The First Record of Aedes vittatus (Diptera: Culicidae) in the Dominican Republic: Public Health Implications of a Potential Invasive Mosquito Species in the Americas.

Aedes vittatus Bigot is distributed throughout Africa, tropical Asia, and southern Europe and occurs in sylvatic as well as peridomestic environments where it readily feeds on humans. Although the vectorial capacity of Ae. vittatus is not well understood, this species is known to play a role in the maintenance and transmission of yellow fever, Zika, chikungunya, and dengue virus within its native range. In October 2019, after a routine inspection of mosquito-breeding containers in Jarabacoa, Dominican Republic, two Ae. vittatus females were captured via human landing catch method. After this finding, a CDC miniature light trap was deployed at the point of initial detection from 18:00 to 08:00 h, 2 d/wk from 3 to 31 October 2019. Potential larval habitats were also sampled via traditional dip method once per week spanning a 150 m radius from point of initial detection. In addition to the 2 adult females, 10 female and 2 male Ae. vittatus were captured. One Ae. vittatus larva also was found in a small puddle formed by an animal hoof print. Conventional PCR and Sanger sequencing were used to confirm morphological identification of collected specimens. This is the first detection of Ae. vittatus in the Dominican Republic as well as the Americas. Therefore, enhanced surveillance is needed to better understand the range and public health risks this potential invasive mosquito species may pose in the Dominican Republic, other Caribbean Islands, and/or the Americas.

Pathogenicity of Phytophthora lateralis Lineages on Different Selections of Chamaecyparis lawsoniana.

Phytophthora lateralis, the cause of Chamaecyparis lawsoniana root disease, was introduced in North America about 1920, and has since killed trees along roads and streams throughout the tree's range. Recent results suggest an Asian origin for this oomycete and four genetic lineages were identified. This raised questions for the genetic exapted resistance demonstrated in 1989 within the wild population of C. lawsoniana but with only one P. lateralis lineage. The main goal of the present research was to test the durability of the demonstrated resistance and to compare the pathogenicity of isolates representing the four lineages. No breakdown of resistance was observed in five separate tests using different inoculation techniques, resistant and susceptible C. lawsoniana trees, and seedling families. Differences in mortality and lesion length were observed between the lineages. The higher aggressiveness of isolates of the TWJ and PNW lineages and the lower aggressiveness of the TWK lineage are discussed in view of the hypotheses on the history of spread and evolutionary history of the P. lateralis lineages.

Multifocal haemorrhagic brain damage following hypoxia and blood pressure lability: case report and rat model.

AIMS: Haemorrhagic brain damage is frequently encountered as a complication of premature birth. Much less frequently, multifocal petechial haemorrhage is identified in asphyxiated term newborns. Our goal was to develop an experimental rat model to reproduce this pattern of brain damage. METHODS: Neonatal rat pups were exposed to a 24-h period of 10% or 8% hypoxia followed by a single dose of phenylephrine. Acute and subacute changes, as well as long-term outcomes, were investigated by histology, brain magnetic resonance imaging and behavioural assessment. Immunostaining for vascular endothelial growth factor and caveolin-1 was performed in the rat brains as well as in a 17-day human case. RESULTS: Small foci of haemorrhage were identified in almost all regions of the rat brain subjected to hypoxia plus phenylephrine, but not hypoxia alone. Exposure to 8% hypoxia was associated with more haemorrhagic foci than 10% hypoxia. With rare exceptions, the blood deposits were too small to be detected by magnetic resonance imaging. Altered immunohistochemical detection of vascular endothelial growth factor and caveolin-1 in the child and the rat model suggests a role for blood-brain barrier compromise. There were no clear behavioural changes and no residual morphological abnormalities in the 78-day follow-up of the rats. CONCLUSIONS: We conclude that transient hypoxia, in a dose-dependent manner, can weaken the vasculature and predispose to brain haemorrhage in the situation of labile blood pressure. Persistent hypoxia is likely to be important in the genesis of permanent severe brain damage.

Stream Monitoring for Detection of Phytophthora ramorum in Oregon Tanoak Forests.

