Enteric coccidial infections. Isospora, Sarcocystis, Cryptosporidium, Besnoitia, and Hammondia

The authors consider coccidial infections of the genera Isospora, Sarcocystis, Cryptosporidium, Besnoitia, and Hammondia.     

The isolation,separation and preservation ofBabesia bigemina

Summary Experiments were performed in Colombia to separateBabesia bigemina from contaminating organisms.Babesia bigemina was passaged serially through five splenectomized calves. The first calf was inoculated with blood carrying several different organisms, and subsequent subinoculations were done soon after blood smears from each calf were found to be positive forB. bigemina. Five blood passages were carried out in 6.5 days.Babesia argentina, B. major andA. marginale were eliminated as contaminants of theB. bigemina isolated after four passages. A stabilate of the isolatedB. bigemina was established.

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El aislamiento, separación y preservacion de babesia bigemina

Sumario Se realizaron experimentos en Colombia para separarBabesia bigemina de organismos contaminantes.Babesia bigemina fue pasada en forma seriada en cinco terneros esplenectomizados. El primer ternero fue inoculado con sangre conteniendo algunos organismos diferentes, y las siguientes subinoculaciones fueron hechas tan pronto como los frotices de sangre de cada ternero fueron encontrados ser positivos conB. bigemina. Se llevaron a cabo cinco pases de sangre en 6.5 dias.Babesia argentina, B. major, yA. marginale fueron eliminados como contaminantes del aislamiento deB. bigemina despues del cuarto pase. Una forma estable del aislamiento deB. bigemina ha sido obtenida.

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Isolement, séparation et conservation debabesia bigemina

Résumé Des expériences ont été effectuées en Colombie pour séparerBabesia bigemina des organismes infectés.Babesia bigemina fut passé en série sur cinq veaux splénectomisés. Le premier veau a été inoculé avec du sang contenant plusieurs organismes différents. Les subinoculatinos suivantes ne furent faites que lorsque les étalements de sang de chaque veau furent positif avec.B. bigemina. Les cinq passages furent effectués en 6,5 jours.Babesia argentina, B. major etAnaplasma marginale ont été élimines, en tant qu'éléments contaminants, après quatre passages. Un stabilat desB. bigemina isolés a été effectué.

     

Cost comparison of anesthetic regimens in the dog and cat

Comparative costs of anesthetic regimens for the dog and cat were calculated. Various combinations of currently popular sedatives, tranquilizers, and anti-muscarinics (preanesthetic drugs), and anesthetic induction and maintenance drugs were studied. The preanesthetic drug affected overall anesthetic cost through its own cost, its effect on the amount of anesthetic drug necessary for intubation, and its effect on the amount of anesthetic necessary to maintain anesthesia. The combination of acetylpromazine-thiamylal-halothane was the least expensive regimen for both the dog and cat, whereas drug combinations that included isoflurane as the maintenance drug were the most expensive. In the cat, induction of anesthesia by use of N2O, O2, and halothane in a plexiglas chamber was more expensive than by the use of thiamylal.     

Activity of mucosubstances and the goblet cell count in the large intestine of piglets infected with the coccidium, Isospora suis

In a group of conventional and gnotobiotic piglets experimentally infected with the Isospora suis coccidia the quantitative presence of acid and neutral mucous substances in the large intestine and the counts of goblet cells in the surface mucosa and in Lieberkühnis crypts (in the following text called just the crypts) were investigated. In conventional piglets infected with the dose of 200,000 oocysts of I. suis coccidia the lowest content of acid mucous substances was recorded from the eighth to tenth day after infection (DAI). A decrease in the activity of neutral mucous substances was somewhat slower. The lowest count of goblet cells was found on DAI 9, especially on the surface mucosa (4.89 to 4.91 goblet cells per 10 enterocytes). There was observed no difference in the piglets infected the first and fifth day after parturition (DAP). Gnotobiotic piglets infected with the dose of 100,000 oocysts of I. suis coccidia on DAP 1 showed the lowest content of mucous substances in the large intestine from the ninth to tenth day after infection. Unlike the conventional piglets, in gnotobiotic piglets there was recorded a decrease in the content of acid and neutral mucous substances. The gnotobiotic piglets had the lowest counts of goblet cells in the surface mucosa (10:4.57) and in the crypts (10:7.71) on DAI 9. As to the quantitative proportions, in the conventional and gnotobiotic piglets neutral mucous substances prevailed on the other days (DAI 3-7 and DAI 11), similarly like on DAI 8. The results of this investigation revealed a functional disease of the large intestine in conventional and gnotobiotic piglets infected experimentally with the Isospora suis coccidia.     

