Rabbits infested with either Ornithodoros sp. ticks or Psoroptes cuniculi mites were assayed for ... more Rabbits infested with either Ornithodoros sp. ticks or Psoroptes cuniculi mites were assayed for anti-tick antibody by enzyme-linked immunosorbent assay. Titration of rabbit serum against Ornithodoros sp. salivary gland extract (SGE) demonstrated both mite- and tick-infested animals to have elevated anti-tick antibody titers. Western blot analysis demonstrated the anti-mite and anti-tick antisera to contain antibodies with affinities for both common and unique subsets of Ornithodoros SGE proteins.
Between 5 and 8 million people globally are infected with Trypanosoma cruzi, the causative parasi... more Between 5 and 8 million people globally are infected with Trypanosoma cruzi, the causative parasitic agent of Chagas disease. The vast majority of incident infections originate in pockets of Latin America where domestic vector-borne transmission cycles are more common. Since 1955, when the first locally-acquired case was reported, fewer than 30 autochthonous cases have been documented in the United States. We describe the case of an 18-year-old US Air Force trainee, a native Texan with no travel history beyond the continental United States, who screened positive for T cruzi infection on blood donation and was subsequently found to have chronic Chagasic cardiomyopathy. This is the first documented case of Chagas disease in a US military trainee and one of the first known autochthonous cases of Chagasic cardiomyopathy in a Texas resident. Diagnostic, therapeutic, and military implications are discussed.
The pathogenesis of bovine pneumonic pasteurellosis is not completely understood, and studies hav... more The pathogenesis of bovine pneumonic pasteurellosis is not completely understood, and studies have not established that Pasteurella haemolytica A1 (Ph1) virulence is exclusively responsible for the development of acute pulmonary lesions. The purpose of this investigation was to determine if immune complex disease is involved in the pathogenesis of bovine pneumonic pasteurellosis. A retrospective immunohistologic study of lung tissue from natural cases of bovine pneumonic pasteurellosis (44) as performed, and immune complexes were observed in alveloar spaces and walls in 88% of these cases. To study this pathologic mechanism experimentally, groups of mice were immunized with purified Ph1 outer membranes (OMs) or sham immunized on days 0 and 14. Mice were challenged intratracheally on day 24 with either live Ph1 or Ph1 OMs, and pulmonary lesions were assessed 24 h after challenge. Placebo immunized mice developed focal infiltrates of neutrophils and macrophages centered around large caliber bronchi. Mice immunized with Ph1 OMs and challenged with live Ph1 or OMs developed severe bronchointerstitial pneumonia with diffuse neutrophilic infiltration, focal necrosis, hemorrhage and edema, that is histologically similar to bovine pneumonic pasteurellosis. Immunohistology revealed flocculent aggregates of IgG and complement positive material within alveolar spaces and walls from mice challenged with live Ph1, and fine granular deposits of IgG and complement positive material were observed lining the alveolar walls from mice challenged with Ph1 OMs. Immunized mice exhibited high serum IgG antibody titers to Ph1 outer membrane proteins (OMPs). Results of this study suggest that immune complex disease plays a role in the pathogenesis of bovine pneumonic pasteurellosis.
... 1967. The life history of Hepatozoon rarefaciens (Sambon and Seligmann, 1907) from Drymarchon... more ... 1967. The life history of Hepatozoon rarefaciens (Sambon and Seligmann, 1907) from Drymarchon cor?is (Co lubridae), and its experimental transfer to Constrictor constrictor (Boidae). J. Parasitol. 53: 897-909. 2. Barta, JR 1989. ... Pp. 83-89. 5. Frye,FL 1991. ...
