Efficacy of Interferon Alpha-2b and Ribavirin Against West Nile Virus in Vitro

Emerging Infectious Diseases. Jan, 2002  |  Pubmed ID: 11749765

A Systems Approach to Improving Error Reporting

Journal of Healthcare Information Management : JHIM. 2002  |  Pubmed ID: 11813522

The reporting of medical errors or near misses within a healthcare organization is one process that has been long overlooked. With strong leadership as its foundation, Baylor Health Care System addressed multiple systems and processes of technology, education, communication, learning, feedback, rewards, and recognition to demonstrate significant improvements in cost reduction, employee satisfaction, and the number of errors reported.

The Natural History of Ticks

The Medical Clinics of North America. Mar, 2002  |  Pubmed ID: 11982298

Ticks have evolved to become one of the most important groups of arthropod vectors of human pathogens. One or more of the approximately 840 known species of ticks are found in most terrestrial regions of the earth. Ticks are a highly specialized group of obligate, bloodsucking, nonpermanent ectoparasitic arthropods that feed on mammals, birds, and reptiles. They are classified into two major families, Ixodidae (hard-bodies ticks) and Argasidae (soft-bodied ticks). The Ixodidae is the largest and most important family. There are many taxonomic keys for identifying ticks to assist the serious investigator. Their life cycles are often complex, and even though ticks are associated with their parasitic habits, ticks spend most of their life off hosts and in vegetation or soil. Maintenance of water balance during periods of overhydration while feeding and periods of dehydration while fasting is significant in the distribution, survival, activity, and transmission of disease-causing pathogens to humans and animals. Ticks attach to skin of the host by using their hypostome as an anchor and create a feeding lesion to ingest blood or tissue fluids. Soft-bodied ticks feed relatively rapidly (hours or less) and ingest only blood. Hard-bodied ticks take days to complete feeding and feed on blood, lymph, and lysed tissues from a pool that forms around the mouthparts. Feeding causes direct damage to the skin of the host. Disease-causing organisms may be ingested or expelled during feeding. Ingestion of relatively enormous quantities of blood is characteristic of ticks.

Examination of the Borrelia Burgdorferi Transcriptome in Ixodes Scapularis During Feeding

Journal of Bacteriology. Jun, 2002  |  Pubmed ID: 12003955

Borrelia burgdorferi gene expression within the guts of engorging Ixodes scapularis ticks was examined by use of differential immunoscreening and differential expression with a customized amplified library. Fourteen chromosomal genes involved in energy metabolism, substrate transport, and signal transduction and 10 (4 chromosomal and 6 plasmid) genes encoding putative lipoproteins and periplasmic proteins were preferentially expressed in engorging ticks. These data demonstrate a new approach to the global analysis of B. burgdorferi genes that are preferentially expressed within the vector during feeding.

A Recombinant Envelope Protein-based Enzyme-linked Immunosorbent Assay for West Nile Virus Serodiagnosis

Vector Borne and Zoonotic Diseases (Larchmont, N.Y.). 2002  |  Pubmed ID: 12653304

Recombinant West Nile virus envelope (E) protein was examined in enzyme-linked immunosorbent assay (ELISA) to detect antibodies elicited during West Nile virus infection. Horses (nine of 10) and humans (six of six) with confirmed West Nile virus infection had IgG and/or IgM antibodies to the E protein. Antibodies to the recombinant West Nile virus membrane and nonstructural 1 proteins were not detected in any of these sera. An E protein-based ELISA may aid in the serological diagnosis of West Nile virus infection.

Drug Treatment Courts Are Popular but Do They Work and Are They Ethical and Appropriate for Canada?

Health Law in Canada. May, 2003  |  Pubmed ID: 12845932

Currently Canada is experimenting with the implementation of drug treatment courts. Pilot projects are underway in both Toronto and Vancouver. In the U.S., drug courts emerged as a response to the overcrowding of the prison system, the end product of the revolving door of substance dependent people moving through the court system. However, this expansion was not accompanied by any rigorous evaluation or critical reflection as to whether drug treatment courts can achieve their desired outcomes or if they are appropriate for dealing with substance dependent offenders. The purpose of this article is to take a critical look at this phenomenon and to discuss whether the drug court model is suitable for Canada.

IFN-gamma-producing Gamma Delta T Cells Help Control Murine West Nile Virus Infection

Journal of Immunology (Baltimore, Md. : 1950). Sep, 2003  |  Pubmed ID: 12928402

West Nile (WN) virus causes fatal meningoencephalitis in laboratory mice, thereby partially mimicking human disease. Using this model, we have demonstrated that mice deficient in gammadelta T cells are more susceptible to WN virus infection. TCRdelta(-/-) mice have elevated viral loads and greater dissemination of the pathogen to the CNS. In wild-type mice, gammadelta T cells expanded significantly during WN virus infection, produced IFN-gamma in ex vivo assays, and enhanced perforin expression by splenic T cells. Adoptive transfer of gammadelta T cells to TCRdelta(-/-) mice reduced the susceptibility of these mice to WN virus, and this effect was primarily due to IFN-gamma-producing gammadelta T cells. These data demonstrate a distinct role for gammadelta T cells in the control of and prevention of mortality from murine WN virus infection.

Transstadial Transfer of West Nile Virus by Three Species of Ixodid Ticks (Acari: Ixodidae)

Journal of Medical Entomology. Jul, 2003  |  Pubmed ID: 14680122

Larvae and/or nymphs of four species of ixodid ticks, Ixodes scapularis Say, Amblyomma americanum (L.), Dermacentor andersoni Stiles, and Dermacentor variabilis Say, were fed to completion on laboratory hamsters or mice which had been inoculated with a West Nile (WN) virus isolate from Culex pipiens L. captured in Connecticut USA. Maximum titers in mice and hamsters were approximately 5 and two logs, respectively, lower than recorded (10 logs) in a naturally infected American crow, Corvus brachyrhynchos Brehm. WN virus was isolated in Vero cell culture from ticks and detected by TaqMan RT-polymerase chain reaction (PCR) in ticks that had completed their feeding as larvae or nymphs, and in I. scapularis, D. andersoni, and D. variabilis that had molted into the next stage of development. Naive hosts, fed upon by nymphs that as larvae had fed on viremic hosts, did not become infected. WN virus was isolated in Vero cell culture from one female I. scapularis and was detected by TaqMan RT-PCR in 24 adult I. scapularis, one D. andersoni, and two D. variabilis adults that had fed to completion as larvae on viremic hosts and as nymphs on naive mice or hamsters. Three species of ixodid ticks acquired WN virus from viremic hosts and transstadially passed the virus, but vector competency was not demonstrated.

OspC Facilitates Borrelia Burgdorferi Invasion of Ixodes Scapularis Salivary Glands

The Journal of Clinical Investigation. Jan, 2004  |  Pubmed ID: 14722614

Outer surface protein C (OspC) is a differentially expressed major surface lipoprotein of Borrelia burgdorferi. ospC is swiftly upregulated when spirochetes leave the Ixodes scapularis tick gut, migrate to the salivary gland, and exit the arthropod vector. Here we show that OspC strongly binds to the tick salivary gland, suggesting a role for OspC in spirochete adherence to this tissue. In vivo studies using a murine model of Lyme borreliosis showed that while OspC F(ab)(2) fragments did not influence either the viability of spirochetes or ospC gene expression, they did interfere with B. burgdorferi invasion of tick salivary glands. We then generated ospC knockout spirochetes in an infectious clone of B. burgdorferi and examined them within the vector. OspC-deficient or wild-type spirochetes persisted equally within the gut of unfed ticks and multiplied during the tick engorgement; however, unlike wild-type B. burgdorferi, the mutants were unable to invade salivary glands. Salivary gland colonization of OspC-deficient spirochetes was completely restored when this mutant was complemented in trans with a plasmid harboring the wild-type ospC gene. These studies conclusively demonstrate the importance of OspC in the invasion of tick salivary glands by B. burgdorferi, a critical step in the transmission of spirochetes from the arthropod vector to the mammalian host.

Disruption of Ixodes Scapularis Anticoagulation by Using RNA Interference

Proceedings of the National Academy of Sciences of the United States of America. Feb, 2004  |  Pubmed ID: 14745044

Ixodes scapularis ticks transmit many pathogens, including Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti. Vaccines directed against arthropod proteins injected into the host during tick engorgement could prevent numerous infectious diseases. Salp14, a salivary anticoagulant, poses a key target for such intervention. Salp14 is the prototypic member of a family of potential I. scapularis anticoagulants, expressed and secreted in tick saliva during tick feeding. RNA interference was used to assess the role of Salp14 in tick feeding. Salp14 and its paralogs were silenced, as demonstrated by the reduction of mRNA and protein specific for these antigens. Tick salivary glands lacking Salp14 had reduced anticoagulant activity, as revealed by a 60-80% reduction of anti-factor Xa activity. Silencing the expression of salp14 and its paralogs also reduced the ability of I. scapularis to feed, as demonstrated by a 50-70% decline in the engorgement weights. Because ticks have several anticoagulants, it is likely that the expression of multiple anticoagulants in I. scapularis saliva would have to be ablated simultaneously to abolish tick feeding. These studies demonstrate that RNA interference can silence I. scapularis genes and disrupt their physiologic function in vivo, and they identify vaccine candidates that can alter vector engorgement.

