Rapid Diagnosis of Babesia gibsoni by Point-of-Need Testing by Insulated Isothermal PCR in Dogs at High Risk of Infection.

BACKGROUND: Dogs seized by law enforcement agencies during dogfighting investigations are at increased risk of Babesia gibsoni infection. A rapid and cost-effective diagnostic test would increase the feasibility of mass screening of dogs for infection and monitoring treatment efficacy in B. gibsoni-infected dogs. OBJECTIVE: To determine the performance of a point-of-need insulated isothermal PCR (iiPCR) test for diagnosis of B. gibsoni in dogs rescued in dogfighting investigations. ANIMALS: Two hundred and thirty-three dogs seized in dogfighting investigations. METHODS: Cross-sectional study. Whole blood samples were tested for B. gibsoni and Babesia spp. by iiPCR. Results were compared to a reference standard comprised of concordant results from real-time PCR in a commercial diagnostic laboratory and antibody titers. RESULTS: The iiPCR system was quick to learn, portable, and had a short processing time of <2 hours. Sensitivity and specificity of the iiPCR assay for B. gibsoni were 90% (95% confidence interval [CI] 81-95%) and 99% (CI, 95-100%), respectively. Sensitivity and specificity of the iiPCR assay for Babesia spp. were 87% (CI, 78-93%) and 98% (CI, 0.94-99%), respectively. CONCLUSIONS AND CLINICAL IMPORTANCE: The iiPCR system produced few false-positive results, indicating that positive results are likely to represent true infections when used in high-risk animals. The iiPCR system can fail to identify 10-15% of truly infected dogs. However, the portability, speed, and economy of the iiPCR system compared to testing through a reference laboratory can allow rescue groups to screen and identify infection in more dogs.

Efficacy of Azithromycin and Compounded Atovaquone for Treatment of Babesia gibsoni in Dogs.

BACKGROUND: Approximately one-third of dogs confiscated during dogfighting investigations are infected with Babesia gibsoni. Traditional management of B. gibsoni with polymerase chain reaction (PCR)-screening, treatment with commercially available azithromycin and atovaquone, and PCR testing after 60 and 90 days is costly and impractical for large numbers of dogs at a time. HYPOTHESIS/OBJECTIVES: To assess the efficacy of an alternative protocol in which commercial atovaquone was replaced by compounded medication and PCR monitoring was initiated at 30 days after the end of treatment to decrease the total management time. METHODS: Prospective observational study. Forty-two pit bull-type dogs confiscated as part of an investigation of dogfighting, diagnosed with B. gibsoni infection, and judged to be suitable for adoption were treated with azithromycin (10 mg/kg PO q24h) and compounded atovaquone (13.4 mg/kg PO q8h with a fatty meal) for 10 days. PCR testing was repeated at 30 and 60 days after end of treatment if dogs with positive PCR tests at either time were tested at 90 days. Treatment was considered successful; 2 PCR tests 30 days apart were negative. RESULTS: Treatment was successful in 39 dogs (93%) as defined by 2 consecutive PCR-negative test results 30 days apart. In 38 dogs (90%), PCR results were the same at 30 and 60 days. CONCLUSIONS AND CLINICAL IMPORTANCE: Use of compounded atovaquone and a reduced monitoring period can reduce costs and holding times without compromising treatment efficacy. This more economical protocol can remove barriers to mass screening and management of B. gibsoni infections in dogfighting cases.

Infectious diseases in dogs rescued during dogfighting investigations.