The goals of this study were to explore the range of vectors of reptilian hemosporozoa, to charac... more The goals of this study were to explore the range of vectors of reptilian hemosporozoa, to characterize pathologic and immunologic aspects of hemogregarine-unnatural host relationships, and to devise a molecular assay to type isolates. These studies are important in zoological medicine because of the high potential for inter-specific transmission between captive specimens. Surveys of wild and captive colubrid, boid, and viperid snakes by blood film examination demonstrated a high prevalence of hemogregarine infections. Blood stages were consistent with hemogregarine gametocytes, with morphometrically similar, non-dividing, fusiform, intraerythrocytic protozoa with single round compact nuclei. Erythrocytic infection was associated with variable hypertrophy, alterations in sedimentation coefficients, cytoplasmic pallor, and erythrocytic protein depletion. Hemogregarine parasites from several colubrid and viperid snakes completed sporogony within Aedes aegypti mosquitoes in 14-18 days, and produced oocysts typical of Hepatozoon sp. All lizards experimentally inoculated per os with mosquito-borne oocysts developed Hepatozoon sp. infections. No sporogonic development was demonstrated in any Ornithodoros turicata ticks (Argasidae) fed on parasitemic snakes. None of the lizards inoculated by tick developed Hepatozoon sp. infections. Molecular characterization of several hemogregarine isolates, two lizard Plasmodium species, and several species of Leishmania and Trypanosoma at the ribosomal DNA (rDNA) locus by PCR amplification and restriction enzyme analysis demonstrated detectable molecular differences between genera, species and isolates. Within unnatural lizard hosts multifocal accumulations of heterophilic leukocytes with intralesional Hepatozoon meronts were observed within the liver and lungs. The accompanying clinicopathologic changes in unnatural hosts included a marked leukocytosis with heterophilia, increases in plasma liver enzyme activities, and mild increases in the serum gamma globulin fraction. No meront-associated inflammation or significant clinico-pathologic changes were noted in naturally infected snakes despite numerous pulmonary and hepatic meronts. Assay of unnatural host serum for anti-Hepatozoon antibody by ELISA, IFA, and Western blot demonstrated post-infection increases in lizards sampled at 38 days post-infection and beyond. Stage-specific antibody directed against surface antigens of infected erythrocytes with a moderate orthochromatic erythroblastosis, was demonstrated only in parasitemic lizards. Molecularly distinct hemogregarines in different snake species and evidence of parasite-associated disease in unnatural hosts, suggest host-parasite coadaptation in nature
New Zealand White rabbits were repeatedly infested with Ornithodoros turicata (Duges), Ornithodor... more New Zealand White rabbits were repeatedly infested with Ornithodoros turicata (Duges), Ornithodoros talaje (Guérin-Méneville), and Ornithodoros coriaceus (Koch) at 2-wk intervals. Blood samples were taken from each animal 10 d after each infestation and the titer of anti-tick antibody was determined by enzyme-linked immunosorbent assay. Subsequent cross-reactivity studies demonstrated that the antitick antisera nonspecifically bound to salivary gland extract proteins prepared from several other tick genera and species' Amblyomma maculatum (Koch), Dermacentor andersoni (Stiles), Dermacentor variabilis (Say), and Ornithodoros moubata (Murray). Absorption of the antisera against an immobilized extract of A. maculatum substantially increased specificity at the genus level. Western blots of electrophoretically separated Ornithodoros salivary gland extract samples were used to further compare the specificity of absorbed and nonabsorbed antitick antisera. The blots demonstrated that many of the Ornithodoros salivary gland extract proteins bear genus specific epitopes. Some differences were noted among the Ornithodoros species examined with respect to the degree of antigenic relatedness with the ixodid ticks.
International Journal for Parasitology, Sep 1, 1991
Haemogregarine parasites, derived from the Florida snakes Coluber constrictor and Nerodia fasciat... more Haemogregarine parasites, derived from the Florida snakes Coluber constrictor and Nerodia fasciata and ingested by Aedes aegypti, completed sporogony within the hemocoeles of nearly all fed mosquitoes in 14-18 days, and produced oocysts typical of Hepatozoon. However, mortalities and morbidity were high in the Culex which had fed on the Coluber. Oocysts were not found in any Ornithodoros turicata (Argasidae) which fed upon either snake host, but many sections of fed ticks had gametocyte-like cells within the gut lumen. Most lizards, Anolis carolinensis and Anolis sagrei, infected per os with oocysts derived from both snake species developed infections. Infections in the lizards were largely confined to hepatic schizonts with few parasites found in erythrocytes. Unlike naturally infected snake hosts, Hepatozoon schizonts in livers of lizards were often either surrounded by an unidentified dark pigment or heavily infiltrated with mononuclear inflammatory cells.
