not identify a role for sympatric speciation. CONCLUSIONS: The observed patterns of differentiation support the recognition of C. alternans as distinct from C. tepejilote. A suite of vegetative and reproductive morphological features can be used to help distinguish these taxa in the field, but they can be difficult to differentiate from preserved material. Previous treatments of the variation found within the complex (C. alternans and C. tepejilote) as intraspecific variance is unjustified. Given that the Los Tuxtlas region has suffered from historical and ongoing deforestation and that Chamaedorea includes numerous endangered taxa, retaining conservation status for C. alternans serves to help safeguard individuals of the species as well as a region and larger taxonomic group all under considerable threat from human activities.

Molecular analysis of Salmonella enteritidis isolates from the Caribbean by pulsed-field gel electrophoresis.

Using pulsed-field gel electrophoresis (PFGE), between 1987 and 1996 we analyzed Salmonella enteritidis isolates from gastroenteritis cases in four Caribbean countries: Barbados, Saint Kitts and Nevis, Saint Lucia, and Trinidad and Tobago. We also determined the resistance of the isolates to 12 antimicrobial agents. Of the 129 isolates of S. enteritidis available for testing, DNA digested by XbaI revealed 13 distinctive PFGE patterns. The most prevalent XbaI PFGE patterns were group 1 (88 of 129 isolates, 68.2%) and group 2 (26 of 129, 20.2%). The patterns found among S. enteritidis isolates correlated with the geographical origin of the isolates. Of the 28 isolates from Barbados, 20 of them (71.4%) belonged to XbaI PFGE group 2, and of the 93 isolates from Trinidad and Tobago, 78 of them (83.9%) belonged to group 1. SpeI digestion of S. enteritidis genome was not as discriminatory as XbaI. Overall, of the 129 isolates, 67 of them (51.9%) exhibited resistance to one or more of the 12 antimicrobial agents that we tested. The prevalence of resistance was 53.8% for the S. enteritidis isolates tested from Trinidad and Tobago, 50.0% for those from Barbados, 28.6% for those from Saint Lucia, and 100.0% for one isolate from the island of Saint Kitts. Resistance was highest to triple sulfur (59 of 129 isolates, 45.7%), followed by furadantoin (10 of 129, 7.8%), ampicillin (7 of 129, 5.4%), and carbamycin (5 of 129, 3.9%).

Venezuelan equine encephalomyelitis virus infection in and transmission by the tick Amblyomma cajennense (Arachnida: Ixodidae).

To assess a possible role of ticks as the maintenance host for epizootic strains of Venezuelan equine encephalomyelitis (VEE) virus, laboratory experiments were conducted to determine if ticks could become infected, maintain, and transmit the virus. Larval and nymphal Amblyomma cajennense (F.) and larval Dermacentor nitens Neumann ticks were exposed to epizootic VEE virus (Trinidad donkey strain) by allowing them to feed on viremic guinea pigs (strain 13). In A. cajennense, transstadial transmission was observed from larvae to nymphs and adults. Horizontal viral transmission to a mammalian host was accomplished by nymphs. Infection rates in nymphs and adults were 2% (42/2,750) and 4% (9/244), respectively, afer ingestion of virus as larvae. Virus was detected in A. cajennense adult ticks for up to 171 d after infection in the larval stage. A cajennense, exposed as nymphs, ingested virus but did not become infected (0/164 after 10 d after taking an infective bloodmeal). No virus was detected in D. nitens 7 d after exposure. These findings suggest that A. cajennense potentially could be involved in an interepizootic maintenance cycle of epizootic VEE viral strains.

Identification of Aedes campestris from New Mexico: with notes on the isolation of western equine encephalitis and other arboviruses.

An arbovirus survey was conducted during August 1985 at White Sands Missile Range in southcentral New Mexico following a suspected arboviral disease epizootic among feral horses. A total of 20,566 mosquitoes (18,505 females and 2,061 males) and 8,900 biting gnats were collected and assayed for virus. Female mosquitoes were principally Aedes campestris (54.8%), Aedes dorsalis (30.4%) and Culex tarsalis (13.2%). Arboviruses were not isolated from biting gnats, but mosquitoes yielded a total of 37 viral isolates, including western equine encephalitis (WEE) (18), California serogroup (15), Cache Valey (1), and Hart Park (1) viruses in addition to 2, as yet unidentified, rhabdoviruses. Isolates of WEE virus were from 9 pools of Ae. campestris, 6 of Cx. tarsalis and 3 of Ae. dorsalis. California serogroup viruses, including 2 subtypes, were obtained from 7 pools of females and 1 pool of males of Ae. campestris and from 4 pools of Ae. dorsalis. Cache Valley and Hart Park viruses were isolated from single pools of Ae. dorsalis and Cx. tarsalis, respectively, and the rhabdoviruses were obtained from Ae. campestris and Psorophora signipennis.