Dogs used for dogfighting often receive minimal preventive health care, and the potential for spread of infectious diseases is high. The purpose of this study was to describe the prevalence of infectious diseases in dogs rescued from fighting operations to guide medical protocols for their immediate and long-term care. A total of 269 pit bull-type dogs were seized in a multi-state investigation. Fleas were present on most dogs, but few ticks were observed. Testing performed at intake included packed cell volume (PCV), serology and PCR for vector-borne pathogens, and fecal analysis. The most common infections were Babesia gibsoni (39%), 'Candidatus Mycoplasma haematoparvum' (32%), Mycoplasma haemocanis (30%), Dirofilaria immitis (12%), and Ancylostoma (23%). Anemia was associated with B. gibsoni infection (63% of infected dogs, odds ratio = 2.5, P <0.001), but not with hemotropic mycoplasmas or Ancylostoma. Pit bull heritage and dogfighting are known risk factors for B. gibsoni infection, possibly via blood transmission from bites and vertical transmission. Hemotropic mycoplasmas have a similar risk pattern. Empirical care for dogs from dogfighting cases should include broad-spectrum internal and external parasiticides and monitoring for anemia. Dogfighting case responders should be prepared for mass screening and treatment of B. gibsoni and heartworm infections and should implement protocols to prevent transmission of infectious and zoonotic diseases in the shelter and following adoption. Former fighting dogs and dogs with possible dog bite scars should not be used as blood donors due to the risk of vector-borne pathogens that can escape detection and for which curative treatment is difficult to document.

Infectious diseases in large-scale cat hoarding investigations.

Animal hoarders accumulate animals in over-crowded conditions without adequate nutrition, sanitation, and veterinary care. As a result, animals rescued from hoarding frequently have a variety of medical conditions including respiratory infections, gastrointestinal disease, parasitism, malnutrition, and other evidence of neglect. The purpose of this study was to characterize the infectious diseases carried by clinically affected cats and to determine the prevalence of retroviral infections among cats in large-scale cat hoarding investigations. Records were reviewed retrospectively from four large-scale seizures of cats from failed sanctuaries from November 2009 through March 2012. The number of cats seized in each case ranged from 387 to 697. Cats were screened for feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) in all four cases and for dermatophytosis in one case. A subset of cats exhibiting signs of upper respiratory disease or diarrhea had been tested for infections by PCR and fecal flotation for treatment planning. Mycoplasma felis (78%), calicivirus (78%), and Streptococcus equi subspecies zooepidemicus (55%) were the most common respiratory infections. Feline enteric coronavirus (88%), Giardia (56%), Clostridium perfringens (49%), and Tritrichomonas foetus (39%) were most common in cats with diarrhea. The seroprevalence of FeLV and FIV were 8% and 8%, respectively. In the one case in which cats with lesions suspicious for dermatophytosis were cultured for Microsporum canis, 69/76 lesional cats were culture-positive; of these, half were believed to be truly infected and half were believed to be fomite carriers. Cats from large-scale hoarding cases had high risk for enteric and respiratory infections, retroviruses, and dermatophytosis. Case responders should be prepared for mass treatment of infectious diseases and should implement protocols to prevent transmission of feline or zoonotic infections during the emergency response and when transferring the rescued cats to other shelters or to adopters.

Serological evidence of H3N8 canine influenza-like virus circulation in USA dogs prior to 2004.

H3N8 canine influenza virus (H3N8 CIV) was first reported as a novel canine respiratory pathogen in racing greyhounds and shelter dogs in the U.S.A. in 2004. Phylogenetic analyses determined that this host-adapted pathogen originated from interspecies transmission of an equine influenza virus (EIV), but it is unknown when the transmission occurred prior to discovery in 2004. The objective of this study was to determine if racing greyhound and shelter dog sera collected from 1984 to 2004 had serological evidence of exposure to H3N8 CIV or EIV. Archived sera from 702 racing greyhounds and 1568 shelter dogs were tested for H3 antibodies to the original 2004 CIV isolate, as well as EIV isolates from 1991 to 1999. None of the racing greyhounds from 1984 and 1985 had detectable H3 antibodies. One of the shelter dogs, which entered a north Florida shelter in 2004, was seropositive. For racing greyhounds sampled from 1999 to 2004, 133/520 (26%) dogs had antibodies to both CIV and EIV H3 proteins. The annual seroprevalence was 27% in 1999, 28% in 2000, 10% in 2001, 1% in 2002, 41% in 2003, and 28% in 2004. The odds of H3 seropositivity were greater among dogs that raced > or =6 months, raced on > or =2 tracks, and raced in 1998, 2002, and 2003. Many of the seropositive dogs raced at tracks that were involved in 'kennel cough' epidemics in 1998-1999 and 2002-2003. Based on serological evidence, a H3N8 canine influenza-like virus was circulating in racing greyhounds in the U.S.A. as early as 1999.

