Expansion of zoonotic babesiosis and reported human cases, Connecticut, 2001-2010.

To document the expansion of human babesiosis in Connecticut, we analyzed reservoir host sera for seroreactivity to Babesia microti Franca and reviewed Connecticut human surveillance case data collected during 2001-2010. Sera from white-footed mice, Peromyscus leucopus Rafinesque, from 10 towns in 5 counties, collected at 4-7-yr periods between 2001 and 2010, were tested for total immunoglobulins. The prevalence of B.

Serum antibodies to Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti in recaptured white-footed mice.

A mark-release-recapture study was conducted during 2007 through 2010 in six, tick-infested sites in Connecticut, United States to measure changes in antibody titers for Borrelia burgdorferi sensu stricto, Anaplasma phagocytophilum, and Babesia microti in Peromyscus leucopus (white-footed mice). There was an overall recapture rate of 40%, but only four tagged mice were caught in ≥2 yr. Sera from 561 mice were analyzed for total antibodies to B.

Seroprevalence of Powassan virus in New England deer, 1979-2010.

Powassan virus and its subtype, deer tick virus, are closely related tick-borne flaviviruses that circulate in North America. The incidence of human infection by these agents appears to have increased in recent years. To define exposure patterns among white-tailed deer, potentially useful sentinels that are frequently parasitized by ticks, we screened serum samples collected during 1979-2010 in Connecticut, Maine, and Vermont for neutralizing antibody by using a novel recombinant deer tick virus-West Nile virus chimeric virus.

Serum antibodies to whole-cell and recombinant antigens of Borrelia burgdorferi in cottontail rabbits.

Archived serum samples, from 95 eastern cottontail rabbits (Sylvilagus floridanus) captured in New York, New York, USA and Millbrook, New York, USA, during 1985-86, were analyzed in solid-phase enzyme-linked immunosorbent assays (ELISA) for total and class-specific immunoglobulin (Ig) M antibodies to whole-cell or recombinant antigens of Borrelia burgdorferi sensu stricto. Using a polyvalent conjugate, rabbit sera contained antibodies to whole-cell and recombinant antigens (protein [p]35, p37, or VlsE) during different seasons, but there was no reactivity to outer surface protein (Osp)A or OspB. Seventy-six of the 102 sera (75%) analyzed were reactive with one or more of the antigens; 61 of the positive samples (80%) reacted to whole-cell antigens, followed by results for the p35 (58%, 44/76), VlsE (43%, 33/76), and p37 (29%, 22/ 76) antigens.

Seasonal prevalence of serum antibodies to whole cell and recombinant antigens of Borrelia burgdorferi and Anaplasma phagocytophilum in white-tailed deer in Connecticut.

Whole-blood samples were obtained from 214 white-tailed deer (Odocoileus virginianus) representing 44 sites in Connecticut (USA) during 1992, 1993, 1996, 1999, and 2000 through 2006. Sera were analyzed for total antibodies to whole-cell or recombinant antigens of Borrelia burgdorferi sensu stricto and Anaplasma phagocytophilum, the respective causative agents of Lyme borreliosis and human granulocytic anaplasmosis. Deer sera contained antibodies to both bacteria during different seasons and throughout the 11-yr study.

Serum antibodies to West Nile virus in naturally exposed and vaccinated horses.

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.

Biology of ticks.

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.

Antibodies to whole-cell or recombinant antigens of Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti in white-footed mice.

Serum samples were obtained from white-footed mice (Peromyscus leucopus) in tick-infested areas of Connecticut during the period 2001 through 2003 and analyzed for antibodies to Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti. Emphasis was placed on the evaluations of highly specific recombinant VlsE or protein (p) 44 antigens of B. burgdorferi and A.

Detection of antibodies to Francisella tularensis in cats.

Blood samples were obtained from privately owned cats in Connecticut and New York State, USA in 1985-1990, and analyzed for evidence of Francisella tularensis, the etiologic agent of tularemia. Of the 91 sera tested by microagglutination (MA) methods, 11 (12%) contained antibodies to F. tularensis.

