This study aimed to determine the seroprevalence and to identify risk factors associated with Neospora spp. infection in horses in Jordan. Management related data were collected from each farm and individual horses. Sera from 227 horses from 5 of 6 climatic regions in Jordan were analyzed for the presence of antibodies to Neospora spp. by ELISA kit. The study was performed during spring of 2010. The association between seropositivity and risk factors was analyzed. A total of 7 (3%) of 227 sera had antibodies for Neospora spp. There was a significant regional difference (P=0.018) between the 5 climatic regions. Positive cases were located in Amman and Irbid, while the other regions (Zarqa, Jordan Valley, and Wadi Mousa) had zero prevalence. The use of anthelmintics at least once a year resulted in a significant reduction of the seroprevalence to Neospora spp. (1.6% vs 9.8%). However, this might be a phenomenon by chance and a better hygiene since owners can invest in anthelmintics. Other risk factors such as age, gender, breed, usage, body condition score, grazing, presence of other animals mixed with the horses in the same property, and a history of previous diseases were not significantly associated with the seroprevalence to Neospora spp. infection. This is the first study to report on the presence of Neospora seropositive horses in Jordan. Further studies are warranted to better understand the role of certain risk factors in the transmission of Neospora spp. among horse population and to determine which Neospora spp. are responsible for the infection.
Animals ; Antibodies, Protozoan/*blood ; Asymptomatic Diseases/epidemiology ; Coccidiosis/blood/epidemiology/*veterinary ; Female ; Horse Diseases/blood/*epidemiology/parasitology ; Horses ; Jordan/epidemiology ; Male ; Neospora/*immunology/physiology ; Seroepidemiologic Studies

The effects of diclazuril on the bursa of Fabricius (BF) structure and secretory IgA (SIgA) expression in chickens infected with Eimeria tenella were examined. The morphology of the BF was observed by hematoxylin and eosin staining, while ultrastructural changes were monitored by transmission electron microscopy. E. tenella infection caused the BF cell volumes to decrease, irregularly arranged, as well as, enlargement of the intercellular space. Diclazuril treatment alleviated the physical signs of damages associated with E. tenella infection. The SIgA expression in BF was analyzed by immunohistochemistry technique. The SIgA expression increased significantly by 350.4% (P<0.01) after E. tenella infection compared to the normal control group. With the treatment of diclazuril, the SIgA was relatively fewer in the cortex, and the expression level was significantly decreased by 46.7% (P<0.01) compared with the infected and untreated group. In conclusion, E. tenella infection in chickens induced obvious harmful changes in BF morphological structure and stimulated the expression of SIgA in the BF. Diclazuril treatment effectively alleviated the morphological changes. This result demonstrates a method to develop an immunological strategy in coccidiosis control.
Animals ; Bursa of Fabricius/anatomy & histology/*parasitology ; Chickens ; Coccidiosis/drug therapy/metabolism/parasitology/*veterinary ; Coccidiostats/administration & dosage/*adverse effects ; Eimeria tenella/*physiology ; Female ; Immunoglobulin A, Secretory/*genetics/metabolism ; Male ; Nitriles/administration & dosage/*adverse effects ; Poultry Diseases/*drug therapy/genetics/metabolism/parasitology ; Triazines/administration & dosage/*adverse effects

In this study a disperse dye immunoassay method was standardized and evaluated for detection of antibodies against Neospora caninum in cattle. Sera from 150 cattle with a recent history of abortion were collected and tested by commercial ELISA kit and a standardized in-house dye immunoassay system. The positivity rate for the sera used in this study was 34.6% for the disperse dye immunoassay (DDIA) compared to 32% obtained by ELISA kit. This study showed no significant difference between DDIA and ELISA. The results indicated that the DDIA provide an economic, simple, rapid and robust test for detection of N. caninum infection in cattle.
Animals ; Antibodies, Protozoan/*blood ; Cattle ; Cattle Diseases/*diagnosis/parasitology ; Coccidiosis/diagnosis/parasitology/*veterinary ; Diagnostic Tests, Routine/*methods ; Female ; Immunoassay/methods ; Neospora/*immunology ; Staining and Labeling/methods ; Veterinary Medicine/*methods

