Early investigations into the infection courts used by <Emphasis Type="Italic">Neonectria fuckeliana</Emphasis> to enter <Emphasis Type="Italic">Pinus radiata</Emphasis> stems

Nectria flute canker is a disease of Pinus radiata stems caused by the pathogen Neonectria fuckeliana occurring in the southern parts of New Zealand. In Northern Hemisphere countries where N. fuckeliana is endemic, it is commonly found in Picea and Abies spp. Open wounds, dead attached branches and branch stubs have been identified as the primary infection courts. Although in New Zealand the development of Nectria flute canker disease is associated with pruned branch stubs, recent studies suggest that this is not the only possible entry method as the fungus has been found in trees prior to pruning. Three field trials were established to examine the potential infection mechanisms for N. fuckeliana in P. radiata in New Zealand; including stem wounds and branch stubs. The difference between inoculations into the stem and into branch wood was clear. Inoculation of deep stem wounds resulted in the greatest fluting with 76% of trees inoculated developing cankers. Inoculation directly into stubs resulted in only small stem depressions that occurred in 17% of cases and the fungus was largely contained within the branch trace. Tree response to inoculation with either ascospores or conidia of the Acremonium anamorph gave similar results in terms of canker development and fungal spread within the stem. Tree response to inoculation was highly variable however: in one study 6% of trees did not respond to inoculation at all, while 26% produced severe cankers regardless of inoculation method. A more thorough understanding of the infection mechanisms of N. fuckeliana will contribute to the development of better disease management protocols to prevent infection and disease development in future plantation stock.     

Effects of repeated hCG injections on reproductive efficiency in mares

Thirty reproductively sound mares were divided into treatment and control groups. In the treatment group, consisting of 14 mares, 2500 I.U. of human chorionic gonadotropin (hCG) was administered intravenously during estrus, in the presence of a 35 mm follicle over five successive cycles in 1987, and at least two cycles in 1988. Beginning with the second cycle of treatment in 1988, these mares were bred to a fertile stallion. The control group, consisting of 16 mares, was followed for two to five cycles in either the 1987 or 1988 season and six of these mares were bred to fertile stallions. Throughout the study period, blood was collected from the mares in the treatment group for analysis of anti-hCG antibodies and cross reactivity of the antibody to purified equine lutenizing hormone (eLH) and equine chorionic gonadotropin (eCG).In 1987, after the first three injections of hCG, mean duration of estrus in treated mares tended to be shorter than in control mares (P<.10). After all five hCG injections in 1987, mean ovulation time for treated mares was shorter than in control mares (P<0.01). However, after two to five hCG injections in 1987, seven treated mares (50%) had some individual ovulation times that did not differ from the control mares.Initially, following the first three injections of hCG in 1988, mean duration of estrus tended to be shorter (P<0.1) in treated mares compared to control mares. A reduction in mean ovulation time was observed after the first two hCG injections of 1988 (P<0.01). However, after one to four hCG injections in 1988, eight treated mares (57.1%) had some individual ovulation times that did not differ from controls.In 1987, all 14 treated mares developed significant levels of antibodies to hCG after one to four injections, and again in 1988, were positive for anti-hCG antibodies after one to three injections. However, no correlation was observed between magnitude of the immune response and duration of ovulation time or pregnancy rate. In cross reactivity studies, no significant binding of plasma anti-hCG antibodies to either eLH or eCG was observed in vitro.Overall, pregnancy and foaling rates of treated (85.7%) and control mares (83.3%) did not differ. Additionally, no difference was observed in number of inseminations per estrus between treated and control mares. In this study, with successive injections of hCG, the expected shortened time to ovulation was not elicited consistently in all mares. However, mares continued to ovulate, conceive and foal in the presence of significant levels of anti-hCG antibodies.     

