Evaluation of clinical, laboratory, imaging findings and outcome in 99 dogs with leptospirosis.

OBJECTIVE: To report clinical, laboratory and diagnostic imaging features and prognostic factors in dogs with leptospirosis from North-East Germany. MATERIALS AND METHODS: Medical records of dogs diagnosed with leptospirosis from 2006 to 2013 were evaluated retrospectively. RESULTS: The study included 99 dogs. At initial presentation, the most common clinical signs were lethargy (96%), anorexia (88%), vomiting (85%), painful abdomen (39%), diarrhoea (38%), oliguria (27%) and tachypnoea (26%). Abnormal laboratory findings included anaemia (63%), thrombocytopenia (63%), leucocytosis (57%), increase of plasma urea (84%) and creatinine concentrations (81%), increased liver enzyme activities (80%), hyperbilirubinaemia (69%), hyperphosphataemia (67%), hyponatraemia (64%), hypoalbuminaemia (55%) and hypokalaemia (29%). Radiological pulmonary changes were detected in 57% of the dogs initially or during the course of disease. Severe dyspnoea, oliguria, azotaemia, hyperbilirubinaemia and severe radiological pulmonary changes were more often found in dogs that did not survive. There was renal, hepatic and pulmonary involvement in 95, 92 and 58% of the dogs, respectively, and multi-organ lesions in 98 dogs (98%); 32 dogs died or were euthanased. CONCLUSION: Several clinical and laboratory abnormalities were associated with a negative outcome; severe lung involvement was specifically associated with high mortality.

Pulmonary abnormalities in dogs with leptospirosis.

BACKGROUND: Leptospirosis in dogs is a multiorgan disease affecting mostly kidneys and liver. OBJECTIVES: The objective was to characterize prevalence, clinical, and radiological features and outcome of dogs with leptospirosis and pulmonary abnormalities. ANIMALS: Fifty dogs with leptospirosis. METHODS: Medical records of dogs diagnosed with leptospirosis at the Small Animal Clinic, Berlin, were reviewed. Diagnosis was based on microscopic agglutination test, blood or urine polymerase chain reaction, and histopathology. Based on clinical and/or radiological signs, patients were grouped into dogs with lung abnormalities (group 1) or without (group 2). Severity of respiratory distress was scored as mild to moderate (grade 1) or severe (grade 2). Thoracic radiographs were scored based on pulmonary changes and location as grade 1 (caudal interstitial pattern), 2 (generalized mild to moderate reticulonodular interstitial pattern), or 3 (generalized severe reticulonodular interstitial pattern with patchy alveolar consolidations). Results of CBC and biochemistry were compared between groups. RESULTS: Thirty-five dogs had radiological pulmonary changes (grade 1: 5; grade 2: 14; grade 3: 16); 31 of them had pulmonary distress (grade 1: 13, grade 2: 18). Sixty-seven percent of the dogs with dyspnea grade 2 were mainly euthanized because of respiratory distress. Fifteen percent of the dogs with dyspnea grade 1 and 21% without clinical respiratory signs were euthanized because of acute renal failure or sepsis. CONCLUSIONS AND CLINICAL IMPORTANCE: In 70% of dogs with leptospirosis pulmonary changes were detected. Lung involvement represented a severe complication causing increased case fatality depending on the severity of respiratory distress.

Polystyrene as an affinity chromatography matrix for the purification of antibodies.

Affinity chromatography is used for the purification of diagnostic polyclonal antibodies in order to ensure specificity. Most commonly, activated bead-formed agarose or its derivatives are used as gel matrices. Alternative matrix materials have been described, but as yet they do not appear to offer important advantages. In this study, pulverized polystyrene (PS 158K, BASF, Mannheim, Germany) was used as a solid phase for the immobilisation of bovine immunoglobulins (Ig). Affinity chromatography was performed using these coated polystyrene beads as the column matrix material in the purification of anti-bovine Ig. The polystyrene binding capacity for the different bovine Ig classes was compared using the Mancini single radial immunodiffusion technique, and ELISA procedures were used to monitor the antibody reactivity of purified and unpurified antibodies. The degree of purification was comparable to the most commonly used procedure using gel matrices from activated bead-formed agarose (e.g. CNBr-activated Sepharose 4B, Pharmacia/LKB Biotechnology, Uppsala, Sweden), but the antibody yield per ml column volume was distinctly lower. In order to raise the yield, such polystyrene bead columns with immobilized antigen can be re-used without loss of activity or larger column volumes can be used to raise the binding capacity. The polystyrene material is quite durable, chemically and immunologically inert and has a long shelf life. We conclude that polystyrene based affinity chromatography is efficient, simple and cheap.

