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

G. S. Z. Ssenyonga  I. Kakoma  Sonia Montenegro-James  R. Hansen 《Tropical animal health and production》1992,24(1):2-8

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.

---

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.

---

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.

  相似文献   

Onchocerca gutturosa and Onchocerca lienalis in cattle: variation in length of microfilariae by site of recovery     

T M Harty  S A Ferenc  D B Copeman  C H Courtney 《American journal of veterinary research》1989,50(1):169-172

Dermal microfilariae recovered from specimens obtained from umbilical and cervical sites of cattle infected with adult Onchocerca gutturosa alone or with adults of O gutturosa and O lienalis were measured and compared with uterine microfilariae obtained directly from gravid female worms of each species. Uterine microfilariae of O gutturosa were longer than dermal microfilariae obtained from cattle harboring only adults of O gutturosa. Dermal microfilariae were recovered from umbilical and cervical sites in these cattle. Those found at the cervical site had lengths equal to or greater than lengths of microfilariae recovered from the umbilical site. There was a significant (P less than 0.0001) shift in length across populations of microfilariae of O gutturosa from various sites in its bovine host, with a progressive decrease in length between microfilariae recovered from the worm's uterus, microfilariae from the cervical dermis, and microfilariae from the umbilical dermis, respectively. A similar direct comparison was not possible for microfilariae of O lienalis, because none of the cattle was infected with only adult worms of this species. In an indirect comparison, microfilariae of O lienalis were identified at the umbilicus, but their presence in the cervical region could not be determined unequivocally because of confounding of microfilariae length by concurrent infection with O gutturosa. Uterine microfilariae from O lienalis were longer than uterine microfilariae of O gutturosa, although a degree of overlap in the range of measurements existed between species.  相似文献   

Chemotherapy of T b brucei infection: Use of DFMO, diminazene aceturate, alone and in combination     

P. A. Onyeyili  S. M. Anika 《The Journal of small animal practice》1989,30(9):505-510

The therapeutic activity of diminazene aceturate, difluoromethylornithine (DFMO) and a combination of the two agents was investigated in experimental Trypanosoma brucei brucei infections in mongrel dogs. The criteria used in the assessment of the trypanocidal effect of these compounds included the examination of the blood for the parasite, as well as clinical and haematological changes at intervals following treatment. Diminazene aceturate (7 mg/kg intramuscularly), DFMO (300 mg/kg/day orally in three divided doses for six days) and the combination of diminazene aceturate (7 mg/kg intramuscularly) and DFMO (300 mg/kg/day orally for six days) produced an intermittent aparasitemia in the dogs. Relapse infection occurred in all the three groups, but the period of aparasitemia produced by the combination of the agents was longest. The packed cell volume, haemoglobin concentration and red cell count values decreased after the dogs were inoculated with the parasite. The values improved slightly following the treatments with the agents or their combination. The total white blood cell counts in the infected dogs indicated leucocytosis, but this improved with drug treatment.  相似文献   

A Potential Screening System for Identifying Sorghum Ecotypes with Increased Lysine in the Seeds     

S. Vernaillen    F. Laureys  M. Jacobs 《Plant Breeding》1993,111(4):295-305

Classical “high-lysine” sorghum lines are characterized by smaller seeds than average, due to a decrease in prolamin synthesis and a subsequent decrease in yield. To exploit the natural variation in lysine content and to identify ecotypes with a seed lysine content higher than average, characterized by plump seeds, a method was developed based on root-growth inhibition of seeds growing on a medium containing aminoethylcysteine (AEC), a lysine analogue. By using a collection of sorghum mutants and ecotypes a correlation coefficient of 0.926 between root length and lysine content was established. This method, which uses the root length of plants growing on aec to indicate which lines have a potential elevated lysine content, can be applied for the screening of sorghum germplasm. Since this is a non-destructive method it can also be used at the individual seed level, for example for screening progenies of regenerated plants from in vitro culture to exploit the somaclonal variation.  相似文献   

Analysis and pharmacokinetics of cimaterol in growing Holstein steers.     

T M Byrem  T F Robinson  Y R Boisclair  A W Bell  W S Schwark  D H Beermann 《Journal of animal science》1992,70(12):3812-3819