Stream monitoring using leaf baits for early detection of Phytophthora ramorum has been an important part of the Oregon Sudden Oak Death (SOD) program since 2002. Sixty-four streams in and near the Oregon quarantine area in the southwest corner of the state were monitored in 2008. Leaves of rhododendron (Rhododendron macrophyllum) and tanoak (Lithocarpus densiflorus) were placed in mesh bags, and bags were floated in streams. Leaf baits were exchanged every 2 weeks throughout the year. Leaves were assayed by isolation on selective medium and by multiplex rDNA internal transcribed spacer polymerase chain reaction (ITS PCR). The two methods gave comparable results, but multiplex PCR was more sensitive. P. ramorum was regularly recovered at all seasons of the year from streams draining infested sites 5 years after eradication treatment. In streams with lower inoculum densities, recovery was much higher in summer than in winter. P. ramorum was isolated from streams in 23 watersheds. When P. ramorum was detected, intensive ground surveys located infected tanoaks or other host plants an average of 306 m upstream from the bait station. P. ramorum was isolated from stream baits up to 1,091 m from the probable inoculum source.

Phytophthora Species Causing Tanoak Stem Cankers in Southwestern Oregon.

Tanoak (Lithocarpus densiflorus) is a principal host of Phytophthora ramorum, cause of sudden oak death (SOD), in the western United States (1). In the course of SOD surveys in southwestern Oregon, other Phytophthora species were encountered to be causing stem cankers on tanoak that were indistinguishable from those caused by P. ramorum. In Oregon, SOD is subject to quarantine and eradication. Aerial surveys are flown two or more times a year to locate symptomatic tanoaks, which are then examined from the ground to determine the cause of death. Isolations on selective media were attempted from all trees with stem cankers typical of Phytophthora. Phytophthora species were identified by morphological features and DNA sequencing of either internal transcribed spacer (ITS) or the mitochondrial COX spacer region. ITS sequences were compared with validated GenBank records, and COX spacer sequences were compared with known reference isolates in the OSU collection. From 2001 through 2006, Phytophthora spp. were isolated from 482 of 1,057 tanoak stem cankers sampled. P. ramorum was isolated from 359 cankers, P. nemorosa was isolated from 102 cankers, P. gonapodyides was isolated from six cankers, P. cambivora was isolated from four cankers (all A1 mating type), P. siskiyouensis was isolated from four cankers, P. pseudosyringae was isolated from two cankers, P. cinnamomi was isolated from one canker (mating type A2), and P. taxon "Pgchlamydo" was isolated from one canker. Three cankers yielded isolates that were not identified but were closely related to P. pseudosyringae based on ITS sequence. No Phytophthora spp. were cultured from the remaining cankers. One isolate from each species identified (except P. ramorum and P. pseudosyringae) was tested for pathogenicity on tanoak stems (11.4 to 16.0 cm DBH) in the field. A 5-mm-diameter plug from the margin of a V8 agar culture was placed in a hole in the bark, covered with wet cheesecloth, and sealed with aluminum foil and duct tape. Each isolate was inoculated into five different stems. Each stem received three different isolates and an agar control. After 4 weeks, bark was removed to reveal lesion development. Lesions were measured (length by width), and pieces from four points on the lesion margin were plated in selective media to reisolate. P. cambivora, P. cinnamomi, P. gonapodyides, P. nemorosa, P. siskiyouensis and P. taxon "Pgchlamydo" all caused substantial lesions in inoculated tanoak trees (average area 11.5 to 18.6 cm2). In all cases, the species used for inoculation was recovered on reisolation from lesion margins. Control inoculations caused necrotic areas averaging 0.2 cm2. Isolations from these areas were clean. Prior to the recent SOD epidemic, no species of Phytophthora were known as pathogens of tanoak. The discovery of P. ramorum as a pathogen of tanoak in California was quickly followed by the discovery that P. nemorosa and P. pseudosyringae were also associated with tanoak cankers (2). Six years of diagnostic support for survey and detection of P. ramorum in tanoak forests of southwest Oregon has revealed the occurrence, at very low frequency, of at least five additional species of Phytophthora causing stem cankers in tanoak. References: (1) D. M. Rizzo et al. Ann. Rev. Phytopathol. 43:309, 2005. (2) A. C. Wickland et al. For. Pathol. Online publication. DOI:10.1111/j.1439-0329.2008.00552.x), 2008.

