Novel approaches to genetic analysis of fertility traits in New Zealand dairy cattle.

The fertility of dairy cattle in New Zealand is well below industry targets, and the current New Zealand fertility breeding value (BV) could potentially be improved using additional information and traits. Data from 169 herds were analyzed to determine the benefits of using alternative phenotypic measures in the calculation of the fertility BV. The heritability of calving season day (CSD; calving season day as an integer day of the year) and the probability of an animal calving within 42 d of the planned start of calving (CR42) increased modestly (from 0.0206 to 0.0213 and 0.0087 to 0.0092, respectively) after accounting for the use of intravaginal progesterone-releasing devices for treatment of anestrous cows (anestrum treatment) and induced calvings. Incidence of either anestrum treatment or calving induction as a single binomial trait (AT/IND) had a heritability of 0.0223 and showed moderate genetic correlation with the probability of an animal being mated within 21 d of the planned start of mating (PM21; -0.4473), but much higher with CSD (0.8445). The use of pregnancy diagnosis data allowed fertility information that would otherwise be discarded to be included in analyses; when used to assign a prolonged CSD and a value of 0 for CR42 to animals that failed to calve, it increased the heritabilities of both of these traits (to 0.0278 and 0.0114, respectively). Because CSD was found to be more than twice as heritable as its binary counterpart, it shows potential to replace CR42 as the calving trait used in the fertility BV. Postpartum anestrous interval (PPAI), derived using incomplete premating estrous recording in some herds, had a heritability of 0.0813 and hence has potential as a trait to be included in genetic improvement programs but would require more rigorous recording of estrous during the premating period to be an effective trait. Based on selection index theory, the modifications made to current selection criteria using novel fertility traits increased the accuracy of prediction of fertility merit by more than 12%. Because of the increasing economic importance of fertility traits, and low heritabilities requiring large numbers of recorded daughters to get accurate fertility BV predictions on sires, data recorded on farm will become increasingly important in the genetic improvement of fertility.

The impact of lead poisoning from ammunition sources on raptor populations in Europe.

Poisoning caused by ingestion of spent lead (Pb) ammunition in food items is a common cause of death of raptors. However, there has been no previous attempt to assess the impact of lead poisoning on populations of raptors throughout Europe or examine how this relates to the prevalence of hunting. We used measurements of lead concentration in the liver from over 3000 raptors of 22 species found dead or dying in the wild in 13 countries and a lead poisoning threshold of 20 ppm (dry weight) to assess the proportion of these in which lead poisoning caused or contributed to death. The prevalence of lead poisoning as a cause of death of raptors varied substantially among European countries and was positively correlated across countries with the reported number of hunters per unit area. Ten species had a non-zero proportion of individuals with concentrations exceeding the lead poisoning threshold ranging between 0.3% and 16.5%. The estimated annual conditional death rate from lead poisoning for these ten species averaged 0.44% (range 0.06-0.85%). Scavenging species feeding regularly on carcasses of game animals,tended to have a high annual probability of death from lead poisoning. So too did some predators which only sometimes scavenge, but prey on frequently hunted birds, such as gamebirds, waterfowl and pigeons, which may contain ingested or embedded lead shot. Small-bodied predators had a low annual probability of death from lead poisoning. Modelling indicated that European populations of adult raptors of the ten focal species averaged 6.0% smaller (range 0.2-14.4%) than they would be without the effects of lead poisoning. A given rate of lead poisoning mortality resulted in greater expected population reductions for species with high annual survival rate and late age at first breeding.

Targeted resequencing of a genomic region influencing tameness and aggression reveals multiple signals of positive selection.

