Eimeria tenella: Host Specificity in Gallinaceous Birds

SYNOPSIS. Eight species representing 8 genera of gallinaceous birds were used: Alectoris graeca; Colinus virginianus; Coturnix coturnix; Gallus gallus; Meleagris gallopavo; Numidia meleagris; Pavo cristatus; Phasianus colchicus. Three-week-old birds were dosed with sporulated oocysts of Eimeria tenella Beltsville strain. At 4, 24, 48, 72, 96, 120, 144, and 168 hr after inoculation, 1-3 infected birds and uninoculated controls of each species were killed by cardiac exsanguination. Pieces of intestines were fixed and examined for stages of E. tenella as stained paraffin sections or indirect fluorescent antibody preparations. Oocyst counts were made in droppings collected for the first 6 days of the patent period. Sporozoites were found in the lamina propria of some birds of 5 species at 4 hr postinoculation, but no stages were found thereafter except in the breeds of G. gallus and A. graeca. At 144 and 168 hr postinoculation, a few macrogametes were found in the ceca of 2 A. graeca , but no oocysts were found in the feces. No statistical difference was found between the number of oocysts produced/bird in the breeds of G. gallus examined. It is evident from these observations that E. tenella did not complete its life cycle in several close phylogenetic relatives of G. gallus , even though in other studies this parasite was found to complete its life cycle in cell cultures derived from the same birds.     

Eimeria tenella: host specificity in gallinaceous birds.

Eight species representing 8 genera of gallinaceous birds were used: Alectoris graeca; Colinus virginianus; Coturnix coturnix; Gallus gallus; Meleagris gallopavo; Numidia meleagris; Pavo cristatus; Phasianus colchicus. Three week-old birds were dosed with sporulated oocysts of Eimeria tenella Beltsville strain. At 4, 24, 48, 72, 96, 120 and 144, and 168 hr after inoculation, 1-3 infected birds and uninoculated controls of each species were killed by cardiac exsanguination. Pieces of intestines were fixed and examined for stages of E. tenella as stained paraffin sections or indirect fluorescent antibody preparations. Oocyst counts were made in droppings collected for the first 6 days of the patent period. Sporozoites were found in the lamina propria of some birds of 5 species at 4 hr postinoculation, but no stages were found thereafter except in the breeds of G. gallus and A. gracea. At 144 and 168 hr postinoculation, a few macrogametes were found in the ceca of 2 A. gracea, but no oocysts were found in the feces. No statistical difference was found between the number of oocysts produced/bird in the breeds of G. gallus examined. It is evident from these observations the E. tenella did not complete its life cycle in several close phylogenetic relatives of G. gallus, even though in other studies this parasite was found to complete its life cycle in cell cultures derived from the same birds.     

Effect of dietary zinc level on serum carotenoid levels,body and shank pigmentation of chickens after experimental infection with coccidia

Abstract

Two experiments were conducted to test the effects of a dietary zinc amino acid complex (Zn-AA) and an anticoccidial drug on Eimeria acervulina or Eimeria tenella infections. In each experiment, 288 day-old Three-Yellow-Chickens were used in a 2 × 3 factorial experimental design. Six groups were arranged randomly to receive three levels of Zn-AA (0, 40, or 80 mg/kg) alone or with salinomycin (60 mg/kg). Additionally an uninfected group was set as negative control. At the age of 21 days birds in Exp. 1 were inoculated with 3 · 10⁴ sporulated E. acervulina oocysts, while birds in Exp. 2 were inoculated with 1.5 · 10⁴ sporulated E. tenella oocysts. In Exp. 1, E. acervulina did not suppress growth performance significantly, but in groups without salinomycin it significantly reduced serum carotenoid levels on day 7 after inoculation and body and shank pigmentation on day 42. Salinomycin medication maintained serum carotenoids and visual colour of inoculated birds, but Zn-AA did not influence these parameters. In Exp. 2, growth performances of infected and uninfected chickens were similar. Infection decreased to only serum carotenoid levels on day 14 after infection, and colour scores on day 42 in the inoculated group without salinomycin and Zn-AA supplementation. The birds that received Zn-AA had significantly higher serum carotenoid levels and colour scores than those that did not. Although supplementation of Zn-AA cannot avoid coccidial damage of caecum, it prevents the reduction of serum carotenoids and pigmentation of Three-Yellow-Chicken infected with E. tenella, but not after infection with E. avervulina. The interactive effects between Zn-AA and salinomycin on growth performance and pigmentation were not significant.     

