Immunodeficiency in canine demodectic mange. I. Experimental production of lesions using antilymphocyte serum

Two experiments were conducted to further clarify the importance of cell-mediated immunity in cases of canine demodectic mange. Pups obtained from pregnant pedigreed beagle bitches were exposed to mites (Demodex canis Leydig, 1859) and then injected with rabbit anticanine lymphocyte serum (ALS). Littermates received either mites alone, ALS alone, or were retained as negative controls. Squamous lesions of demodicosis developed in all pups receiving both ALS and mites as well as in two pups receiving only mites. No lesions were observed in any of the remaining pups. A marked decline in the number of peripheral lymphocytes developed in pups receiving ALS. These cells returned to normal numbers 3 weeks after treatment was discontinued. The squamous lesions on affected pups continued to coalesce until generalized demodicosis became established. Spontaneous resolution of mite-infested areas was never observed and pustular lesions did not develop.It was concluded that the cellular immune system of the dog, functioning as a defense mechanism against the establishment and proliferation of D. canis, is of paramount importance in preventing the development of demodectic mange.     

Immunodeficiency in canine demodectic mange. II. Skin reactions to phytohemagglutinin and concanavalin A

The T-lymphocyte mitogens phytohemagglutinin (PHA) and concanavalin A (Con A) were used as skin test agents to assess the cellular immune competence of dogs with demodectic mange. The intradermal injection of either mitogen into the abdomen of dogs produced a local erythematous and indurated papule characteristic of a delayed hypersensitive response. The mean diameter of the reaction at 24 and 48 following injection was significantly smaller in eight demodectic dogs than in eight dogs free from mite infestation (P < 0.05). This suppressed response suggested that dogs with demodicosis may have a hypoactive cellular immune system rendering them less capable of combating invading demodicids.The intradermal injection of PHA and Con A also produced an immediate wheal reaction. The mean diameter of the wheal after 15 min was significantly larger in demodectic dogs than in the control animals (P < 0.05). A possible reason for this difference and a potential mechanism for the reaction are discussed.     

Bovine reaginic antibody III. Cross-reaction of antihuman IgE and antibovine reaginic immunoglobulin antisera with sera from several species of mammals.

Using antisera specific for the heavy chain of human IgE and bovine reaginic immunoglobulin, the degree of cross-reaction amongst sera from pig, rat, rabbit, guinea pig, goat, cow, horse, dog, cat and human was tested. Antihuman IgE antiserum gave strong reactions with pig, rabbit, cow, goat and human sera (100% to 15.1%) and weak reactions with rat, guinea pig, horse, dog and cat sera (10.1% to 3.22%). Antibovine reagin antiserum produced a considerable amount of cross-reaction with sera from pig, rat, rabbit, goat, horse and human (43.6% to 20.1%) with limited reactions with guinea pig, dog and cat sera (13.9% to 9.3%).     

Unusual manifestation of a concurrent demodectic and sarcoptic mange in a Zebu-Friesian cross-bred heifer

We describe a rare case of a concurrent demodectic and sarcoptic mange in a 2-year-old heifer in Khartoum, Sudan. The lesions were massive lumps of granulomatous tumour-like dermatitis with thick, nodular folds mainly covering the head, neck and shoulders. Histopathological examination of the lesions revealed the presence of both Demodex bovis and Sarcoptes scabiei. The animal died regardless of the anti-parasitic treatment it received.     

Differential serological testing by simultaneous indirect immunofluorescent antibody test in canine leishmaniosis and ehrlichiosis

A mixed indirect fluorescence antibody test (IFAT), based on cultured promastigotes Leishmania infantum and formol-inactivated suspension of cells infected with the bacteria Ehrlichia canis, was applied to make a differential diagnosis between canine ehrlichiosis and leishmaniosis. A titre greater than 80 was considered positive for antibodies to E. canis and suggestive of antibodies to L. infantum. Positive sera were titrated subsequently by serial dilutions to confirm antibodies positive to Leishmania and establishing the antibody titre of both pathogens. Fluorescence was absent with negative control sera and background staining was minimal. No serological cross-reactions between positive sera for L. infantum or E. canis were detected. Results obtained by mixed IFAT did not differ when the same serum IFAT standard was compared. The test showed equivalent sensitivity (100%). The specifities were 100% for L. infantum and 98.5% for E. canis. The equivalence in sensitivity was confirmed by calculating the correlation coefficient between IFAT standards and mixed IFAT (r>or=0.99 for both pathogens). The results of our investigations demonstrated that mixed IFAT is a specific means of establishing serological differential diagnosis of canine leishmaniosis and ehrlichiosis.     

