Anaplasmosis in Uganda. I. Use of dried blood on filter papers and serum samples for serodiagnosis of anaplasmosis—A comparative study

Summary The suitability of blood collected on filter papers in comparison with corresponding conventional serum samples in the diagnosis of bovine anaplasmosis was studied using the complement fixation test, DOT-ELISA, Western immunoblot and rapid card agglutination test. Dried blood on Whatman filter paper no. 1 was eluted in PBS 0·05% Tween 20 giving an initial dilution of 1∶10. The reactivity of the eluted samples in both DOT-ELISA and Western immunoblotting were similar to those obtained with the corresponding straight serum sample dilutions. Filter paper samples gave lower reactivity in the remaining tests when compared with corresponding serum samples. There was no significant difference in the reactivity between the eluates from filter papers stored at temperatures ranging between 15·5 and 24°C and those kept refrigerated. Storage at 15·5 to 24°C did not significantly affect reactivity for up to six months. Eluates from filter papers stored for six months at 15·5 to 24°C continued to give similar reactivity as those from freshly prepared filter papers in both DOT-ELISA and Western blot, and in the rapid card agglutination test. It is concluded that collecting blood on filter papers is a suitable technique for large scale seroepidemiological studies on anaplasmosis and offers many advantages in developing countries where transport and cold chain facilities are a major constraint.

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Resumen Se estudió la efectividad de muestras de sangre colectadas en papel filtro, en comparación con las correspondientes muestras convencionales du suero, en el diagnóstico de anaplasmosis bovina, utilizando fijación de complemento, DOT-ELISA, Western, immunoblot y la prueba rápida de la tarjeta. La sangre seca en papel Whatman N^o 1 fue removida con PBS 0·05% entre 20, dando una dilución inicial de 1∶10. La reactividad de las muestras removidas de papel filtro, en la prueba de DOT-ELISA y Western immunoblotting, fueron similares a la obtenida con la correspondiente muestra de suero. Las muestras de papel filtro reaccionaron menos en las otras pruebas, cuando se compararon con las correspondients muestras du suero. No hubo diferencia significativa en la reactividad entre los lavados del papel filtro guardados a temperaturas entre 15·5 y 24°C y aquellos guardados en refrigeración. El almacenaje entre 15·5 y 24°C, no afectó la reactividad hastas seis meses. Los lavados de papel filtro guardados por seis meses entre 15·5 y 24°C, dieron la misma reactividad como los lavados frescos, en la prueba DOT-ELISA y Western blot, lo mismo que en la prueba de aglutinación rápida de tarjeta. Se concluye, qu la colección de sangre en papel filtro, es una buena técnica para estudios epidemiológicos de cierta magnitud, sobre anaplasmosis, ofreciendo ventajas considerables en paises en desarrollo en donde las cadenas de frío son deficientes.

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Résumé La fiabilité du sang récolté sur papier filtre comparée à celle des prélèvements conventionnels de sérum pour le diagnostic de l'anaplasmose a été étudiée à l'aide des tests suivants: fixation du complément, ELISA, immunoblot de Western, test rapide d'agglutination sur carte. Du sang séché sur papier filtre Whatman N^o 1 a fait l'object d'une élution dans une solution de PBS à 0,05 p. 100 (Tween 20) pour donner une dilution de base au dilution de base au 1∶10. Le réactivité des échantillons, autant avec le test ELISA que l'immunoblot Western, a été identique à celle obtenue par dilution directe de sérums homologue. Les échantillons sur papier filtre ont donné une réactivité plus faible pour les autres tests, comparée à celle des échantillons de sérum semblables. Aucune différence significative n'a été décelée quant à leur réactivité les éluats provenant de papiers filtres stockés à des températures comprises entre 15,5 et 24°C at ceux conservés au réfrigérateur. Le stockage entre 15,5 et 24°C n'a pas non plus affecté la réactivité de fa?on significative; les éluats conservés à partir des papiers filtres, à cette même température durant 6 mois, ont montré des réactions identiques que ceux provenant de papiers filtres fra?chement préparés, à la fois avec le test ELISA, celui de Western Blot et le test d'agglutination rapide sur carte. On peut conclure que la collecte du sang sur papier filtre est une technique adaptée à l'étude épidémiologique de l'anaplasmose à grande échelle. Elle offre de nombreux avantages dans les pays en développement où offre de nombreux avantages dans les pays en développement où les moyens de transports et la cha?ne du froid constituent des contraintes majeures.