OspB Antibody Prevents Borrelia Burgdorferi Colonization of Ixodes Scapularis

Infection and Immunity. Mar, 2004  |  Pubmed ID: 14977984

Borrelia burgdorferi outer surface protein OspB is expressed by spirochetes in the Ixodes scapularis gut. ospB is transcribed from a bicistronic operon with ospA, a known spirochete adhesion gene in the tick gut. Here we examine whether OspB also has a specific function in ticks. OspB specifically binds to a protein or protein complex within the tick gut. We also assessed whether selected nonborreliacidal OspB antibodies or F(ab)(2) fragments interfere with B. burgdorferi-tick attachment in vivo. We examined engorged ticks that fed on B. burgdorferi N40-infected scid mice that had been treated with OspB F(ab)(2) fragments. Control F(ab)(2) fragments did not interfere with B. burgdorferi colonization of the tick gut, whereas OspB F(ab)(2) fragments significantly inhibited the attachment of spirochetes to the tick gut. These studies show that nonbactericidal OspB antibodies interfere with B. burgdorferi colonization of I. scapularis, highlighting a specific role for OspB in spirochete- arthropod interactions and suggesting new antibody-mediated strategies for interfering with B. burgdorferi transmission.

Client Retention in the British Columbia Methadone Program, 1996-1999

Canadian Journal of Public Health. Revue Canadienne De Santé Publique. Mar-Apr, 2004  |  Pubmed ID: 15074899

Methadone treatment for heroin addiction has been available for 40 years, but there is relatively little research on the effectiveness of Canadian programs. This paper describes one-year retention among the client cohorts entering the British Columbia Methadone Program during expansion between 1996 and 1999, and examines some factors previously shown to influence retention.

Babesiosis in Fairfield County, Connecticut

Emerging Infectious Diseases. Mar, 2004  |  Pubmed ID: 15116708

Prevalence of West Nile Virus in Tree Canopy-inhabiting Culex Pipiens and Associated Mosquitoes

The American Journal of Tropical Medicine and Hygiene. Jul, 2004  |  Pubmed ID: 15238699

Culex pipiens was the dominant mosquito captured in a West Nile virus (WNV) focus in Stratford, Connecticut. More Cx. pipiens were captured in Centers for Disease Control miniature light traps baited with CO(2), quail/hamster traps, and mosquito magnet experimental (MMX) traps placed in the tree canopy than in similar traps placed near the ground. Significantly more Cx. pipiens were captured in MMX traps placed in the canopy than in the other traps tested. Ninety-two percent and 85% of the 206 and 68 WNV isolations were from Cx. pipiens in 2002 and 2003, respectively; 5% and 12% were from Cx. salinarius. Eighty-five percent and 87% of the isolates were from mosquitoes captured in the canopy in each of the two years. The significantly larger numbers of WNV isolates from Cx. pipiens captured in the canopy are attributed to the significantly larger numbers of Cx. pipiens captured in the canopy in comparison to those captured in traps near the ground.

TROSPA, an Ixodes Scapularis Receptor for Borrelia Burgdorferi

Cell. Nov, 2004  |  Pubmed ID: 15537536

The Lyme disease agent Borrelia burgdorferi naturally persists in a cycle that primarily involves ticks and mammals. We have now identified a tick receptor (TROSPA) that is required for spirochetal colonization of Ixodes scapularis. B. burgdorferi outer surface protein A, which is abundantly expressed on spirochetes within the arthropod and essential for pathogen adherence to the vector, specifically bound to TROSPA. TROSPA mRNA levels in ticks increased following spirochete infestation and decreased in response to engorgement, events that are temporally linked to B. burgdorferi entry into and egress from the vector. The blockade of TROSPA by TROSPA antisera or by the repression of TROSPA expression via RNA interference reduced B. burgdorferi adherence to the I. scapularis gut in vivo, thereby preventing efficient colonization of the vector and subsequently reducing pathogen transmission to the mammalian host. Identification of an I. scapularis receptor for B. burgdorferi is the first step toward elucidating arthropod ligands that are required for survival of spirochetes in nature.

Toll-like Receptor 3 Mediates West Nile Virus Entry into the Brain Causing Lethal Encephalitis

Nature Medicine. Dec, 2004  |  Pubmed ID: 15558055

West Nile virus (WNV), a mosquito-borne single-stranded (ss)RNA flavivirus, causes human disease of variable severity. We investigated the involvement of Toll-like receptor (Tlr) 3, which recognizes viral double-stranded (ds)RNA, on WNV infection. Tlr3-deficient (Tlr3(-/-)) mice were more resistant to lethal WNV infection and had impaired cytokine production and enhanced viral load in the periphery, whereas in the brain, viral load, inflammatory responses and neuropathology were reduced compared to wild-type mice. Peripheral WNV infection led to a breakdown of the blood-brain barrier and enhanced brain infection in wild-type but not in Tlr3(-/-) mice, although both groups were equally susceptible upon intracerebroventricular administration of the virus. Tumor necrosis factor-alpha receptor 1 signaling is vital for blood-brain barrier compromise upon Tlr3 stimulation by dsRNA or WNV. Collectively, WNV infection leads to a Tlr3-dependent inflammatory response, which is involved in brain penetration of the virus and neuronal injury.

Epidemiology of West Nile Virus in Connecticut: a Five-year Analysis of Mosquito Data 1999-2003

Vector Borne and Zoonotic Diseases (Larchmont, N.Y.). 2004  |  Pubmed ID: 15682518

Two hundred and ten isolations of West Nile virus (WNV) were obtained from 17 mosquito species in six genera in statewide surveillance conducted in Connecticut from June through October, 1999-2003. Culex pipiens (86), Culex salinarius (32), Culex restuans (26), Culiseta melanura (32), and Aedes vexans (12) were implicated as the most likely vectors of WNV in the region based on virus isolation data. Culex pipiens was abundant from July through September and is likely involved in early season enzootic transmission and late season epizootic amplification of the virus in wild bird populations. Epidemic transmission of WNV to humans in urban locales is probable. The abundance of Cx. restuans in June and July and isolations of WNV in early July suggest that this species may play an important role as an enzootic vector involved in early amplification of WNV virus among wild birds. Its involvement as a bridge vector to humans is unlikely. Culex salinarius was the most frequently captured Culex species and was abundant in August and September when virus activity was at its height. Frequent isolations of WNV from this species in September when the majority of human cases were reported in union with its abundance at this time of the year, demonstrated vector competence, and broad feeding habits, make Cx. salinarius a likely bridge vector to humans, horses and other mammals. Multiple isolations WNV from Cs. melanura collected in more rural locales in late August and September, provide supportive evidence to suggest that this predominant avian feeder may play a significant role in epizootic amplification of the virus among wild bird populations in these environs. Aedes vexans was the only species of Aedes or Ochlerotatus from which multiple isolations of WNV were made in more than one year and was among the most frequently trapped and abundant species throughout the season. Since Ae. vexans predominately feeds on mammals it is unlikely to play a significant role in epizootic amplification of WNV, however, because of its abundance and aggressive mammalian and human biting behavior it must receive strong consideration as a bridge vector to humans and horses. The occasional virus isolations obtained from Aedes cinereus (4), Uranotaenia sapphirina (3), Ochlerotatus canadensis (2), Ochlerotatus trivittatus (2), Ochlerotatus sollicitans (2), Ochlerotatus sticticus (2), Psorophora ferox (2), Anopheles punctipennis, Anopheles walkeri, Ochlerotatus cantator, Ochlerotatus taeniorhynchus, and Ochlerotatus triseriatus in conjunction with their inefficient vector competency and host feeding preferences indicate that these species likely play a very minor role in either the enzootic maintenance or epizootic transmission of WNV in this region. The principal foci of WNV activity in Connecticut were identified as densely populated (>3,000 people/mi2) residential communities in coastal Fairfield and New Haven Counties, and in the case of 2002, similar locales in proximity of the city of Hartford in central Hartford County. In almost all instances we observed a correlation both temporally and spatially between the isolation of WNV from field-collected mosquitoes and subsequent human cases in these locales. In most years the incidence of human cases closely paralleled the number of virus isolations made from mosquitoes with both peaks falling in early September. We conclude that the isolation of WNV from field-collected mosquitoes is a sensitive indicator of virus activity that is associated with the risk of human infection that habitually extends from early August through the end of October in Connecticut.

Drs. John F. Anderson, David J. Ballard, Carl E. Couch, and Peter A. Dysert II Discuss Clinical Transformation with the Editor. Interview by William Clifford Roberts

Proceedings (Baylor University. Medical Center). Jul, 2004  |  Pubmed ID: 16200111

Protective and Therapeutic Capacity of Human Single-chain Fv-Fc Fusion Proteins Against West Nile Virus

Journal of Virology. Dec, 2005  |  Pubmed ID: 16282460

West Nile virus has spread rapidly across the United States, and there is currently no approved human vaccine or therapy to prevent or treat disease. Passive immunization with antibodies against the envelope protein represents a promising means to provide short-term prophylaxis and treatment for West Nile virus infection. In this study, we identified a panel of 11 unique human single-chain variable region antibody fragments (scFvs) that bind the envelope protein of West Nile virus. Selected scFvs were converted to Fc fusion proteins (scFv-Fcs) and were tested in mice for their ability to prevent lethal West Nile virus infection. Five of these scFv-Fcs, 11, 15, 71, 85, and 95, protected 100% of mice from death when given prior to infection with virus. Two of them, 11 and 15, protected 80% of mice when given at days 1 and 4 after infection. In addition, four of the scFv-Fcs cross-neutralized dengue virus, serotype 2. Binding assays using yeast surface display demonstrated that all of our scFvs bind to sites within domains I and II of West Nile virus envelope protein. These recombinant human scFvs are potential candidates for immunoprophylaxis and therapy of flavivirus infections.