Copro-antibody in calves from dams vaccinated against Salmonella typhimurium.

Calves received colostrum either with (positive colostrum) or without (negative colostrum) anti-Salmonella typhimurium antibodies. Regarding the detectability of copro-antibodies, the following observations have been made. In calves that had been given positive colostrum on day 4 post natum (p.n.), copro-antibodies were detectable over 7 days, while in those that had received positive colostrum on day 1 p.n. copro-antibodies were detectable over 45 days. After supplying highly positive colostrum, copro-antibodies were found over a period of 8 weeks, and after supplying weakly positive colostrum, they were found over a period of 5 weeks. It is concluded that serum antibodies are transferred to the intestine for maximum local protection, and that there exists a preference for the intestinal system regarding the distribution of salmonella antibodies. Challenge infection on day 5 p.n. of calves that had received highly positive colostrum resulted in a copro-antibody gap that reached the limit of detectability in two calves that were excreting salmonellas. Challenge infection of calves that had received negative colostrum provoked a local IgM and IgA antibody response.

[Immunologic memory in the course of udder immunization for the protection of calves against Salmonella infections]. / Immunologisches Gedächtnis (Memory) im Verlauf von Euter-Immunisierung zum Schutz des Kalbes gegenüber Salmonella-Infektionen.

Cows were vaccinated once or twice intracisternally in order to protect calves against salmonella infections. The amount of antibodies measured in the colostrum of re-vaccinated and not re-vaccinated cows indicated that a sensitive immunological memory has been provoked by this vaccination technique. This immunological memory showed a distinct synchronism with the state of pregnancy in order to provide a maximum amount of antibodies in the colostrum.

[Udder immunization for the protection of calves from salmonella infections. 2. Estimation of serological findings of colostrum with regard to its protective activity]. / Euter-Immunisierung zum Schutz des Kalbes gegenüber Salmonella-Infektionen. 2. Beurteilung serologischer Befunde des Kolostrums hinsichtlich dessen Schutzwirkung.

Colostrum from cows which had been mammary gland vaccinated against salmonellosis were tested for antibodies by four different serological tests. Results from these tests were related to the protective activity for artificially infected calves. In opposition to agglutinating antibodies and antibodies detected by ELISA, complement fixing antibodies of the IgG1 class were shown to transfer early protection in artificially infected calves.

[A simple method for the isolation of immunoglobulin (Y) from the eggs of immunized hens]. / Einfache Methode zur Isolierung von Immunglobulin (Y) aus Eiern immunisierter Hühner.

Hens were vaccinated with Salmonella typhi-murium and with serum-IgG from a goat. Antibodies (IgY) were isolated from the eggs by ammonium sulphate precipitation. The concentration of egg antibodies was equal or superior to the concentration of serum antibodies of the hen. The longlasting antibody plateau of 9 and 28 weeks, respectively, provides evidence of the high yields of specific IgY which can be collected from a single immunized hen.

[Udder immunization for the protection of calves against salmonella infections. 1. Detection of specific Ig classes in the mother, colostrum and calf and the course of infection]. / Euter-Immunisierung zum Schutz des Kalbes gegenüber Salmonella-Infektionen. 1. Nachweis spezifischer Ig-Klassen im Muttertier, Kolostrum und Kalb sowie Verlauf der Infektion.

Pregnant cows were vaccinated at ablactation by infusion of heat inactivated S. dublin or S. typhimurium into the mammary gland in order to protect their offsprings via colostrum against salmonellosis. This method of vaccination is based upon statements according to which specific IgA and IgM play a prominent role in respect of protection. Both Ig classes are produced locally and are not channelled from the mother's system into the mammary gland. In comparison to calves from non-vaccinated control animals, calves from vaccinated cows did not exhibit clinical symptoms after challenge infection, and had a reduced salmonella excretion rate in respect of quantity but not in respect of the duration of shedding salmonella. Serological quantitation of specific Ig-classes (DIG-ELISA) did not allow to identify a specific Ig-class responsible for protection.