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1.
J Parasitol ; 97(1): 140-3, 2011 Feb.
Article in English | MEDLINE | ID: mdl-21348621

ABSTRACT

Dogs are reservoir hosts for Trypanosoma cruzi , the causative agent of American trypanosomiasis. A rapid immunochromatographic dipstick test (ICT) is available commercially for canine serological testing. The ICT was developed with the use of sera from South American dogs, but it is not routinely used in the United States. We evaluated the utility of the ICT in detecting anti-T. cruzi antibodies in dogs from the United States. Dogs (N  =  64) were experimentally infected with United States' isolates of T. cruzi from an opossum (Didelphis virginiana), an armadillo (Dasypus novemcinctus), and a domestic dog (Canis familiaris), and were tested after experimental infection. Sera from uninfected United States dogs (n  =  79; hemaculture negative) were used as negative controls. In a blind study, sera were tested by the ICT and compared to the indirect immunofluorescent antibody test with the use of Brazil-strain epimastigotes as antigen. The sensitivity of the ICT was 91% and the specificity was 98% in dogs experimentally infected with United States isolates. Our study indicates that the ICT could be a useful screening tool for serological surveillance of canine T. cruzi exposure in the United States.


Subject(s)
Antibodies, Protozoan/blood , Armadillos/parasitology , Chagas Disease/veterinary , Dog Diseases/diagnosis , Opossums/parasitology , Trypanosoma cruzi/immunology , Animals , Chagas Disease/diagnosis , Chagas Disease/immunology , Chromatography/methods , Cross Reactions , Disease Reservoirs , Dog Diseases/immunology , Dog Diseases/parasitology , Dogs , Fluorescent Antibody Technique, Indirect/veterinary , Sensitivity and Specificity
2.
J Parasitol ; 96(6): 1119-22, 2010 Dec.
Article in English | MEDLINE | ID: mdl-21158620

ABSTRACT

We examined the prevalence of antibodies to zoonotic protozoan parasites ( Trypanosoma cruzi, Toxoplasma gondii, and Encephalitozoon cuniculi) and protozoans of veterinary importance ( Neospora caninum, Sarcocystis neurona, and Besnoitia darlingi) in a population of North American opossums ( Didelphis virginiana) from Louisiana. Samples from 30 opossums were collected as part of a survey for T. cruzi in Louisiana. Frozen sera from these 30 opossums were examined using an indirect immunofluorescent antibody test (IFAT) against in vitro-produced antigenic stages of these protozoans. Additionally, 24 of the 30 samples were examined using hemoculture, and all 30 were examined in the modified direct agglutination test (MAT) for antibodies to To. gondii. The prevalences of reactive IFAT samples were as follows: 60% for T. cruzi, 27% for To. gondii, 23% for E. cuniculi, 17% for S. neurona, 47% for B. darlingi, and 0% for N. caninum. Hemoculture revealed that 16 (67%) of 24 samples were positive for T. cruzi, compared to 18 of 30 (60%) by IFAT. The sensitivity and specificity for the IFAT compared to hemoculture was 100% for each. The modified direct agglutination test revealed that 9 (30%) of the 30 samples from opossums had antibodies to To. gondii , compared to 8 (27%) using the IFAT. The sensitivity and specificity of the IFAT compared to the MAT was 100% and 72%, respectively.


Subject(s)
Antibodies, Fungal/blood , Antibodies, Protozoan/blood , Didelphis/parasitology , Encephalitozoonosis/veterinary , Protozoan Infections, Animal/epidemiology , Agglutination Tests/veterinary , Animals , Chagas Disease/epidemiology , Chagas Disease/transmission , Chagas Disease/veterinary , Coccidiosis/epidemiology , Coccidiosis/transmission , Coccidiosis/veterinary , Encephalitozoon cuniculi/immunology , Encephalitozoonosis/epidemiology , Encephalitozoonosis/transmission , Fluorescent Antibody Technique, Indirect/veterinary , Louisiana/epidemiology , Neospora/immunology , Protozoan Infections, Animal/transmission , Sarcocystidae/immunology , Sarcocystis/immunology , Sarcocystosis/epidemiology , Sarcocystosis/transmission , Sarcocystosis/veterinary , Sensitivity and Specificity , Seroepidemiologic Studies , Toxoplasma/immunology , Toxoplasmosis, Animal/epidemiology , Toxoplasmosis, Animal/transmission , Trypanosoma cruzi/immunology , Zoonoses
3.
Am J Vet Res ; 71(11): 1294-304, 2010 Nov.
Article in English | MEDLINE | ID: mdl-21034320