Effects of differential supplementation of fatty acids during the peripartum and breeding periods of Holstein cows: II. Neutrophil fatty acids and function, and acute phase proteins.

The objectives were to evaluate the effects of differential supplementation of Ca salts (CS) of fatty acids (FA) on plasma acute phase proteins and both FA composition and function (i.e., activity and cytokine production) of neutrophils, during the peripartum and breeding periods. Holstein cows were assigned randomly to receive either CS of palm (PO) or safflower (SO) oils from 30 d prepartum until 35 d postpartum (dpp) and CS of PO or fish oil (FO) from 35 to 160 dpp. Supplementation of CS of FA was at 1.5% of dietary dry matter. Cows (n=32) were sampled three times weekly from parturition to 35 dpp for analyses of plasma concentrations of haptoglobin and fibrinogen. Cows (n=47) were sampled for neutrophil phagocytic and oxidative burst activities toward Escherichia coli and Staphylococcus aureus, and neutrophil abundances of L-selectin and ß(2)-integrin assessed by flow cytometry at 32 d prepartum, within 7h after parturition, and 4 and 7 dpp. Profiles of FA in neutrophils and cytokine production (i.e., tumor necrosis factor alpha, TNF-α, and IL-1ß) were assessed prepartum (n=14), 35 (PO vs. SO; n=26) and 85 dpp (PO vs. FO; n=28). Plasma concentrations of haptoglobin and fibrinogen were greater for cows fed SO compared with PO. The percentage of neutrophils with phagocytic and oxidative burst activities was not affected by transition diets, but activities per neutrophil were greater in SO compared with PO diets at 4 (phagocytosis and oxidative burst) and 7 dpp (oxidative burst). Neutrophil abundance of L-selectin, but not ß(2)-integrin, was greater in SO compared with PO at 4 and 7 dpp. Neutrophil productions of TNF-α and IL-1ß were increased at 35 dpp in SO compared with PO diets, but production of TNF-α was attenuated in FO compared with PO at 85 dpp. Neutrophil ratios of n-6:n-3 FA were greater at 35 dpp in the SO diet and less at 85 dpp in FO compared with PO diets. In conclusion, cows supplemented with CS of SO had improved innate immunity (i.e., acute phase response and neutrophil function) to better cope with the bacterial challenges in the postpartum period. Conversely, CS of FO attenuated neutrophil cytokine production.

Canine H3N8 influenza virus infection in dogs and mice.

An H3N8 influenza virus closely related to equine influenza virus was identified in racing greyhound dogs with respiratory disease in 2004 and subsequently identified in shelter and pet dogs. Pathologic findings in dogs spontaneously infected with canine influenza virus were compared with lesions induced in beagle and mongrel dogs following experimental inoculation with influenza A/canine/Florida/43/2004. BALB/c mice were inoculated with canine influenza virus to assess their suitability as an experimental model for viral pathogenesis studies. All dogs inoculated with virus developed necrotizing and hyperplastic tracheitis and bronchitis with involvement of submucosal glands as well as mild bronchiolitis and pneumonia. Viral antigen was identified in bronchial and tracheal epithelial cells of all dogs and in alveolar macrophages of several dogs. Many dogs that were spontaneously infected with virus also developed bacterial pneumonia, and greyhound dogs with fatal spontaneous infection developed severe pulmonary hemorrhage with hemothorax. Virus-inoculated BALB/c mice developed tracheitis, bronchitis, bronchiolitis, and mild pneumonia in association with viral antigen in airway epithelial cells and in type 2 alveolar epithelial cells. Virus was not detected in extrarespiratory sites in any animals. The results indicate that canine influenza virus infection consistently induces acute tracheitis and bronchitis in dogs. Mice may be a useful model for some pathogenesis studies on canine influenza virus infection.

Differentiation of feline immunodeficiency virus vaccination, infection, or vaccination and infection in cats.