The identification of the causative genetic variants in quantitative trait loci (QTL) influencing phenotypic traits is challenging, especially in crosses between outbred strains. We have previously identified several QTL influencing tameness and aggression in a cross between two lines of wild-derived, outbred rats (Rattus norvegicus) selected for their behavior towards humans. Here, we use targeted sequence capture and massively parallel sequencing of all genes in the strongest QTL in the founder animals of the cross. We identify many novel sequence variants, several of which are potentially functionally relevant. The QTL contains several regions where either the tame or the aggressive founders contain no sequence variation, and two regions where alternative haplotypes are fixed between the founders. A re-analysis of the QTL signal showed that the causative site is likely to be fixed among the tame founder animals, but that several causative alleles may segregate among the aggressive founder animals. Using a formal test for the detection of positive selection, we find 10 putative positively selected regions, some of which are close to genes known to influence behavior. Together, these results show that the QTL is probably not caused by a single selected site, but may instead represent the joint effects of several sites that were targets of polygenic selection.

Global mapping of ecosystem services and conservation priorities.

Global efforts to conserve biodiversity have the potential to deliver economic benefits to people (i.e., "ecosystem services"). However, regions for which conservation benefits both biodiversity and ecosystem services cannot be identified unless ecosystem services can be quantified and valued and their areas of production mapped. Here we review the theory, data, and analyses needed to produce such maps and find that data availability allows us to quantify imperfect global proxies for only four ecosystem services. Using this incomplete set as an illustration, we compare ecosystem service maps with the global distributions of conventional targets for biodiversity conservation. Our preliminary results show that regions selected to maximize biodiversity provide no more ecosystem services than regions chosen randomly. Furthermore, spatial concordance among different services, and between ecosystem services and established conservation priorities, varies widely. Despite this lack of general concordance, "win-win" areas-regions important for both ecosystem services and biodiversity-can be usefully identified, both among ecoregions and at finer scales within them. An ambitious interdisciplinary research effort is needed to move beyond these preliminary and illustrative analyses to fully assess synergies and trade-offs in conserving biodiversity and ecosystem services.

Viability of OPS vitrified sheep embryos after direct transfer.

The aim of this study was to evaluate the viability in the effect of open pulled straw (OPS) vitrification procedure of sheep embryos after direct transference. Embryos were produced in vivo and cryopreserved in slow freezing or OPS vitrification. The survival rates of cryopreserved embryos were compared to non-frozen standard pattern. In a first set of experiments, embryos at morula and blastocyst stages were dived in ethylene glycol (1.5 M) and frozen in an automatic freezer. After being thawed, they were directly or indirectly transferred to ewes recipient. A second group of embryos were drawn into OPS and plunged into liquid nitrogen after being exposed at room temperature for 1 min and 45 s in 10% EG plus 10% dimethyl sulphoxide (DMSO), then again for 30 s in 20% EG + 20% DMSO + 0.5 M sucrose. After being warmed, embryos were also directly transferred using a French mini straw as the catheter for the transplantation process or after in vitro dilution of cryoprotectants (two-step-process). No significant difference was observed among fresh, frozen or vitrified embryos on pregnancy rate (50.0%, 38.6% and 55.8%). However, when we evaluated only the direct transference, the pregnancy rate of OPS vitrified embryos was higher than that of frozen embryos (57.1% vs 34.8%) (p = 0.07). In addition, vitrified morulae had a higher pregnancy rate than the one with frozen embryos (64.0% vs 38.9%) (p = 0.07). Finally, our results indicate that OPS vitrification technique in association with direct transference improves the viability of sheep embryos with potential applications to field conditions.

Movement of bromacil in a Hawaii soil under pineapple cultivation - a field study.

Recent discovery of low concentrations of bromacil in drinking water prompted the State of Hawaii to examine the leaching behavior of bromacil in pineapple fields. This study is a follow up to earlier work on bromacil concentrations in soil profiles in a pineapple field in central Oahu, Hawaii. Soil samples were collected for bromacil analysis at different times prior to and after application from a pineapple field that was previously surveyed by other research workers. The leaching pattern of bromacil was further investigated at two different application rates (2.25 and 1.8 kg ha(-1)). The concentration of bromacil in the topsoil about 100 days after bromacil application (1.8 kg ha(-1)) was substantially higher in 2002 compared to 1999. The distribution profiles were generally consistent with the one presented in the previous study. Residual bromacil was present in the entire sampled zone (3m deep) about 18 months after the previous bromacil application. Over a period of 9 months, there was substantial dissipation of bromacil residue present in the topsoil. The residual concentration of bromacil in the area that received the reduced application rate (1.8 kg ha(-1)) were lower than those receiving the current application rate (2.25 kg ha(-1)) and the depth of penetration of the bromacil front was shallower at the reduced application rate. Because of the common practice of placing plastic mulch around the base of the pineapple plants to retain volatile nematicides, the applied bromacil was found to be concentrated in the areas between the plastic mulch, transported by runoff from the plastic. The study results encourage the use of less than the label led rate of application of bromacil for pineapple fields.