Using synchrotron-based FTIR microspectroscopy to reveal chemical features of feather protein secondary structure: comparison with other feed protein sources

Studying the secondary structure of proteins leads to an understanding of the components that make up a whole protein. An understanding of the structure of the whole protein is often vital to understanding its digestive behavior in animals and nutritive quality. Usually protein secondary structures include alpha-helix and beta-sheet. The percentages of these two structures in protein secondary structures may influence feed protein quality and digestive behavior. Feathers are widely available as a potential protein supplement. They are very high in protein (84%), but the digestibility of the protein is very low (5%). The objective of this study was to use synchrotron-based Fourier transform infrared (FTIR) microspectroscopy to reveal chemical features of feather protein secondary structure within amide I at ultraspatial resolution (pixel size = 10 x 10 microm), in comparison with other protein sources from easily digested feeds such as barley, oat, and wheat tissue at endosperm regions (without destruction of their inherent structure). This experiment was performed at beamline U2B of the Albert Einstein Center for Synchrotron Biosciences at the National Synchrotron Light Source (NSLS) in Brookhaven National Laboratory (BNL), U.S. Dept of Energy (NSLS-BNL, Upton, NY). The results showed that ultraspatially resolved chemical imaging of feed protein secondary structure in terms of beta-sheet to alpha-helix peak height ratio by stepping in pixel-sized increments was obtained. Using synchrotron FTIR microspectroscopy can distinguish structures of protein amide I among the different feed protein sources. The results show that the secondary structure of feather protein differed from those of other feed protein sources in terms of the line-shape and position of amide I. The feather protein amide I peaked at approximately 1630 cm(-1). However, other feed protein sources showed a peak at approximately 1650 cm(-1). By using multicomponent peak modeling, the relatively quantitative amounts of alpha-helix and beta-sheet in protein secondary structure were obtained, which showed that feather contains 88% beta-sheet and 4% alpha-helix, barley contains 17% beta-sheet and 71% alpha-helix, oat contains 2% beta-sheet and 92% alpha-helix, and wheat contains 42% beta-sheet and 50% alpha-helix. The difference in percentage of protein secondary structure may be part of the reason for different feed protein digestive behaviors. These results demonstrate the potential of highly spatially resolved infrared microspectroscopy to reveal feed protein secondary structure. Information from this study by the infrared probing of feed protein secondary structure may be valuable as a guide for feed breeders to improve and maintain protein quality for animal use.     

Imaging molecular chemistry of Pioneer corn

Synchrotron Fourier transform infrared (FTIR) microspectroscopy as a rapid, direct, and nondestructive analytical technique can explore molecular chemical features of the microstructure of biological samples. The objective of this study was to use synchrotron FTIR microspectroscopy to image the molecular chemistry of corn (cv. Pioneer 39P78) to reveal spatial intensity and distribution of chemical functional groups in corn tissue. This experiment was performed at the U2B station of the National Synchrotron Light Source in Brookhaven National Laboratory (NSLS-BNL, Upton, NY). The Pioneer corn tissue was imaged from the pericarp, seed coat, aleurone, and endosperm under peaks at 1736 (carbonyl C=O ester), 1510 (aromatic compound), 1650 (amide I), 1550 (amide II), 1246 (cellulosic material), 1160 (CHO), 1150 (CHO), 1080 (CHO), 929 (CHO), 860 (CHO), 3350 (OH and NH stretching), 2929 (CH(2) stretching band), and 2885 cm(-1) (CH(3) stretching band). The results showed that with synchrotron FTIR microspectroscopy, the images of the molecular chemistry of Pioneer corn could be generated. Such information on the microstructural-chemical features of grain corn can also be used for corn breeding programs for selecting superior varieties of corn for targeted food and feed purposes and for prediction of corn quality and nutritive value for humans and animals.     