Isolations of Potosi Virus from Mosquitoes (Diptera: Culicidae) Collected in Connecticut

Journal of Medical Entomology. Sep, 2005  |  Pubmed ID: 16363172

Potosi virus (POTV) (Bunyaviridae: Orthobunyavirus) was first isolated from Aedes albopictus (Skuse) collected in Potosi, MO, in 1989, and subsequent isolations were reported from Illinois, Michigan, Ohio, and the Carolinas. To determine whether the distribution of this virus extends into the northeastern United States, we analyzed arboviruses acquired from mosquitoes collected in Connecticut from 1998 to 2004. In 2001, a bunyavirus was isolated from Aedes vexans (Meigen) that was different from other arboviruses known to occur in Connecticut by cross-neutralization and reverse transcription-polymerase chain reaction (RT-PCR) assays. Nucleotide and encoded amino acid sequences of a portion of the G2 envelope gene were 99 and 100% similar to POTV, respectively, yet distinct from indigenous strains of Jamestown Canyon (JCV), Cache Valley (CVV), and Trivittatus virus (TVTV). Viral isolates obtained from the statewide surveillance program were retested by RT-PCR coupled with restriction enzyme analysis to distinguish POTV from other bunyaviruses. POTV isolates, previously typed by neutralization, were correctly identified by RT-PCR; however, many isolates classified as JCV or CVV by enzyme-linked immunosorbent assay proved to be POTV by molecular assays. In total, 92 strains of POTV were isolated from 12 mosquito species in 2000, 2001, and 2003, whereas POTV was not detected in mosquitoes sampled during 1998, 1999, 2002, and 2004. Viral isolation rates were highest for Anopheles punctipennis (Say) (3.2-11.3 infection rate per 1,000 mosquitoes), whereas the greatest number of isolates came from Ochlerotatus trivittatus (Coquillett) (8-16 isolates). This finding represents the first detection of POTV in the northeastern United States where it infects a diverse array of mosquito species.

A Recombinant Envelope Protein Vaccine Against West Nile Virus

Vaccine. Jun, 2005  |  Pubmed ID: 15917113

West Nile (WN) virus is a flavivirus that first appeared in North America in 1999. Since then, more than 600 human deaths and 22,000 equine infections have been attributed to the virus in the United States. We expressed a truncated form of WN virus envelope (E) protein in Drosophila S2 cells. This soluble recombinant E protein was recognized by antibodies from naturally infected horses, indicating that it contains native epitopes. Mice and horses produced high-titer antibodies when immunized with recombinant E protein combined with aluminum hydroxide. Immunized mice were resistant to challenge with a lethal viral dose. Sera from immunized horses, administered to naive mice, conferred resistance against a lethal WN viral challenge. In addition, sera of immunized horses neutralized West Nile virus in vitro, as demonstrated by plaque reduction assays. This recombinant form of E protein, combined with aluminum hydroxide, is a candidate vaccine that may protect humans and horses against WN virus infections.

The Lyme Disease Agent Exploits a Tick Protein to Infect the Mammalian Host

Nature. Jul, 2005  |  Pubmed ID: 16049492

The Lyme disease agent, Borrelia burgdorferi, is maintained in a tick-mouse cycle. Here we show that B. burgdorferi usurps a tick salivary protein, Salp15 (ref. 3), to facilitate the infection of mice. The level of salp15 expression was selectively enhanced by the presence of B. burgdorferi in Ixodes scapularis, first indicating that spirochaetes might use Salp15 during transmission. Salp15 was then shown to adhere to the spirochaete, both in vitro and in vivo, and specifically interacted with B. burgdorferi outer surface protein C. The binding of Salp15 protected B. burgdorferi from antibody-mediated killing in vitro and provided spirochaetes with a marked advantage when they were inoculated into naive mice or animals previously infected with B. burgdorferi. Moreover, RNA interference-mediated repression of salp15 in I. scapularis drastically reduced the capacity of tick-borne spirochaetes to infect mice. These results show the capacity of a pathogen to use a secreted arthropod protein to help it colonize the mammalian host.

Experimental Measles in the Monkey: a Supplemental Note. 1911

Public Health Reports (Washington, D.C. : 1974). 2006  |  Pubmed ID: 16550765

Typhus Fever. 1915

Public Health Reports (Washington, D.C. : 1974). 2006  |  Pubmed ID: 16550770

Host Feeding Patterns of Culex Mosquitoes and West Nile Virus Transmission, Northeastern United States

Emerging Infectious Diseases. Mar, 2006  |  Pubmed ID: 16704786

To evaluate the role of Culex mosquitoes as enzootic and epidemic vectors for WNV, we identified the source of vertebrate blood by polymerase chain reaction amplification and sequencing portions of the cytochrome b gene of mitochondrial DNA. All Cx. restuans and 93% of Cx. pipiens acquired blood from avian hosts; Cx. salinarius fed frequently on both mammals (53%) and birds (36%). Mixed-blood meals were detected in 11% and 4% of Cx. salinarius and Cx. pipiens, respectively. American robin was the most common source of vertebrate blood for Cx. pipiens (38%) and Cx. restuans (37%). American crow represented <1% of the blood meals in Cx. pipiens and none in Cx. restuans. Human-derived blood meals were identified from 2 Cx. salinarius and 1 Cx. pipiens. Results suggest that Cx. salinarius is an important bridge vector to humans, while Cx. pipiens and Cx. restuans are more efficient enzootic vectors in the northeastern United States.

An Ixodes Scapularis Protein Required for Survival of Anaplasma Phagocytophilum in Tick Salivary Glands

The Journal of Experimental Medicine. Jun, 2006  |  Pubmed ID: 16717118

Anaplasma phagocytophilum is the agent of human anaplasmosis, the second most common tick-borne illness in the United States. This pathogen, which is closely related to obligate intracellular organisms in the genera Rickettsia, Ehrlichia, and Anaplasma, persists in ticks and mammalian hosts; however, the mechanisms for survival in the arthropod are not known. We now show that A. phagocytophilum induces expression of the Ixodes scapularis salp16 gene in the arthropod salivary glands during vector engorgement. RNA interference-mediated silencing of salp16 gene expression interfered with the survival of A. phagocytophilum that entered ticks fed on A. phagocytophilum-infected mice. A. phagocytophilum migrated normally from A. phagocytophilum-infected mice to the gut of engorging salp16-deficient ticks, but up to 90% of the bacteria that entered the ticks were not able to successfully infect I. scapularis salivary glands. These data demonstrate the specific requirement of a pathogen for a tick salivary protein to persist within the arthropod and provide a paradigm for understanding how Rickettsia-like pathogens are maintained within vectors.

Gamma Delta T Cells Facilitate Adaptive Immunity Against West Nile Virus Infection in Mice

Journal of Immunology (Baltimore, Md. : 1950). Aug, 2006  |  Pubmed ID: 16849493

West Nile (WN) virus causes fatal meningoencephalitis in laboratory mice, and gammadelta T cells are involved in the protective immune response against viral challenge. We have now examined whether gammadelta T cells contribute to the development of adaptive immune responses that help control WN virus infection. Approximately 15% of TCRdelta(-/-) mice survived primary infection with WN virus compared with 80-85% of the wild-type mice. These mice were more susceptible to secondary challenge with WN virus than the wild-type mice that survived primary challenge with the virus. Depletion of gammadelta T cells in wild-type mice that survived the primary infection, however, does not affect host susceptibility during secondary challenge with WN virus. Furthermore, gammadelta T cells do not influence the development of Ab responses during primary and at the early stages of secondary infection with WN virus. Adoptive transfer of CD8(+) T cells from wild-type mice that survived primary infection with WN virus to naive mice afforded partial protection from lethal infection. In contrast, transfer of CD8(+) T cells from TCRdelta(-/-) mice that survived primary challenge with WN virus failed to alter infection in naive mice. This difference in survival correlated with the numeric and functional reduction of CD8 memory T cells in these mice. These data demonstrate that gammadelta T cells directly link innate and adaptive immunity during WN virus infection.

West Nile Virus from Female and Male Mosquitoes (Diptera: Culicidae) in Subterranean, Ground, and Canopy Habitats in Connecticut

Journal of Medical Entomology. Sep, 2006  |  Pubmed ID: 17017241

In total, 93,532 female mosquitoes (Diptera: Culicidae) were captured in traps placed in subterranean (catch basin), ground (approximately 1.5 m above ground), and canopy (approximately 7.0 m above ground) habitats in Stamford and Stratford, CT, during 2003-2005. Culex pipiens L. was the most abundant (64.8%) of the 31 species identified. Significantly greater numbers of Cx. pipiens were captured in canopy-placed mosquito magnet experimental traps, and significantly greater numbers were collected in catch basin-placed (Centers for Disease Control) CDC traps than in CDC traps placed elsewhere. Culex restuans Theobald was captured in significantly greater numbers in traps placed in catch basins. Aedes vexans (Meigen), Aedes cinereus Meigen, and Aedes cantator (Coquillett) were significantly more abundant in ground traps. In total, 429 isolations of West Nile virus (WNV) were made from seven species of mosquitoes from late June through the end of October during 2003 through 2005. Three hundred ninety-eight (92.8%) isolates were from Cx. pipiens. Others were from Cx. restuans (n = 16), Culex salinarius Coquillett (n = 5), Ae. vexans (n = 4), Ae. cantator (n = 3), Aedes triseriatus (Say) (n = 2), and Ae. cinereus (n = 1). Multiple isolates from Cx. pipiens were made each week, primarily during the later part of July through the end of September. Weekly minimum infection rates (MIRs) were lower in 2004 (highest weekly MIR = 7.1) when no human cases were reported in Connecticut in comparison with 2003 and 2005 (highest weekly MIR = 83.9) when human cases were documented. Frequencies of infected pools were significantly higher in Cx. pipiens captured in traps in the canopy and significantly higher in catch basin placed traps than in traps at ground level. The physiological age structure of Cx. pipiens captured in the canopy was significantly different from that of Cx. pipiens collected in catch basins. Invariably, Cx. pipiens captured in the canopy were nulliparous or parous with ovaries in Christophers' stage 2, whereas 58.7% of the females captured in catch basins possessed ovaries filled with mature oocytes in Christophers' stage 5. Our results suggest that females in the canopy are seeking hosts, and after digestion of the bloodmeal and development of mature oocytes, they descend to catch basins for shelter and deposition of eggs. WNV was isolated from three, one, and two pools of male Cx. pipiens captured in catch basin-, ground-, and canopy-placed traps, respectively, and from six nulliparous Cx. pipiens females collected in the canopy. Weekly MIR ranged from 1.2 to 31.1 per 1,000 male specimens. These data show that mosquitoes become infected by means other than by blood feeding, possibly by transovarial transmission. The placement of traps in tree canopies and in catch basins can be used to augment current practices of placement of traps near the ground for surveillance of mosquitoes infected with WNV and for studies of the ecology of WNV.