ABSTRACT

OBJECTIVE: To evaluate the influence of treatment with ultralow-dose aspirin (ULDAsp) on platelet aggregation, P-selectin (CD62P) expression, and formation of platelet-leukocyte aggregates in clinically normal dogs. ANIMALS: 18 clinically normal dogs. PROCEDURES: Studies were conducted before and 24 hours after ULDAsp administration (0.5 mg/kg, PO, q 24 h, for 2 days). Whole blood impedance aggregometry for the assessment of platelet function was performed with sodium citrate-anticoagulated blood and aggregation agonists (ADP at 20, 10, and 5 µmol/L; collagen at 10, 5, and 2 µg/mL). Onset, maximum response, and rate of platelet aggregation were recorded. Flow cytometric assays were configured to detect thrombin-induced CD62P expression and platelet-leukocyte aggregates in EDTA-anticoagulated whole blood. Externalized platelet CD62P and constitutive CD61 (GPIIIa) were labeled with antibodies conjugated to phycoerythrin (PE) and fluorescein isothiocyanate (FITC), respectively. Red blood cell-lysed paraformaldehyde-fixed EDTA-anticoagulated whole blood was dual labeled with CD61-FITC and a panleukocyte antibody (CD18-PE) to characterize platelet-leukocyte aggregates. RESULTS: ULDAsp significantly delayed platelet aggregation onset with ADP at 20 µmol/L by 54% to 104%, attenuated maximum aggregation with various concentrations of ADP and collagen by ≥ 41%, and slowed aggregation rate with the highest ADP and collagen concentrations by ≥ 39%. Depending on the parameter tested, up to 30% of dogs failed to have an ULDAsp effect. Thrombin stimulation significantly increased CD62P expression in platelets and platelet-leukocyte aggregates, but ULDAsp did not alter basal or thrombin-stimulated CD62P expression. CONCLUSIONS AND CLINICAL RELEVANCE: ULDAsp treatment of clinically normal dogs impaired platelet aggregation in most dogs, but did not influence CD62P platelet membrane expression.


Subject(s)
Aspirin/pharmacology , P-Selectin/genetics , Platelet Aggregation Inhibitors/pharmacology , Platelet Aggregation/drug effects , Adenosine Diphosphate/pharmacology , Animals , Aspirin/administration & dosage , Blood Platelets/drug effects , Blood Platelets/physiology , Body Composition/drug effects , Body Weight , Dogs , Dose-Response Relationship, Drug , Female , Flow Cytometry , Gene Expression Regulation , Male , Orchiectomy , Ovariectomy , P-Selectin/drug effects , Platelet Aggregation/physiology , Platelet Aggregation Inhibitors/administration & dosage , Reference Values
5.
Vet Clin North Am Small Anim Pract ; 39(6): 1055-64, v-vi, 2009 Nov.
Article in English | MEDLINE | ID: mdl-19932362

ABSTRACT

Chagas' disease, or American trypanosomiasis, caused by the hemoflagellated protozoan Trypanosoma cruzi (class Zoomastigophorea and family Trypanosomatidae), is the leading cause of dilated cardiomyopathy in man. In dogs in North America, Chagas disease mainly occurs in working dogs in southeastern Texas. It is likely that most dogs become infected by eating infected vectors, causing the release of the organisms into the mouth of the host. Most dogs are diagnosed during the chronic stage of the disease, which is typified by dilated cardiomyopathy and malignant ventricular-based arrhythmias. This article reviews the etiology, epidemiology, pathogenesis, diagnosis, and available therapy for Chagas' disease in dogs.