     

A device for determining the sorptivity of soil aggregates

A device that infiltrated water over a small circular surface area having a radius ranging between 1.45 mm and 2.5 mm was used to measure the sorptivity of initially air-dry aggregates of size greater than 20 mm. The small infiltration area caused the water uptake to be dominated by capillarity that allowed the use of a simple formula to obtain the sorptivity from the steady-state flow rate that occurred very early after the start of imbibition. The results of measurements of sorptivity made on a fine sand agreed with those obtained from measurements on one-dimensional water infiltration into columns of the sand. Sorptivity measurements on stabilized aggregates of a clay soil and on air-dried field aggregates of a clay soil were easily made. The simplicity and rapidity of the method allowed measurements to be conveniently replicated.     

Serum biochemical values of rusa deer (Cervus timorensis russa) in New Caledonia

Blood samples were collected from 91 rusa deer (Cervus timorensis russa), immediately after being shot. Serum mean biochemical values from shot deer are presented for blood urea nitrogen, creatinine, creatine kinase, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, total bilirubin, total protein, albumin, calcium, and phosphorus. Mean total protein and albumin increased with age. There was an age-associated increase of gamma globulins. Mean creatine kinase activity and creatinine, albumin and phosphorus concentrations were higher in stags than in hinds. Pregnant hinds had lower mean creatine kinase activity and phosphorus and higher mean alanine aminotransferase and total protein than non-pregnant hinds. Mean calcium concentration increased when deer were agitated before bleeding.     

The Effect of Nitrogen and Phosphorus Fertilization on the Nitrogen Absorption by Sunflowers

Little is known about the effect of fertilization on the N uptake of sunflowers. A 4² factorial trial with 0, 60, 120 and 180 kg N ha⁻¹ and 0, 15, 30 and 45 kg P ha⁻¹ was conducted over three years. The N content and concentration of leaves, stems and capitula were determined at three growth stages. High N levels increased the N content and concentration of all plant parts at all growth stages sharply. High P levels increased the N content of all plant components through better growth. P has an inconsistent effect on N concentration but tended to decrease it. After flowering the crop assimilated 20 to 25 % of the total N. This implies that N applied can still be applied and utilized by the crop at a late stage. This should be substantiated by further research.     

MAGNETIC RESONANCE, ULTRASOUND AND HISTOPATHOLOGIC CORRELATION OF ACUTE AND HEALING EQUINE TENDON INJURIES

Ultrasonography and MRI have become valuable tools for imaging of tendon injuries. The current study examines the histopathologic basis for the imaging abnormalities. Five injured equine forelimbs and two normal contralateral limbs were studied with high resolution real time ultrasound and MRI. Histologic sections were made and correlated with the diagnostic images. All lesions were readily seen by both modalities. Lesions characterized by hemorrhage, edma, and cellular infiltration were sonolucent on ultrasound and bright on MRI images. MRI returned to normal as fibrogenesis ensued. Ultrasound images remained abnormal until fibrillar reallgnment occured with completion of the healing process. High resolution real time ultrasound and MRI both accurately reflect the tissue abnormalities in acute tendon injuries. The injured horse does appear to be an effective model for correlative imaging studies of tendon injuries.     

The diagnosis of nephrogenic diabetes insipidus in the dog]

Nephrogenic diabetes insipidus is characterized by polyuria, hyposthenuria and compensatory polydipsia. With conventional clinical and laboratory examinations the diagnosis can be established and the differential diagnostic diseases excluded. In order to verify the diagnosis, differentiate the condition from central and nephrogenic diabetes insipidus and rule out psychogenic polydipsia, additional laboratory studies are required: of these, the concentration test, modified Carter-Robbins test and AVP stimulation test are described.     

Intradermal testing of horses with chronic obstructive pulmonary disease and recurrent urticaria.