The host antibody response of CD1 mice resulting from sequential exposures to two species of Orni... more The host antibody response of CD1 mice resulting from sequential exposures to two species of Ornithodoros ticks was isotyped (ELISA) in an effort to determine the mechanism for the development of acquired resistance. In addition, salivary gland proteins from two argasid and one ixodid species were examined (RIA) for cross-reactivity. Significant antibody responses, detectable for at least 90 d after last exposure, were shown to occur. Isotyping showed that the initial response was of the IgM class with a secondary class switch to the IgG1 subclass occurring. Evidence that cross-reactive proteins exist between argasid and ixodid ticks is also discussed.
The purpose of this study was to immunologically characterize soft tick salivary antigens. BALB/c... more The purpose of this study was to immunologically characterize soft tick salivary antigens. BALB/c mice hyperimmunized with salivary gland extract prepared from Ornithodoros talaje (Guérin-Méneville) were observed to develop high titers of antitick salivary antigen antibodies. Subsequent fusion of splenic lymphocytes from the hyperimmunized mice with SP-2/0-AG14 myeloma cells resulted in the production of 10 antitick IgM-producing hybridoma clones. Partial characterization of the respective tick antigens by gel filtration and SDS-PAGE demonstrated all 10 monoclonal antitick antibodies to be reactive with a salivary gland extract fraction containing proteins 50-110 kDa in molecular weight. Cross-reactivity assays and electrophoretic comparison of salivary gland extract specimens demonstrated similar proteins in several ixodid tick genera and species.
<p>The asterisk and arrowhead represent the molecular masses for native GlpQ and BipA, resp... more <p>The asterisk and arrowhead represent the molecular masses for native GlpQ and BipA, respectively (A). rBipA was produced as a thioredoxin fusion protein and is 15 kDa larger than the native protein. Immunoblots were also probed with a serum sample from an uninfected patient (B) and a monoclonal antibody against the six-histidine fusion tag (C and D). Molecular masses are shown to the left of each immunoblot.</p
Journal of Herpetological Medicine and Surgery, 2000
ABSTRACT The common snake mite, Ophionyssus natricis (Parasitiformes: Macronyssidae), is a blood ... more ABSTRACT The common snake mite, Ophionyssus natricis (Parasitiformes: Macronyssidae), is a blood feeding, mesostigmatid mite that parasitizes reptiles. Anemia, dehydration, dermatitis, and several blood-borne infectious diseases have been linked to infestations (Camin, 1948Camin, 1953, Chiodini, et al, 1983). Severe pruritic dermatitis has been reported in humans bitten by Ophionyssus natricis (Schultz, 1975, Beck, 1996). All levels of Ophionyssus natricis infestation should be considered a serious problem worthy of prompt treatment. This article describes the general morphology, biology, and behavior of each life stage and discusses the components of an integrated control and prevention program for the maintenance of mite-free snake collections.
Journal of Herpetological Medicine and Surgery, 2000
ABSTRACT A group of seven juvenile California mountain kingsnakes, Lampropeltis zonata multicinct... more ABSTRACT A group of seven juvenile California mountain kingsnakes, Lampropeltis zonata multicincta, developed severe acute gastroenteritis characterized by emesis, dehydration and sudden death. Histopathology demonstrated segmental mucosal necrosis and hyperplasia, villus blunting and fusion. Abundant basophilic Cowdry type-A intranuclear inclusions were demonstrated in enterocytes peripheral to the zone of acute necrosis throughout the small intestine. Electron microscopy demonstrated intranuclear crystalline arrays of non-enveloped virus-like particles that measured 65 to 70 nm in diameter. Clusters of smaller non-enveloped virus-like particles that measured 15 to 18 nm in diameter were found in association with the larger particles. The size, morphology and location of the two virus-like particles were consistent with an adenovirus and a parvovirus. The presence of the parvovirus-like particles in the same nuclei and in association with adenovirus-like particles suggested the parvovirus-like agent to b...