Subject(s)
Chagas Disease/veterinary , Dog Diseases/parasitology , Aging , Animals , Antiprotozoal Agents/therapeutic use , Chagas Disease/drug therapy , Chagas Disease/epidemiology , Chagas Disease/transmission , Disease Reservoirs , Dogs , Insect Vectors , North America/epidemiology , Psychodidae
6.
Vet Clin North Am Small Anim Pract ; 39(6): 1065-74, vi, 2009 Nov.
Article in English | MEDLINE | ID: mdl-19932363

ABSTRACT

Canine leishmaniasis is a fatal zoonotic visceralizing disease usually associated with tropical areas. The etiologic agent is an obligate intracellular protozoan, Leishmania infantum. In 1999, an outbreak of a canine leishmaniasis was reported in a Foxhound kennel in New York, and since that report, several other outbreaks have occurred across the United States in additional Foxhound kennels. Because of the high mortality and transmissibility associated with these outbreaks, it is essential that clinicians be aware of this disease to permit its rapid recognition and institution of control measures. Cases with a travel history may suggest imported disease; these are mainly observed from Southern Europe (eg, south of France, Spain, and Italy). Breeds from these and other endemic areas may be at higher risk of infection with Leishmania because of vertical transmission. The purpose of this report is to discuss the clinical signs, epidemiology, diagnosis, control, and treatment of canine leishmaniasis with focus on the aspects of this disease within North America.


Subject(s)
Communicable Diseases, Emerging/veterinary , Dog Diseases/epidemiology , Leishmaniasis/veterinary , Animals , Communicable Diseases, Emerging/epidemiology , Demography , Dog Diseases/parasitology , Dogs , Endemic Diseases/veterinary , Insect Vectors , Leishmaniasis/epidemiology , Leishmaniasis/parasitology , North America/epidemiology , Psychodidae
7.
J Am Anim Hosp Assoc ; 45(2): 93-6, 2009.
Article in English | MEDLINE | ID: mdl-19258422

ABSTRACT

An adult domestic shorthair (DSH) cat was presented with acute vomiting, anorexia, lethargy, and dyspnea. The cat's clinical status worsened over 24 hours with conservative medical management. An exploratory celiotomy was performed. Acute intestinal obstruction resulting from infection with Taenia (T.) taeniaeformis was diagnosed. Surgical removal of the cestodes via multiple enterotomies resolved the obstruction. This paper reports, for the first time, small intestinal obstruction caused by T. taeniaeformis infection in a cat.


Subject(s)
Cat Diseases/parasitology , Intestinal Obstruction/veterinary , Taeniasis/veterinary , Animals , Anthelmintics/administration & dosage , Australia , Cat Diseases/diagnostic imaging , Cat Diseases/surgery , Cats , Cestoda/isolation & purification , Intestinal Obstruction/diagnostic imaging , Intestinal Obstruction/parasitology , Intestinal Obstruction/surgery , Male , Radiography, Abdominal/veterinary , Taenia/isolation & purification , Taeniasis/complications , Taeniasis/diagnostic imaging , Taeniasis/surgery , Treatment Outcome
8.
Microbes Infect ; 11(2): 230-7, 2009 Feb.
Article in English | MEDLINE | ID: mdl-19070678

ABSTRACT

Leptospiral immunoglobulin-like protein (LigB) was truncated into conserved (LigBcon) and variable (varB1, varB2) fragments and expressed as GST/His-tag fusion proteins. Four-week-old hamsters were immunized with equal amounts of each fragment individually or combined in alum adjuvant at days 0 and 21 and subsequently challenged three weeks after the booster with 2.5 LD(50) live virulent Leptospira interrogans serovar Pomona. Our results demonstrate that immunization with LigB produced strong humoral immune responses as revealed by high titers against each fragment and significant enhancement in Th2 cytokines (IL-4, IL-10). A significant activation of CMI is revealed by enhanced proliferation of lymphocytes and up regulation of Th1 cytokines (IL-12p40, IFN-gamma) was also noted. Of the peptides studied, rLigBcon was able to impart maximum protection (71%), followed by rVarB1 (54%), whereas rVarB2 was not able to impart a significant level of protection (33%) against lethal infection as revealed by enhanced survival and reduced severity of histopathological lesions in vital organs (viz. kidney, liver, spleen) of the immunized animals. Moreover, concurrent administration of all three fragments significantly enhanced the protective efficacy of the vaccine (83%). Overall, our results clearly demonstrate that LigB has emerged as novel protective antigen that can be used in future subunit vaccines against leptospirosis.