Efficacy of a novel formulation of metaflumizone plus amitraz for the treatment of demodectic mange in dogs

A novel spot-on formulation containing metaflumizone plus amitraz (ProMeris/ProMeris Duo for Dogs, Fort Dodge Animal Health, Overland Park, KS) was evaluated for efficacy against demodectic mange mites in naturally infested dogs. Sixteen dogs were allocated to two equal groups and individually housed. Eight of the dogs were treated topically with metaflumizone plus amitraz at the proposed minimum dose rate (20mg/kg of each of metaflumizone and amitraz, 0.133ml/kg) on Days 0, 28, and 56. The other eight were treated with metaflumizone plus amitraz at the proposed minimum dose rate on Days 0, 14, 28, 42, 56, and 70. Mite numbers were estimated from skin scrapings taken on Days -3 to -1, 28, 56, and 84. Clinical signs of mange and the extent of demodectic lesions on each dog were evaluated when skin scrapings were conducted. Efficacy of the treatment was based on a reduction in mite numbers and an assessment of the clinical signs associated with canine demodectic mange. Treatment at monthly or two-weekly intervals for 3 months resulted in a rapid reduction in mite numbers (>94 and >99% for the monthly and two-weekly treatments, respectively) and an improvement in clinical signs. Success rates, based on zero mite counts in skin scrapings at Day 84 were 42.9 and 62.5% of dogs for the monthly and two-weekly regimens, respectively.     

Evaluation of an enzyme-linked immunosorbent assay (ELISA) for the serological diagnosis of canine sarcoptic mange

