Seroprevalence of Ehrlichia canis infections in police dogs in Spain

A seroepidemiological study of Ehrlichia canis was made in police dogs in Madrid (Spain). AntiEhrlichia canis antibodies were detected by indirect fluorescent antibody test for the detection of IgG.

The results obtained (three positive dogs out of a population of 131 animals) represents a seroprevalence of canine ehrlichiosis of 2.29%. This seroprevalence is one of the lowest described for this type of population.

The seroprevalence obtained from hunting dogs kennelled in Madrid in 1993 was 66.7% (24 positive out of a population of 36). Environmental conditions (temperature, humidity, rainfall, etc.) in both populations were very similar. We suggest that these conflicting results are due to the different prophylactic programmes used in these two populations.     

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

OBJECTIVE: To test serum samples of dogs and horses by use of class-specific recombinant-based ELISA for establishing a diagnosis of granulocytic ehrlichiosis attributable to infection with organisms from the Ehrlichia phagocytophila genogroup. SAMPLE POPULATION: Serum samples from 43 client-owned dogs and 131 horses (81 with signs of acute illness and 50 without signs of disease). PROCEDURE: Serum samples were analyzed, using ELISA with a recombinant 44-kd protein antigen for IgM and IgG antibodies to the human granulocytic ehrlichiosis (HGE) agent (NCH-1 strain). Western blot analyses, using infected human promyelocytic leukemia cells, were conducted on 38 serum samples of horses and 11 serum samples of dogs to verify reactivity to the 44-kd peptide. RESULTS: IgM or IgG antibodies to the HGE agent were detected in 5 to 28% of dog serum samples and 5 to 37% of horse serum samples. Thirty-five of 38 (92%) horse serum samples had corresponding results on both tests (2 positive results for 26 samples and 2 negative results for 9 samples), using an ELISA for IgG antibodies or immunoblotting for total immunoglobulins. All 11 serum samples of dogs had positive results for both methods. CONCLUSION AND CLINICAL RELEVANCE: These ELISA with recombinant 44-kd antigen are suitable for detecting IgM or IgG antibodies to the HGE agent in serum samples of dogs and horses. Positive results for serum samples of horses from Connecticut, New York, Virginia, and Georgia indicate that the HGE agent is widely distributed in tick-infested areas of the eastern United States.     

Evaluation of canine serum for the presence of antiretinal autoantibodies in sudden acquired retinal degeneration syndrome

OBJECTIVE: To determine the presence of serum antiretinal antibodies in sudden acquired retinal degeneration syndrome (SARDS) affected dogs and the size of the antigen to which these antibodies bind via the use of enzyme-linked immunosorbent assay (ELISA) and Western blot immunoassays. ANIMALS STUDIED: Serum was collected from 13 dogs affected by SARDS and five dogs with normal ocular examinations. PROCEDURES: All serum samples were subjected to ELISA with saline-soluble canine retinal tissue and Western blot analyses with SDS solubilized normal canine retinal tissue as the antigen. Antirecoverin (23 kDa) and antiheat shock cognate (65 kDa) antibodies were used as positive controls for both procedures. Affinity-purified goat antidog IgG and IgM labeled with horseradish peroxidase were used for all clinical samples and goat antirabbit IgG was used as the secondary antibody for the positive controls. RESULTS: ELISA demonstrated antibody reaction with all samples. Western blot immunoassays identified multiple bands in all canine serum samples, as well as in negative controls. Approximate sizes of the bands were 25 and 50 kDa, corresponding to IgG light and heavy chains, respectively. CONCLUSION: No antiretinal autoantibodies were identified in the serum of dogs affected by SARDS as compared to normal canine patients.     