Statistically designed survey of polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans, and co-planar polychlorinated biphenyls in U. S. meat and poultry, 2002-2003: results, trends, and implications.

To obtain information on dioxin levels in the human diet, the Food Safety and Inspection Service of the United States Department of Agriculture recently determined levels of dioxin-like compounds (dioxins/dibenzofurans/PCBs) in four major slaughter classes (steers and heifers, market hogs, young chickens, and young turkeys) that comprise over 90% of the meat and poultry production in the United States. The data were analyzed and compared to data from smaller surveys carried out from 1994 to 1996. These surveys were conducted by different laboratories nearly 10 years apart, so a direct comparison of the data was not straightforward. Three approaches were taken: (1) comparison with nondetects set to zero, (2) comparison with nondetects set to half the limit of detection, and (3) comparison applying the earlier surveys' limits of detection to the newer data. The data analyses indicated that dioxin levels appear to have declined in three of the four slaughter classes, with young chickens, market hogs, and young turkeys declining 20-80%, while any declines in cattle dioxin levels, if real, are less than those observed in the other slaughter classes. Further study is needed to examine factors that might explain the differences in dioxin levels and distribution profiles in the four slaughter classes. A small number of market hog and steers/ heifers samples had dioxin toxic equivalency levels (TEQs) greater than 2 pg/g lipid weight. Follow-up investigations for those samples indicated a common source for the market hog samples (a dioxin-contaminated mineral supplement), but no commonality was found for the steers/ heifers samples.

Susceptibility of Oregon Forest Trees and Shrubs to Phytophthora ramorum: A Comparison of Artificial Inoculation and Natural Infection.

Phytophthora ramorum is an invasive pathogen in some mixed-hardwood forests in California and southwestern Oregon, where it causes sudden oak death (SOD) on some members of Fagaceae, ramorum shoot dieback on some members of Ericaceae and conifers, and ramorum leaf blight on diverse hosts. We compared symptoms of P. ramorum infection resulting from four different artificial inoculation techniques with the symptoms of natural infection on 49 western forest trees and shrubs; 80% proved susceptible to one degree or another. No single inoculation method predicted the full range of symptoms observed in the field, but whole plant dip came closest. Detached-leaf-dip inoculation provided a rapid assay and permitted a reasonable assessment of susceptibility to leaf blight. Both leaf age and inoculum dose affected detached-leaf assays. SOD and dieback hosts often developed limited leaf symptoms, although the pattern of midrib and petiole necrosis was distinctive. Stem-wound inoculation of seedlings correlated with field symptoms for several hosts. The results suggested that additional conifer species may be damaged in the field. Log inoculation provided a realistic test of susceptibility to SOD, but was cumbersome and subject to seasonal variability. Pacific rhododendron, salmonberry, cascara, and poison oak were confirmed as hosts by completing Koch's postulates. Douglas-fir was most susceptible to shoot dieback shortly after budburst, with infection occurring at the bud.

First Report of A1 Mating Type of Phytophthora ramorum in North America.