Abstract The purpose of this study was to evaluate a serodiagnostic test (enzyme-linked immunosorbent assay; ELISA) for sarcoptic mange in dogs and to characterize the assay antigen, based on the mite Sarcoptes scabiei var. vulpes. The ELISA, applied to sera from 359 dogs suspected of having sarcoptic mange, showed a sensitivity and specificity of 92 and 96%, respectively. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) of the antigen employed in the ELISA revealed polypeptide bands with molecular weights ranging between 14 and 164 kDa. In Western blot analyses antigens of molecular weights between 62 and 64 kDa dominated. Particularly dominant were antigens of 164 and 147 kDa. These were found to have isoelectrical points in the range of 5.7–6.9. Sera from dogs infected with Cheyletiella sp., Demodex canis, Linognathus setosus and Otodectes cynotis, as well as from dogs allergic to fleas, were negative in the ELISA. Résumé— Le but de cette étude est d'évaluer un test sérologique ELISA pour le diagnostic de la gale sarcoptique chez le chien et de caractériser l'antigène révélateur, extrait de l'acarien Sarcoptes scabiei var. vulpes. Le test ELISA, lors d'une étude conduite avec les sérums de 359 chiens suspects de gale sarcoptique a démontré une sensibilité et une spécificité de 92 et 96%, respectivement. L'électrophorèse en gel polyacrilamide dodécyl sulfate de sodium de l'antigène utilisé dans l'ELISA a révélé des bandes polypeptidiques de poids moléculaire compris entre 14 et 164 kDa. Dans l'analyse en Western blot, les antigènes de poids moléculaire compris entre 62 et 164 kDa étaient les plus abondants, notamment ceux de 164 et 147 kDa. Ces derniers ont des points isoélectriques compris entre 5.7 et 6.9. Les sérums de chiens infectés par des Cheyletiella sp. Demodex canis, Linognathus setosus et Otodectes cynotis, ou par des chiens allergiques aux puces, se sont révélés négatifs en ELISA. [Bornstein, S., Thebo, P., Zakrisson, G. Evaluation of an enzyme-linked immunosorbent assay (ELISA) for the serological diagnosis of canine sarcoptic mange (Evaluation d'un test ELISA pour le diagnostic sérologique de la gale sarcoptique canine). Veterinary Dermatology 1996; 7 : 21–28.] Resumen El objetivo de este estudio fue el de evaluar una pruba serodiagnóstica (prueba de inmunoadsorción ligada a enzima; ELISA) para la sarna sarcóptica en el perro y caracterizar el antigeno prueba, basado en el ácaro Sarcoptes scabei, var. vulpes. El ELISA, aplicado a sueros de 359 perros sospechosos de padecer sarna sarcóptica, mostró una sensibilidad y especificidad del 92 y 96%, respectivamente. La electroforesis en gel de poliacrilamida dodecil sulfato sódico (SDS-PAGE) del antigeno usado en el ELISA reveló bandas de polipétidos con peso molecular entre 14 y 164 kDa. En el análisis Western blot, predominaron los antigenos de pesos moleculares entre 62 y 164 kDa. Los antigenos entre 164 y 147 kDa fueron especialmente predominantes. Estos tuvieron puntos isoeléctricos entre 5.7 y 6.9. Los sueros de perros infectados por Cheyletiella sp., Demodex canis, Linognathus setosus y Otodectes cynotis, asi como el de perros alérgicos a las pulgas fueron negativos en el ELISA. [Bornstein, S., Thebo, P., Zakrisson, G. Evaluation of an enzyme-linked immunosorbent assay (ELISA) for the serological diagnosis of canine sarcoptic mange (Evaluation de una prueba de immunoadsorcion ligada a enzima (ELISA) para el diagnostico serologico de la sarna sarcóptica canina). Veterinary Dermatology 1996; 7 : 21–28.] Zusammenfassung— Ziel dieser Studie war, einen Serodiagnostiktest (Enzyme-Linked-Immunosorbent-Assay, ELISA) für Sarkoptesräude des Hundes zu überprüfen und das Testantigen zu charakterisieren, das auf der Milbe Sarcoptes scabiei var. vulpes basiert. Der ELISA-Test, der bei den Sera von 359 Hunden mit Sarkoptesverdacht angewendet wurde, zeigte eine Sensitivität von 92% bzw. 96%. Die Natriumdodecylsul-fatpolyacrylamid-Gelelektrophorese (SDS-PAGE) des Antigen, das im ELISA verwendet wurde, zeigte Polypeptid-Banden mit Molekulargewichten zwischen 14 und 164 kDa. In der Wester-blot-Analyse dominierten Antigene mit einem Molekulargewicht zwischen 62 und 164 kDa. Besonders dominierend waren Antigene von 164 und 147 kDa. Bei diesen stellte man isoelektrische Punkte im Bereich von 5,7 bis 6,9 fest. Die Sera von Hunden, die mit Cheyletiella sp., Demodex canis, Linognathus setosus und Otodectes cynotis infiziert waren, fielen ebenso wie die Hunde mit Allergie auf Flöhe im ELISA negativ aus. [Bornstein, S., Thebo, P., Zakrisson, G. Evaluation of an enzyme-linked immunosorbent assay (ELISA) for the serological diagnosis of canine sarcoptic mange (Die Auswertung eines Enzym-Linked-Immunosorbent-Assay (ELISA) für die serologische Diagnose der kaninen Sarkoptesräude). Veterinary Dermatology 1996; 7 : 21–28.]     