Subject(s)
Antigens, Bacterial/immunology , Leptospira interrogans serovar pomona/immunology , Leptospirosis/prevention & control , Adjuvants, Immunologic/administration & dosage , Adjuvants, Immunologic/pharmacology , Alum Compounds/administration & dosage , Alum Compounds/pharmacology , Animals , Antibodies, Bacterial/blood , Antigens, Bacterial/genetics , Cell Proliferation , Cricetinae , Immunization, Secondary , Interleukin-10/metabolism , Interleukin-4/metabolism , Kidney/pathology , Leptospira interrogans serovar pomona/genetics , Liver/pathology , Lymphocytes/immunology , Recombinant Proteins/genetics , Recombinant Proteins/immunology , Spleen/pathology , Survival Analysis , Vaccines, Subunit/genetics , Vaccines, Subunit/immunology
9.
J Feline Med Surg ; 10(4): 384-7, 2008 Aug.
Article in English | MEDLINE | ID: mdl-18313344

ABSTRACT

A young adult male domestic shorthair cat was presented for physical examination, routine vaccinations, and a fecal examination. Physical examination revealed no significant abnormalities. Eggs of the raccoon pancreatic fluke Eurytrema procyonis were detected by fecal flotation. Results of a complete blood count and serum biochemistry panel were normal. Abdominal sonography revealed an enlarged hypoechoic pancreas with a hyperechoic rim, and a distended and thickened pancreatic duct. Serum pancreatic lipase immunoreactivity (PLI) was increased. These findings supported the possibility of fluke-associated pancreatitis. Treatment with praziquantel/pyrantel/febantel was associated with resolution of sonographic abnormalities and normalization of PLI.


Subject(s)
Cat Diseases/diagnosis , Dicrocoeliidae/isolation & purification , Pancreas/parasitology , Pancreatitis/veterinary , Trematode Infections/veterinary , Animals , Anthelmintics/therapeutic use , Cat Diseases/drug therapy , Cat Diseases/parasitology , Cats , Feces/parasitology , Male , Pancreas/pathology , Pancreatitis/diagnosis , Pancreatitis/drug therapy , Pancreatitis/parasitology , Treatment Outcome , Trematode Infections/diagnosis , Trematode Infections/drug therapy
10.
J Vet Intern Med ; 20(3): 489-94, 2006.
Article in English | MEDLINE | ID: mdl-16734079

ABSTRACT

The purpose of this study was to review recent cases of leptospirosis seen at referral centers in New York State and to identify differences in clinical or clinicopathologic aspects of the disease among different suspected infecting serogroups. Medical records at the Cornell University Hospital for Animals and the Animal Medical Center in New York City were reviewed to identify dogs diagnosed with leptospirosis from September 1996 to August 2002. Records of 55 dogs met the inclusion criteria for the study. The suspected infecting serogroups included 21 occurrences of Grippotyphosa, 12 of Pomona, 6 of Autumnalis, 5 of Bratislava, 2 of Hardjo, and 1 of Canicola. Five dogs had equal titers to serogroups Grippotyphosa and Pomona, and 3 had equal titers to 2 other serogroups. Common clinical signs included lethargy, anorexia, and vomiting. Common clinicopathologic findings included anemia, thrombocytopenia, azotemia, hyperphosphatemia, high liver enzyme activity, and hyperbilirubinemia. Forty-three of 55 dogs were discharged from the hospital. Serogroup-specific analysis indicated that dogs with suspected serogroup Pomona infection were more likely to suffer from vomiting (P = .01), thrombocytopenia (P = .009), severe azotemia (P = .04), and hyperphosphatemia (P = .006) than dogs with other serogroups and were less likely to be discharged alive from the hospital (P = .03). This study suggests that only minor clinically relevant differences exist among serogroups. Leptospira serogroup Pomona caused more severe renal disease and was associated with a worse outcome compared with disease caused by other serogroups.


Subject(s)
Dog Diseases/epidemiology , Dog Diseases/microbiology , Leptospira/classification , Leptospirosis/veterinary , Animals , Dog Diseases/diagnostic imaging , Dog Diseases/etiology , Dog Diseases/genetics , Dog Diseases/pathology , Dogs , Female , Leptospirosis/epidemiology , Leptospirosis/microbiology , Male , New York City/epidemiology , Pedigree , Radiography, Thoracic/veterinary , Records/veterinary , Referral and Consultation , Retrospective Studies , Seasons , Seroepidemiologic Studies
11.
Vet Parasitol ; 135(3-4): 303-14, 2006 Feb 18.
Article in English | MEDLINE | ID: mdl-16289566