Importance of Vertical and Horizontal Transmission of West Nile Virus by Culex Pipiens in the Northeastern United States

The Journal of Infectious Diseases. Dec, 2006  |  Pubmed ID: 17083043

West Nile virus (WNV) has become established in the northeastern United States, where mosquitoes are inactive during winter. There have been no documented studies to explain how this virus survives winter and reinitiates infection in spring. We report that WNV was vertically transmitted to 2 F(1) female Culex pipiens from a naturally infected female collected in Stratford, Connecticut. One vertically infected F(1) female, which was 168 days old, fed on a hamster that died 8 days later of West Nile disease. This suggests that WNV survives winter in unfed, vertically infected C. pipiens with amplification initiated in spring by horizontal transmission.

Antiviral Peptides Targeting the West Nile Virus Envelope Protein

Journal of Virology. Feb, 2007  |  Pubmed ID: 17151121

West Nile virus (WNV) can cause fatal murine and human encephalitis. The viral envelope protein interacts with host cells. A murine brain cDNA phage display library was therefore probed with WNV envelope protein, resulting in the identification of several adherent peptides. Of these, peptide 1 prevented WNV infection in vitro with a 50% inhibition concentration of 67 muM and also inhibited infection of a related flavivirus, dengue virus. Peptide 9, a derivative of peptide 1, was a particularly potent inhibitor of WNV in vitro, with a 50% inhibition concentration of 2.6 muM. Moreover, mice challenged with WNV that had been incubated with peptide 9 had reduced viremia and fatality compared with control animals. Peptide 9 penetrated the murine blood-brain barrier and was found in the brain parenchyma, implying that it may have antiviral activity in the central nervous system. These short peptides serve as the basis for developing new therapeutics for West Nile encephalitis and, potentially, other flaviviruses.

The Lyme Disease Agent Borrelia Burgdorferi Requires BB0690, a Dps Homologue, to Persist Within Ticks

Molecular Microbiology. Feb, 2007  |  Pubmed ID: 17181780

Borrelia burgdorferi survives in an enzootic cycle, and Dps proteins protect DNA against damage during starvation or oxidative stress. The role of a Dps homologue encoded by Borrelia in spirochaete survival was assessed. Dps-deficient spirochaetes were infectious in mice via needle-inoculation at the dose of 10(5) spirochaetes. Larval ticks successfully acquired Dps-deficient spirochaetes via a blood meal on mice. However, after extended periods within unfed nymphs, the Dps-deficient spirochaetes failed to be transmitted to a new host when nymphs fed. Our data suggest that Dps functions to protect the spirochaetes during dormancy in unfed ticks, and in its absence, the spirochaetes become susceptible during tick feeding. dps is differentially expressed in vivo- low in mice and high in ticks - but constitutively expressed in vitro, showing little change during growth or in response to oxidative stress. Borrelia Dps forms a dodecameric complex capable of sequestering iron. The Dps-deficient spirochaetes showed no defect in starvation and oxidative stress assays, perhaps due to the lack of iron in spirochaetes grown in vitro. Dps is critical for spirochaete persistence within ticks, and strategies to interfere with Dps could potentially reduce Borrelia populations in nature and thereby influence the incidence of Lyme disease.

Immunity Against Ixodes Scapularis Salivary Proteins Expressed Within 24 Hours of Attachment Thwarts Tick Feeding and Impairs Borrelia Transmission

PloS One. 2007  |  Pubmed ID: 17505544

In North America, the black-legged tick, Ixodes scapularis, an obligate haematophagus arthropod, is a vector of several human pathogens including Borrelia burgdorferi, the Lyme disease agent. In this report, we show that the tick salivary gland transcriptome and proteome is dynamic and changes during the process of engorgement. We demonstrate, using a guinea pig model of I. scapularis feeding and B. burgdorferi transmission, that immunity directed against salivary proteins expressed in the first 24 h of tick attachment - and not later - is sufficient to evoke all the hallmarks of acquired tick-immunity, to thwart tick feeding and also to impair Borrelia transmission. Defining this subset of proteins will promote a mechanistic understanding of novel I. scapularis proteins critical for the initiation of tick feeding and for Borrelia transmission.

Role of Outer Surface Protein D in the Borrelia Burgdorferi Life Cycle

Infection and Immunity. Sep, 2007  |  Pubmed ID: 17620358

Borrelia burgdorferi preferentially induces selected genes in mice or ticks, and studies suggest that ospD is down-regulated in response to host-specific signals. We now directly show that ospD expression is generally elevated within Ixodes scapularis compared with mice. We then assessed the importance of OspD throughout the spirochete life cycle by generating OspD-deficient B. burgdorferi and examining the mutant in the murine model of tick-transmitted Lyme borreliosis. The lack of OspD did not influence B. burgdorferi infectivity in mice or the acquisition of spirochetes by I. scapularis. OspD adhered to tick gut extracts in vitro, and the OspD-deficient B. burgdorferi strain had a threefold decrease in colonization of the tick gut in vivo. This decrease, however, did not alter subsequent spirochete transmission during a second blood meal. These data suggest that B. burgdorferi can compensate for the lack of OspD in both ticks and mice and that OspD may have a nonessential, secondary, role in B. burgdorferi persistence within I. scapularis.

A Tick Antioxidant Facilitates the Lyme Disease Agent's Successful Migration from the Mammalian Host to the Arthropod Vector

Cell Host & Microbe. Jul, 2007  |  Pubmed ID: 18005713

The tick Ixodes scapularis is an efficient vector for microbes, including the Lyme disease agent Borrelia burgdorferi. Ticks engorging on vertebrates induce recruitment of inflammatory cells to the bite site. For efficient transmission to the vector, pathogens have to traffic through this complex feeding site while avoiding the deleterious effects of immune cells. We show that a tick protein, Salp25D, plays a critical role-in the mammalian host-for acquisition of Borrelia burgdorferi by the vector. Silencing salp25D in tick salivary glands impaired spirochete acquisition by ticks engorging on B. burgdorferi-infected mice. Immunizing mice against Salp25D also decreased Borrelia acquisition by I. scapularis. Salp25D detoxified reactive oxygen species at the vector-pathogen-host interface, thereby providing a survival advantage to B. burgdorferi at the tick feeding site in mice. These data demonstrate that pathogens can exploit arthropod molecules to defuse mammalian responses in order to successfully enter the vector.

Nocturnal Activity of Mosquitoes (Diptera: Culicidae) in a West Nile Virus Focus in Connecticut

Journal of Medical Entomology. Nov, 2007  |  Pubmed ID: 18047212

Six species of mosquitoes (Diptera: Culicidae) were collected in sufficient numbers for analysis in segregating traps set at 2-h intervals by using CO2 and light as attractants in a West Nile virus (family Flaviviridae, genus Flavivirus, WNV) focus in Stratford, CT. The Kolmogorov-Smirnov one-sided test for two samples was used to analyze the data. Mosquito activity began shortly before sunset and continued until shortly after sunrise the next morning. All species had geometric means that were significantly higher during the 2-h period shortly after sunset compared with the 2-h collection before sunset. Species, known to be naturally infected with WNV, were often attracted to these traps in about equal numbers at 2-h intervals during an 8- to 10-h period commencing shortly after sunset. Differences of geometric means were not significant among the four or five 2-h collection periods commencing at sunset for Aedes vexans (Meigen), Culex salinarius Coquillett, and Aedes cinereus Meigen. Aedes cantator (Coquillett) had a significantly higher geometric mean for the 2-h period commencing at sunset, and Coquillettidia perturbans (Walker) was captured in significantly greater numbers during the 2-h period starting at sunset compared with periods commencing 6 h after sunset. Culex pipiens L. tended to have an activity pattern that was primarily nocturnal. Time of night, not meteorological conditions, was the most important factor in determining the nightly variation in the number of trapped mosquitoes. Parity rates of Cx. pipiens collected during specific periods of the night were not significant. In total, 39 isolations of WNV were made from seven species collected primarily during periods of total darkness. Humans are at risk of being bitten by infected mosquitoes throughout the night.