Pharmacokinetic parameters for the beta 2-adrenergic agonist, cimaterol (CIM), were determined in growing Holstein steers. Compartmental analysis was used after measurement of CIM in body fluids by affinity chromatography and HPLC using UV detection. Recoveries from spiked plasma and urine standards were 70 +/- 1.2% and 68 +/- 1.1%, respectively. The minimum detection level in plasma was 1 ng/mL and the average CV was 5.1% for concentrations that ranged from 1 to 30 ng/mL. Four steers (276 +/- 24 kg) received 15 mg of CIM by bolus intravenous injection. Plasma CIM levels declined in a biphasic manner with half-lives of 2.5 min for the distribution phase and 54 min for the elimination phase. A two-compartment open model was used to describe the disappearance of CIM and the following pharmacokinetic parameters were obtained: central compartment volume (Vc) = .76 L/kg, apparent volume of distribution (Vd) = 4.1 L/kg, and transfer rate constants from the central to peripheral compartment (k12) = .177/min, from the peripheral to central compartment (k21) = .054/min and elimination from the central compartment (kel) = .074/min. After 8 h, total urinary CIM accounted for only 18.3% of the administered dose. Results suggest that circulating concentrations of CIM in growing steers are influenced by its accumulation in an unidentified peripheral pool and its conversion into unknown metabolite(s) before elimination.  相似文献   

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

S.E. Bienhoff  G.K. Allen  J.N. Berg 《Veterinary immunology and immunopathology》1992,30(4):341-357

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.  相似文献   

Eastern equine encephalitis in a flock of emus (Dromaius novaehollandiae).     

T N Tully  S M Shane  R P Poston  J J England  C C Vice  D Y Cho  B Panigrahy 《Avian diseases》1992,36(3):808-812

Eastern equine encephalitis (EEE) was diagnosed in a flock of emus in southeastern Louisiana. The outbreak involved juvenile and adult breeders ranging in age from 20 to 36 months, with an attack rate of 76% and a case fatality rate of 87%. The diagnosis was confirmed by isolation and characterization of the viral agent, and by detection of EEE antibody in two recovered emus. High mortality was preceded by marked depression, hemorrhagic diarrhea, and emesis of blood-stained ingesta. On postmortem examination, hemorrhagic enteritis and multiple petechia of viscera were observed. Microscopic changes included severe necrosis of hepatocytes, intestinal mucosa, and necrotizing vasculitis of the spleen and lamina propria of the intestine. No nervous system lesions were observed. This outbreak occurred concurrently with EEE in horses and was attributed to unseasonably heavy rainfall with an abundance of arthropod vectors and proximity to free-living reservoir host species.  相似文献   

On the genetic basis of equine allergic diseases: II. Insect bite dermal hypersensitivity.     

E Marti  H Gerber  S Lazary 《Equine veterinary journal》1992,24(2):113-117

The horses studied were of the Swiss Warmblood breed and most were ELA-typed to assess a possible association of dermal hypersensitivity to insect bites with the major histocompatibility complex. Firstly, the occurrence of the condition was examined in 304 half-siblings sired by six stallions (A to F). Fourteen cases of dermal hypersensitivity were recognized and all were in the 153 offspring of Stallions C, E and F. Most animals of this group were also investigated for chronic hypersensitivity bronchitis: none of the sires displayed clinical signs of dermal hypersensitivity, but Stallions D, E and F were affected by chronic bronchitis. Among the animals investigated for both conditions only one horse showed coincidence of the two diseases as can be expected when the diseases are not correlated. The frequency of manifest dermal hypersensitivity and/or chronic hypersensitivity bronchitis varied in the half-sibling groups of individual sires. These findings suggest that the allergic conditions are independent entities. Secondly, the occurrence of dermal hypersensitivity was studied in three generations of horses at a stud at which Stallion C had exerted a particularly strong influence. A total of 302 animals, all born and raised at this stud, were surveyed over a period of 12 years. The descendants of Stallion C showed a significantly higher incidence (P less than 0.01) of dermal hypersensitivity (two daughters out of 19; eight second generation offspring out of 103; one third generation offspring out of 85) than did the controls of the same age classes but unrelated to Stallion C at the same stud (0 out of 95).(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

Potential of a recombinant antigen as a prophylactic vaccine for day-old broiler chickens against Eimeria acervulina and Eimeria tenella infections.   总被引：5，自引：0，他引：5  

B S Bhogal  G A Miller  A C Anderson  E J Jessee  S Strausberg  R McCandliss  J Nagle  R L Strausberg 《Veterinary immunology and immunopathology》1992,31(3-4):323-335

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.  相似文献   

Nutritional support of hospitalized patients     

S Donoghue 《Veterinary Clinics of North America: Small Animal Practice》1989,19(3):475-495

Effective nutritional support requires sound knowledge of both basic and clinical nutrition of dogs and cats as well as familiarity with products and delivery systems. Case management includes assessment of nutritional status and estimation of fuel sources. Most starved or stressed patients use fatty acids for over 70 per cent kcalME and protein for over 20 per cent kcalME. Approximate kcal needs are calculated from maintenance energy equations. Most patients respond best to enteral nutrition. Meat-based pet foods, liquid enteral products, and nutrient modules are offered in slurries or are tube-fed. Management includes careful monitoring of patients and gradual transitions to diets with more complex nutrient sources.  相似文献