Observations on Haemogregarina balli sp. n. from the Common Snapping Turtle,Chelydra serpentina*

SYNOPSIS. Haemogregarina balli sp. n. is described from the blood and organs of the common snapping turtle Chelydra serpentina serpentina and from the gastric and intestinal ceca of the presumed invertebrate hosts, the leeches Placobdella parasitica and Placobdella ornata. In the peripheral blood of the turtle, male and female gametocytes and immature erythrocytic schizonts are found within erythrocytes. The maturation of erythrocytic schizonts containing 6–8 merozoites is recorded from liver imprints. Schizonts with 13–25 merozoites are found in various cells of the liver, lung and spleen. In the gastric ceca of the leeches the host erythrocytes are digested, releasing the gametocytes and immature erythrocytic schizonts. Immature erythrocytic schizonts degenerate. Association of the gametocytes occurs in the intestinal ceca. The microgametocyte apparently gives rise to 4 nonmotile microgametes, one of which fertilizes the macrogamete while the other remain as condensed, residual nuclei on the periphery of the developing oocyst. The oocyst increases in size with maturity. A mature oocyst produces 8 sporozoites from a single germinal center. Sporozoites liberated from the oocyst are found in the tissues of the leech. Transovarial transmission of the parasite does not occur in the turtle. Attempts at experimental transmission failed. Previously unfed (control) leeches were negative for the parasite. Haemogregarina balli is compared with other haemogregarines described from C. serpentina. Features of species of Haemogregarina and Hepatozoon as well as the taxonomy of these genera are discussed.     

Virus-like particles in Eimeria tenella are associated with multiple RNA segments

Total nucleic acids from sporulated oocysts of Eimeria tenella isolated from Changchun in China were found to contain three extrachromosomal double-stranded RNA segments (dsRNAs) of 1.4, 2.4 and 3.6 kb in sizes. These RNAs were resistant to RNase A digestion under high salt concentration (0.3 M NaCl). RNA-dependent RNA polymerase (RDRP) activity was detected in crude extracts of E. tenella sporulated oocysts containing these nucleic acid species. Virus-like particles (VLPs) were shown to have a diameter of approximately 38 nm under Electron Microscopy (EM) after purification by sucrose density gradient centrifugation. In keeping with the nomenclature generally adopted for protozoan viruses, we have named this isolate as E. tenella virus (ETV) which is the first virus isolated from E. tenella.     

Excystation of Eimeria tenella sporozoites impaired by antibody recognizing gametocyte/oocyst antigens GAM22 and GAM56

Eimeria tenella is the causative agent of coccidiosis in poultry. Infection of the chicken intestine begins with ingestion of sporulated oocysts releasing sporocysts, which in turn release invasive sporozoites. The monoclonal antibody E2E5 recognizes wall-forming body type II (WFBII) in gametocytes and the WFBII-derived inner wall of oocysts. Here we describe that this antibody also binds to the stieda body of sporocysts and significantly impairs in vitro excystation of sporozoites. Using affinity chromatography and protein sequence analysis, E2E5 is shown to recognize EtGAM56, the E. tenella ortholog of the Eimeria maxima gametocyte-specific GAM56 protein. In addition, this antibody was used to screen a genomic phage display library presenting E. tenella antigens as fusion proteins with the gene VIII product on the surfaces of phagemid particles and identified the novel 22-kDa histidine- and proline-rich protein EtGAM22. The Etgam22 mRNA is expressed predominantly at the gametocyte stage, as detected by Northern blotting. Southern blot analysis in combination with data from the E. tenella genome project revealed that Etgam22 is an intronless multicopy gene, with approximately 12 to 22 copies in head-to-tail arrangement. Conspicuously, Etgam56 is also intronless and is localized adjacent to another gam56-like gene, Etgam59. Our data suggest that amplification is common for genes encoding oocyst wall proteins.     

The Life Cycle of Eimeria bateri (Protozoa, Eimeriidae) in the Japanese Quail Coturnix coturnix japonicum

SYNOPSIS. Eimeria bateri, a parasite of the Indian quail, was described from laboratory infections in Japanese quail. First generation schizonts developed in the glands of the duodenum and upper intestine. Second, 3rd and 4th generation schizonts and gametocytes occurred in the villous epithelium. There was a gradual spread along the small intestine until the whole organ was affected. The prepatent period was 4 days and the patent period 6 days. Graded doses from 5,000 to 1,280,000 oocysts produced very little pathogenic effect in young quail. E. bateri did not infect young pheasants or chicks.     