ABSTRACT

Canine dirofilariasis caused by Dirofilaria immitis is usually diagnosed by specific antigen testing and/or identification of microfilariae. However, D. immitis and at least six other filariae can produce canine microfilaremias with negative heartworm antigen tests. Discriminating these can be of clinical importance. To resolve discordant diagnoses by two diagnostic laboratories in an antigen-negative, microfilaremic dog recently imported into the US from Europe we developed a simple molecular method of identifying different microfilariae, and subsequently validated our method against six different filariae known to infect dogs by amplifying ribosomal DNA spacer sequences by polymerase chain reaction using common and species-specific primers, and sequencing the products to confirm the genotype of the filariae. We identified the filaria in this dog as D. repens. This is the first case of D. repens infection in the United States. Additionally, we examined microfilariae from five additional antigen-negative, microfilaremic dogs and successfully identified the infecting parasite in each case. Our diagnoses differed from the initial morphological diagnosis in three of these cases, demonstrating the inaccuracy of morphological diagnosis. In each case, microfilariae identified morphologically as A. reconditum were identified as D. immitis by molecular methods. Finally, we demonstrated that our PCR method should amplify DNA from at least two additional filariae (Onchocerca and Mansonella), suggesting that this method may be suitable for genotyping all members of the family Onchocercidae.


Subject(s)
DNA, Helminth/analysis , Dirofilaria/isolation & purification , Dirofilariasis/diagnosis , Dog Diseases/diagnosis , Microfilariae/isolation & purification , Polymerase Chain Reaction/veterinary , Animals , Base Sequence , Diagnosis, Differential , Dirofilaria/classification , Dirofilaria immitis/classification , Dirofilaria immitis/isolation & purification , Dirofilariasis/parasitology , Dog Diseases/parasitology , Dogs , Female , Genotype , Male , Microfilariae/classification , Molecular Sequence Data , Phenotype , Polymerase Chain Reaction/methods , Sequence Homology, Nucleic Acid , Species Specificity
12.
Am J Vet Res ; 66(10): 1780-4, 2005 Oct.
Article in English | MEDLINE | ID: mdl-16273911

ABSTRACT

OBJECTIVE: To evaluate serum titers obtained by use of the microscopic agglutination test (ie, MAT titers) to Leptospira interrogans serovar pomona and autumnalis and Leptospira kirschneri serovar grippotyphosa in dogs given a commercial vaccine against serovars pomona and grippotyphosa. ANIMALS: Forty 12-week-old puppies and 20 mature Beagles. PROCEDURE: Puppies received a commercial vaccine against serovars pomona and grippotyphosa at 12 weeks of age, then received a booster vaccine and 3 weeks later; mature dogs received the vaccine once. Serum MAT titers to serovars pomona, autumnalis, and grippotyphosa were measured before vaccination and at 2, 4, 6, 10, and 16 weeks after the first or only vaccination. RESULTS: Of the 40 puppies vaccinated, 40, 0, and 40 developed MAT titers of > 100 after vaccination to serovars pomona, grippotyphosa, and autumnalis, respectively. Microscopic agglutination test titers to serovar autumnalis were higher than MAT titers to serovars pomona and grippotyphosa and persisted in some dogs for 16 weeks (6 weeks longer than for titers to serovar pomona). Of the 20 mature dogs, 13, 5, and 20 developed MAT titers of > 100 at 2 weeks to serovars pomona, grippotyphosa, and autumnalis, respectively. Titers to serovar pomona were higher and persisted in some dogs beyond 16 weeks after vaccination, compared with titers to serovars pomona and grippotyphosa, which persisted for 10 and 6 weeks, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Subunit vaccines against serovars pomona and grippotyphosa induce MAT titers not only to homologous antigens but also to serovar autumnalis, which could lead to a misdiagnosis of leptospirosis caused by serovar autumnalis.


Subject(s)
Antibodies, Bacterial/blood , Bacterial Vaccines/immunology , Dog Diseases/blood , Dog Diseases/prevention & control , Leptospira interrogans serovar pomona/immunology , Leptospirosis/veterinary , Agglutination Tests/veterinary , Animals , Dogs , Leptospirosis/blood , Leptospirosis/prevention & control
13.
J Vet Intern Med ; 19(4): 553-9, 2005.
Article in English | MEDLINE | ID: mdl-16095173