The pathogenesis of bovine pneumonic pasteurellosis is not completely understood, and studies hav... more The pathogenesis of bovine pneumonic pasteurellosis is not completely understood, and studies have not established that Pasteurella haemolytica A1 (Ph1) virulence is exclusively responsible for the development of acute pulmonary lesions. The purpose of this investigation was to determine if immune complex disease is involved in the pathogenesis of bovine pneumonic pasteurellosis. A retrospective immunohistologic study of lung tissue from natural cases of bovine pneumonic pasteurellosis (44) as performed, and immune complexes were observed in alveloar spaces and walls in 88% of these cases. To study this pathologic mechanism experimentally, groups of mice were immunized with purified Ph1 outer membranes (OMs) or sham immunized on days 0 and 14. Mice were challenged intratracheally on day 24 with either live Ph1 or Ph1 OMs, and pulmonary lesions were assessed 24 h after challenge. Placebo immunized mice developed focal infiltrates of neutrophils and macrophages centered around large caliber bronchi. Mice immunized with Ph1 OMs and challenged with live Ph1 or OMs developed severe bronchointerstitial pneumonia with diffuse neutrophilic infiltration, focal necrosis, hemorrhage and edema, that is histologically similar to bovine pneumonic pasteurellosis. Immunohistology revealed flocculent aggregates of IgG and complement positive material within alveolar spaces and walls from mice challenged with live Ph1, and fine granular deposits of IgG and complement positive material were observed lining the alveolar walls from mice challenged with Ph1 OMs. Immunized mice exhibited high serum IgG antibody titers to Ph1 outer membrane proteins (OMPs). Results of this study suggest that immune complex disease plays a role in the pathogenesis of bovine pneumonic pasteurellosis.
Rabbits infested with either Ornithodoros sp. ticks or Psoroptes cuniculi mites were assayed for ... more Rabbits infested with either Ornithodoros sp. ticks or Psoroptes cuniculi mites were assayed for anti-tick antibody by enzyme-linked immunosorbent assay. Titration of rabbit serum against Ornithodoros sp. salivary gland extract (SGE) demonstrated both mite- and tick-infested animals to have elevated anti-tick antibody titers. Western blot analysis demonstrated the anti-mite and anti-tick antisera to contain antibodies with affinities for both common and unique subsets of Ornithodoros SGE proteins.
The objective of this study was to develop a highly specific enzyme-linked immunosorbent assay (E... more The objective of this study was to develop a highly specific enzyme-linked immunosorbent assay (ELISA) for the serological detection of anti-Ornithodoros tick antibodies in animals. Affinity-purified rabbit anti-Ornithodoros IgG antibodies were employed in indirect competitive inhibition ELISA assays designed to measure the anti-Ornithodoros antibody titers in other animal species using the domestic goat (Capra hircus) as a large animal model. Repeated infestation of goats with Ornithodoros coriaceus was found to elicit the formation of antibodies capable of inhibiting the binding of the Ornithodoros-specific rabbit IgG. Western blot analysis of goat and rabbit anti-tick antisera demonstrated both animal species to respond immunologically to a set of 9 major protein bands in O. coriaceus salivary gland extracts. The results of these experiments demonstrate that a history of animal exposure to O. coriaceus may be detected serologically by competitive inhibition ELISA.
Ophionyssus natricis (Parasitiformes: Macronyssidae) is a mesostigmatid mite which commonly paras... more Ophionyssus natricis (Parasitiformes: Macronyssidae) is a mesostigmatid mite which commonly parasitizes captive snakes. The mites feed on the host's blood by lacerating the soft skin between scales with their chelicerae. Heavy infestations are common in captive specimens and have been shown to result in severe anemia, dehydration, and death. Additionally, the mites have been shown to transmit several bacterial, protozoal, and filariid pathogens. While well recognized as a significant pest in captive snake collections, the snake mite has been difficult to control. The inability to control O. natricis is related to its complex life cycle, which includes both blood feeding and non-feeding stages. In order to develop a proper treatment regimen for O. natricis, it is critical to understand the natural history of the organism. This paper will describe the life cycle and ethology of O. natricis and summarize current therapies utilized in controlling and attempting to eradicate this pest.