Eimeria colchici sp. nov. (Protozoa: Eimeriidae), the Cause of Cecal Coccidiosis in English Covert Pheasants

SYNOPSIS. Eimeria colchici sp. n. is described from English pheasants Phasianus colchicus. It has cocysts measuring 19–33.5 (27.4) by 13–21 (16.7) μ, having both micropyle and polar granule. The large 1st and 2nd generation schizonts occur in the glands of the small intestine, 3rd generation schizonts occur deep in the glands of the ceca, and gametocytes occur in the epithelial cells of the ceca. The prepatent period is 6 days. In young pheasants the parasite causes coccidiosis characterized by formation of white cores in the ceca. Mortality is high in experimental infections. Zoalene (0.0125%) in the feed almost completely suppressed the infection; Amprolium (0.0125%) also gave good control, but sulfaquinoxaline (0.0125%) was not so effective. Sulfaquinoxaline, Saquadil and Sulfamezathine in the drinking water controlled mortality, but treatment had to be applied early to avoid weight losses.     

Digestive and metabolic utilization of lauric,myristic and stearic acid in cows,and associated effects on milk fat quality

A study was carried out to examine the effect of dietary supplementation of oregano essential oil on performance of broiler chickens experimentally infected with Eimeria tenella at 14 days of age. A total of 120 day-old Cobb-500 chicks separated into 4 equal groups with three replicates each, were used in this study. Two groups, one infected with 5·10⁴ sporulated oocysts of E. tenella and the other not, were given a basal diet and served as controls. The other two groups also infected with E. tenella were administered diets supplemented with oregano essential oil at a level of 300 mg/kg, or with the anticoccidial lasalocid at 75 mg/kg. Following this infection, survival rate, bloody diarrhoea and oocysts excretion as well as lesion score were determined. Throughout the experimental period of 42 days, body weight gain and feed intake were recorded weekly, and feed conversion ratios were calculated. Two weeks after the infection with E. tenella supplementation with dietary oregano oil resulted in body weight gains and feed conversion ratios not differing from the non-infected group, but higher than those of the infected control group and lower than those of the lasalocid group. These parameters correspond with the extent of bloody diarrhoea, survival rate, lesion score and oocyst numbers and indicated that oregano essential oil exerted an anticoccidial effect against E. tenella, which was, however, lower than that exhibited by lasalocid.     

Structure, Cytochemistry, and Locomotion of Haemogregarina sp. from Rana berlandieri

Intra- and extracellular gametocytes of Haemogregarina sp. from Rana berlandieri were studied by light and electron microscopy. Locomotion in free gametocytes appears to be related to series of horizontal “peristaltic” waves of constriction, passing from anterior to posterior along the body. Intracellular gametocytes lie within a vacuole in the erythrocyte cytoplasm. The pellicle of the parasite consists of a trilaminar plasmalemma and an inner electron dense layer, beneath which lies a ring of 80 microtubules. The inner dense layer becomes thickened and modified in the apical region, to form a cap-like structure. The gametocytes contain a prominent nucleus, several mitochondria, and many granular inclusions. One type of inclusion consists of elliptical, electron-dense, profeinaceous bodies scattered throughout the cytoplasm, while other inclusions are larger and electron-opaque, polysaccharide in nature, and occur predominantly in the pre- and post-nuclear regions. In the electron microscope, pronounced pellicular folds were observed in longitudinally sectioned extracellular gametocytes. These folds are thought to represent the waves of constriction seen in motile specimens by light microscopy. The mechanism of movement of the parasite is discussed and compared with that in haemosporidian ookinetes, as well as in gregarines.     

Serum and liver zinc,copper, and iron in chicks infected withEimeria acervulina orEimeria tenella