Phytophthora ramorum is known in Europe and the western United States (1). In Europe, it is found in nurseries and landscape plantings. In the United States, it has been confined to coastal forests, and in California, it is found in a few horticultural nurseries. All European isolates tested have been A1 mating type, while all North American isolates were A2 mating type (2). Amplified fragment length polymorphism markers also indicated that the populations on the two continents are distinct, and nearly all North American isolates are from one clone (Kelly Ivors, unpublished). In June 2003, P. ramorum was isolated from diseased Viburnum and Pieris spp. cultivars from a Clackamas County nursery in northern Oregon and diseased Camellia sp. cultivar from a Jackson County nursery in southern Oregon. Representative isolates were submitted to the American Type Culture Collection, Manassas, VA. As part of the effort to determine the origin of these new infestations, we tested the nursery isolates for mating type. Seven Oregon nursery isolates, three Oregon forest isolates (from the predominant North American clone), and two European isolates were paired. Agar plugs from 3-day-old colonies were placed in close proximity on carrot agar plates, and then the plates were examined for oogonia after 3 and 10 days as advised by C. M. Brasier (personal communication). Oogonia and antheridia typical of P. ramorum (2) formed when isolates from the Clackamas County nursery were paired with the Oregon forest isolates and also when isolates from the Jackson County nursery were paired with the European isolates. Gametangia also formed in pairings between Oregon forest isolates and European isolates, but not in any other combinations. We developed polymerase chain reaction (PCR) primers for four microsatellite loci and determined allele sizes for the same set of isolates (unpublished). Microsatellite alleles of the Clackamas County isolates were identical to the European tester isolates, and alleles of the Jackson County isolates were identical to the Oregon forest isolates. These results indicate that the recent Oregon nursery infestations are of separate origins. The Clackamas County isolates are A1 mating type and have microsatellite alleles like the European testers, but according to shipping records, the nursery has received no host nursery stock directly from Europe. However, host nursery stock has been received from a Canadian nursery. The Jackson County isolates are of A2 mating type and have microsatellite alleles like the forest isolates of Oregon, which is consistent with the reported origin of these plants from a California nursery. These preliminary microsatellite results need to be validated against a larger isolate set but are congruent with the mating type results. The Oregon nursery infestations highlight the dangers of unregulated or underregulated transport of host nursery stock from infested areas to noninfested areas. All host plants from infested nursery blocks at the affected Oregon nurseries have been destroyed by incineration, and a monitoring program has been implemented. Other host nursery stock on site has been taken "off-sale" pending verification that it is disease free, per the United States Department of Agriculture, APHIS requirements. References: (1) J. M. Davidson et al. On-line publication. doi:10.1094/PHP-2003-0707-01-DG. Plant Health Progress, 2003. (2) S. Werres et al. Mycol. Res. 105:1155, 2001.

Sudden Oak Death Caused by Phytophthora ramorum in Oregon.

Sudden oak death, caused by Phytophthora ramorum (1,2), has been found for the first time in Oregon, killing tanoak, Lithocarpus densiflorus, trees. To our knowledge, this is the first report of the disease outside of the San Francisco to Monterey area in California, (300 km to the south). Nine areas of infestation, all within a 24-km2 area, were discovered on forest lands near Brookings, in southwest Oregon. Mortality centers ranged in size from 0.2 to 4.5 ha and included 5 to approximately 40 diseased trees. P. ramorum was isolated from stem cankers using Phytophthora-selective medium. Isolates had distinctive morphological features characteristic of P. ramorum, including abundant production of chlamydospores and caducous, semipapillate sporangia on solid media. Internal transcribed spacer (ITS) sequences of isolates of P. ramorum from Oregon were identical to ITS sequences of isolates from California (1). The pathogen also was isolated from necrotic lesions on leaves and stems of native Rhododendron macrophyllum and Vaccinium ovatum growing beneath diseased tanoaks. In July 2001, the disease was located by an aerial survey conducted cooperatively by the USDA Forest Service and Oregon Department of Forestry. All lands within 1.6 km (1 mile) of the mortality centers are subject to Oregon quarantine, which bars the transport of any host plant materials. An eradication effort is currently underway. Symptomatic plants and all known host plants within 15 to 30 m of symptomatic plants are being cut and burned in the first phase of this operation. The total treated area is approximately 16 ha. References: (1) D. M. Rizzo et al. Plant Dis. In press. (2) S. Werres et al. Mycol. Res. 105:1155, 2001.

Macaque blood-derived antigen-presenting cells elicit SIV-specific immune responses.

Natural blood-borne antigen-presenting cells (APCs) were tested for their ability to augment antigen presentation for SIV vaccines. Fibrocytes and monocyte-derived dendritic cells (DCs) were isolated from multiple Macaca fascicularis. Macaque fibrocytes displayed the characteristic cellular morphology and stained positive for CD34 and collagen, as observed in human and murine fibrocytes. Macaque DCs were generated from monocytes by culturing in granulocyte-macrophage colony stimulating factor and interleukin-4 (IL-4). Two days after maturation, cells were enriched for the DC marker CD83. Fibrocytes and DCs were each transfected with green fluorescence protein expression plasmids or DNA expression vectors encoding all of the SIVmne structural and regulatory genes. Autologous DCs were re-infused into macaques subcutaneously (sc) following transfection; mixing with recombinant SIV antigens or inactivated whole SIV in vitro; or mock-treatment. Autologous monocyte-derived DCs pulsed with whole inactivated SIV were re-infused and elicited cellular and/or humoral responses in vivo in eight of ten vaccinated macaques.