ABSTRACT

The objective of this investigation was to determine whether or not herpesvirus (herpes-), adenovirus (adeno-), or canine parvovirus DNA is present in the brains of dogs with necrotizing meningoencephalitis (NME), necrotizing leukoencephalitis (NLE), and granulomatous meningoencephalitis (GME). Paraffin-embedded brain specimens from 12 histopathologically confirmed dogs with NME, 3 with NLE, and 7 with GME were screened for viral DNA with degenerate herpes- and adenovirus polymerase chain reaction (PCR) and a canine parvovirus-specific PCR. Positive-control specimens included genomic viral DNA and paraffin-embedded tissues from dogs with confirmed herpes-, adeno-, or canine parvovirus infections. Herpes-, adeno-, or canine parvovirus DNA was amplified by PCR from the corresponding positive-control specimens. Negative controls included 7 dogs with various brain disorders and produced no viral amplicons. The 22 dogs with NME, NLE, and GME were negative for viral DNA. Additional studies testing for other viruses or inherited genetic mutations are warranted to gain insight into the etiologies of NME, NLE, and GME. We discuss potential etiologies and provide a clinical and histopathologic overview of these common canine encephalitides.


Subject(s)
Brain/virology , DNA Virus Infections/veterinary , DNA Viruses/isolation & purification , Dog Diseases/virology , Encephalitis, Viral/veterinary , Meningoencephalitis/veterinary , Animals , DNA Virus Infections/diagnosis , DNA Virus Infections/pathology , Dog Diseases/diagnosis , Dogs , Encephalitis, Viral/diagnosis , Encephalitis, Viral/pathology , Female , Granuloma/veterinary , Male , Meningoencephalitis/pathology , Meningoencephalitis/virology , Necrosis/veterinary , Polymerase Chain Reaction/veterinary
14.
J Am Vet Med Assoc ; 226(11): 1869-80, 2005 Jun 01.
Article in English | MEDLINE | ID: mdl-15934255

ABSTRACT

OBJECTIVE: To evaluate prognostic factors, survival, and treatment protocols for immune-mediated hemolytic anemia (IMHA) in dogs. DESIGN: Retrospective study. ANIMALS: 151 dogs with IMHA not associated with underlying infectious or neoplastic disease. PROCEDURE: lnformation recorded from review of medical records included signalment at the time of initial evaluation; vaccination history; 30-, 60-, and 365-day follow-up outcomes; laboratory data; results of imaging studies; and necropsy findings. Dogs were grouped according to the presence of spherocytes, autoagglutination, a regenerative erythrocyte response, and treatments received (azathioprine, azathioprine plus ultralow-dose aspirin, azathioprine plus mixed-molecular-weight heparin [mHEP], or azathioprine plus ultralow-dose aspirin plus mHEP) for comparisons. All dogs received glucocorticoids. RESULTS: Cocker Spaniels, Miniature Schnauzers, neutered dogs, and female dogs were overrepresented. Alterations in certain clinicopathologic variables were associated with increased mortality rate. Rates of survival following treatment with azathioprine, azathioprine plus ultralow-dose aspirin, azathioprine plus mHEP, and azathioprine plus ultralow-dose aspirin plus mHEP were 74%, 88%, 23%, and 70%, respectively, at hospital discharge; 57%, 82%, 17%, and 67%, respectively, at 30 days; and 45%, 69%, 17%, and 64%, respectively, at 1 year. In comparison, mean survival rates at discharge and at 30 days and 1 year after evaluation collated from 7 published reviews of canine IMHA were 57%, 58%, and 34%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Treatment with a combination of glucocorticoids, azathioprine, and ultralow-dose aspirin significantly improved short- and long-term survival in dogs with IMHA.


Subject(s)
Anemia, Hemolytic, Autoimmune/veterinary , Dog Diseases/mortality , Anemia, Hemolytic, Autoimmune/drug therapy , Anemia, Hemolytic, Autoimmune/mortality , Animals , Aspirin/therapeutic use , Azathioprine/therapeutic use , Breeding , Dog Diseases/drug therapy , Dog Diseases/immunology , Dogs , Female , Glucocorticoids/therapeutic use , Immunosuppressive Agents/therapeutic use , Male , Prognosis , Retrospective Studies , Sex Factors , Survival Rate
16.
J Vet Diagn Invest ; 17(6): 565-8, 2005 Nov.
Article in English | MEDLINE | ID: mdl-16475515

ABSTRACT

This report describes a 3-year-old male castrated Mastiff dog that died unexpectedly with locally extensive, acute, necrotizing myocarditis and myocardial infarction. Intralesional protozoal tachyzoites in the affected myocardium were confirmed to be Neospora caninum by a novel multiplex polymerase chain reaction (PCR) and immunohistochemistry. Protozoal organisms were not identified in other tissues by histology, immunohistochemistry, or PCR. The multiplex PCR assay was used to quickly provide preliminary results on fresh myocardium to differentiate N. caninum and Toxoplasma gondii. Neosporosis is an uncommon cause of myocarditis in adult dogs and differential diagnoses for myocarditis in this population of dogs are reviewed.