Two-wk-old broiler chicks were inoculated via crop intubation withEimeria acervulina at two doses: 10⁵ or 10⁶ sporulated oocysts/bird or withEimeria tenella at a dose of 10⁵ sporulated oocysts/bird. Serum and liver samples were collected on days 3 and 6 post-inoculation (PI). There were no significant changes in serum or liver zinc, copper, and iron concentrations in any of the infected groups by 3 d PI. However, on d 6, PI serum protein was significantly reduced in all of the infected groups compared to their pair-fed controls. The chicks infected withE. tennella had significantly reduced serum zinc (1.20 vs 1.77 μg/mL) and iron (0.44 vs 1.28 μg/mL) concentrations and significantly elevated serum copper (0.28 vs 0.17 μg/mL) and ceruloplasmin levels (20.33 vs 11.11 μg/mL) compared to their pair-fed counterparts. Those chicks infected withE. acervulina (10⁶ oocysts/bird) exhibited significantly reduced serum iron concentration by 6 days PI (0.90 vs 1.14 μg/mL). Liver zinc was significantly increased in the chicks infected withE. tenella (349 vs 113 μg/g dry liver wt), as was copper (24 vs 19 μg/g), whereas liver iron concentration was significantly reduced (172 vs 243 μg/g) compared to pair-fed controls. At both dose levels, the chicks infected withE. acervulina exhibited a significant reduction in liver iron by 6 d PI. Hepatic cytosol metals generally reflected whole tissue levels. Metallothionein (MT)-bound zinc was significantly elevated in the chicks infected withE. tenella. Iron bound to a high molecular weight, heat-stable protein fraction (presumably cytoplasmic ferritin) was significantly reduced in chicks infected withE. acervulina, as well as those infected withE. tenella. Collectively, the changes in serum zinc, copper, and iron concentrations, as well as the changes in hepatic zinc and MT-zinc concentrations in the chicks infected withE. tenella were similar to changes evoked during an acute phase response to infection. It is possible that a secondary bacterial infection or inflammation stemming from erosion of the lining of the cecum may play a role in the response of trace element metabolism to theE. tenella infection. Mentions of a trademarkr, proprietary product or specific equipment does not consitute a guarantee or warranty by the US Department of Agriculture and does not imply its approval to the exclusion of other products.     

Comparative study of the digestive system of three species of tinamou. I. Crypturellus tataupa,Nothoprocta cinerascens,and Nothura maculosa (Aves: Tinamidae)

The morphology and histology, as well as the cytochemistry of complex carbohydrates, of the digestive system of Crypturellus tataupa (tataupa tinamou), Nothoprocta cinerascens (brushland tinamou), and Nothura maculosa (spotted tinamou) are described. The general morphology of the digestive system of these birds follows the basic model of the avian alimentary canal, although statistical analysis shows that the lengths of the organs are significantly different among the species. From cephalic to caudal regions the alimentary tract consists of esophagus, ingluvies or crop, proventriculus or glandular stomach, ventriculus or muscular stomach, small intestine, well-developed ceca, and rectum. Histologically, each section of the tract consists of four primary tissue layers: mucosa, submucosa, muscularis, and adventitia. Variations are found in the thickness of the esophageal epithelium, which shows the highest value in C. tataupa. In the proventriculus, the depth of the compound glands is greatest in N. cinerascens. The villi of the epithelial cells in the small intestine are most extensively developed in C. tataupa. Heterogeneity of mucins is detected not only in the surface coat of the alimentary tract but in the cellular content of the glands as well. Comparisons with the morphology of the digestive system of closely related and more advanced birds are made, and the possible relationship between morphological and cytochemical variation and the diet is discussed. © 1996 Wiley-Liss, Inc.     

Isospora papionis n. sp. (Eimeriidae) of the Chacma Baboon Papio ursinus (Kerr, 1792)

SYNOPSIS. Isospora papionis n. sp. is described from the chacma baboon Papio ursinis (Kerr, 1792). Histopathologic examination revealed 3 of 20 baboons to be infected, but no organisms were found on examination of the feces. Macrogametes and sporulated oocysts were observed in the small intestine, mainly in the jejunum and ileum, invariably in an epithelial or subepithelial location at the proximal portions of the crypts of Lieberkühn. Intact sporulated oocysts averaged 17 by 11 μ and had very thin walls which were often partially collapsed between the sporocysts. The sporocysts were somewhat flattened on one side and averaged 11 by 8.5 μ. A large sporocyst residuum was present.     

Fever and Blood Sugar in Pinottii Malaria of Pigeons*

SYNOPSIS. Plasmodium pinottii is an avian malaria parasite originally isolated from the Brazilian toucan, and maintained chiefly in the pigeon, in which it is extremely pathogenic. Infections must be transferred by blood inoculation, since gametocytes are no longer produced (and, in any event, the vector is unknown). Unlike most forms of avian malaria, pinottii malaria in the pigeon is characterized by fever, often high, with peaks at the time of parasite reproduction. Blood sugar levels vary considerably even in healthy birds, in which the mean was found to be 267 ± 9.2 SEm mg per 100 ml plasma (range 208–335, based on examination of 14 individuals. Values during fasting approximated those when feeding was ad libitum. Levels tended to rise in the early stages of the infection, reaching a mean of 305 ± 14.5 mg per 100 ml plasma (based on 10 cases) while parasitemia was 20-49%. At death the mean (determined from 46 birds) had dropped to 155.8 ± 15.5 mg per 100 ml. However there is little reason to think that parasite demand on the host's glycogen reserves was a significant factor in causing death, despite the very high parasitemias often reached.     