Rabbits infested with either Ornithodoros sp. ticks or Psoroptes cuniculi mites were assayed for ... more Rabbits infested with either Ornithodoros sp. ticks or Psoroptes cuniculi mites were assayed for anti-tick antibody by enzyme-linked immunosorbent assay. Titration of rabbit serum against Ornithodoros sp. salivary gland extract (SGE) demonstrated both mite- and tick-infested animals to have elevated anti-tick antibody titers. Western blot analysis demonstrated the anti-mite and anti-tick antisera to contain antibodies with affinities for both common and unique subsets of Ornithodoros SGE proteins.
Between 5 and 8 million people globally are infected with Trypanosoma cruzi, the causative parasi... more Between 5 and 8 million people globally are infected with Trypanosoma cruzi, the causative parasitic agent of Chagas disease. The vast majority of incident infections originate in pockets of Latin America where domestic vector-borne transmission cycles are more common. Since 1955, when the first locally-acquired case was reported, fewer than 30 autochthonous cases have been documented in the United States. We describe the case of an 18-year-old US Air Force trainee, a native Texan with no travel history beyond the continental United States, who screened positive for T cruzi infection on blood donation and was subsequently found to have chronic Chagasic cardiomyopathy. This is the first documented case of Chagas disease in a US military trainee and one of the first known autochthonous cases of Chagasic cardiomyopathy in a Texas resident. Diagnostic, therapeutic, and military implications are discussed.
The pathogenesis of bovine pneumonic pasteurellosis is not completely understood, and studies hav... more The pathogenesis of bovine pneumonic pasteurellosis is not completely understood, and studies have not established that Pasteurella haemolytica A1 (Ph1) virulence is exclusively responsible for the development of acute pulmonary lesions. The purpose of this investigation was to determine if immune complex disease is involved in the pathogenesis of bovine pneumonic pasteurellosis. A retrospective immunohistologic study of lung tissue from natural cases of bovine pneumonic pasteurellosis (44) as performed, and immune complexes were observed in alveloar spaces and walls in 88% of these cases. To study this pathologic mechanism experimentally, groups of mice were immunized with purified Ph1 outer membranes (OMs) or sham immunized on days 0 and 14. Mice were challenged intratracheally on day 24 with either live Ph1 or Ph1 OMs, and pulmonary lesions were assessed 24 h after challenge. Placebo immunized mice developed focal infiltrates of neutrophils and macrophages centered around large caliber bronchi. Mice immunized with Ph1 OMs and challenged with live Ph1 or OMs developed severe bronchointerstitial pneumonia with diffuse neutrophilic infiltration, focal necrosis, hemorrhage and edema, that is histologically similar to bovine pneumonic pasteurellosis. Immunohistology revealed flocculent aggregates of IgG and complement positive material within alveolar spaces and walls from mice challenged with live Ph1, and fine granular deposits of IgG and complement positive material were observed lining the alveolar walls from mice challenged with Ph1 OMs. Immunized mice exhibited high serum IgG antibody titers to Ph1 outer membrane proteins (OMPs). Results of this study suggest that immune complex disease plays a role in the pathogenesis of bovine pneumonic pasteurellosis.
... 1967. The life history of Hepatozoon rarefaciens (Sambon and Seligmann, 1907) from Drymarchon... more ... 1967. The life history of Hepatozoon rarefaciens (Sambon and Seligmann, 1907) from Drymarchon cor?is (Co lubridae), and its experimental transfer to Constrictor constrictor (Boidae). J. Parasitol. 53: 897-909. 2. Barta, JR 1989. ... Pp. 83-89. 5. Frye,FL 1991. ...