Rural Hospitals and the "5 Million Lives Campaign". Catholic Health Initiatives is Encouraging Its Smaller Facilities to Join a Patient-safety Effort

Health Progress (Saint Louis, Mo.). Nov-Dec, 2007  |  Pubmed ID: 18062375

A Rift Valley Fever Risk Surveillance System for Africa Using Remotely Sensed Data: Potential for Use on Other Continents

Veterinaria Italiana. Jul-Sep, 2007  |  Pubmed ID: 20422546

The authors developed a monitoring and risk mapping system using normalized difference vegetation index (NDVI) times series data derived from the advanced very high resolution radiometer (AVHRR) instrument on polar orbiting national oceanographic and atmospheric administration (NOAA) satellites to map areas with a potential for a Rift Valley fever (RVF) outbreaks in sub-Saharan Africa. This system is potentially an important tool for local, national and international organisations involved in the prevention and control of animal and human disease, permitting focused and timely implementation of disease control strategies several months before an outbreak. We are currently developing a geographic information system (GIS)-based remotely sensed early warning system for potential RVF vectors in the United States. Forecasts of the potential emergence of mosquito vectors will be disseminated throughout the United States, providing several months' warning in advance of potentially elevated mosquito populations. This would allow timely, targeted implementation of mosquito control, animal quarantine and vaccine strategies to reduce or prevent animal and human disease.

A Differential Role for BB0365 in the Persistence of Borrelia Burgdorferi in Mice and Ticks

The Journal of Infectious Diseases. Jan, 2008  |  Pubmed ID: 18171298

Borrelia burgdorferi, the etiologic agent of Lyme disease, persists in both an arthropod vector and vertebrate hosts, usually wild rodents. Analysis of the B. burgdorferi transcriptome in vivo indicates that the bb0365 gene is markedly induced as spirochetes enter the feeding ticks from infected mice. To understand the importance of the bb0365 gene product in the spirochete life cycle, we inactivated this gene in an infectious isolate of B. burgdorferi B31. BB0365-deficient spirochetes were fully pathogenic in mice and survived in diverse murine tissues. When naive ticks engorged on spirochete-infected mice, the B. burgdorferi bb0365 mutant entered ticks but had a markedly decreased survival rate compared with wild type B. burgdorferi. BB0365 therefore is not necessary for B. burgdorferi persistence in the vertebrate host but is required for survival of the Lyme disease agent within the feeding arthropod vector, and strategies for interfering with this gene may potentially interrupt the B. burgdorferi life cycle.

Antibiotic Resistance of Enterococci in American Bison (Bison Bison) from a Nature Preserve Compared to That of Enterococci in Pastured Cattle

Applied and Environmental Microbiology. Mar, 2008  |  Pubmed ID: 18245252

Enterococci isolated from a bison population on a native tall-grass prairie preserve in Kansas were characterized and compared to enterococci isolated from pastured cattle. The species diversity was dominated by Enterococcus casseliflavus in bison (62.4%), while Enterococcus hirae was the most common isolate from cattle (39.7%). Enterococcus faecalis was the second most common species isolated from bison (16%). In cattle, E. faecalis and Enterococcus faecium were isolated at lower percentages (3.2% and 1.6%, respectively). No resistance to ampicillin, chloramphenicol, gentamicin, or high levels of vancomycin was detected from either source. Tetracycline and erythromycin resistance phenotypes, encoded by tetO and ermB, respectively, were common in cattle isolates (42.9% and 12.7%, respectively). A significant percentage of bison isolates (8% and 4%, respectively) were also resistant to these two antibiotics. The tetracycline resistance genes from both bison and cattle isolates resided on mobile genetic elements and showed a transfer frequency of 10(-6) per donor, whereas erythromycin resistance was not transferable. Resistance to ciprofloxacin was found to be higher in enterococci from bison (14.4%) than in enterococci isolated from cattle (9.5%). The bison population can serve as a sentinel population for studying the spread and origin of antibiotic resistance.

West Nile Virus Attenuates Activation of Primary Human Macrophages

Viral Immunology. Mar, 2008  |  Pubmed ID: 18355125

West Nile virus (WNV) is a mosquito-borne flavivirus that has spread rapidly throughout the U.S. and there is currently no effective treatment. Understanding the pathogenesis of WNV infection in humans is critical for development of a potent therapy. In this study, we examined the activation of primary human macrophages in response to WNV infection, and showed that WNV interacts with human macrophages at multiple levels. While infection with WNV induced production of interleukin (IL)-8, production of IL-1beta, and type I interferon was inhibited. Infection with WNV interferes with the downstream JAK/STAT pathway, which is important for macrophage activation. In comparison to other related flaviviruses, the differential response of proinflammatory cytokines is distinct to WNV.

Four-dimensional Computed Tomography Scan Analysis of Tumor and Organ Motion at Varying Levels of Abdominal Compression During Stereotactic Treatment of Lung and Liver

International Journal of Radiation Oncology, Biology, Physics. Apr, 2008  |  Pubmed ID: 18374231

To investigate the effectiveness of different abdominal compression levels on tumor and organ motion during stereotactic body radiotherapy of lower lobe lung and liver tumors using four-dimensional (4D)-CT scan analysis.

Isolations of Jamestown Canyon Virus (Bunyaviridae: Orthobunyavirus) from Field-collected Mosquitoes (Diptera: Culicidae) in Connecticut, USA: a Ten-year Analysis, 1997-2006

Vector Borne and Zoonotic Diseases (Larchmont, N.Y.). Apr, 2008  |  Pubmed ID: 18386967

Jamestown Canyon virus (JCV) (Bunyaviridae: Orthobunyavirus) is a mosquito-borne zoonosis belonging to the California serogroup. It has a wide geographic distribution, occurring throughout much of temperate North America. White-tailed deer, Odocoileus virginianus are the principal amplification hosts, and boreal Aedes and Ochlerotatus mosquitoes are the primary vectors. A 10-year study was undertaken to identify potential mosquito vectors in Connecticut, quantify seasonal prevalence rates of infection, and define the geographic distribution of JCV in the state as a function of land use and white-tailed deer populations, which have increased substantially over this period. Jamestown Canyon virus was isolated from 22 mosquito species. Five of them, Ochlerotatus canadensis, Oc. cantator, Anopheles punctipennis, Coquillettidia perturbans, and Oc. abserratus were incriminated as the most likely vectors, based on yearly isolation frequencies and the spatial geographic distribution of infected mosquitoes. Jamestown Canyon virus was isolated from Oc. canadensis more consistently and from a greater range of collection sites than any other species. Frequent virus isolations were also made from Aedes cinereus, Aedes vexans, and Oc. sticticus, and new North American isolation records were established for Anopheles walkeri, Culex restuans, Culiseta morsitans, Oc. sticticus, Oc. taeniorhynchus, and Psorophora ferox. Other species from which JCV was isolated included C. melanura, Oc. aurifer, Oc. communis, Oc. excrucians, Oc. provocans, Oc. sollicitans, Oc. stimulans, Oc. triseriatus, and Oc. trivittatus. Jamestown Canyon virus was widely distributed throughout Connecticut and found to consistently circulate in a diverse array of mosquito vectors. Infected mosquitoes were collected from June through September, and peak infection rates paralleled mosquito abundance from mid-June through mid-July. Infection rates in mosquitoes were consistent from year to year, and overall virus activity was directly related to local mosquito abundance. Infected mosquitoes were equally distributed throughout the state, irrespective of land use, and infection rates were not directly associated with the abundance of white-tailed deer, possibly because of their saturation throughout the region.

Biology of Ticks

Infectious Disease Clinics of North America. Jun, 2008  |  Pubmed ID: 18452797

Ticks are among the most significant blood-sucking arthropods worldwide. They transmit various pathogens that can cause disease and death in people, domesticated animals, and wildlife. Ticks have several morphologic features and physiologic mechanisms that facilitate host selection, ingestion of vertebrate blood, mating, survival, and reproduction. Although the natural history of ticks varies considerably among species, these arthropods are well-adapted to survive in tropical, temperate, and even subarctic habitats. Key factors, including the reversion of agricultural lands to forests and a close association between people and ticks, have greatly increased the risk of tick bite and human disease.

Role of Two Distinct Gammadelta T Cell Subsets During West Nile Virus Infection

FEMS Immunology and Medical Microbiology. Jul, 2008  |  Pubmed ID: 18513355

gammadelta T cells respond rapidly following West Nile virus (WNV) infection, limiting viremia and invasion of the central nervous system and thereby protecting the host from lethal encephalitis. Here, we investigated the role of two major subpopulations of peripheral gammadelta T cells, Vgamma1(+) and Vgamma4(+) cells, in host immunity against WNV infection. We found initially that aged mice were more susceptible to WNV infection than young mice. Following WNV challenge, Vgamma1(+) cells in young mice expanded significantly whereas Vgamma4(+) cells expanded modestly. In contrast, aged mice exhibited a slower and reduced response of Vgamma1(+) cells but maintained a higher content of Vgamma4(+) cells. Vgamma1(+) cells were the major gammadelta subset producing IFN-gamma during WNV infection. Mice depleted of Vgamma1(+) cells had an enhanced viremia and higher mortality to WNV encephalitis. Vgamma4(+) cells had a higher potential for producing tumor necrosis factor-alpha (TNF-alpha), a cytokine known to be involved in blood-brain barrier compromise and WNV entry into the brain. Depletion of Vgamma4(+) cells reduced TNF-alpha level in the periphery, accompanied by a decreased viral load in the brain and a lower mortality to WN encephalitis. These results suggest that Vgamma1(+) and Vgamma4(+) cells play distinct roles in protection and pathogenesis during WNV infection.