Incidence and Impact of Swiss Needle Cast in Forest Plantations of Douglas-fir in Coastal Oregon.

An epidemic of Swiss needle cast, caused by the ascomycete Phaeocryptopus gaeumannii, is causing defoliation and growth reductions in Douglas-fir forest plantations along the Oregon Coast. The area of symptomatic plantations has been monitored annually since 1996 by aerial survey; in spring 1999, 119,500 ha were affected. Pathogen and symptom development have also been monitored on nine permanent plots in stands of differing disease severity. Infection levels and symptom severity are greatest in low elevation plantations close to the coast. In areas of severe disease, trees retain only current year needles. Defoliation is proportional to the number of stomata occluded by pseudothecia of the fungus, with needles being shed when about 50% of stomata are occupied, regardless of needle age. Fungus sporulation and premature needle abscission are greatest on the upper branches of trees. Annual application of fungicides increases needle retention significantly. Tree height and diameter growth and total tree volume are reduced by disease, and tree volume is significantly correlated with needle retention on our plot trees. The epidemic continues to be most severe in Douglas-fir plantations established on sites where Sitka spruce and western hemlock or red alder predominated in earlier times.

Plasma viremia in macaques infected with simian immunodeficiency virus: plasma viral load early in infection predicts survival.

A reliable method for the quantitation of plasma viremia in nonhuman primates infected with simian immunodeficiency virus (SIV) and related viruses is described. This method is based on an established quantitative-competitive PCR format and includes a truncated control for internal assay calibration. Optimization of assay conditions has significantly improved amplification specificity, and interassay variability is comparable to that of commercially available assays for human immunodeficiency virus (HIV) quantitation. This procedure was used to monitor viral loads in a group of Macaca mulatta animals that were infected with SIVsmE660 for over 2 years. Highly diverse profiles of plasma viremia were observed among animals, and high viral loads were associated with more rapid disease progression. Spearman rank correlation analyses were done for survival versus three parameters of viral load: plasma viremia, p27 core antigen, and frequency of infected peripheral blood mononuclear cells. Plasma viremia had the strongest overall correlation and was significantly (P < 0.05 to P < 0.01) associated with survival at 10 of the 13 time points examined. Plasma viremia did not correlate with survival during the primary viremia phase; however, the strength of this correlation increased with time postinfection and, remarkably, viremia levels as early as week 6 postinfection were highly predictive (P < 0.01) of relative survival. These findings are consistent with the available clinical data concerning viral load correlates early in HIV infection, and they provide further support for the view that disease outcome in lentiviral infection may be largely determined by events that occur shortly after infection.

Variation in phosphoric acid concentration and treatment time and HEMA diffusion through dentin.

PURPOSE: The effects of phosphoric acid concentration and time of dentin treatment on the movement of 2-hydroxyethylmethacrylate (HEMA) from a bonding resin-resin composite combination through dentin were investigated in vitro. MATERIALS AND METHODS: Freshly extracted human third molar teeth were divided into seven groups each of 10 teeth. A closed chamber was attached to the CEJ of each tooth to contain 1 ml distilled water. An occlusal cavity of 6 mm diameter and remaining dentin thickness of 1.0-1.5 mm was prepared in each tooth. Dentin was treated with either 10% phosphoric acid for 15, 30 or 60 seconds or with 37% phosphoric acid for 15, 30 or 60 seconds. A control group not treated with acid was also prepared. The cavities were rinsed, dried and then treated with the HEMA-containing Scotchbond Multi-Purpose bonding resin which was light-cured for 10 seconds. The cavities were then restored with Z100 and light-cured for 30 seconds. Water samples were retrieved from the chambers over a time course (4.32, 14.4, 43.2, 144 & 432 minutes; 1, 3 and 10 days) and analyzed by high performance liquid chromatography. RESULTS: HEMA was detected in the pulp chambers of all teeth from 4.32 minutes after resin placement. Mean total (cumulative) release at 10 days for all groups was in the 2-4 mumol range. The highest HEMA diffusion rate was 4.32 minutes after placement and were in the range 6-13 times greater than control in the 10%-15 seconds, 10%-30 seconds, 10%-60 seconds, 37%-15 seconds, and 37%-30 seconds groups. Unexpectedly, by far the lowest early rate at 4.32 minutes was in the 37%-60 seconds group (0.6 times of no-acid control).