BACKGROUND: Serodiagnosis of feline immunodeficiency virus (FIV) is complicated by the use of a formalin-inactivated whole-virus FIV vaccine. Cats respond to immunization with antibodies indistinguishable from those produced during natural infection by currently available diagnostic tests, which are unable to distinguish cats that are vaccinated against FIV, infected with FIV, or both. HYPOTHESIS: An enzyme-linked immunosorbent assay (ELISA) detecting antibodies against formalin-treated FIV whole virus and untreated transmembrane peptide will distinguish uninfected from infected cats, regardless of vaccination status. ANIMALS: Blood samples were evaluated from uninfected unvaccinated cats (n = 73 samples), uninfected FIV-vaccinated cats (n = 89), and FIV-infected cats (n = 102, including 3 from cats that were also vaccinated). METHODS: The true status of each sample was determined by virus isolation. Plasma samples were tested for FIV antibodies by a commercial FIV diagnostic assay and an experimental discriminant ELISA. RESULTS: All samples from uninfected cats were correctly identified by the discriminant ELISA (specificity 100%). Of the samples collected from FIV-infected cats, 99 were correctly identified as FIV-infected (sensitivity 97.1%). CONCLUSIONS AND CLINICAL IMPORTANCE: With the exception of viral isolation, the discriminant ELISA is the most reliable assay for diagnosis of FIV. A practical strategy for the diagnosis of FIV infection would be to use existing commercial FIV antibody assays as screening tests. Negative results with commercial assays are highly reliable predictors for lack of infection. Positive results can be confirmed with the discriminant ELISA. If the discriminant ELISA is negative, the cat is probably vaccinated against FIV but not infected. Positive results are likely to represent infection.

Infectious diseases of dogs and cats on Isabela Island, Galapagos.

BACKGROUND: Vaccination and importation of dogs and cats are prohibited in the Galapagos, resulting in a uniquely isolated population. The purpose of this study was to determine the prevalence of infectious diseases of dogs and cats that impact their health, could spill over to native wildlife, or sentinel diseases of concern to humans. HYPOTHESIS: The isolation of dogs and cats in the Galapagos protects them from diseases common in mainland populations. ANIMALS: Ninety-five dogs and 52 cats presented during a neutering campaign. METHODS: A prospective cross-sectional study was performed. Blood was collected for serological and DNA evaluation of a panel of infectious diseases. RESULTS: Antibodies against parvovirus (100%), parainfluenza virus (100%), adenovirus 1/2 (66-67%), and distemper virus (22%) were present in dogs. Dirofilaria immitis was also common in dogs (34%), with lower prevalences of Wolbachia pipiens (22%), Bartonella sp. (13%), Ehrlichia/Anaplasma spp. (1%), and Mycoplasma haemocanis (1%) observed. Antibodies against panleukopenia virus (67%), Toxoplasma gondii (63%), calicivirus (44%), and herpesvirus 1 (10%) were detected in cats. Feline leukemia virus antigen, feline immunodeficiency virus antibody, or coronavirus antibodies were not detected. Bartonella sp. (44%) infections were common in cats, but only one was infected with M. haemofelis. CONCLUSIONS AND CLINICAL IMPORTANCE: Despite their relative seclusion from the rest of the world, cats and dogs of Isabela were exposed to many pathogens found in mainland South America. Parasite prophylaxis, neutering, and strict enforcement of animal movement restrictions would control a majority of the diseases. In the absence of vaccination, a reservoir of susceptible animals remains vulnerable to new disease introductions.

Transmission of equine influenza virus to dogs.

Molecular and antigenic analyses of three influenza viruses isolated from outbreaks of severe respiratory disease in racing greyhounds revealed that they are closely related to H3N8 equine influenza virus. Phylogenetic analysis indicated that the canine influenza virus genomes form a monophyletic group, consistent with a single interspecies virus transfer. Molecular changes in the hemagglutinin suggested adaptive evolution in the new host. The etiologic role of this virus in respiratory disease was supported by the temporal association of rising antibody titers with disease and by experimental inoculation studies. The geographic expansion of the infection and its persistence for several years indicate efficient transmission of canine influenza virus among greyhounds. Evidence of infection in pet dogs suggests that this infection may also become enzootic in this population.

Suppurative rhinitis associated with Haemophilus species infection in a cat.