The use of toxicokinetics and exposure studies to show that carprofen in cattle tissue could lead to secondary toxicity and death in wild vultures.

Veterinary medicines can be extremely damaging to the environment, as seen with the catastrophic declines in Gyps vulture in South Asia due to their secondary exposure to diclofenac in their primary food source. Not surprisingly, concern has been raised over other similar drugs. In this study, we evaluate the toxicity of carprofen to the Gyps vulture clade through plasma pharmacokinetics evaluations in Bos taurus cattle (their food source) and Gyps africanus (a validated model species); tissue residues in cattle; and the effect of carprofen as a secondary toxicant as both tissue-bound residue or pure drug at levels expected in cattle tissues. Carprofen residues were highest in cattle kidney (7.72 ± 2.38 mg/kg) and injection site muscle (289.05 ± 98.96 mg/kg of dimension of 5 × 5 × 5 cm). Vultures exposed to carprofen as residues in the kidney tissue or pure drug equivalents showed no toxic signs. When exposed to average injection site concentrations (64 mg/kg) one of two birds died with evidence of severe renal and liver damage. Toxicokinetic analysis revealed a prolonged drug half-life of 37.75 h in the dead bird as opposed to 13.99 ± 5.61 h from healthy birds dosed intravenously at 5 mg/kg. While carprofen may generally be harmless to Gyps vultures, its high levels at the injection site in treated cattle can result in lethal exposure in foraging vultures, due to relative small area of tissue it is found therein. We thus suggest that carprofen not be used in domesticated ungulates in areas where carcasses are accessible or provided to vultures at supplementary feeding sites.

Diclofenac residues in carcasses of domestic ungulates available to vultures in India.

Gyps vulture populations across the Indian subcontinent are declining rapidly and evidence indicates that veterinary use of the non-steroidal anti-inflammatory drug (NSAID) diclofenac is the major cause. Exposure of vultures to diclofenac is likely to arise from the consumption of livestock carcasses that have been treated shortly before death, however, detailed information regarding the prevalence and residual levels of diclofenac in carcasses available to vultures in India remains unreported. Here, we present data on diclofenac residues in 1848 liver samples taken from carcasses of dead livestock sampled at 67 sites in 12 states within India, between May 2004 and July 2005. Diclofenac residues were detected in carcasses in all states except Orisa, where only one site was sampled. The overall prevalence of detectable diclofenac (>10 microg kg(-1)) across all states was 10.1% and varied significantly among states, with up to 22.3% prevalence determined in Bihar. The geometric mean concentration of diclofenac found in samples in which the drug was detected was 352 microg kg(-1). The prevalence of carcasses containing diclofenac is similar to that previously proposed to be required to have caused the observed Gyps vulture declines in India. On the 11th of May 2006, the Drug Controller General (India) ordered the withdrawal of all licenses granted for the manufacture of diclofenac for veterinary use within India. However, if Gyps vultures are to be protected, potentially substantial existing stocks now need to be quickly and effectively removed from the Indian veterinary market.

Diclofenac disposition in Indian cow and goat with reference to Gyps vulture population declines.