Extrinsic Incubation Periods for Horizontal and Vertical Transmission of West Nile Virus by Culex Pipiens Pipiens (Diptera: Culicidae)

Journal of Medical Entomology. May, 2008  |  Pubmed ID: 18533438

Culex pipiens pipiens L. (Diptera: Culicidae), infected per os from a membrane feeder, transmitted West Nile virus (family Flaviviridae, genus Flavivirus, WNV) at 26 degrees C horizontally during feeding to hamsters and suckling mice and vertically to F1 progeny during egg deposition. Horizontal transmission rates increased with extrinsic incubation, with 75-100% of the females transmitting on days 16 through 25 postinfection (pi). No females deposited eggs infected with WNV after the first bloodmeal on 3-8 d pi. Females vertically transmitted WNV during egg laying after their second, third, and fourth bloodmeals on days 13-33 pi. The vertical transmission rate was 4.7%. The vertical minimal infection rate was 0.52 infected F1 specimens/1,000 specimens tested from females feeding during their second and later bloodmeals on hamsters or suckling mice. The sequence of horizontal and vertical transmission is reported. A female may transmit WNV 1) horizontally to a host during feeding and subsequently vertically to her offspring during egg laying, 2) vertically to her offspring during oviposition without prior horizontal transmission to a host, and 3) horizontally to a host without vertically transmitting the virus. These two means of transmission by Cx. p. pipiens contribute to the relatively high minimum infection rates that are reached in late summer and to the survival of virus during winter and initiation of amplification in the spring in the northeastern United States.

Drak2 Contributes to West Nile Virus Entry into the Brain and Lethal Encephalitis

Journal of Immunology (Baltimore, Md. : 1950). Aug, 2008  |  Pubmed ID: 18641347

Death-associated protein kinase-related apoptosis-inducing kinase-2 (Drak2), a member of the death-associated protein family of serine/threonine kinases, is specifically expressed in T and B cells. In the absence of Drak2, mice are resistant to experimental autoimmune encephalomyelitis due to a decrease in the number of cells infiltrating the CNS. In the present study, we investigated the role of Drak2 in West Nile virus (WNV)-induced encephalitis and found that Drak2(-/-) mice were also more resistant to lethal WNV infection than wild-type mice. Although Drak2(-/-) mice had an increase in the number of IFN-gamma-producing T cells in the spleen after infection, viral levels in the peripheral tissues were not significantly different between these two groups of mice. In contrast, there was a reduced viral load in the brains of Drak2(-/-) mice, which was accompanied by a decrease in the number of Drak2(-/-) CD4(+) and CD8(+) T cells in the brain following WNV infection. Moreover, we detected viral Ags in T cells isolated from the spleen or brain of WNV-infected mice. These results suggest that following a systemic infection, WNV might cross the blood brain barrier and enter the CNS by being carried by infected infiltrating T cells.

Tracking Eastern Equine Encephalitis Virus Perpetuation in the Northeastern United States by Phylogenetic Analysis

The American Journal of Tropical Medicine and Hygiene. Aug, 2008  |  Pubmed ID: 18689638

Epidemics and epizootics of eastern equine encephalitis virus (EEEV) occur sporadically in temperate regions where transmission is seasonal from late summer to early fall. These outbreaks may derive from virus that overwinters locally or perhaps results from reintroduction of virus from other sites. To evaluate these possibilities, we compared the phylogenetic relationships of EEEV isolates obtained from mosquitoes collected during statewide arbovirus surveillance in Connecticut, in addition to isolates from concurrent outbreaks in southern New Hampshire and upstate New York. In Connecticut, viral isolates grouped into temporally discrete clades by year of isolation or over 2 years of sampling. Two or more clades arose in 2000, 2001, 2003, 2004, and 2006, possibly the result of separate introduction events into the state, whereas viruses from upstate New York and New Hampshire segregated into single clades that persisted for 2 or more years. New Hampshire viruses shared recent common ancestry to those isolated in Connecticut suggesting viral dispersal among these regions. These results provide additional evidence for independent episodes of EEEV overwintering in northern foci.

Borrelia Burgdorferi Complement Regulator-acquiring Surface Protein 2 Does Not Contribute to Complement Resistance or Host Infectivity

PloS One. 2008  |  Pubmed ID: 18714378

Borrelia burgdorferi, the pathogen of Lyme disease, cycles in nature through Ixodes ticks and mammalian hosts. At least five Complement Regulator-Acquiring Surface Proteins (BbCRASPs) are produced by B. burgdorferi, which are thought to assist spirochetes in host immune evasion. Recent studies established that BbCRASP-2 is preferentially expressed in mammals, and elicits robust antibody response in infected hosts, including humans. We show that BbCRASP-2 is ubiquitously expressed in diverse murine tissues, but not in ticks, reinforcing a role of BbCRASP-2 in conferring B. burgdorferi defense against persistent host immune threats, such as complement. BbCRASP-2 immunization, however, fails to protect mice from B. burgdorferi infection and does not modify disease, as reflected by the development of arthritis. An infectious BbCRASP-2 mutant was generated, therefore, to examine the precise role of the gene product in spirochete infectivity. Similar to wild type B. burgdorferi, BbCRASP-2 mutants remain insensitive to complement-mediated killing in vitro, retain full murine infectivity and induce arthritis. Quantitative RT-PCR assessment indicates that survivability of BbCRASP-2-deficient B. burgdorferi is not due to altered expression of other BbCRASPs. Together, these results suggest that the function of a selectively expressed B. burgdorferi gene, BbCRASP-2, is not essential for complement resistance or infectivity in the murine host.

Serum Antibodies to West Nile Virus in Naturally Exposed and Vaccinated Horses

Journal of Medical Microbiology. Sep, 2008  |  Pubmed ID: 18719177

A polyvalent ELISA and plaque reduction neutralization tests (PRNTs) were used to measure serum antibodies to West Nile virus (WNV) in horses naturally exposed to or vaccinated against this flavivirus in Connecticut and New York State, USA. Relying on a PRNT as a 'gold standard', the main objective was to validate a modified ELISA containing a recombinant WNV envelope protein antigen. It was also important to assess specificity by testing sera from horses that had other, undiagnosed illnesses. Sera for the latter study were obtained from 43 privately owned horses during 1995-1996. Analyses by an ELISA and a PRNT confirmed the presence of WNV antibodies in 21 (91%) of 23 sera from naturally exposed horses and in 85% of the 20 vaccinated subjects; overall results for both study groups were highly concordant (91% agreement). Humoral responses of naturally exposed and immunized horses were similar. Both serological tests were useful in confirming past infections with WNV, but there was no evidence that horses with undiagnosed illnesses were exposed to WNV prior to a 1999 outbreak in Connecticut, USA.

A Recombinant West Nile Virus Envelope Protein Vaccine Candidate Produced in Spodoptera Frugiperda ExpresSF+ Cells

Vaccine. Jan, 2009  |  Pubmed ID: 18996430

In this study, a recombinant truncated West Nile virus envelope protein antigen (rWNV-E) was produced in serum-free cultures of the expresSF+ insect cell line via baculovirus infection. This production system was selected based on its use in the production of candidate human and animal vaccine antigens. A defined fermentation and purification process for the rWNV-E antigen was established to control for purity and immunogenicity of each protein batch. The material formulated with aluminum hydroxide was stable for greater than 8months at 4 degrees C. The recombinant vaccine candidate was evaluated for immunogenicity and protective efficacy in several animal models. In mouse and hamster WNV challenge models, the vaccine candidate induced viral protection that correlated with anti-rWNV-E immunogenicity and WNV neutralizing antibody titers. The rWNV-E vaccine candidate was used to boost horses previously immunized with the Fort Dodge inactivated WNV vaccine and also to induce WNV neutralizing titers in naïve foals that were at least 14weeks of age. Furthermore, the vaccine candidate was found safe when high doses were injected into rats, with no detectable treatment-related clinical adverse effects. These observations demonstrate that baculovirus-produced rWNV-E can be formulated with aluminum hydroxide to produce a stable and safe vaccine which induces humoral immunity that can protect against WNV infection.

Toll-like Receptor 7 Mitigates Lethal West Nile Encephalitis Via Interleukin 23-dependent Immune Cell Infiltration and Homing

Immunity. Feb, 2009  |  Pubmed ID: 19200759

West Nile virus (WNV), a mosquito-transmitted single-stranded RNA (ssRNA) flavivirus, causes human disease of variable severity. We investigated Toll-like receptor 7-deficient (Tlr7(-/-)) and myeloid differentiation factor 88-deficient (Myd88(-/-)) mice, which both have defective recognition of ssRNA, and found increased viremia and susceptibility to lethal WNV infection. Despite increased tissue concentrations of most innate cytokines, CD45(+) leukocytes and CD11b(+) macrophages failed to home to WNV-infected cells and infiltrate into target organs of Tlr7(-/-) mice. Tlr7(-/-) mice and macrophages had reduced interleukin-12 (IL-12) and IL-23 responses after WNV infection, and mice deficient in IL-12 p40 and IL-23 p40 (Il12b(-/-)) or IL-23 p19 (Il23a(-/-)), but not IL-12 p35 (Il12a(-/-)), responded similarly to Tlr7(-/-) mice, with increased susceptibility to lethal WNV encephalitis. Collectively, these results demonstrate that TLR7 and IL-23-dependent WNV responses represent a vital host defense mechanism that operates by affecting immune cell homing to infected target cells.