Comparison of the value of measurement of serum galactomannan and Aspergillus-specific antibodies in the diagnosis of canine sino-nasal aspergillosis

Serology is currently used for the diagnosis of canine sino-nasal aspergillosis (SNA). However, the accuracy of serological testing using commercially available, standardized purified antigen preparations of Aspergillus (CAPurAspAg) has only been poorly documented. The aim of the present study was to assess the diagnostic value of an agar-gel double immunodiffusion (AGDD) test and an anti-Aspergillus IgG ELISA, using CAPurAspAg and the commercially available Platelia test for the detection of serum galactomannan. Sera from 17 dogs with SNA, 18 dogs with a nasal tumour (NT), 11 dogs with lymphoplasmacytic rhinitis (LPR) and 33 control dogs were tested with the 3 methods. AGDD result was positive in 76.5% of dogs with SNA, whereas all sera from dogs with non-fungal nasal disease and control dogs were negative. A positive IgG ELISA result was obtained in 88% of dogs with SNA and in 18% of dogs with LPR. All patients with NT and control dogs had a negative IgG ELISA result. The Platelia test was positive in 24% of dogs with SNA, 11% of dogs with NT, 9% of dogs with LPR and 24% of control dogs. The results of this study suggest that (1) the detection of serum Aspergillus-specific antibodies with AGDD or ELISA, using CAPurAspAg, provides excellent specificity and good sensitivity, (2) the specificity is higher for AGDD (100%) than for ELISA (96.8%) while sensitivity is higher for ELISA (88.2%) than for AGDD (76.5%) and (3) serum galactomannan quantification with the Plateliat test is unreliable for the diagnosis of canine SNA.     

ELISA法检测犬腹泻粪样中的犬冠状病毒

用ＦＥ细胞增殖犬冠状病毒（ＣＣＶ）参考株，分别免疫家兔和ＢＡＬＢ／ｃ小鼠制备ＣＣＶ多抗和单抗，建立了夹心ＥＬＩＳＡ及Ｄｏｔ－ＥＬＩＳＡ诊断方法。在检测的８４例犬腹泻粪样中，多抗、单抗夹心法显示ＣＣＶ阳性１６例，Ｄｏｔ－ＥＬＩＳＡ阳性１３例，后１３例包括在前１６例中。从８４例腹泻犬粪样中随机取３８例作ＣＣＶ、犬细小病毒（ＣＰＶ）双项检测，ＣＣＶ阳性１６例，ＣＰＶ阳性６例，ＣＣＶ、ＣＰＶ混合感染４例。结果显示，在南京地区流行的犬腹泻中，ＣＣＶ感染比例有超过ＣＰＶ的趋势。     

Enzyme-linked immunosorbent assay for detection of antibody to Pseudomonas aeruginosa and measurement of antibody titer in horse serum

Enzyme-linked immunosorbent assay (ELISA) was used for detection of immunoglobulin (Ig) M and IgG antibodies against a serologically common antigen (original endotoxin protein), protease, and elastase of Pseudomonas aeruginosa. The P aeruginosa antibody in horse sera was measured, using ELISA. Horseradish peroxidase-labeled rabbit anti-horse IgM and IgG antibodies were used for enzyme-labeled antibody conjugate. 5-Aminosalicylic acid and H2O2 were used for substrate. Sera collected from a vaccinated horse, a newborn foal, and 72 healthy racehorses were investigated for antibodies against P aeruginosa by ELISA and passive hemagglutination procedure. Changes in IgM and IgG antibody titers with vaccination were clear by ELISA. In the newborn foal, significant amounts of IgM and IgG antibodies from colostrum were present on the 1st day after birth. It was shown by ELISA that the level of antibodies in the newborn decreased initially and then increased. Some antibodies against original endotoxin protein, protease, and elastase of P aeruginosa were detected in almost all the healthy racehorses investigated.     

Seroprevalence of Toxoplasma gondii antibodies in clinically ill cats in the United States

OBJECTIVE: To determine regional seroprevalence estimates of Toxoplasma gondi-specific IgM and IgG in clinically ill cats throughout the United States. Sample Population-Sera from 12,628 clinically ill, client-owned cats. PROCEDURE: Toxoplasma gondii-specific IgM and IgG antibodies were detected by use of ELISAs. Sera from clinically ill cats previously submitted for T. gondii antibody testing were sequentially selected from our serum bank and the sample submission paperwork reviewed. The country was divided into 12 geographic regions. Overall prevalence as well as prevalence for each region, age group, season, sex (male vs female), and breed (domestic shorthair vs other) was calculated. Data were analyzed by logistic regression analysis. RESULTS: Overall, 31.6% of the cats were seropositive for T. gondii-specific IgM, IgG, or both. Percentage of cats seropositive for T. gondii antibodies ranged from 16.1% (southwestern United States) to 43.5% (northeastern United States). As age increased, odds of positive T. gondii antibody assay results (IgM alone, IgG alone, and any combination of IgM or IgG) increased. Males were more likely than females to be seropositive for T. gondii antibodies (IgG alone and any combination of IgM or IgG). Domestic shorthair cats were more likely than other breeds to be seropositive for T. gondii antibodies (IgM alone, IgG alone, and any combination of IgM or IgG). CONCLUSIONS AND CLINICAL RELEVANCE: Toxoplasma gondii-specific antibodies are common in serum samples of clinically ill cats from all regions of the United States. Seroprevalence increases as cats age and is higher in male and domestic shorthair cats, compared with females and other breeds.     