Besnoitia cysts in the adrenal gland of a cow

Summary

An unusual finding of Besnoitia besnoiti was discovered during histopathological examination of the adrenal gland of a cow brought to slaughter.     

The immune responses to antigenic variants of Babesia bigemina in the bovine.

Four Babesia bigemina stabilates were used to determine the immune response of cattle to acute and chronic blood- and tick-borne infections. Thirty-two intact calves were divided into 16 groups of two and each calf was inoculated with infective B bigemina erythrocytic stabilates. Twenty-eight days later they were challanged with homologous and heterologous stabilates, and monitored for an additional 20 days. The hosts' apparently reduced response to homologous challenge but marked immune response to heterologous challenge indicated antigenic differences between the isolates and confirmed the conclusions reached by examination of the serological data.     

Immunodiagnosis of Besnoitia besnoiti infection by ELISA and Western blot

Besnoitia besnoiti, an obligate intracellular apicomplexan protozoan parasite, is the causative agent of bovine besnoitiosis. This infection may dramatically affect body condition and lead to irreversible infertility in males, resulting in important economical losses in livestock production. Identification of serologically positive animals is of major relevance to elaborate appropriate measures of control. While identification of clinical cases is relatively easy to carry out, the finding of subclinical forms of infection is more difficult, thus serology is considered as an appropriate diagnostic tool. In view to improve and validate immunodiagnosis, we evaluated an enzyme-linked immunosorbent assay (ELISA), complemented with a Western blot (both using a somatic B. besnoiti-tachyzoite antigen) to detect anti-B. besnoiti antibodies in bovine sera. The comparative evaluation of the 2 methods, using 13 sera from animals affected by the chronic phase of besnoitiosis and 10 asymptomatic carriers, yielded a diagnostic sensitivity of 87% for ELISA and 91% for Western blot analyses. Specificity was tested with sera from animals with confirmed Toxoplasma gondii (n=5) and Neospora caninum (n=12) infection, and with 64 negative sera from either an endemic or a non-endemic area. The ELISA specificity ranged between 96.4% and 98%, the Western blot specificity between 96.4% and 100%. The present study demonstrated that ELISA and Western blot, using in vitro generated somatic B. besnoiti antigen, is a useful tool combination to reliably detect animals that have been exposed to B. besnoiti infection, including both asymptomatic and symptomatic courses of disease.     

双芽巴贝斯虫在长角血蜱肠内发育的形态学研究

以耳袋法将长角血蜱(Haemaphysalis longicornis)幼虫饲于实验感染双芽巴贝斯虫(Babesia bigemina)牛,幼虫饱血后24h内,其肠管内容物中红细胞内、外见有单梨子型(3.5～4.5μm×1.2～2.6μm)和双梨子型(4.1～4.8μm×1.8～3.0μm)两种裂殖体.饱血后24～48h,随着裂殖体细胞膜及核变性而发生形态变化.48～72h,绝大多数裂殖体出现溶解.72h后,这些裂殖体从肠道消失.其后,在蜱肠上皮及血淋巴中也未能找到双芽巴贝斯虫体.本实验从形态学上证明双芽巴贝斯虫在长角血蜱若虫肠道内不能发育.     