Six horses with chronic obstructive pulmonary disease (COPD) and 8 horses with recurrent urticaria were skin tested with 67 extracts from 58 allergens, including pollens, epidermals, cultivated farm plants, dusts, molds, and insects. Reactions were evaluated 3 times over a 24-hour period immediately after the injections. Results were compared with those obtained from 11 clinically normal horses. All horses had positive skin test reactions. Significant differences was evident between horses with COPD and clinically normal horses for only 3.0% of the possible extract reactions, and between horses with urticaria and clinically normal horses for only 4.5% of the possible extract reactions. Horses with COPD or urticaria had greater total percentage of allergen extract reactions than did clinically normal horses. Positive reactions were observed at all 3 evaluation periods, and late-onset reactions were not always preceded by positive reaction at earlier periods. All horses with COPD or urticaria had at least 1 skin test reaction that exceeded the mean +/- 2 SD, as calculated for each of the 67 extracts for the group of clinically normal horses.     

Actinobacillus (Haemophilus) pleuropneumoniae: use of coagglutination and complement fixation to determine the relationship between presence of organism and antibody titer in slaughterhouse pigs

The conventional culture method was compared to coagglutination for detection of Actinobacillus (Haemophilus) pleuropneumoniae in 425 sets of pig lungs. Sera from the same animals were evaluated for antibodies to A. pleuropneumoniae by the complement fixation (CF) test. All samples were collected at 2 packing plants in Iowa. In 2 nonvaccinated herds with no history of respiratory disease, the difference between standard culture results and coagglutination was highly significant (P less than 0.001). None of the 57 pigs in this group were positive for A. pleuropneumoniae by conventional culture, but 7 were positive by the coagglutination test. There were 15 animals with CF titers between 1:8 and 1:32. Animals from 6 herds vaccinated for A. pleuropneumoniae and without recent respiratory problems were evaluated. One out of 118 animals tested was positive for A. pleuropneumoniae by standard culture as compared to 9 positive by coagglutination. The difference in positive results between culture and coagglutination was highly significant (P less than 0.001). Twenty-eight animals had CF titers to A. pleuropneumoniae (1:4 to greater than or equal to 1:128). Two hundred fifty lungs and sera samples were collected from 7 herds which had recently experienced varying degrees of respiratory disease. Thirty-nine lungs were positive for A. pleuropneumoniae by culture and 182 were positive by coagglutination. The number of positives detected by coagglutination was significantly different (P less than 0.001) from the number positive by culture. There were 172 animals with antibody titers ranging from suspect to greater than or equal to 1:128. There were significantly fewer positive animals detected by standard culture than with the CF test (P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)     

Release of tumor necrosis factor-alpha from bovine alveolar macrophages stimulated with bovine respiratory viruses and bacterial endotoxins

The release of tumor necrosis factor-alpha (TNF-) from cultured bovine alveolar macrophages (BAM) was evaluated following stimulation of BAM with bovine herpesvirus-1 (BHV-1), parainfluenza-3 (PI-3) virus, bovine respiratory syncytial virus (BRSV), Escherichia coli 0111:B4 endotoxin, Pasteurella haemolytica type 1 endotoxin, Pasteurella multocida endotoxin, and virus/endotoxin combinations. A cytotoxic assay system using Georgia bovine kidney cells as targets was used to measure TNF- activity. The cytotoxic activity was neutralized by an anti-human TNF- monoclonal antibody.

Stimulation of BAM with 1 median tissue culture infectious dose (TCID₅₀) of live or ultraviolet (UV)-inactivated PI-3 virus/cell resulted in release of TNF- in significantly (P<0.05) higher amounts than sham-induced BAM. The quantities of TNF- released after live or UV-inactivated BHV-1 or BRSV induction were not significantly higher than sham-induced BAM. E. coli 0111:B4, P. haemolytica type 1 and P. multocida endotoxins stimulated TNF- release in a dose-dependent manner. Sequential exposure of BAM to 1 TCID₅₀ per cell of either live BHV-1, PI-3 virus or BRSV and then 5 μg ml⁻¹ of either E. coli 0111:B4, P. haemolytica type 1 or P. multocida endotoxin caused a significant (P<0.05) reduction in detectable TNF- in seven of nine virus/endotoxin combinations tested, when compared with 5 μg ml⁻¹ of endotoxin alone. Parainfluenza-3 virus/endotoxin combinations stimulated higher TNF- release when compared with other virus/endotoxin combinations. Five out of six test animals had serum-neutralizing antibodies to PI-3 virus, one out of six had serum-neutralizing antibodies to BHV-1, and two out of six had serum-neutralizing antibodies to BRSV, suggesting a possible relationship between serum neutralizing antibodies and TNF- release from in vitro cultivated BAM.