Subject(s)
Coccidiosis/diagnosis , Coccidiosis/veterinary , Myocardial Infarction/veterinary , Myocarditis/veterinary , Neospora/isolation & purification , Toxoplasma/isolation & purification , Toxoplasmosis, Animal/diagnosis , Animals , Coccidiosis/complications , Coccidiosis/parasitology , Dogs , Fatal Outcome , Heart/parasitology , Male , Myocardial Infarction/complications , Myocardial Infarction/parasitology , Myocarditis/complications , Myocarditis/diagnosis , Myocarditis/parasitology , Myocardium/pathology , Neospora/genetics , Polymerase Chain Reaction , Toxoplasma/genetics , Toxoplasmosis, Animal/complications , Toxoplasmosis, Animal/parasitology
17.
J Med Microbiol ; 53(Pt 10): 975-984, 2004 Oct.
Article in English | MEDLINE | ID: mdl-15358819

ABSTRACT

The search for novel antigens suitable for improved vaccines and diagnostic reagents against leptospirosis led to the identification of LigA and LigB. LigA and LigB expression were not detectable at the translation level but were detectable at the transcription level in leptospires grown in vitro. Lig genes were present in pathogenic serovars of Leptospira, but not in non-pathogenic Leptospira biflexa. The conserved and variable regions of LigA and LigB (Con, VarA and VarB) were cloned, expressed and purified as GST-fusion proteins. Purified recombinant LigA and LigB were evaluated for their diagnostic potential in a kinetic ELISA (KELA) using sera from vaccinated and microscopic agglutination test (MAT)-positive dogs. Sera from vaccinated dogs showed reactivity to whole-cell antigens of leptospires but did not show reactivity in the KELA assay with recombinant antigens, suggesting a lack of antibodies to Lig proteins in the vaccinated animals. The diagnostic potential of recombinant Lig antigens in the KELA assay was evaluated by using 67 serum samples with MAT > or =1600, which showed reactivity of 76, 41 and 35% to rConA, rVarA and rVarB, respectively. These findings suggest that recombinant antigen to the conserved region of LigA and LigB can differentiate between vaccinated and naturally infected animals.


Subject(s)
Antigens, Bacterial/immunology , Leptospira interrogans/immunology , Leptospirosis/diagnosis , Amino Acid Sequence , Animals , Antibodies, Bacterial/blood , Bacterial Vaccines/immunology , Dogs , Enzyme-Linked Immunosorbent Assay , Molecular Sequence Data , Protein Biosynthesis , Serologic Tests , Transcription, Genetic , Vaccination , Vaccines, Synthetic/immunology
18.
Am J Vet Res ; 65(4): 480-4, 2004 Apr.
Article in English | MEDLINE | ID: mdl-15077691

ABSTRACT

OBJECTIVE: To perform respiratory chain enzymatic activity assays on canine skeletal muscle biopsy specimens and establish reference range values of skeletal muscle enzyme activities for dogs. SAMPLE POPULATION: Biopsy specimens from the vastus lateralis muscle were obtained from 24 dogs (8 sexually intact males and 14 sexually intact females) ranging from 15 months to 6 years of age. PROCEDURE: Mean values of citrate synthase, cytochrome-c oxidase, succinate dehydrogenase, succinate dehydrogenase-cytochrome-c reductase, nicotinamide adenine dinucleotide (NADH) dehydrogenase, and NADH dehydrogenase-cytochrome-c reductase activities were established by use of 6 standard spectrophotometric assays for respiratory chain enzyme analysis. RESULTS: Compared with published data for skeletal muscle enzyme activities in humans, skeletal muscle enzyme activities in dogs were 2- to 4-fold higher. Additionally, citrate synthase activity, a marker for mitochondrial volume, was positively correlated with age in dogs, suggesting that mitochondrial volume increases with age, although no apparent change in respiratory chain enzymatic activity with an increase in age was found. CONCLUSIONS AND CLINICAL RELEVANCE: Reference range values for skeletal muscle enzyme activities of dogs are needed to accurately interpret results of respiratory chain enzymatic activity assays. During investigation of metabolic myopathies, if skeletal muscle biopsy specimens are evaluated for respiratory chain enzyme kinetics, they should be performed and evaluated in concert with skeletal muscle biopsy specimens from clinically normal animals of the same species.