The goals of this study were to explore the range of vectors of reptilian hemosporozoa, to charac... more The goals of this study were to explore the range of vectors of reptilian hemosporozoa, to characterize pathologic and immunologic aspects of hemogregarine-unnatural host relationships, and to devise a molecular assay to type isolates. These studies are important in zoological medicine because of the high potential for inter-specific transmission between captive specimens. Surveys of wild and captive colubrid, boid, and viperid snakes by blood film examination demonstrated a high prevalence of hemogregarine infections. Blood stages were consistent with hemogregarine gametocytes, with morphometrically similar, non-dividing, fusiform, intraerythrocytic protozoa with single round compact nuclei. Erythrocytic infection was associated with variable hypertrophy, alterations in sedimentation coefficients, cytoplasmic pallor, and erythrocytic protein depletion. Hemogregarine parasites from several colubrid and viperid snakes completed sporogony within Aedes aegypti mosquitoes in 14-18 days, and produced oocysts typical of Hepatozoon sp. All lizards experimentally inoculated per os with mosquito-borne oocysts developed Hepatozoon sp. infections. No sporogonic development was demonstrated in any Ornithodoros turicata ticks (Argasidae) fed on parasitemic snakes. None of the lizards inoculated by tick developed Hepatozoon sp. infections. Molecular characterization of several hemogregarine isolates, two lizard Plasmodium species, and several species of Leishmania and Trypanosoma at the ribosomal DNA (rDNA) locus by PCR amplification and restriction enzyme analysis demonstrated detectable molecular differences between genera, species and isolates. Within unnatural lizard hosts multifocal accumulations of heterophilic leukocytes with intralesional Hepatozoon meronts were observed within the liver and lungs. The accompanying clinicopathologic changes in unnatural hosts included a marked leukocytosis with heterophilia, increases in plasma liver enzyme activities, and mild increases in the serum gamma globulin fraction. No meront-associated inflammation or significant clinico-pathologic changes were noted in naturally infected snakes despite numerous pulmonary and hepatic meronts. Assay of unnatural host serum for anti-Hepatozoon antibody by ELISA, IFA, and Western blot demonstrated post-infection increases in lizards sampled at 38 days post-infection and beyond. Stage-specific antibody directed against surface antigens of infected erythrocytes with a moderate orthochromatic erythroblastosis, was demonstrated only in parasitemic lizards. Molecularly distinct hemogregarines in different snake species and evidence of parasite-associated disease in unnatural hosts, suggest host-parasite coadaptation in nature
New Zealand White rabbits were repeatedly infested with Ornithodoros turicata (Duges), Ornithodor... more New Zealand White rabbits were repeatedly infested with Ornithodoros turicata (Duges), Ornithodoros talaje (Guérin-Méneville), and Ornithodoros coriaceus (Koch) at 2-wk intervals. Blood samples were taken from each animal 10 d after each infestation and the titer of anti-tick antibody was determined by enzyme-linked immunosorbent assay. Subsequent cross-reactivity studies demonstrated that the antitick antisera nonspecifically bound to salivary gland extract proteins prepared from several other tick genera and species&#39; Amblyomma maculatum (Koch), Dermacentor andersoni (Stiles), Dermacentor variabilis (Say), and Ornithodoros moubata (Murray). Absorption of the antisera against an immobilized extract of A. maculatum substantially increased specificity at the genus level. Western blots of electrophoretically separated Ornithodoros salivary gland extract samples were used to further compare the specificity of absorbed and nonabsorbed antitick antisera. The blots demonstrated that many of the Ornithodoros salivary gland extract proteins bear genus specific epitopes. Some differences were noted among the Ornithodoros species examined with respect to the degree of antigenic relatedness with the ixodid ticks.
International Journal for Parasitology, Sep 1, 1991
Haemogregarine parasites, derived from the Florida snakes Coluber constrictor and Nerodia fasciat... more Haemogregarine parasites, derived from the Florida snakes Coluber constrictor and Nerodia fasciata and ingested by Aedes aegypti, completed sporogony within the hemocoeles of nearly all fed mosquitoes in 14-18 days, and produced oocysts typical of Hepatozoon. However, mortalities and morbidity were high in the Culex which had fed on the Coluber. Oocysts were not found in any Ornithodoros turicata (Argasidae) which fed upon either snake host, but many sections of fed ticks had gametocyte-like cells within the gut lumen. Most lizards, Anolis carolinensis and Anolis sagrei, infected per os with oocysts derived from both snake species developed infections. Infections in the lizards were largely confined to hepatic schizonts with few parasites found in erythrocytes. Unlike naturally infected snake hosts, Hepatozoon schizonts in livers of lizards were often either surrounded by an unidentified dark pigment or heavily infiltrated with mononuclear inflammatory cells.