Seroconversion of pigs in contact with dogs exposed to canine coronavirus.

In order to determine if canine coronavirus (CCV) could be transmitted to pigs, two dogs were inoculated orally with virulent CCV. After 24 h, the dogs were moved to an isolation room that contained three three-day-old pigs. A wire mesh fence, allowing close contact between the animals, separated the dogs from the pigs. The dogs and pigs were observed for 14 days for clinical signs of disease. Samples of blood were obtained from dogs and pigs immediately before the dogs were inoculated with virus and 14 and 28 days later. The dogs developed mild clinical signs of an infection, but the pigs remained normal throughout the observation period. The dogs shed CCV for eight days after exposure. All three pigs developed neutralizing antibodies against CCV and transmissible gastroenteritis virus by 14 days after they were exposed to the dogs.     

Persistence of antibodies to Borrelia burgdorferi in dogs of New York and Connecticut

Multiple blood samples were obtained from privately owned dogs living in tick-infested areas of New York (Westchester County) and Connecticut, where Lyme disease in human beings has been reported. Of the 175 dogs examined, 127 (72.6%) had limb/joint disorder, whereas the remaining 48 dogs were considered healthy. Results of analysis of 419 serum samples revealed IgM antibody to Borrelia burgdorferi in healthy and lame dogs during all seasons. Prevalence of seropositivity was significantly (P less than 0.01) greater, using a polyvalent ELISA (89.5%) than using a class-specific ELISA for IGM antibody (57.8%). Mean antibody titers obtained by use of polyvalent ELISA were likewise higher than IgM titers. Analysis of paired serum samples from dogs with limb/joint disorder indicated that 118 (92.9%) remained positive for IgM or IgG antibodies when retested weeks or months after initial testing. In 48 dogs without history of joint involvement or other signs of disease, 43 (89.6%) had antibody to B burgdorferi 2 or more times. Serotest results also revealed little or no change in antibody titer for lame dogs given antibiotics or for healthy dogs 2 or more months after initial sample collection.     

Determination of IgM and IgG antibodies to Toxoplasma using the IFA test, ELISA, and Dot-ELISA procedures

The dot enzyme-linked immunosorbent assay (Dot-ELISA) and the enzyme-linked immunosorbent assay (ELISA) were compared with the immunofluorescent antibody test (IFA) for detection of IgM- and IgG-specific antibodies to human toxoplasmosis. Reciprocal titers were determined in all three assays using sera from 56 patients with suspected toxoplasmosis or with symptoms and diseases requiring exclusion of toxoplasmosis and control sera from 56 healthy persons. Using the Dot-ELISA, six patient sera (10.7%) were positive at titers of greater than equal to 1024 for IgM antibodies (titer range 1024-16 384) and 47 sera (84%) were positive for IgG antibodies (titer range 16-262 144) at a titer of greater than or equal to 16. One control serum was reactive for IgM (titer 1024) and 10 control sera (18%) were positive for IgG in the Dot-ELISA. In the ELISA, at titers of greater than or equal to 128, five sera (9%) were reactive for IgM (titer range 128-512) and 52 sera (92.8%) were reactive for IgG (titer range 32-8192) at a titer of greater than or equal to 32; no control sera gave positive reactions for IgM while 10 sera (18%) were positive for IgG in the ELISA. Using the IFA test at reciprocal titers of greater than or equal to 16, four sera (7.1%) were positive for IgM (titer range 32-512), and 51 sera (91%) were positive for IgG (titer range 16-8192). None was reactive for IgM, and eight sera (14%) were positive for IgG (titer range 32-128) in the IFA test. The Dot-ELISA correlated well with the IFA test (correlation coefficient = 0.895) and the ELISA correlated slightly higher with the IFA test (correlation coefficient = 0.910) for detection of IgG antibodies to Toxoplasma gondii.     