Life Cycle of Eimeria christenseni Levine,Ivens & Fritz, 1962 from the Domestic Goat,Capra hircus L.1

The endogenous development of Eimeria christenseni was studied in 10 two- to four-week-old kids inoculated with 10⁶-10⁷ sporulated oocysts. They were killed at intervals from two to 26 days after inoculation, and their tissues were examined for endogenous stages of the coccidian by light microscopy. Such stages were found in the small intestine and mesenteric lymph nodes. In the sexual cycle, two generations of meronts were found. The first generation developed in endothelial cells of lacteals in the jejunun and ileum and mesenteric lymph nodes, and mature meronts were first seen 14 days after inoculation. The second generation developed in epithelial cells of the glands of Lieberkuehn in the jejunum and ileum and in mesenteric lymph nodes, and its mature meronts were first seen by 16 days. Sexual stages were present mostly in epithelial cells of the tips and sides of the villi and less frequently in crypt cells of the jejunum and ileum. Mature macrogametes and microgamonts and oocysts were also first seen by 16 days. The prepatent period was 17 (14-23) days; the patent ranged from 8 to more than 30 days. Sporulation time was 3-4 days at 30°C. E. christenseni was found to be pathogenic, kids inoculated with 1-5 × 10⁵ sporulated oocysts exhibited the following signs: severe diarrhea, anorexia, polydipsia, poor hair coat, and extreme weakness. They recovered about a month later, but their growth rates appeared to be lower than those of uninoculated animals kept under the same conditions. One kid died 20 days after inoculation with 10⁷ oocysts.     

Eimeria acervulina and E. tenella:effect on methionine absorption by the avian intestine.

Absorption of l-methionine in the duodenum of intestine of chicks infected with Eimeria acervulina was markedly less than in uninoculated controls or birds infected with E. tenella. Absorption of methionine in the jejunal area of E. acervulina-infected birds was reduced although not as drastically as in the duodenum. There was no difference in the rate of methionine absorption by the ileum. The kinetics of methionine absorption showed that V_max (maximum velocity) in the duodenum and jejunum of E. acervulina-infected birds was reduced when compared with the V_max found in uninoculated controls or E. tenella-infected birds. There was no difference in the K_t (transport constant) regardless of the infection or the region of the intestine examined. No major consistent effect of decreased pH on the rate of methionine absorption could be shown.The broadly spatulate villi seen, using the scanning electron microscope, in control and E. tenella infected duodenum were absent in E. acervulina-infected duodenum. Instead, the villi were reduced in height and noticeably thickened. This reduction in villar height suggests that a portion of the reduction in methionine absorption was related to the change in surface area and loss of transport loci due to damage of the mucosal surface.     

Life Cycles of Leucocytozoon dubreuili Mathis and Leger, 1911 and L. fringillinarum Woodcock, 1910 (Haemosporidia: Leucocytozoidae)*

SYNOPSIS The development of Leucocytozoon dubreuili and L. fringillinarum was studied on successive days in simuliid and avian hosts. Sporogony of both parasites is completed in at least 5 species of sylvatic Simuliidae in a minimum of 4-5 days at 21 C. The pattern of development of the 2 species is similar but the size of the oocysts and the number of sporozoites differ. Sporozoites of L. dubreuili and L. fringillinarum were injected into uninfected robins (Turdus m. migratorius) and grackles (Quiscalus quiscula versicolor), respectively. Hepatic biopsies were performed on some of the injected birds. These and others were killed at intervals following inoculation and their tissues examined to detect stages of schizogony. Blood and macerated tissues from birds injected with sporozoites were transferred to uninfected birds to determine whether asexual stages would develop in the latter as a result of the inoculations. The 1st asexual cycle of L. dubreuili is completed in hepatic parenchymal cells in a minimum of 84 hr. Merozoites produced by the hepatic schizonts apparently follow one of 3 courses: invade hepatic parenchymal cells to initiate another cycle; penetrate blood cells and become gametocytes; penetrate tubular cells of the kidneys and grow into renal schizonts. The minimum prepatent period in infections with L. fringillinarum is 76 hr. The 1st asexual cycle occurs in hepatic parenchymal cells and in tubular cells of the kidney. A schizogonic cycle is completed in a minimum of 72 hr in the former and 96 hr in the kidney. Merozoites from the primary hepatic schizonts apparently give rise to (a) gametocytes; (b) secondary hepatic schizonts; (c) renal schizonts. Thus the schizogonic cycles of L. dubreuili and L. fringillinarum differ from each other and from those of L. simondi in ducks.