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.     

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.     

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.     

Potential of a recombinant antigen as a prophylactic vaccine for day-old broiler chickens against Eimeria acervulina and Eimeria tenella infections.

A genetically engineered Eimeria tenella antigen (GX3262), produced as a fusion protein with beta-galactosidase and identified with a monoclonal antibody, induced partial but significant protection in young broiler chickens against experimental E. tenella and Eimeria acervulina infections. The antigen appears to share a T-helper cell epitope with the parasite as evidenced by (a) booster inoculation with either the recombinant antigen or with a small number of live oocysts enhanced the protective immunity in GX3262 primed chickens, and (b) ability of the antigen to induce in vitro stimulation of T-cells from chickens immunized with antigen or parasite. These observations suggest the feasibility of a single vaccination of 1 or 2-day-old broilers with GX3262 to induce an acceptable degree of protective immunity. The implications of the observations reported here are far reaching in terms of a practical coccidiosis vaccine for poultry, and show for the first time that 1-day-old broiler chickens can be efficiently vaccinated with a recombinant antigen against one or more species of Eimeria.     

Plasma triglyceride concentration during intravenous infusions of propofol and Intralipid in sheep

Plasma triglyceride concentrations were measured in sheep given Intralipid or propofol, which is carried in a vehicle very similar to 10% Intralipid. A bolus dose was administered followed immediately by an infusion of the same agent for 2 h. In the animals that received propofol, the measured concentration increased by a mean amount of 3.39 mmol/l when the infusion rate was l ml/min (Group Pl) and by 7.13 mmol/l when it was 2 ml/min (Group P2). When 10% Intralipid was administered and infused at 1 ml/min (Group I10), the measured concentration increased only by 0.95 mmol/l. One hour after stopping the infusion, the excess of measured concentration over baseline had decreased in the Pl and I 10 groups to 0.52 and 0.13, respectively, of the corresponding maximum excess. The method adopted for measuring plasma triglycerides is widely used in hospitals; however, an incidental observation revealed that it is inappropriate in the presence of injections of propofol or Intralipid. Despite this, evidence and argument are presented to support the conclusion that, with propofol, plasma triglyceride concentrations increased more rapidly during the infusions and returned to baseline more slowly than with a corresponding amount of Intralipid.     

High-resolution computed tomography of the mammalian lung.

High-resolution computed tomography (HRCT) was performed in 21 isolated animal lungs, from 4 mammalian species (pigs, rabbits, dogs, sheep). Gross and subgross central and peripheral lung morphology was determined by HRCT. Three distinct types of lungs can be identified, principally based on the extent of interlobular septal development; the relationship of major vessels to airways; and the thickness of the visceral pleura. Type-I lung is found in pigs, sheep, and cattle; type-II lung is found in rabbits, dogs, cats, and monkeys; and type-III lung is found in human beings and horses. These mammalian lungs were compared with human lungs. The potential use of HRCT to investigate specific human lung diseases in the aforementioned species also was considered.