A young cat with signs of chronic rhinitis was evaluated for underlying anatomical, inflammatory, or infectious disease. Initial diagnostics were significant for the isolation of an unusual pathogen, Haemophilus species. Isolation using a human RapID NH system erroneously identified the isolate as H. segnis, a human pathogen. No database of veterinary pathogens (Haemophilus) are included in the system and animal pathogens will either be erroneously identified or yield a unique biocode not listed. Because of the unique nature of the pathogen we explored the possibility of immunosuppression as a contributory factor to infection. A variety of laboratory tests were employed to evaluate immune function. The clinical indications and utility of immune function testing are discussed. No immune dysfunction was identified.

Use of adult cat serum to correct failure of passive transfer in kittens.

OBJECTIVE: To evaluate the use of adult cat serum as an immunoglobulin supplement in kittens with failure of passive transfer. DESIGN: Randomized controlled study. ANIMALS: 11 specific pathogen-free queens and their 43 kittens. PROCEDURE: Kittens were removed from the queens at birth, prior to suckling colostrum, and randomly assigned to 1 of 4 groups: colostrum-deprived, colostrum-fed, colostrum-deprived and administration of pooled adult cat serum i.p., and colostrum-deprived and administration of pooled adult serum s.c.. Colostrum-fed kittens were returned to the queen and allowed to suckle normally. Colostrum-deprived kittens were isolated from the queen and fed a kitten milk replacer for 2 days to prevent absorption of colostral IgG. All colostrum-deprived kittens were returned to the queen on day 3. Serum IgG concentrations were measured by radial immunodiffusion in the kittens at birth and 2 days and 1, 2, 4, 6, and 8 weeks after birth. RESULTS: None of the kittens had detectable serum IgG at birth. Both i.p. and s.c. administration of adult cat serum resulted in peak serum IgG concentrations equivalent to those in kittens that suckled normally. Untreated colostrum-deprived kittens did not achieve serum IgG concentrations comparable to those for kittens in the other groups until 6 weeks of age. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that adult cat serum may be used as an immunoglobulin supplement in colostrum-deprived kittens. Although the minimum concentration of IgG necessary to protect kittens from infection is unknown, concentrations achieved were comparable to those in kittens that suckled normally.

Tissue dynamics of CD8 lymphocytes that suppress viral replication in cats infected neonatally with feline immunodeficiency virus.

The purpose of this study was to determine the tissue distribution and antiviral activity of the CD8 lymphocytes that suppress the replication of feline immunodeficiency virus (FIV). Cell-associated FIV load, CD8alpha(+)beta(low) cells, and CD8 cell-mediated suppression of FIV were measured serially in the blood, thymus, and peripheral lymph nodes after neonatal inoculation. Between 6 and 10 weeks, relative numbers of CD8alpha(+)beta(low) cells increased, whereas CD8alpha(+)beta(high) cells declined in the thymus and blood of infected cats. By 12-16 weeks, the lymph nodes were enlarged because of an absolute expansion of all CD8beta subpopulations. The strength of CD8 cell-mediated FIV suppression in vitro, but not CD8alpha(+)beta(low) cell content, was correlated inversely with virus load in the thymus and blood. Thus, after neonatal FIV inoculation, CD8alpha(+)beta(low) cells first occupy the thymus and blood, where strong CD8 cell-mediated antiviral activity is linked to reduced virus load in multiple lymphoid tissues.

Unique susceptibility of the fetal thymus to feline immunodeficiency virus infection: an animal model for HIV infection in utero.

PROBLEM: Human infants infected in utero with HIV develop thymus insufficiency and progress to AIDS sooner than infants infected peripartum. However, direct analysis of the thymus is difficult due to limited tissue access and variable timing of vertical transmission. METHOD OF STUDY: Fetal and neonatal cats were inoculated with feline immunodeficiency virus (FIV) at an equivalent infectious dose. The thymus, blood, and lymph nodes were harvested and compared at 23 and 46 days post-inoculation (p.i.) and also compared to sham-inoculated, age-matched controls. Lymphocyte phenotypes were analyzed by flow cytometry and virus burden was quantified in histologic sections and by virus isolation from plasma. RESULTS: Fetal cats inoculated with FIV had acute thymus atrophy at birth, which coincided with peak viremia. At 46 days p.i., thymus size and cell composition rebounded and supported increased productive infection. In contrast, neonatal cats inoculated with FIV developed chronic thymus atrophy and degeneration, which was associated with decreasing productive infection and low-level viremia. CONCLUSIONS: The fetal thymus is uniquely vulnerable to acute, transient depletion and high-level productive infection. The neonatal thymus is less vulnerable to acute changes, and responds through progressive atrophy and declining productive infection. Reduced immune competence, as reflected by the failure to control virus replication, may contribute to the accelerated progression of FIV and HIV infections in utero.