Gyps vultures across India are declining rapidly and the NSAID diclofenac has been shown to be the major cause. Vultures scavenge livestock carcasses that have been treated with diclofenac within the days preceding death. We present data on diclofenac disposition in Indian cow and goat, and field data on the prevalence of diclofenac in carcases in the environment. In the disposition experiment, animals were treated with a single intramuscular injection of diclofenac at 1000 microg kg-1 bw. In cow, diclofenac was detectable in liver, kidney and intestine up to 71 h post-treatment; in plasma, half-life was 12.2 h. In goat, tissue residues were undetectable after 26 h. Prevalence of diclofenac in liver from 36 dead livestock collected in the field was 13.9%. Data suggest that diclofenac residues in Indian cow and goat are short-lived, but diclofenac prevalence in carcasses available to vultures may still be very high.

Methylation of the protein phosphatase 2A catalytic subunit is essential for association of Balpha regulatory subunit but not SG2NA, striatin, or polyomavirus middle tumor antigen.

Binding of different regulatory subunits and methylation of the catalytic (C) subunit carboxy-terminal leucine 309 are two important mechanisms by which protein phosphatase 2A (PP2A) can be regulated. In this study, both genetic and biochemical approaches were used to investigate regulation of regulatory subunit binding by C subunit methylation. Monoclonal antibodies selectively recognizing unmethylated C subunit were used to quantitate the methylation status of wild-type and mutant C subunits. Analysis of 13 C subunit mutants showed that both carboxy-terminal and active site residues are important for maintaining methylation in vivo. Severe impairment of methylation invariably led to a dramatic decrease in Balpha subunit binding but not of striatin, SG2NA, or polyomavirus middle tumor antigen (MT) binding. In fact, most unmethylated C subunit mutants showed enhanced binding to striatin and SG2NA. Certain carboxy-terminal mutations decreased Balpha subunit binding without greatly affecting methylation, indicating that Balpha subunit binding is not required for a high steady-state level of C subunit methylation. Demethylation of PP2A in cell lysates with recombinant PP2A methylesterase greatly decreased the amount of C subunit that could be coimmunoprecipitated via the Balpha subunit but not the amount that could be coimmunoprecipitated with Aalpha subunit or MT. When C subunit methylation levels were greatly reduced in vivo, Balpha subunits were found complexed exclusively to methylated C subunits, whereas striatin and SG2NA in the same cells bound both methylated and unmethylated C subunits. Thus, C subunit methylation is critical for assembly of PP2A heterotrimers containing Balpha subunit but not for formation of heterotrimers containing MT, striatin, or SG2NA. These findings suggest that methylation may be able to selectively regulate the association of certain regulatory subunits with the A/C heterodimer.

Frontotemporal correlates of impulsivity and machine learning in retired professional athletes with a history of multiple concussions.

The frontotemporal cortical network is associated with behaviours such as impulsivity and aggression. The health of the uncinate fasciculus (UF) that connects the orbitofrontal cortex (OFC) with the anterior temporal lobe (ATL) may be a crucial determinant of behavioural regulation. Behavioural changes can emerge after repeated concussion and thus we used MRI to examine the UF and connected gray matter as it relates to impulsivity and aggression in retired professional football players who had sustained multiple concussions. Behaviourally, athletes had faster reaction times and an increased error rate on a go/no-go task, and increased aggression and mania compared to controls. MRI revealed that the athletes had (1) cortical thinning of the ATL, (2) negative correlations of OFC thickness with aggression and task errors, indicative of impulsivity, (3) negative correlations of UF axial diffusivity with error rates and aggression, and (4) elevated resting-state functional connectivity between the ATL and OFC. Using machine learning, we found that UF diffusion imaging differentiates athletes from healthy controls with significant classifiers based on UF mean and radial diffusivity showing 79-84 % sensitivity and specificity, and 0.8 areas under the ROC curves. The spatial pattern of classifier weights revealed hot spots at the orbitofrontal and temporal ends of the UF. These data implicate the UF system in the pathological outcomes of repeated concussion as they relate to impulsive behaviour. Furthermore, a support vector machine has potential utility in the general assessment and diagnosis of brain abnormalities following concussion.

Probing the mechanism of transport and compartmentalisation of polyamines in mammalian cells.