Black cohosh acts as a mixed competitive ligand and partial agonist of the serotonin receptor

Extracts of the rhizome of black cohosh [Actaea racemosa L., formerly called Cimicifuga racemosa (L.) Nutt.] were evaluated for potential mechanisms of action in the alleviation of menopausal hot flashes. Ovariectomized Sprague-Dawley rats were administered a 40% 2-propanol extract of black cohosh [4, 40, and 400 mg/(kg.day)] by gavage for 2 weeks with or without estradiol [50 microg/(kg.day)] to determine if black cohosh could act as an estrogen or antiestrogen on the basis of an increase in uterine weight or vaginal cellular cornification. No effects were observed on uterine weight or on vaginal cellular cornification in rats treated with black cohosh alone or in combination with 17beta-estradiol, indicating this black cohosh extract had no estrogenic or antiestrogenic properties in the ovariectomized rat model. To evaluate other potential pathways by which black cohosh might reduce menopausal hot flashes, serotonin activity was first assessed by the inhibition of radioligand binding to cell membrane preparations containing recombinant human serotonin receptor (5-HT) subtypes. A 40% 2-propanol extract of black cohosh was tested against 10 subtypes of the serotonin receptor, revealing the presence of compounds with strong binding to the 5-HT(1A), 5-HT(1D), and 5-HT(7) subtypes. Subsequent binding studies were carried out using 5-HT(1A) and 5-HT(7) receptors because of their association with the hypothalamus, which has been implicated in the generation of hot flashes. The black cohosh 40% 2-propanol extract inhibited [(3)H]lysergic acid diethylamide (LSD) binding to the human 5-HT(7) receptor (IC(50) = 2.4 +/- 0.4 microg/mL) with greater potency than binding of [(3)H]-8-hydroxy-2-(di-N-propylamino)tetralin to the rat 5-HT(1A) receptor (IC(50) = 13.9 +/- 0.6 microg/mL). Analysis of ligand binding data indicated that components of a black cohosh methanol extract functioned as a mixed competitive ligand of the 5-HT(7) receptor. In addition, a black cohosh methanol extract elevated cAMP levels in 293T-5-HT(7)-transfected HEK cells, suggesting the extract acted as a partial agonist at the receptor. The elevation in cAMP mediated by the black cohosh extract could be reversed in the presence of the antagonist methiothepin, indicating a receptor-mediated process. These data suggest that reductions in hot flashes in some women taking black cohosh may not be due to estrogenic properties. This study identifies other possible biological targets of black cohosh that could account for reported biological effects.     

Variant of SCN5A sodium channel implicated in risk of cardiac arrhythmia

Every year, approximately 450,000 individuals in the United States die suddenly of cardiac arrhythmia. We identified a variant of the cardiac sodium channel gene SCN5A that is associated with arrhythmia in African Americans (P = 0.000028) and linked with arrhythmia risk in an African-American family (P = 0.005). In transfected cells, the variant allele (Y1102) accelerated channel activation, increasing the likelihood of abnormal cardiac repolarization and arrhythmia. About 13.2% of African Americans carry the Y1102 allele. Because Y1102 has a subtle effect on risk, most carriers will never have an arrhythmia. However, Y1102 may be a useful molecular marker for the prediction of arrhythmia susceptibility in the context of additional acquired risk factors such as the use of certain medications.     

Occurrence of antimicrobial resistant bacteria in healthy dogs and cats presented to private veterinary hospitals in southern Ontario: A preliminary study

The prevalence and patterns of antimicrobial susceptibility of fecal Escherichia coli, Salmonella spp., extended β-lactamase producing E. coli (ESBL-E. coli), methicillin-resistant Staphylococcus aureus (MRSA), and methicillin-resistant Staphylococcus pseudintermedius (MRSP) were determined for healthy dogs (n = 188) and cats (n = 39) from veterinary hospitals in southern Ontario that had not had recent exposure to antimicrobials. The prevalence of antimicrobial resistance in E. coli was as follows: streptomycin (dogs — 17%, cats — 2%), ampicillin (dogs — 13%, cats — 4%), cephalothin (dogs — 13%, cats — < 1%), and tetracycline (dogs — 11%, cats — 2%). Eleven percent of dogs and 15% of cats had isolates that were resistant to at least 2 antimicrobials. Cephamycinase (CMY)-2 producing E. coli was cultured from 2 dogs. No Salmonella spp., ESBL-E. coli, MRSA, or MRSP isolates were recovered. The observed prevalence of resistance in commensal E. coli from this population was lower than that previously reported in companion animals, but a small percentage of dogs may be a reservoir for CMY-2 E. coli.     