Evaluation of a dot ELISA kit for measuring immunoglobulin M antibodies to canine parvovirus and distemper virus

A dot ELISA for the detection of immunoglobulin M (IgM) antibodies to canine distemper virus (CDC) and canine parvovirus (CPV) was assessed. The titres of IgM antibodies to CDV and CPV in 100 dogs were measured by the Immunocomb ELISA kit and compared with the results derived from the immunofluorescence assay (IFA). There was a strong correlation between the results of the dot ELISA technique and the IFA (P < 0.001). The dot ELISA kit was also used to assess the changes in the levels of immunoglobulin G (IgG) and IgM antibodies to CPV and CDV in 10 puppies vaccinated with a polyvalent vaccine. High levels of IgM antibodies to CPV were first detected seven days after they were vaccinated, and after nine days all the pups had high titres of IgG antibodies to CPV. High levels of IgM antibodies to CDV were detected after nine days and the highest average titres were recorded after 12 days. IgG antibodies to CDV were present from nine days after vaccination.     

Serosurvey for canine distemper virus exposure in dogs in communal lands in Zimbabwe

Sera from 173 apparently healthy, unvaccinated dogs from 4 widely separated communal lands in Zimbabwe were tested by ELISA for antibodies against canine distemper virus. Overall, 82% were positive with high prevalences found in each communal land. The highest seroprevalence was in dogs between 1 and 2 years of age (91%; 49/54). These results show dogs in the communal lands of Zimbabwe are commonly exposed to canine distemper virus and that a substantial number survive infection. The role that the virus might play in the high mortality rate of the dog population on communal land warrants further investigation.     

Profile of anti-Leishmania antibodies related to clinical picture in canine visceral leishmaniasis

This research investigated the profile of anti-Leishmania antibodies in different clinical forms of canine visceral leishmaniasis (CVL). Naturally infected dogs were divided into two groups: subclinical dogs (SD, n=10) and clinical dogs (CD, n=68). Non-infected dogs (ND, n=7) comprised the negative control group. The humoral response was evaluated by the profile of total IgG, IgG1, IgG2, IgM, IgA and IgE, determined by ELISA. Infected animals showed increased levels of total IgG, IgA and IgE in addition to IgG1 and IgG2 in groups SD and CD, when compared with group ND. Furthermore, it was observed that IgG2 and IgM were correlated with symptomatology, while total IgG, IgG1 and IgA were negatively correlated and IgE showed no correlation. It follows that serum levels of IgG2 anti-Leishmania are correlated with typical clinical signs of disease. Furthermore the determination of specific anti-Leishmania antibodies could be an important tool in monitoring CVL clinical picture.     

Anti-insulin antibodies in diabetic dogs before and after treatment with different insulin preparations

Background: Anti‐insulin antibodies (AIA) occur in diabetic dogs after insulin therapy, although their clinical significance is unclear. Hypothesis: Treatment of diabetic dogs with heterologous insulin is more likely to stimulate production of AIA than is treatment with homologous insulin. Animals: Diabetic dogs sampled before insulin therapy (n = 40), diabetic dogs sampled following treatment with porcine (homologous) insulin (n = 100), bovine (heterologous) lente insulin (n = 100), or bovine protamine zinc (PZI) insulin (n = 20), and nondiabetic control dogs (n = 120). Methods: Prospective observational study. Sera were analyzed by ELISA for antibodies against porcine insulin, bovine insulin, insulin A, B, or C peptides, and control antigens; canine distemper virus (CDV) and canine thyroglobulin (TG). Canine isotype‐specific antibodies were used to determine total and anti‐insulin IgG1 : IgG2 ratios. Results: There was no difference in CDV or TG reactivity among the groups. AIA were detected in 5 of 40 newly diagnosed (untreated) diabetic dogs. There was no significant difference in AIA (ELISA optical density reactivity) comparing control and porcine insulin‐treated diabetic dogs (P > .05). Anti‐insulin reactivity was most prevalent in bovine PZI insulin‐treated dogs (90%; P < .01), and bovine lente insulin‐treated dogs (56%; P < .01). AIA induced by treatment were enriched for the IgG1 isotype. Conclusions and Clinical Importance: This study indicates that bovine insulin is more immunogenic than porcine insulin when used for treatment of diabetic dogs.     