BACKGROUND: Many mammalian cells possess an active polyamine uptake system but little is known about the molecular mechanism of this transporter. The fate of polyamines taken up from the medium and the relationship to polyamine homeostasis remains to be fully established. The aim of this study was to develop a range of modified polyamines, particularly ligands incorporating a fluorophore, to explore the structural tolerances of the polyamine transport system and to probe the intracellular location of polyamines acquired from the medium. RESULTS: We synthesised a wide range of polyamine analogues incorporating cytotoxic agents, fluorescent chromophores and bulky substituents. All of these analogues have been shown to be good competitive inhibitors of spermidine uptake in a range of mammalian cells. Direct evidence for uptake of the fluorescent polyamine analogues and their subcellular distribution was obtained from confocal laser scanning fluorescence microscopy, which showed that they accumulated in granular structures within the cytoplasm and not in the nucleus. We demonstrated that their uptake is through the polyamine transport system by showing that pretreatment with DFMO, a potent inhibitor of polyamine biosynthesis, led to enhanced uptake, and cells deficient in the polyamine transport system did not accumulate these polyamine analogues. CONCLUSIONS: The polyamine transport system has a surprisingly broad structural tolerance. Fluorophore-containing polyamine analogues derived from the extracellular pool are located in granular structures within the cytoplasm and not to any great extent in the nuclei of mammalian cells. These observations might be consistent with a mechanism involving receptor-mediated endocytosis, and the granular 'structures' seen might reflect polyamine compartmentalisation within vesicles.

Neuropsychologic impairment after coronary bypass surgery: effect of gaseous microemboli during perfusionist interventions.

OBJECTIVE: Neuropsychologic impairment is a common complication of coronary bypass surgery. Cerebral microemboli during cardiopulmonary bypass are the principal cause of cognitive deficits after coronary bypass grafting. We have previously demonstrated that the majority of cerebral emboli occur during perfusionist interventions (ie, during the injection of air into the venous side of the cardiopulmonary bypass circuit). The purpose of this study was to determine whether an increase in perfusionist interventions is associated with an increased risk of postoperative cognitive impairment. METHODS: Patients undergoing elective coronary artery bypass grafting (n = 83) underwent a battery of neuropsychologic tests preoperatively and 3 months postoperatively. Patients were divided into 2 groups according to the median value of perfusionist interventions during cardiopulmonary bypass. Group 1 patients (n = 42) had fewer than 10 perfusionist interventions, and group 2 patients (n = 41) had 10 or more interventions. RESULTS: The 2 groups of patients were similar for all preoperative, intraoperative, and postoperative variables, with the exception of longer cardiopulmonary bypass times in group 2 patients (P <.001). Group 2 patients had lower mean scores on 9 of 10 neuropsychologic tests, with 3 (Rey Auditory Verbal Learning, Digit Span, and Visual Span) being statistically significant. Group 2 patients had worse cognitive test scores, even when controlling for increased bypass times. Group 2 patients had a nonsignificant trend toward an increased prevalence of neuropsychologic impairment 3 months postoperatively. CONCLUSIONS: Introduction of air into the cardiopulmonary bypass circuit by perfusionists, resulting in cerebral microembolization, may contribute to postoperative cognitive impairment.

Automatic platelet counting with the Couler particle counter.

A method for accurately counting platelets is described using the Coulter counter model B fitted with a standard 100mu aperture tube. This enables the counter to be used for red and white cell as well as platelet counts using the same aperture tube. The method uses standard equipment except for a small inexpensive electronic speed controller.

ABO blood groups in hematologic malignancies.

This article examines the genetic predisposition of individuals to lymphoma and leukemia with regard to the ABO blood groups. Blood samples from 558 patients suffering from various forms of lymphoma and leukemia were collected and typed for ABO blood groups. The ABO blood group phenotype frequencies of lymphoma patients were similar to those in control samples. Among leukemia patients, a significant increase in the frequency of the A2 phenotype was found in chronic lymphocytic leukemia. Possible mechanisms underlying the predisposition of individuals with the A2 blood group to chronic lymphocytic leukemia suggested by these preliminary results are discussed.

Disseminated Nocardia brasiliensis infection.