Characterization of magnesium-induced urinary disease in the cat and comparison with feline urologic syndrome

Aggregates of struvite crystals caused urethral obstruction in a high percentage of cats fed moist and dry diets supplemented with Mg oxide. Some of the diets were associated with cystolith formation as well. The percentage of Mg in the experimental diets was a misleading indicator of Mg intake because of differences between moist and dry diets in their caloric density. Magnesium homeostasis was maintained in cats ingesting large quantities of Mg. Tissue (kidney, muscle, and rib) concentrations of Mg were the same in cats fed high Mg and control diets. Plasma Mg concentration was increased only in cats ingesting the largest amount of Mg. Magnesium homeostasis was maintained by a marked increase in urine Mg excretion. However, urine Mg concentration was not directly related to Mg intake, apparently because of differences between diets in intestinal absorption of Mg. Urethral obstruction of experimental cats was not associated with a transient increase in Mg intake, nor did obstructing cats have higher urine Mg concentrations than did nonobstructing cats fed the same diet. This observation indicates that factor(s) other than urine Mg concentration are important in urethral obstruction. Cats with urethral obstruction due to naturally occurring disease, feline urological syndrome (FUS), had markedly lower urine Mg concentrations than cats fed high Mg diets. This finding refutes the theory that cats develop FUS because of primary Mg hyperabsorptive phenomena or because of a primary urinary leak of Mg. It also indicates that factors other than urine Mg concentration are involved in the genesis of naturally occurring urethral obstruction. Another difference between the natural and the induced disease was related to the character of the urinary precipitates. Experimental diets higher in Mg concentration caused urolith formation, which is uncommon with FUS. Lower Mg diets caused obstruction with aggregates of crystals, but mucus was not observed. However, in the experimental disease induced in the present study, urinary precipitates were predominantly or exclusively struvite, as has been reported in the natural disease. Many similarities were seen between the diet-induced disease and FUS, but factors in addition to Mg intake are involved in the natural disease. The importance of Mg, compared with the undefined factors, remains to be established.     

The sporulation time of Isospora suis oocysts from different sources

Feces containing Isospora suis oocysts were collected from naturally- and experimentally-infected pigs from four different areas of the United States. The unsporulated oocysts were cleaned, concentrated, mixed with 2.5% aqueous potassium dichromate solution, poured into petri dishes to a depth of 5 mm, and incubated at 25 degrees C. The oocysts were examined with a microscope at 12 h intervals and the stages of sporulation present were counted. Although a few oocysts were completely sporulated after 12 h of incubation, in most fecal samples the majority of the oocysts were not completely sporulated until 24 or 36 h. In the present study, the sporulation time of I. suis oocysts was considered to be less than or equal to 48 h. There were no major differences in the sporulation times of I. suis oocysts from the different sources.     

A modified agglutination test for the diagnosis of Besnoitia besnoiti infection

Bovine besnoitiosis is caused by Besnoitia besnoiti, an obligate intracellular apicomplexan parasite. Affected animals present cutaneous and systemic manifestations and the disease may lead to considerable economic losses. Although generally associated to tropical and subtropical areas, bovine besnoitiosis is now considered an emergent disease in Europe, due to the increasing number of new cases and apparent geographical expansion. In this study we evaluated the performance of a modified agglutination test (B-MAT) in the serodiagnosis of bovine besnoitiosis in comparison to the indirect immunofluorescent-antibody test (IFAT). To establish optimal protocol conditions we used bovine sera with a known infection status for B. besnoiti infection. Positive animals (n=36) presented B. besnoiti dermal cysts and anti-B. besnoiti specific antibodies, as determined by the indirect immunofluorescence test (IFAT). Negative animals (n=103) were from non-endemic areas in Portugal and negative by the IFAT. From here, we evaluated the sensitivity and specificity of the B-MAT relative to the IFAT with a panel of sera from herds with history of bovine besnoitiosis in Portugal, Spain and France (n=402), using three serum dilutions (1:80, 1:160, 1:320). Considering the positive cut-off at 1:160 serum dilution, the B-MAT showed an almost perfect test agreement with the IFAT; (κ=0.968; 95% CI: 0.941-0.996) with a relative sensitivity of 97.2% (95% CI: 94.1-100%) and a relative specificity of 99.3% (95% CI: 98.4-100%). As a simple and inexpensive technique the B-MAT represents a valuable tool for the diagnosis and study of the epidemiology of bovine besnoitiosis.