Clinical,clinicopathologic, and radiographic findings in dogs with coccidioidomycosis: 24 cases (1995-2000)

OBJECTIVE: To determine clinical, clinicopathologic, and radiographic abnormalities in dogs with coccidioidomycosis. DESIGN: Retrospective case series. ANIMALS: 24 dogs. PROCEDURE: Clinical information and results of clinicopathologic testing were obtained from medical records. Thoracic radiographs were reviewed to characterize abnormalities. RESULTS: Dogs ranged from 1 to 10 years old at the time of diagnosis, with 12 dogs being between 1 and 3 years old. Historical complaints included cough, lameness, signs of head or neck pain, and difficulty breathing. Mild anemia, neutrophilia, and monocytosis were common. All dogs had hypoalbuminemia, and 8 of 15 had hyperglobulinemia. Thoracic radiographs of 19 dogs were reviewed. Pulmonary infiltrates were seen in 13 dogs, with an interstitial pattern of infiltration being most common. Hilar lymphadenopathy was seen radiographically in 10 dogs. Serum from 20 dogs was tested for antibodies against Coccidioides immitis. One dog was positive for IgM antibodies, 5 were positive for IgM and IgG antibodies, and 14 were positive for IgG antibodies. Quantitative IgG titers measured in 14 dogs ranged from 1:2 to 1:128 (median and mode, 1:32). In 6 dogs, histologic examination of biopsy samples revealed fungal spherules ranging from 8 to 70 microm in diameter. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that in dogs, coccidioidomycosis may be associated with a wide spectrum of nonspecific respiratory and musculoskeletal abnormalities. The chronic nature of the disease makes diagnosis difficult, even in regions in which the organism is endemic.     

Pitfalls in immunofluorescence testing in canine dermatology

Indirect immunofluorescence testing for pemphigus-like antibodies was performed on 100 dogs: 50 dogs with various nonpemphigus dermatologic diseases, 25 dogs with various nondermatologic diseases, and 25 normal dogs. One dog (generalized demodicosis) was positive for pemphigus-like antibodies at a titer of 1:10. It was concluded that canine pemphigus-like antibodies are a potential source of misinterpretation and misdiagnosis in indirect immunofluorescence testing. Direct immunofluorescence testing for IgG, IgA, IgM, and C3 was performed on the footpads of 11 normal dogs. Granular deposition of IgM at the basement membrane zone was demonstrated in 5 of the 11 dogs. It was concluded that direct immunofluorescence testing of canine footpads using only polyvalent immunoglobulin antisera or anti-IgM antisera may lead to misinterpretation and misdiagnosis in up to 45% of all dogs tested.     

Differentiation of canine coronavirus and porcine transmissible gastroenteritis virus by neutralization with canine,porcine and feline sera

Monospecific antisera were prepared in rabbits against canine coronavirus (CCV) and transmissible gastroenteritis virus of pigs (TGEV), and in 24 pigs and 3 cats against TGEV alone. Neutralizing antibody titres were higher for the immunizing than the heterologous virus, although cross-neutralization usually was detected. This confirmed that CCV and TGEV are distinct, but antigenically related coronaviruses. In sera from 41 dogs, CCV-neutralizing titres were on average 2.7 fold higher than TGEV-neutralizing titres, suggesting that CCV was the causal agent. Sera from 29 cats in colonies with feline infectious peritonitis (FIP) and known to contain TGEV-neutralizing antibody, were found to have titres 12.3 fold higher against CCV. The FIP virus (FIPV) is probably more closely related to CCV than TGEV as judged by antigens involved in virus neutralization.Antisera to two isolates of bovine coronavirus, three isolates of haemagglutinating encephalomyelitis virus, seven strains of avian infectious bronchitis virus and the 229E strain of human coronavirus all failed to neutralize CCV and TGEV. Thus CCV, TGEV and probably FIPV fall into a group of antigenically related agents, separable from other members of the family Coronaviridae, by both virus neutralization and immunofluorescence tests.     