Seroprevalence of antibodies against Borrelia burgdorferi and Anaplasma phagocytophilum in cats.

Objective: To determine whether cats in the northeastern United States develop serum antibodies against antigens of Borrelia burgdorferi and Anaplasma phagocytophilum and whether coinfection with the 2 organisms occurs.

Sample Population: Serum samples from 84 healthy cats and 9 cats with lameness, fever, anorexia, or fatigue.

Procedure: Serum antibodies against B.

Detection of antibodies to Borrelia burgdorferi in naturally infected horses in the USA by enzyme-linked immunosorbent assay using whole-cell and recombinant antigens.

Blood samples were collected from 98 horses suspected of having borreliosis or granulocytic ehrlichiosis in Connecticut and New York State, USA during 1985, 1995, and 1996. Serum antibodies to Borrelia burgdorferi were detected by an enzyme-linked immunosorbent assay (ELISA), based on whole-cell and recombinant antigens, in 82 (84%) horses. Of the 181 sera tested, 59% were positive, using whole-cell antigens, compared to 48% with protein (p)37 and 35% with VlsE antigens.

A comparison of serologic tests for the detection of serum antibodies to whole-cell and recombinant Borrelia burgdorferi antigens in cattle.

Serum samples from healthy dairy and beef cattle, living in tick-infested areas of Connecticut, USA, were analyzed by polyvalent enzyme-linked immunosorbent assays (ELISA), indirect fluorescent antibody (IFA) staining methods, or Western blot procedures to detect antibodies to tick-borne agents. Of the 80 sera tested by ELISA with whole-cell or 10 separate recombinant antigens (fusion proteins) of Borrelia burgdorferi sensu stricto, 57 (71%) were positive to 1 or more antigens, while 36 (45%) reacted to whole-cell antigens by IFA staining methods. Three (4%) of 80 samples had antibodies to Anaplasma phagocytophilum.

Use of recombinant antigens of Borrelia burgdorferi and Anaplasma phagocytophilum in enzyme-linked immunosorbent assays to detect antibodies in white-tailed deer.

Serum samples obtained from white-tailed deer (Odocoileus virginianus) in Connecticut (n=218) and South Carolina (n=20) (USA) during the period 1992-2002 were analyzed for antibodies to whole-cell or recombinant antigens (i.e., fusion proteins) of Borrelia burgdorferi sensu stricto and Anaplasma phagocytophilum, etiologic agents of Lyme borreliosis and granulocytic ehrlichiosis, respectively.

A recombinant envelope protein-based enzyme-linked immunosorbent assay for West Nile virus serodiagnosis.

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.

Comparative reactivity of human sera to recombinant VlsE and other Borrelia burgdorferi antigens in class-specific enzyme-linked immunosorbent assays for Lyme borreliosis.

A comparative study of human sera was conducted to determine which purified preparations of 11 recombinant antigens of Borrelia burgdorferi sensu stricto were diagnostically most important in enzyme-linked immunosorbent assays (ELISAs). To assess sensitivity, 20 serum samples obtained 1-6 weeks after onset of illness from 20 persons who had physician-diagnosed erythema migrans (EM) were tested for IgM and IgG antibodies. In tests for IgM antibody, seropositivity of > or = 25% was recorded when ELISAs had separate preparations of protein (p) 37, p41-G, outer-surface protein (Osp) C, OspE, OspF or VlsE antigens.

Antibodies to granulocytic ehrlichiae in cattle from Connecticut.

Enzyme-linked immunosorbent assays (ELISA) with a purified recombinant 44-kDa protein and indirect fluorescent antibody (IFA) staining methods incorporating whole-cell antigens of the human granulocytic ehrlichiosis (HGE) agent were used to detect antibodies to Ehrlichia phagocytophila genogroup organisms in cattle sera. The cattle lived in tick-infested areas of Connecticut, USA and were healthy at the times blood samples were collected in 1990, 1999 and 2000. Of the 339 serum samples analysed, 40 (12%) and 15 (4%) were positive by ELISA and IFA, respectively.

West Nile virus envelope protein: role in diagnosis and immunity.