Subject(s)
Dogs/metabolism , Electron Transport Chain Complex Proteins/metabolism , Mitochondria, Muscle/enzymology , Muscle, Skeletal/enzymology , Animals , Spectrophotometry
19.
Am J Vet Res ; 65(1): 53-8, 2004 Jan.
Article in English | MEDLINE | ID: mdl-14719702

ABSTRACT

OBJECTIVE: To determine the gene sequences of canine and feline cardiac troponin I (cTnI), express the protein from the cloned gene in vitro, and validate the use of a commercial cTnI serum analyzer in these species via detection of the expressed protein or comparison of sequence homology. SAMPLE POPULATION: Samples of ventricular myocardium from 5 healthy adult mixed-breed dogs and 5 healthy adult domestic shorthair cats. PROCEDURE: The RNA was extracted from myocardial samples, and cDNA was synthesized via reverse transcriptase polymerase chain reaction and sequenced. The canine cDNA for the coding region was expressed in cell culture and analyzed by western blot and sandwich enzyme-linked immunosorbent assays. RESULTS: Canine and feline cTnI genes were cloned and sequenced. Homology of the nucleotide and amino acid sequences of the canine and feline cTnI genes with human and rodent cTnI genes were high; the greatest homology was detected between canine and feline genes (95% and 96%, respectively). Recombinant canine cTnI protein was detected by a commercial serum cTnI analyzer and by western blot analysis. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that commercial cTnI analyzers can be used to measure serum cTnI concentration from dogs and cats. Additionally, our preliminary characterization of the feline cTnI gene may facilitate further investigation of cTnI and its role in familial hypertrophic cardiomyopathy in cats.


Subject(s)
Cats/genetics , Dogs/genetics , Troponin I/genetics , Animals , Base Sequence , Blotting, Western , DNA Primers , DNA, Complementary/genetics , Enzyme-Linked Immunosorbent Assay , Molecular Sequence Data , Phylogeny , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNA , Sequence Homology , Spectrophotometry
20.
Am J Vet Res ; 64(12): 1507-13, 2003 Dec.
Article in English | MEDLINE | ID: mdl-14672429

ABSTRACT

OBJECTIVE: To develop a multiplex polymerase chain reaction (PCR) assay for the detection of Toxoplasma gondii and Neospora caninum DNA in canine and feline biological samples. SAMPLE POPULATION; Biological samples from 7 cats with systemic (n = 4) or CNS (3) toxoplasmosis, 6 dogs with neospora- or toxoplasma-associated encephalitis, and 11 animals with nonprotozoal disease. PROCEDURE: Primers for T gondii, N caninum, and the canine ferritin gene (dogs) or feline histone 3.3 gene (cats) were combined in a single PCR assay. The DNA was extracted from paraffin-embedded brain tissue, CSF, or skeletal muscle. The PCR products with positive results were cloned, and sequence identity was confirmed. RESULTS: Of 7 cats and 4 dogs with immunohistochemical or serologic evidence of toxoplasmosis, PCR results were positive for all cats and 3 dogs for T gondii, and positive for T gondii and N caninum for 1 dog. Another dog had negative PCR results for both parasites. Of 2 dogs with immunohistochemical or serologic evidence of neosporosis, PCR results were positive for 1 for N caninum and positive for the other for T gondii. All negative-control samples yielded negative results for T gondii and N caninum on the PCR assay. CONCLUSIONS AND CLINICAL RELEVANCE: Standard tests for toxoplasmosis or neosporosis associated with the CNS rely on serologic, histologic, or immunohistochemical analysis and can be difficult to interpret. The multiplex PCR assay with built-in control reactions could be a complementary clinical tool for the antemortem diagnosis of toxoplasmosis or neosporosis associated with the CNS.


Subject(s)
Coccidiosis/diagnosis , Coccidiosis/veterinary , Neospora/genetics , Polymerase Chain Reaction/methods , Toxoplasma/genetics , Toxoplasmosis, Animal/diagnosis , Animals , Cats , Central Nervous System/parasitology , DNA Primers , Dogs , Electrophoresis, Agar Gel , Sequence Analysis, DNA
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