The host antibody response of CD1 mice resulting from sequential exposures to two species of Orni... more The host antibody response of CD1 mice resulting from sequential exposures to two species of Ornithodoros ticks was isotyped (ELISA) in an effort to determine the mechanism for the development of acquired resistance. In addition, salivary gland proteins from two argasid and one ixodid species were examined (RIA) for cross-reactivity. Significant antibody responses, detectable for at least 90 d after last exposure, were shown to occur. Isotyping showed that the initial response was of the IgM class with a secondary class switch to the IgG1 subclass occurring. Evidence that cross-reactive proteins exist between argasid and ixodid ticks is also discussed.
The purpose of this study was to immunologically characterize soft tick salivary antigens. BALB/c... more The purpose of this study was to immunologically characterize soft tick salivary antigens. BALB/c mice hyperimmunized with salivary gland extract prepared from Ornithodoros talaje (Guérin-Méneville) were observed to develop high titers of antitick salivary antigen antibodies. Subsequent fusion of splenic lymphocytes from the hyperimmunized mice with SP-2/0-AG14 myeloma cells resulted in the production of 10 antitick IgM-producing hybridoma clones. Partial characterization of the respective tick antigens by gel filtration and SDS-PAGE demonstrated all 10 monoclonal antitick antibodies to be reactive with a salivary gland extract fraction containing proteins 50-110 kDa in molecular weight. Cross-reactivity assays and electrophoretic comparison of salivary gland extract specimens demonstrated similar proteins in several ixodid tick genera and species.
<p>The asterisk and arrowhead represent the molecular masses for native GlpQ and BipA, resp... more <p>The asterisk and arrowhead represent the molecular masses for native GlpQ and BipA, respectively (A). rBipA was produced as a thioredoxin fusion protein and is 15 kDa larger than the native protein. Immunoblots were also probed with a serum sample from an uninfected patient (B) and a monoclonal antibody against the six-histidine fusion tag (C and D). Molecular masses are shown to the left of each immunoblot.</p
Journal of Herpetological Medicine and Surgery, 2000
ABSTRACT The common snake mite, Ophionyssus natricis (Parasitiformes: Macronyssidae), is a blood ... more ABSTRACT The common snake mite, Ophionyssus natricis (Parasitiformes: Macronyssidae), is a blood feeding, mesostigmatid mite that parasitizes reptiles. Anemia, dehydration, dermatitis, and several blood-borne infectious diseases have been linked to infestations (Camin, 1948Camin, 1953, Chiodini, et al, 1983). Severe pruritic dermatitis has been reported in humans bitten by Ophionyssus natricis (Schultz, 1975, Beck, 1996). All levels of Ophionyssus natricis infestation should be considered a serious problem worthy of prompt treatment. This article describes the general morphology, biology, and behavior of each life stage and discusses the components of an integrated control and prevention program for the maintenance of mite-free snake collections.
Journal of Herpetological Medicine and Surgery, 2000
ABSTRACT A group of seven juvenile California mountain kingsnakes, Lampropeltis zonata multicinct... more ABSTRACT A group of seven juvenile California mountain kingsnakes, Lampropeltis zonata multicincta, developed severe acute gastroenteritis characterized by emesis, dehydration and sudden death. Histopathology demonstrated segmental mucosal necrosis and hyperplasia, villus blunting and fusion. Abundant basophilic Cowdry type-A intranuclear inclusions were demonstrated in enterocytes peripheral to the zone of acute necrosis throughout the small intestine. Electron microscopy demonstrated intranuclear crystalline arrays of non-enveloped virus-like particles that measured 65 to 70 nm in diameter. Clusters of smaller non-enveloped virus-like particles that measured 15 to 18 nm in diameter were found in association with the larger particles. The size, morphology and location of the two virus-like particles were consistent with an adenovirus and a parvovirus. The presence of the parvovirus-like particles in the same nuclei and in association with adenovirus-like particles suggested the parvovirus-like agent to b...