Evaluation of enzyme-linked immunosorbent assay (ELISA) and immunofluorescent antibody (IFA) test for the detection of porcine reproductive and respiratory syndrome virus (PRRSV) antibody in pigs from conventional farms

To evaluate the immunofluorescent antibody (IFA) test and enzyme-linked immunosorbent assay (ELISA) for detecting the porcine reproductive and respiratory syndrome virus (PRRSV) antibody, conventional pigs in PRRSV-positive and -negative commercial farms were examined. Antibody development patterns in ELISA and IFA tests were compared in 3 week old piglets experimentally infected with the PRRSV. The virus was detected from 2 days post infection (PI) and then the antibody titers and S/P ratios rose by both methods. A total of 208 serum samples were collected from 4 PRRSV-negative farms and 210 samples from PRRSV-positive farms, and were tested for the PRRSV antibody by IFA and ELISA. The titer of 64 should be set as the cut-off point in IFA for field sera. Similarly, the cut-off S/P ratio should be set at 0.4 in ELISA. A high degree of correlation was observed between antibody titers by the two methods in these 418 samples, with a correlation coefficient of 0.84. The coincidence rate between the two tests was 84.7% (354/418). In non-coincident cases, ELISA was able to detect the antibody with a low titer in the serum samples which were negative in IFA but from PRRSV positive farms. ELISA was more sensitive than IFA to detect PRRSV infected animals or farms.     

An outbreak of canine tracheobronchitis (kennel cough) in Zaria

The clinical and epidemiological features of the first recorded outbreak of canine tracheobronchititis (kennel cough) in Zaria are described. During the outbreak which occurred between October and December 1980, 21 per cent of dogs presented to the Small Animal Clinic were diagnosed clinically as suffering from kennel cough, compared with 0.2 per cent in the preceding year. Bordetella bronchiseptica was among the several microorganisms isolated from affected dogs.     

免疫荧光技术快速检测鸡轮状病毒的研究

用分离的轮状病毒与福氏完全佐剂和福氏不完全佐剂免疫家兔制备高免血清，分离球蛋白，用荧光素标记，建立荧光抗体快速诊断鸡轮状病毒技术。阴性试验、阳性试验、类症试验表明，建立的荧光抗体诊断技术特异性强、敏感性高，与新城疫病毒、禽流感病毒、传染性法氏囊病病毒、大肠杆菌不发生交叉反应。人工感染试验结果表明，20日龄SPF鸡感染轮状病毒后12h，肠粘膜深层涂片可出现较强荧光。自然感染检测结果表明，荧光抗体诊断结果与病毒的分离鉴定结果一致，符合率达100％。     

FC-44 The therapeutic effect of selamectin and ivermectin regimens in canine sarcoptic mange

Worldwide, sarcoptic mange in cats is seldom reported, and then only in sporadic individual cases. We describe an epidemic in a household with a dog and 25 cats. From September 2002, the dog was repeatedly treated with ivermectin for sarcoptic mange. The diagnosis was confirmed by skin scrapings. Fifteen months later, cats from the same household were diagnosed with severe sarcoptic mange. Twenty-one of the cats were euthanized and necropsies were performed. Skin samples were taken from all cats from different body sites for histology, and skin scrapings were examined for ectoparasites. Samples for bacterial and dermatophyte culture were taken from six cats. Smears for cytology were made from lesions on four cats with severe mange. Sera from 21 cats and the dog were analysed for specific antibodies to Sarcoptes scabiei . Molecular characterizations of six individual mites were done. Large numbers of S. scabiei were isolated from the infected skin of most of the cats. Two-thirds of the cats showed skin lesions compatible with chronic sarcoptic mange. Macroscopically, internal organs exhibited no obvious pathology. Yeast organisms and coccoid bacteria were found in the smears; penicillinase-negative Staphylococcus aureus was isolated from all samples and Malassezia pachydermatis was identified from four cats. Sarcoptes scabiei was seen histologically in all cats showing chronic skin lesions. No other ectoparasites were found. All analysed cats had specific antibodies against S. scabiei . Twenty-one cats tested negatively for FeLV and FIV. The mites had DNA sequences identical to S. scabiei from naturally infected dogs and Swedish wildlife.
Funding: Self-funded.     

ELISA方法检测犬瘟热抗体初探

本实验以细胞培养犬瘟热病毒作为包被抗原,建立了检测犬瘟热病毒血清抗体的间接ELISA方法。该检测方法抗原最佳包被浓度为5μg/mL,血清最佳稀释度为1∶100。阻断实验、敏感性实验等结果表明该方法特异性强、重复性较好,可用于犬瘟热血清抗体的定量和定性检测。