Thymic lesions in cats infected with a pathogenic molecular clone or an ORF-A/2-deficient molecular clone of feline immunodeficiency virus.

Previous studies using feline immunodeficiency virus (FIV) molecular clones lacking the putative transactivator gene (ORF-A/2) failed to address the issue of thymus pathogenesis or investigate the levels of viral replication in separate lymphoid compartments (Y. Inoshima, et al., J. Virol. 70:8518-8526, 1996; E. E. Sparger, et al., Virology 205:546-553, 1994). Using a highly pathogenic molecular clone of FIV, JSY3, and an ORF-A/2-deficient mutant, JSY3DeltaORF-A/2, we compared viral replication and the extent of thymic dysfunction as measured by the formation of lymphoid follicles and alteration of the thymocyte subsets. Viral replication was reduced in JSY3DeltaORF-A/2-infected cats as measured by lymphocyte coculture, immunohistochemistry, and quantitative PCR. Cell-associated viral load measured by lymphocyte coculture varied in a tissue-dependent manner with replication highest in lymphocytes isolated from the thymus, lower in those from the peripheral blood, and lowest in those from lymph node. Thymic proviral load and the number of viral p24 Gag-positive cells within the thymus detected by immunohistochemistry were also reduced. In addition, the onset of a reduced peripheral blood CD4/CD8 ratio was delayed in JSY3DeltaORF-A/2-infected cats. The formation and extent of thymic lymphoid follicular hyperplasia were similar in JSY3 and JSY3DeltaORF-A/2-infected cats as measured by anticytokeratin immunohistochemistry and flow cytometry for percent pan T-negative, immunoglobulin G-positive cells within the thymus. In contrast, comparison of thymocyte subpopulations demonstrated a reduced expansion of single-positive CD4(-) CD8(+) thymocytes in JSY3DeltaORF-A/2-infected cats. Level of viral replication, therefore, may not correlate with the formation of thymic lymphoid follicles but may correlate with the expansion of the single-positive CD4(-) CD8(+) thymocyte subpopulation.

CD8+ thymic lymphocytes express reduced levels of CD8beta and increased interferon gamma in cats perinatally infected with the JSY3 molecular clone of feline immunodeficiency virus.

Biological isolates of feline immunodeficiency virus (FIV) cause a relative expansion of activated single-positive CD8(+) (SP CD8(+)) lymphocytes within the thymus of infected cats. In this study, thymic SP CD8(+) lymphocytes were analyzed from cats inoculated as neonates with a pathogenic molecular clone of FIV, JSY3, which was previously derived from the wild-type biological isolate FIV(NCSU-1) (NCSU-1). Four cats were inoculated intraperitoneally with NCSU-1 and compared with 11 cats inoculated with JSY3. Five control cats matched in litter and age were administered an intraperitoneal sham inoculum. Between 12 and 16 weeks postinoculation, interferon-gamma (IFN-gamma) mRNA was quantified by RT-PCR in freshly isolated thymocytes and peripheral blood mononuclear cells (PBMCs). The quantity of IFN-gamma mRNA was increased more than 10-fold in thymocytes and PBMCs of 13 of 13 FIV-inoculated cats as compared with the sham-inoculated controls. IFN-gamma mRNA coenriched with magnetically sorted CD8(+) PBMCs and single-positive (SP) CD8(+) thymocytes. Cells expressing IFN-gamma mRNA were located within the thymic perivascular zone, along the corticomedullary junction, and adjacent to lymphoid follicles. The expansion of thymic SP CD8(+) cells was associated with an increase in CD8alpha(+)/beta(neg) and CD8alpha(+)/beta(lo) phenotypes, the latter population resembling a previously reported memory/effector peripheral blood cell with FIV suppressor activity. From these data we conclude that JSY3 and NCSU-1 induce similar phenotypic changes in thymic and peripheral blood CD8(+) cells. Thus, JSY3 is pathogenic for the thymus in vivo and will be useful for defining determinants of the CD8(+) cell response in this pediatric AIDS model.