The role of antibodies to the West Nile virus envelope (E) protein in serodiagnosis and protection was examined. The E protein was expressed and purified in recombinant form. Antibodies to the E protein were detected in patients with West Nile virus infection.

Immunization of mice against West Nile virus with recombinant envelope protein.

West Nile (WN) virus is a mosquito-borne flavivirus that emerged in the United States in 1999 and can cause fatal encephalitis. Envelope (E) protein cDNA from a WN virus isolate recovered from Culex pipiens in Connecticut was expressed in Escherichia coli. The recombinant E protein was purified and used as Ag in immunoblot assays and immunization experiments.

Reactivity of dog sera to whole-cell or recombinant antigens of Borrelia burgdorferi by ELISA and immunoblot analysis.

Enzyme-linked immunosorbent assays (ELISAs) with separate preparations of 10 purified recombinant antigens of Borrelia burgdorferi sensu stricto were used to test sera from 36 dogs not vaccinated with whole cells of this agent and from five dogs vaccinated with whole-cell B. burgdorferi bacteria. All dogs lived in tick-infested areas of Connecticut and south-eastern New York state, USA.

Recombinant protein-44-based class-specific enzyme-linked immunosorbent assays for serologic diagnosis of human granulocytic ehrlichiosis.

Recombinant protein 44, expressed and purified as a maltose-binding protein fusion peptide of the human granulocytic ehrlichiosis (HGE) agent (Ehrlichia phagocytophila genogroup), was used as antigen in enzyme-linked immunosorbent assays (ELISAs) to detect total antibodies, immunoglobulin (Ig) M antibodies, and IgG antibodies. Of the 67 human sera obtained from 64 HGE patients 3-5 weeks after the onset of illness and confirmed as having total immunoglobulins to whole-cell antigen by indirect fluorescent antibody analyses, 63 were positive in a polyvalent ELISA. Fifty-six and 61 sera had IgM or IgG antibodies, respectively.

Reactivity of serum samples of dogs and horses tested by use of class-specific recombinant-based enzyme-linked immunosorbent assays for detection of granulocytic ehrlichiosis.

Objective: To test serum samples of dogs and horses by use of class-specific recombinant-based ELISA for establishing a diagnosis of granulocytic ehrlichiosis attributable to infection with organisms from the Ehrlichia phagocytophila genogroup.

Sample Population: Serum samples from 43 client-owned dogs and 131 horses (81 with signs of acute illness and 50 without signs of disease).

Procedure: Serum samples were analyzed, using ELISA with a recombinant 44-kd protein antigen for IgM and IgG antibodies to the human granulocytic ehrlichiosis (HGE) agent (NCH-1 strain).

Evaluation of a polyvalent enzyme-linked immunosorbent assay incorporating a recombinant p44 antigen for diagnosis of granulocytic ehrlichiosis in dogs and horses.

Objective: To develop and evaluate a polyvalent ELISA incorporating a highly specific recombinant antigen (p44) for diagnosis of granulocytic ehrlichiosis in dogs and horses.

Animals: 32 dogs and 43 horses.

Procedure: Results of the ELISA were compared with results of indirect fluorescent antibody (IFA) staining and western immunoblotting incorporating whole-cell antigen.

Detection of Ehrlichia chaffeensis DNA in Amblyomma americanum ticks in Connecticut and Rhode Island.

Ehrlichia chaffeensis, the causative agent of human monocytic ehrlichiosis, is transmitted by Amblyomma americanum ticks, which are most abundant in the southern United States. Because serologic evidence suggests that residents of Connecticut are exposed to E. chaffeensis, A.

Serologic confirmation of Ehrlichia equi and Borrelia burgdorferi infections in horses from the northeastern United States.

Objective: To determine whether horses living in tick-infested areas of northeastern United States with clinical signs of borreliosis or granulocytic ehrlichiosis had detectable serum antibodies to both Borrelia burgdorferi and Ehrlichia equi.

Design: Prospective study.

Animals: Serum samples from 51 clinically normal horses, 14 horses with clinical signs of borreliosis, and 17 horses with clinical signs of granulocytic ehrlichiosis.