Lateral Inferior Prefrontal Cortex and Anterior Cingulate Cortex Are Engaged at Different Stages in the Solution of Insight Problems

Proceedings of the National Academy of Sciences of the United States of America. Jun, 2009  |  Pubmed ID: 19541657

Two studies used puzzles that required participants to find a word that satisfied a set of constraints. The first study used a remote-association task, where participants had to find a word that would form compound words with 3 other words. The second study required participants to complete a word fragment with an associate of another word. Both studies produced distinct patterns of activity in the lateral inferior prefrontal cortex (LIPFC) and the anterior cingulate cortex (ACC). Activation in the LIPFC rose only as long as the participants were trying to retrieve the solution and dropped off as soon as the solution was obtained. However, activation in the ACC increased upon the retrieval of a solution, reflecting the need to process that solution. The data of the second experiment are fit by an information-processing model that interprets the activity in the LIPFC as reflecting retrieval operations and the activity in the ACC as reflecting subgoal setting.

Urinary Side Effects and Complications After Permanent Prostate Brachytherapy: the MD Anderson Cancer Center Experience

Urology. Sep, 2009  |  Pubmed ID: 19589580

To evaluate acute and long-term urinary morbidity after permanent prostate brachytherapy at a single tertiary care center. To minimize the risk of long-term urinary morbidity, it is important for clinicians to be able to distinguish acute urinary side effects after prostate brachytherapy from longer-term treatment-related urinary complications.

Borrelia Burgdorferi Small Lipoprotein Lp6.6 is a Member of Multiple Protein Complexes in the Outer Membrane and Facilitates Pathogen Transmission from Ticks to Mice

Molecular Microbiology. Oct, 2009  |  Pubmed ID: 19703109

Borrelia burgdorferi lipoprotein Lp6.6 is a differentially produced spirochete antigen. An assessment of lp6.6 expression covering representative stages of the infectious cycle of spirochetes demonstrates that the gene is solely expressed during pathogen persistence in ticks. Deletion of lp6.6 in infectious B. burgdorferi did not influence in vitro growth, or its ability to persist and induce inflammation in mice, migrate to larval or nymphal ticks or survive through the larval-nymphal molt. However, Lp6.6-deficient spirochetes displayed significant impairment in their ability to transmit from infected ticks to naïve mice, which was restored upon genetic complementation of the mutant with a wild-type copy of lp6.6, establishing that Lp6.6 plays a role in pathogen transmission from ticks to mammals. Lp6.6 is a subsurface, yet highly abundant, outer membrane antigen. Two-dimensional blue native/SDS-PAGE coupled with liquid chromatography-mass spectrometry (LC-MS/MS) analysis and protein cross-linking studies independently shows that Lp6.6 exists in multiple protein complexes in the outer membrane. We speculate that the function of Lp6.6 is connected to the physiological processes of these membrane complexes. Further characterization of differentially produced membrane antigens and associated protein complexes will likely aid in our understanding of the molecular details of B. burgdorferi persistence and transmission through a complex enzootic cycle.

Slowing Bacterial Translation Speed Enhances Eukaryotic Protein Folding Efficiency

Journal of Molecular Biology. Mar, 2010  |  Pubmed ID: 20043920

The mechanisms for de novo protein folding differ significantly between bacteria and eukaryotes, as evidenced by the often observed poor yields of native eukaryotic proteins upon recombinant production in bacterial systems. Polypeptide synthesis rates are faster in bacteria than in eukaryotes, but the effects of general variations in translation rates on protein folding efficiency have remained largely unexplored. By employing Escherichia coli cells with mutant ribosomes whose translation speed can be modulated, we show here that reducing polypeptide elongation rates leads to enhanced folding of diverse proteins of eukaryotic origin. These results suggest that in eukaryotes, protein folding necessitates slow translation rates. In contrast, folding in bacteria appears to be uncoupled from protein synthesis, explaining our findings that a generalized reduction in translation speed does not adversely impact the folding of the endogenous bacterial proteome. Utilization of this strategy has allowed the production of a native eukaryotic multidomain protein that has been previously unattainable in bacterial systems and may constitute a general alternative to the production of aggregation-prone recombinant proteins.

The Sacsin Repeating Region (SRR): a Novel Hsp90-related Supra-domain Associated with Neurodegeneration

Journal of Molecular Biology. Jul, 2010  |  Pubmed ID: 20488193

Protein supra-domains are defined as recurring arrangements of two or three domains present adjacent to each other along a polypeptide chain. Such combinations have novel functions beyond those of the individual partner domains that compose them, which can exist in isolation. Here, we describe a new type of large supra-domain (approximately 360 residues) in which one of the component partners (approximately 200 residues) appears to be incapable of existing in a context other than immediately adjacent to the C-terminus of the well-characterized Hsp90-like ATPase domain. We found that this supra-domain has a broad phylogenetic distribution, with examples in Archaea, Bacteria, and Eukarya. There is strong selective pressure for this arrangement to occur as part of repeated regions of unprecedented length. We identified multiple strategies of convergent evolution to attain such configurations. In humans, this supra-domain is present in triplicate at the N-terminus of the protein sacsin (4579 residues), mutated in the neurodegenerative disorder known as spastic ataxia of Charlevoix-Saguenay, and thus, we termed it "sacsin repeating region" (SRR). Biochemical characterization demonstrated that SRRs possess ATPase activity, which appears to be a requirement for sacsin function, as a disease-causing mutation leads to an alternate conformation completely incapable of hydrolyzing ATP. We also found evidence of a convergent evolutionary strategy to place SRRs in proteins containing C-terminal J domains, which we demonstrated here to be capable of stimulating the intrinsic ATPase activity of Hsp70. Our sequence and biochemical analyses indicate that SRRs necessitate nucleotide hydrolysis for their function, provided by the common Hsp90 ATPase domain, which, when coupled to the unique adjacent sequence, may give rise to a novel activity related to protein quality control.

Anaplasma Phagocytophilum Induces Ixodes Scapularis Ticks to Express an Antifreeze Glycoprotein Gene That Enhances Their Survival in the Cold

The Journal of Clinical Investigation. Sep, 2010  |  Pubmed ID: 20739755

In the United States, Ixodes scapularis ticks overwinter in the Northeast and Upper Midwest and transmit the agent of human granulocytic anaplasmosis, Anaplasma phagocytophilum, among other pathogens. We now show that the presence of A. phagocytophilum in I. scapularis ticks increases their ability to survive in the cold. We identified an I. scapularis antifreeze glycoprotein, designated IAFGP, and demonstrated via RNAi knockdown studies the importance of IAFGP for the survival of I. scapularis ticks in a cold environment. Transfection studies also show that IAFGP increased the viability of yeast cells subjected to cold temperature. Remarkably, A. phagocytophilum induced the expression of iafgp, thereby increasing the cold tolerance and survival of I. scapularis. These data define a molecular basis for symbiosis between a human pathogenic bacterium and its arthropod vector and delineate what we believe to be a new pathway that may be targeted to alter the life cycle of this microbe and its invertebrate host.

A C-type Lectin Collaborates with a CD45 Phosphatase Homolog to Facilitate West Nile Virus Infection of Mosquitoes

Cell. Sep, 2010  |  Pubmed ID: 20797779

West Nile virus (WNV) is the most common arthropod-borne flavivirus in the United States; however, the vector ligand(s) that participate in infection are not known. We now show that an Aedes aegypti C-type lectin, mosGCTL-1, is induced by WNV, interacts with WNV in a calcium-dependent manner, and facilitates infection in vivo and in vitro. A mosquito homolog of human CD45 in A. aegypti, designated mosPTP-1, recruits mosGCTL-1 to enable viral attachment to cells and to enhance viral entry. In vivo experiments show that mosGCTL-1 and mosPTP-1 function as part of the same pathway and are critical for WNV infection of mosquitoes. A similar phenomenon was also observed in Culex quinquefasciatus, a natural vector of WNV, further demonstrating that these genes participate in WNV infection. During the mosquito blood-feeding process, WNV infection was blocked in vivo with mosGCTL-1 antibodies. A molecular understanding of flaviviral-arthropod interactions may lead to strategies to control viral dissemination in nature.

Disorders of Protein Biogenesis and Stability

Protein and Peptide Letters. Feb, 2011  |  Pubmed ID: 21121895

The great diversity of structural conformations available to proteins allows this class of molecules to carry out the vast majority of biochemical functions in the cell. In order to function adequately, proteins must be synthesized, folded/assembled and degraded with great temporal and spatial accuracy. Precise coordination of multiple processes, including ribosome assembly and movement along mRNA, charging and recycling of tRNAs, recruitment and action of molecular chaperones, and tight control of the degradation machinery is essential to create and maintain a stable proteome. It has become recently evident that even slight errors in any of these processes may lead to disease states. Accordingly, increasing numbers of human diseases have been identified that are due to mutations in genes encoding proteins involved in this so-called "protein quality control". Since these processes are essential for the production and maintenance of the entire proteome of the cell, the deleterious effects of these mutations often extend far beyond the faulty gene. This review provides an overview of human disorders caused by defects in mechanisms underlying protein biogenesis and stability.