We report a 40-year-old man with advanced Hodgkin disease who received combination chemotherapy and developed disseminated Nocardia brasiliensis infection. We discuss opportunistic infections in the compromised host, with particular reference to nocardial disease and certain biologic characteristics of that organism.

The effects of combining sanctions and rehabilitation for driving under the influence: an evaluation of the New Jersey Alcohol Countermeasures Program.

In contrast to many other state Driving under the Influence (DUI) programs developed in the United States in the 1970s as alternatives to traditional sanctions, the New Jersey Alcohol Countermeasures Program combined sanctions with mandatory education/rehabilitation for offenders. Three components were evaluated: DUI education, "treatment," and Alcoholics Anonymous. For 2,734 first and repeat offenders participating in this program between 1979 and 1982, the program was effective in reducing DUI recidivism for program completers (66% while licensed and 51% while suspended) compared with noncompleters, but it was less effective in reducing subsequent moving violations while licensed (20% compared with noncompleters) and accidents while licensed (18% compared with noncompleters). Completers had higher rates of violations and accidents while suspended (9% compared with noncompleters). A small group of repeat offenders, missassigned to DUI education, had higher post conviction rates of negative driving events than those of comparable offenders assigned to "treatment" or Alcoholics Anonymous, indicating that for these offenders the latter interventions were more effective.

Multiparameter system for investigation of the effects of high-power ultrasound on metals.

A multiparameter system has been developed which, for the first time, permits simultaneous measurement of sufficient quantities to test all of the proposed mechanisms for the influence of high-power ultrasound on metal specimens. Results of measurements using this system give new insight into the mechanical and thermal processes involved. Data are presented which show typical results obtained during tensile elongation of polycrystalline and single-crystal aluminum specimens.

Passive smoking and middle ear effusions in children of British servicemen in West Germany--a point prevalence survey by clinics of outpatient attendance.

Three hundred and twenty eight children of British servicemen serving in West Germany were studied in order to ascertain any relationship between parental smoking and the presence of middle ear effusion. The age range of the children was 18 months to 8 years. A questionnaire was completed by the parents of each child, and clinical examination and audiological tests demonstrated the presence or otherwise of middle ear effusions. One hundred and sixty four patients who had been referred with otological symptoms to the Ear, Nose and Throat Out Patient Department (ENT OPD) were surveyed, as were 164 other patients drawn from the Paediatric, Orthopaedic and Ophthalmic Out Patient Departments (OPDs) at BMH Rinteln. These 2 groups had similar age and sex distributions. The presence of middle ear effusions amongst children attending the ENT OPD was strongly associated with maternal cigarette smoking. This finding implies that mothers should not smoke at all in the same living accommodation as that used by their children. Paternal cigarette smoking showed no significant association with the presence of middle ear effusions.

Two new Eimeria species parasitic in corncrakes (Crex crex) (Gruiformes: Rallidae) in the United Kingdom.

In this study we describe 2 new species of coccidia (Apicomplexa: Eimeriidae) parasites isolated from the feces of corncrake (Crex crex) (Gruiformes: Rallidae), bred in captivity in the U.K. Oocysts of Eimeria crecis n. sp. were approximately spherical and measured 15.3 µm (13-18) × 14.3 (12-16), providing an oocyst shape index of 1.1. A micropyle and oocyst residuum were absent, but a polar granule was present. Oocysts of Eimeria nenei n. sp. were ellipsoidal and measured 23.6 (21-26) × 18.1 (17-20), providing an oocyst shape index of 1.3. A micropyle and polar granule were present. Surveying free-living, wild adult corncrakes in Scotland (U.K.) demonstrated both parasite species to be widespread. These are the first species described to infect the corncrake, and they are distinct from those previously found to infect members of the closely related crane family (Gruiformes: Gruidae). Partial amplification and sequencing of the 18S rRNA gene and internal transcribed spacer 2 indicated a close relationship between the 2 proposed new species as a group distinct from the Eimeria species known to infect cranes. These newly identified parasite species have been associated with enteric disease in corncrakes being prepared for reproduction in captivity and reintroduction into England (U.K.).