The pathogenesis of bovine pneumonic pasteurellosis is not completely understood, and studies hav... more The pathogenesis of bovine pneumonic pasteurellosis is not completely understood, and studies have not established that Pasteurella haemolytica A1 (Ph1) virulence is exclusively responsible for the development of acute pulmonary lesions. The purpose of this investigation was to determine if immune complex disease is involved in the pathogenesis of bovine pneumonic pasteurellosis. A retrospective immunohistologic study of lung tissue from natural cases of bovine pneumonic pasteurellosis (44) as performed, and immune complexes were observed in alveloar spaces and walls in 88% of these cases. To study this pathologic mechanism experimentally, groups of mice were immunized with purified Ph1 outer membranes (OMs) or sham immunized on days 0 and 14. Mice were challenged intratracheally on day 24 with either live Ph1 or Ph1 OMs, and pulmonary lesions were assessed 24 h after challenge. Placebo immunized mice developed focal infiltrates of neutrophils and macrophages centered around large caliber bronchi. Mice immunized with Ph1 OMs and challenged with live Ph1 or OMs developed severe bronchointerstitial pneumonia with diffuse neutrophilic infiltration, focal necrosis, hemorrhage and edema, that is histologically similar to bovine pneumonic pasteurellosis. Immunohistology revealed flocculent aggregates of IgG and complement positive material within alveolar spaces and walls from mice challenged with live Ph1, and fine granular deposits of IgG and complement positive material were observed lining the alveolar walls from mice challenged with Ph1 OMs. Immunized mice exhibited high serum IgG antibody titers to Ph1 outer membrane proteins (OMPs). Results of this study suggest that immune complex disease plays a role in the pathogenesis of bovine pneumonic pasteurellosis.
Rabbits infested with either Ornithodoros sp. ticks or Psoroptes cuniculi mites were assayed for ... more Rabbits infested with either Ornithodoros sp. ticks or Psoroptes cuniculi mites were assayed for anti-tick antibody by enzyme-linked immunosorbent assay. Titration of rabbit serum against Ornithodoros sp. salivary gland extract (SGE) demonstrated both mite- and tick-infested animals to have elevated anti-tick antibody titers. Western blot analysis demonstrated the anti-mite and anti-tick antisera to contain antibodies with affinities for both common and unique subsets of Ornithodoros SGE proteins.
The objective of this study was to develop a highly specific enzyme-linked immunosorbent assay (E... more The objective of this study was to develop a highly specific enzyme-linked immunosorbent assay (ELISA) for the serological detection of anti-Ornithodoros tick antibodies in animals. Affinity-purified rabbit anti-Ornithodoros IgG antibodies were employed in indirect competitive inhibition ELISA assays designed to measure the anti-Ornithodoros antibody titers in other animal species using the domestic goat (Capra hircus) as a large animal model. Repeated infestation of goats with Ornithodoros coriaceus was found to elicit the formation of antibodies capable of inhibiting the binding of the Ornithodoros-specific rabbit IgG. Western blot analysis of goat and rabbit anti-tick antisera demonstrated both animal species to respond immunologically to a set of 9 major protein bands in O. coriaceus salivary gland extracts. The results of these experiments demonstrate that a history of animal exposure to O. coriaceus may be detected serologically by competitive inhibition ELISA.
Ophionyssus natricis (Parasitiformes: Macronyssidae) is a mesostigmatid mite which commonly paras... more Ophionyssus natricis (Parasitiformes: Macronyssidae) is a mesostigmatid mite which commonly parasitizes captive snakes. The mites feed on the host's blood by lacerating the soft skin between scales with their chelicerae. Heavy infestations are common in captive specimens and have been shown to result in severe anemia, dehydration, and death. Additionally, the mites have been shown to transmit several bacterial, protozoal, and filariid pathogens. While well recognized as a significant pest in captive snake collections, the snake mite has been difficult to control. The inability to control O. natricis is related to its complex life cycle, which includes both blood feeding and non-feeding stages. In order to develop a proper treatment regimen for O. natricis, it is critical to understand the natural history of the organism. This paper will describe the life cycle and ethology of O. natricis and summarize current therapies utilized in controlling and attempting to eradicate this pest.
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Papers by Edward Wozniak