Control of Mosquitoes in Catch Basins in Connecticut with Bacillus Thuringiensis Israelensis, Bacillus Sphaericus, [corrected] and Spinosad

Journal of the American Mosquito Control Association. Mar, 2011  |  Pubmed ID: 21476447

Catch basins are a major source of Culex pipiens pipiens, Cx. restuans, and Aedes japonicus in northeastern USA. VectoBac CG (Bacillus thuringiensis israelensis [Bti]), VectoLex CG (Bacillus sphaericus [Bs]), and VectoBac 12AS (Bti), each applied at maximum label rate of 1.8 g, 1.8 g, and 0.193 ml per catch basin, respectively, significantly reduced the numbers of larvae for 1 wk. The dosages on the labels for treatment of mosquito larvae in catch basins, where mosquito breeding is continuous, are not adequate for providing long-term control in the northeastern USA without the need for frequent retreatment. When applied at 3 times the maximum label rate, VectoLex CG, VectoBac 12AS, and VectoBac CG significantly reduced the numbers of larvae for 5, 4, and 2 wk, respectively. A single application of VectoMax WSP (Bti + Bs) (1 pouch containing 10 g) per catch basin significantly reduced the numbers of 3rd and 4th instars and healthy pupae in catch basins in 2008, but numbers of 3rd and 4th instars in treated catch basins at 21 days after treatment had increased to 40% of the numbers in untreated catch basins. A 2nd treatment of 1 pouch per catch basin reduced the numbers of 3rd and 4th instars and healthy pupae to near zero for the next 4 wk, into the middle of September 2008. In 2009, VectoMax applied as 1 pouch per catch basin on July 1 and again on August 18 significantly reduced the numbers of healthy pupae throughout the summer until the end of September. A 2nd application of VectoMax to catch basins is likely needed during summer, when rainfall averages 13.7 in. (approximately 34.25 cm) during June through September, to keep the numbers of Culex and Ae. japonicus significantly reduced to lower risk of human exposure to West Nile virus. The application of 1 Natular XRT tablet, each weighing approximately 40.5 g (6.25% spinosad), to individual catch basins in 2009 significantly reduced the total numbers of larvae for 5 wk.

Molecular Evolution of West Nile Virus in a Northern Temperate Region: Connecticut, USA 1999-2008

Virology. Aug, 2011  |  Pubmed ID: 21723580

West Nile virus (WNV) has become firmly established in northeastern US, reemerging every summer since its introduction into North America in 1999. To determine whether WNV overwinters locally or is reseeded annually, we examined the patterns of viral lineage persistence and replacement in Connecticut over 10 consecutive transmission seasons by phylogenetic analysis. In addition, we compared the full protein coding sequence among WNV isolates to search for evidence of convergent and adaptive evolution. Viruses sampled from Connecticut segregated into a number of well-supported subclades by year of isolation with few clades persisting ≥2 years. Similar viral strains were dispersed in different locations across the state and divergent strains appeared within a single location during a single transmission season, implying widespread movement and rapid colonization of virus. Numerous amino acid substitutions arose in the population but only one change, V→A at position 159 of the envelope protein, became permanently fixed. Several instances of parallel evolution were identified in independent lineages, including one amino acid change in the NS4A protein that appears to be positively selected. Our results suggest that annual reemergence of WNV is driven by both reintroduction and local-overwintering of virus. Despite ongoing evolution of WNV, most amino acid variants occurred at low frequencies and were transient in the virus population.

The Neurodegenerative-disease-related Protein Sacsin is a Molecular Chaperone

Journal of Molecular Biology. Aug, 2011  |  Pubmed ID: 21726565

Various human neurodegenerative disorders are associated with processes that involve misfolding of polypeptide chains. These so-called protein misfolding disorders include Alzheimer's and Parkinson's diseases and an increasing number of inherited syndromes that affect neurons involved in motor control circuits throughout the central nervous system. The reasons behind the particular susceptibility of neurons to misfolded proteins are currently not known. The main function of a class of proteins known as molecular chaperones is to prevent protein misfolding and aggregation. Although neuronal cells contain the major known classes of molecular chaperones, central-nervous-system-specific chaperones that maintain the neuronal proteome free from misfolded proteins are not well defined. In this study, we assign a novel molecular chaperone activity to the protein sacsin responsible for autosomal recessive spastic ataxia of Charlevoix-Saguenay, a degenerative disorder of the cerebellum and spinal cord. Using purified components, we demonstrate that a region of sacsin that contains a segment with homology to the molecular chaperone Hsp90 is able to enhance the refolding efficiency of the model client protein firefly luciferase. We show that this region of sacsin is highly capable of maintaining client polypeptides in soluble folding-competent states. Furthermore, we demonstrate that sacsin can efficiently cooperate with members of the Hsp70 chaperone family to increase the yields of correctly folded client proteins. Thus, we have identified a novel chaperone directly involved in a human neurodegenerative disorder.

An in Vivo Transfection Approach Elucidates a Role for Aedes Aegypti Thioester-containing Proteins in Flaviviral Infection

PloS One. 2011  |  Pubmed ID: 21818390

Mosquitoes transmit pathogens that cause infectious diseases of global importance. Techniques to easily introduce genes into mosquitoes, however, limit investigations of the interaction between microbes and their arthropod vectors. We now show that a cationic liposome significantly enhances delivery and expression of plasmid DNA in Aedes aegypti and Anopheles gambiae mosquitoes. We then introduced the genes for Ae. aegypti thioester-containing proteins (AeTEPs), which are involved in the control of flaviviral infection, into mosquitoes using this technique. In vivo transfection of AeTEP-1 into Ae. aegypti significantly reduced dengue virus infection, suggesting that the approach can further our understanding of pathogen-mosquito interactions.

Widespread Dispersal of Borrelia Burgdorferi-infected Ticks Collected from Songbirds Across Canada

The Journal of Parasitology. Aug, 2011  |  Pubmed ID: 21864130

Abstract Millions of Lyme disease vector ticks are dispersed annually by songbirds across Canada, but often overlooked as the source of infection. For clarity on vector distribution, we sampled 481 ticks (12 species and 3 undetermined ticks) from 211 songbirds (42 species/subspecies) nationwide. Using PCR, 52 (29.5%) of 176 Ixodes ticks tested were positive for the Lyme disease spirochete, Borrelia burgdorferi s.l. Immature blacklegged ticks, Ixodes scapularis, collected from infested songbirds had a B. burgdorferi infection prevalence of 36% (larvae, 48%; nymphs, 31%). Notably, Ixodes affinis is reported in Canada for the first time and, similarly, Ixodes auritulus for the initial time in the Yukon. Firsts for bird-parasitizing ticks include I. scapularis in Quebec and Saskatchewan. We provide the first records of 3 tick species cofeeding on passerines (song sparrow, Swainson's thrush). New host records reveal I. scapularis on the blackpoll warbler and Nashville warbler. We furnish the following first Canadian reports of B. burgdorferi-positive ticks: I. scapularis on chipping sparrow, house wren, indigo bunting; I. auritulus on Bewick's wren; and I. spinipalpis on a Bewick's wren and song sparrow. First records of B. burgdorferi-infected ticks on songbirds include: the rabbit-associated tick, Ixodes dentatus, in western Canada; I. scapularis in Quebec, Saskatchewan, northern New Brunswick, northern Ontario; and Ixodes spinipalpis (collected in British Columbia). The presence of B. burgdorferi in Ixodes larvae suggests reservoir competency in 9 passerines (Bewick's wren, common yellowthroat, dark-eyed junco, Oregon junco, red-winged blackbird, song sparrow, Swainson's thrush, swamp sparrow, and white-throated sparrow). We report transstadial transmission (larva to nymph) of B. burgdorferi in I. auritulus. Data suggest a possible 4-tick, i.e., I. angustus, I. auritulus, I. pacificus, and I. spinipalpis enzootic cycle of B. burgdorferi on Vancouver Island, British Columbia. Our results suggest that songbirds infested with B. burgdorferi-infected ticks have the potential to start new tick populations endemic for Lyme disease. Because songbirds disperse B. burgdorferi-infected ticks outside their anticipated range, health-care providers are advised that people can contract Lyme disease locally without any history of travel.

Borrelia Burgdorferi BBA52 is a Potential Target for Transmission Blocking Lyme Disease Vaccine

Vaccine. Nov, 2011  |  Pubmed ID: 21945261

The surface-exposed antigens of Borrelia burgdorferi represent important targets for induction of protective host immune responses. BBA52 is preferentially expressed by B. burgdorferi in the feeding tick, and a targeted deletion of bba52 interferes with vector-host transitions in vivo. In this study, we demonstrate that BBA52 is an outer membrane surface-exposed protein and that disulfide bridges take part in the homo-oligomeric assembly of native protein. BBA52 antibodies lack detectable borreliacidal activities in vitro. However, active immunization studies demonstrated that BBA52 vaccinated mice were significantly less susceptible to subsequent tick-borne challenge infection. Similarly, passive transfer of BBA52 antibodies in ticks completely blocked B. burgdorferi transmission from feeding ticks to naïve mice. Taken together, these studies highlight the role of BBA52 in spirochete dissemination from ticks to mice and demonstrate the potential of BBA52 antibody-mediated strategy to complement the ongoing efforts to develop vaccines for blocking the transmission of B. burgdorferi.

Horizontal and Vertical Transmission of West Nile Virus Genotype NY99 by Culex Salinarius and Genotypes NY99 and WN02 by Culex Tarsalis

The American Journal of Tropical Medicine and Hygiene. Jan, 2012  |  Pubmed ID: 22232464

Culex tarsalis is a superior horizontal and vertical vector of West Nile virus (WNV) compared with Culex salinarius. Culex salinarius transmitted WNV genotype NY99 (CT 2741-99 strain) horizontally to suckling mice at significantly lower rates than Cx. tarsalis on Days 8, 9, 10, and 12 post-infection, and Cx. salinarius transmitted WNV genotype NY99 to offspring at a lower vertical transmission infection rate than Cx. tarsalis. Culex tarsalis transmitted WNV genotypes NY99 and WN02 (CT S0084-08 strain) with equal efficiency. Daily percent horizontal transmission of genotype NY99 by Cx. tarsalis-infected per os and by intra-thoracic infection was not significantly different from daily transmission of genotype WN02 from Days 5-23 and Days 2-9 post-infection, respectively. Our findings do not support the previously published hypothesis that genotype NY99 was replaced in the New World by WN02 because of a shorter extrinsic incubation of WN02.