Seroprevalence of various infectious agents in dogs with suspected acute canine polyradiculoneuritis

Background: Acute canine polyradiculoneuritis (ACP) is considered to be an animal model of the acute axonal form of Guillain‐Barré syndrome (GBS) in humans. Various antecedent events have been associated with GBS, including bacterial or viral infection. The relationship between ACP and previous infection requires additional attention. Hypothesis: We hypothesized a relationship between ACP and serological evidence of exposure to Ehrlichia canis, Borrelia burgdorferi, Toxoplasma gondii, Neospora caninum, Campylobacter jejuni, and canine distemper virus (CDV). Animals: Eighty‐eight client‐owned dogs, 44 with ACP, 44 age‐matched controls. Methods: Retrospective study with stored serum samples. Serum antibodies against the target organisms were measured with commercially available assays. Sera from dogs with and without ACP that were positive for T. gondii IgG by ELISA were assayed by an IgG heavy chain‐specific, Western blot immunoassay. Results: Dogs with ACP (55.8%) were more likely to have T. gondii IgG serum antibody titers than dogs without ACP (11.4%). Serum antibodies from 8 affected dogs and 11 control dogs bound to T. gondii antigens with apparent molecular masses of 67, 61, 58, 45, 33, 24, 9, and 6 kDa. An antigen with an apparent molecular mass of 36 kDa was recognized by 2 dogs with ACP but none of the control dogs. Conclusions: Results of this study suggest that ACP in some dogs, like GBS in some humans, may be triggered by T. gondii and a prospective study should be performed to further evaluate this potential association.     

Microscopic, serologic and molecular surveys on Dirofilaria immitis in stray dogs, Turkey

The prevalence of Dirofilaria immitis infection was evaluated in stray dogs of Erzurum, Turkey. A total of 123 whole-blood and 93 sera samples were collected from stray dogs older than 6 months were lived in animal shelter. The PCR and direct microscopic examinations were used for the detection of microfilaria and indirect-ELISA was performed for the detection of anti-D. immitis antibodies. The prevalence of D. immitis in the canine population was 8.1% by PCR, 2.1% by ELISA. In addition, microfilaria burdens of Dirofilaria sp. was 4.8% by direct blood smear examination. There was a statistical difference (P=0.05) in the prevalence between males (10.5%) and females (2.3%) by direct blood smear examination. Similarly there was a statistical difference (P<0.05) in the prevalence between males (15.8%) and females (4.7%) by PCR. Dogs belonging to the 0.5-1 years old group showed the highest prevalence than 2-4 ages group with three tests. Among the 93 samples screened by the ELISA, two samples were positive for the D. immitis antibodies. Both positive dogs with ELISA were females.     

Detection of canine coronaviruses genotype I and II in raised Canidae animals in China

An RT-nPCR assay was used for testing fecal samples of dogs, foxes, raccoon dogs and minks for the presence of canine coronavirus (CCV). The animals were raised in homes, dog schools or farms. Seventy out of 81 healthy dog feces from three cities and 21 out of 48 diarrhea feces from pet dogs were positive for type II CCV. From a total of 61 healthy fox feces, 43 were positive for type II and 29 for type I CCV, out of which 25 were simultaneously positive for the two different genotypes. Among 24 raccoon dogs samples, 22 were CCV type II-positive, and from those 16 were additionally type I positive. No CCVs was detected from healthy mink feces. Sequence analysis found that ten type II CCVs fragments of M gene shared a high similarity with reference strain CCV 1-71 (96.5-99.5%), and four type I CCVs shared a high similarity (96.7%-98.1%) with a reported FCV-like CCV strain. The sequence of one particular M gene fragment was found to cluster between the type I and type II CCV branches in phylogenetic analysis, suggesting the existence of a novel strain. Our study confirmed that type II CCVs infection is very common in domestic dog, fox, and raccoon dog populations in China. This is also the first report on the co-existence of two CCV genotypes in healthy foxes and raccoon dogs.