Impacts of the Cretaceous Terrestrial Revolution and KPg extinction on mammal diversification

Previous analyses of relations, divergence times, and diversification patterns among extant mammalian families have relied on supertree methods and local molecular clocks. We constructed a molecular supermatrix for mammalian families and analyzed these data with likelihood-based methods and relaxed molecular clocks. Phylogenetic analyses resulted in a robust phylogeny with better resolution than phylogenies from supertree methods. Relaxed clock analyses support the long-fuse model of diversification and highlight the importance of including multiple fossil calibrations that are spread across the tree. Molecular time trees and diversification analyses suggest important roles for the Cretaceous Terrestrial Revolution and Cretaceous-Paleogene (KPg) mass extinction in opening up ecospace that promoted interordinal and intraordinal diversification, respectively. By contrast, diversification analyses provide no support for the hypothesis concerning the delayed rise of present-day mammals during the Eocene Period.     

Distribution and prevalence of Cytauxzoon felis in bobcats (Lynx rufus), the natural reservoir, and other wild felids in thirteen states

Cytauxzoon felis, a protozoan parasite of wild and domestic felids, is the causative agent of cytauxzoonosis in domestic and some exotic felids in the United States. The bobcat (Lynx rufus) is the natural reservoir for this parasite, but other felids such as Florida panthers (Puma concolor coryii) and domestic cats may maintain long-term parasitemias and serve as reservoirs. Experimentally, two tick species, Dermacentor variabilis and Amblyomma americanum, have demonstrated the ability to transmit C. felis. These two tick species have overlapping distributions throughout much of the southeastern United States. The objective of the current study was to determine the distribution and prevalence of C. felis in free-ranging bobcat populations from 13 states including California, Colorado, Florida, Georgia, Kansas, Kentucky, Missouri, North Carolina, North Dakota, Ohio, Oklahoma, South Carolina, and West Virginia. These states were selected because of differential vector presence; D. variabilis is present in each of these states except for the region of Colorado sampled and A. americanum is currently known to be present only in a subset of these states. Blood or spleen samples from 696 bobcats were tested for C. felis infection by a polymerase chain reaction (PCR) assay which targeted the first ribosomal internal transcribed spacer region (ITS-1). Significantly higher prevalences of C. felis were detected from Missouri (79%, n=39), North Carolina (63%, n=8), Oklahoma (60%, n=20), South Carolina (57%, n=7), Kentucky (55%, n=74), Florida (44%, n=45), and Kansas (27%, n=41) compared with Georgia (9%, n=159), North Dakota (2.4%, n=124), Ohio (0%, n=19), West Virginia (0%, n=37), California (0%, n=26), and Colorado (0%, n=67). In addition to bobcats, seven cougars (Puma concolor) from Georgia, Louisiana, and North Dakota and one serval (Leptailurus serval) from Louisiana were tested for C. felis. Only one cougar from Louisiana was PCR positive, which represents the first report of an infected cougar outside of the Florida panther population. These data also indicate that C. felis is present in North Dakota where infection has not been reported in domestic cats. Based on a nonparametric analysis, prevalence rates were significantly higher in states where there are established populations of A. americanum, which supports recent data on the experimental transmission of C. felis by A. americanum and the fact that domestic cat clinical cases are temporally associated with A. americanum activity. Collectively, these data confirm that bobcats are a common reservoir for C. felis and that A. americanum is likely an epidemiologically important vector.