Cardiopulmonary perfusion.

Open heart surgery is one of the most highly technical of all modern medical techniques, and includes procedures such as coronary artery bypass grafting, cardiac valve repair or replacement, correction of congenital defects, resection of aneurysms, ablation of abnormal pathways of conduction, etc. It relies on the coordinated interaction of a heart surgeon, an anesthesiologist, several nurses and technicians, and a perfusionist. The first successful open heart surgery was performed in Philadelphia forty years ago by Dr. John Gibbon, Jr., whose wife, Mary, was his perfusionist. This historical landmark came after two decades of laboratory exploration and perfection of their extracorporeal circuit and its ability to sustain life. Perfusion, the technology which has evolved from those groundbreaking discoveries, controls, supports and maintains the circulation by application of extracorporeal devices. During open-heart surgery, perfusion (cardiopulmonary bypass - CPB) supplants the functions of the heart and lungs to provide the surgeon with a still, dry operating field. Today, this highly specialized role is performed by individuals conversant in a variety of scientific modalities working in close communication and cooperation with the surgeon. Perfusionists understand the anatomy, pathology, and physiology of the patient, while administering medications, anesthetics, blood, blood components and blood substitutes. Simultaneously, they operate a highly sophisticated electromechanical device to substitute for the human heart and lungs. Today's perfusionists know and utilize aspects of varied pursuits which include a functional comprehension of machines and motors, electronics and electrical safety, plastics and biocompatibility, drugs and pharmacology, blood and its components, hemodynamics and fluid dynamics, hypothermia and hyperthermia, gas exchange and metabolism, electrolytes and blood compatibility, anticoagulation and anesthesia. The logarithmic expansion in these unrelated fields of study have enhanced our ability to provide patient care.

Modulation of Actinomyces viscosus colonization of mouse teeth in vivo by immunization with fimbrial adhesins.

Experiments were performed to determine whether immunization of mice with fimbrial adhesins isolated from Actinomyces viscosus T14V could modulate infection of tooth surfaces in animals challenged with the homologous strain. Saliva and sera from animals immunized in the submandibular gland region contained elevated levels of fimbria-specific immunoglobulin A (IgA) and IgG, whereas saliva and sera from sham-immunized animals did not. There was a statistically significant inverse correlation between the presence of fimbria-specific antibodies in saliva and serum and the levels of bacterial colonization on molar tooth surfaces. These results suggest that fimbrial adhesins may effectively modulate infection of tooth surfaces by periodontopathic bacteria.

Fimbria-specific antibodies in serum and saliva of mice immunized with Actinomyces viscosus T14V fimbriae.

Fimbria-specific antibody responses were compared in mice immunized with purified fimbrial adhesins in the region of the submandibular gland (i.e., local site) or at a remote site in the back. One hundred micrograms of fimbriae isolated from Actinomyces viscosus T14V was used as the vaccine. Four subcutaneous injections of the vaccine in the local site induced greater amounts of fimbria-specific immunoglobulin G (IgG) in serum and saliva than three injections. However, there was no difference in the response of fimbria-specific IgA in serum and saliva. Fimbria-specific IgG in serum and saliva were first detected 21 days after the primary immunization at both the local or remote sites. Fimbria-specific IgA in serum was first detected 28 days after the primary immunization at both the local or remote sites. However, fimbria-specific IgA in saliva occurred only in mice immunized with the fimbrial vaccine at the local site and was first detected 14 days after the primary immunization. Both serum and saliva from mice immunized 4 times with the fimbrial vaccine in the local site inhibited in vitro adsorption of strain T14V cells to hydroxyapatite beads pretreated with normal mouse saliva, whereas adsorption of strain T14V cells suspended in serum and saliva from sham-immunized animals was not inhibited. Collectively, these data suggest that mice immunized locally in the submandibular gland region with a vaccine composed of purified fimbrial adhesins provide a potential model for evaluating the efficacy of fimbria-specific antibodies in saliva to inhibit strain T14V colonization of tooth surfaces.