Passive immune globulin therapy in the SIV/macaque model: early intervention can alter disease profile.

One of the major questions in AIDS is the role that the host immune system and the virus play in the dynamics of infection and the development of AIDS in an infected individual. In order to test the role of antibody in controlling viral infection, high-dose SIV-immune globulin was passively transferred to infected macaques early in infection. Immune globulin purified from the plasma of an SIV-infected long-term non-progressor macaque (SIVIG) or a pool of normal immune globulin (normal Ig) was infused into SIVsmE660-infected macaques (170 mg/kg) at one and fourteen days post infection. Animals were monitored for SIV-specific antibodies, viremia, plasma antigenemia, and clinical course. All animals were infected by SIV. At 16 months post infection, five macaques in the combined control groups have been euthanized, one as a rapid progressor with debilitating disease at 20 weeks post infection. Four macaques from the comparison groups have signs of AIDS, accompanied by high and increasing levels of virus and p27 antigenemia. One of the ten control animals had a very low virus load in plasma and peripheral blood and lymph node mononuclear cells at all times tested and has remained disease-free. In the SIVIG treatment group, two macaques were euthanized at 18-20 weeks due to AIDS, rapid progressors to disease. Three macaques in the SIVIG group had an initial high level of virus in plasma, peripheral blood mononuclear cells (PBMC), and lymph node mononuclear cells (LNMC), which dropped to baseline at 6 weeks post infection and has remained very low or negative for 16 months, a disease profile which has not been observed in untreated animals in this model to date. These macaques have remained clinically healthy. The sixth treated animal is also healthy, with very low virus burden that is detectable only by nested set polymerase chain reaction (PCR). All SIVIG-treated macaques had no detectable p27 plasma antigenemia for the first 10 weeks of infection, demonstrating that the IgG effectively complexed with the virus. The immunological correlates in the treated animals include development of de novo virus-specific antibodies and/or cytotoxic T cell (CTL), both of which are hallmarks of long term non-progressors. The two SIVIG-treated macaques that progress to disease rapidly had no detectable de novo humoral immune responses, as is often seen in rapid HIV disease in humans. Envelope-specific and virus neutralizing antibodies alone were not sufficient to prevent disease progression, as the plasma of both non-progressors as well as progressors had high titers of envelope-specific and neutralizing antibodies against SIVsmE660. Poor clinical prognosis was associated with moderate to high and increasing virus loads in plasma, PBMC, and lymph nodes. Good clinical prognosis correlated with low or undetectable post acute viremia in the peripheral blood and lymph nodes. We hypothesize that SIVIG reduced the spread of virus by eliminating or reducing plasma virus through immune complexes during the first four to 8 weeks of infection and then maintaining this low level of viremia until the host immune response was capable of virus control. Reduction of virus burden early in infection by passive IgG can alter disease outcome in SIV infection of macaques. Modifications of this strategy may lead to effective early treatment of HIV-1 infection in humans.

Displaced supracondylar humeral fractures in children. A comparison of results and costs in patients treated by skeletal traction versus percutaneous pinning.

Clinical outcome and treatment cost were compared in 65 children treated by either percutaneous pinning or skeletal traction for displaced supracondylar humeral fractures. Clinical outcome was evaluated by loss of elbow motion and change in carrying angle. Results of treatment were basically equivalent in the two groups and were satisfactory in 90% or more. To determine treatment cost, the authors analyzed factors that vary according to the type of therapy. Cost of treatment was lowest in those who had percutaneous pinning and subsequent pin removal in the office. Compared to this group, the cost of treatment increased by 23% in those who had percutaneous pinning and removal of the pins as a surgical procedure, by 117% in those treated by traction with the olecranon pin inserted in the emergency room, and by 142% in those treated by traction with the olecranon pin inserted in the operating room.