Congenital Neospora caninum infection was diagnosed in two Saanen goat kids from two distinct herds with a history of abortion and weak newborn goat kids in the Southern region of the State of Minas Gerais, Brazil. The first kid was weak at birth, had difficulty to rise and was unable to nurse. Gross lesions of porencephaly and hydrocephalus ex vacuo were seen. Multifocal necrosis, gliosis and non-supurative encephalitis were observed in the brain. Several parasitic cysts with a thick wall that reacted strongly only with polyclonal antiserum to Neospora caninum were seen in the cerebral cortex, brain stem and cerebellum. The second kid was born from a Neospora caninum seropositive mother that aborted in the last pregnancy. It was born without clinical signs. The diagnosis of neosporosis was based on antibody titer of 1:800 to N. caninum by indirect fluorescence antibody test obtained from blood collected before the goat kid ingested the colostrum and Neospora caninum DNA was detected by polymerase chain reaction and sequenced from placenta. This is the first report of neosporosis in goats in the southeast region of Brazil.
Animals ; Antibodies, Protozoan/immunology ; Brazil ; Coccidiosis/congenital/immunology/parasitology/*veterinary ; Female ; Goat Diseases/congenital/immunology/*parasitology ; Goats ; Molecular Sequence Data ; Neospora/genetics/immunology/*isolation & purification/physiology ; Pregnancy

Neospora caninum is an important cause of abortion in dairy cattle worldwide. Dog is the definitive host for N. caninum and can infect dairy cattle. The aim of this study is to determine the prevalence of Neospora oocysts in feces of dogs from dairy farms. A total of 174 fecal samples was collected from 89 farm dogs and 85 household dogs during 2006 and 2008. Fecal samples of dogs were microscopically examined for detecting Hammondia Neospora-like oocysts (HNLO) by Mini Parasep(R)SF fecal parasite concentrator. HNLO were microscopically detected in 4 fecal samples (2.2%). The fecal samples with HNLO were examined by N. caninum-specific PCR. Two of the samples were positive for N. caninum. The 2 positive fecal samples were selected for inoculation to calves. Two inoculated calves were seronegative by ELISA for 4 months post-infection. This is the first report of finding N. caninum DNA in feces of farm dogs in Mashhad area, Iran.
Animals ; Antibodies, Fungal/blood ; Cattle ; Cattle Diseases/immunology/parasitology ; Coccidiosis/epidemiology/parasitology/*veterinary ; DNA, Fungal/genetics/isolation & purification ; Dog Diseases/*epidemiology/*parasitology ; Dogs ; Enzyme-Linked Immunosorbent Assay/methods ; Feces/*microbiology ; Iran/epidemiology ; Male ; Microscopy/methods ; Neospora/*genetics/*isolation & purification ; Oocysts/cytology ; Polymerase Chain Reaction/methods ; Prevalence

In order to clone and identify differentially expressed genes in the sporogony stage of Eimeria tenella, the cDNAs from unsporulated oocysts and sporulated oocysts of E. tenella were used as driver, respectively, the cDNAs from sporozoites of E. tenella was used tester, Two subtractive cDNA libraries of sporozoites were constructed by using the technique of suppression subtractive hybridization (SSH). the cDNAs from unsporulated oocysts was used driver, the cDNAs from sporulated ooceysts was used tester, one subtractive cDNA library of sporulated oocysts was constructed. PCR amplification revealed that the two subtractive cDNA libraries of sporozoites and one subtractive cDNA library of sporulated oocysts contained approximated 96%, 96% and 98% recombinant clones, respectively. Fifty positive clones were sequenced and analyzed in GenBank with Blast search from three subtractive cDNA libraries, respectively, thirteen unique sequences were found from the subtractive cDNA library of sporulated oocysts, eight ESTs shared significant identity with previously described. A total of forty unique sequences were obtained from the two subtractive cDNA libraries, nine ESTs shared significant identity with previously described, the other sequences represent novel genes of E. tenella with no significant homology to the proteins in Genbank. These results have provided the foundation for cloning new genes of E. tenella and further studying new approaches to control coccidiosis.
Animals ; Chickens ; parasitology ; Coccidiosis ; parasitology ; veterinary ; DNA, Protozoan ; genetics ; Eimeria tenella ; genetics ; physiology ; Gene Expression Regulation ; Gene Library ; Nucleic Acid Hybridization ; methods ; Oocytes ; metabolism ; Poultry Diseases ; parasitology ; Spores

The purpose of this study was to investigate whether sporulated Neospora caninum oocysts, which had been stored for 46 mo in a 2% sulfuric acid solution at 4 degrees C, remain morphologically viable and infective to gerbils (Meriones unguiculatus). Six gerbils were orally inoculated with doses of 400 or 1,200 oocysts. Two mo after inoculation, the animals did not show any clinical signs, had no histological lesions, and were seronegative for N. caninum at 1: 50 in an immunofluorescent antibody test. PCR using the brain from each gerbil did not reveal N. caninum specific DNA. We conclude that oocysts preserved for 46 mo are not infective, despite being morphologically intact.
Acids ; Animals ; Brain/parasitology/pathology ; Cattle/parasitology ; Coccidiosis/parasitology/pathology/*veterinary ; Feces/parasitology ; Female ; Gerbillinae/*parasitology ; Neospora/genetics/growth & development/*pathogenicity ; Oocysts/*growth & development ; Polymerase Chain Reaction/methods ; Refrigeration ; Virulence

Eimeria (E.) tenella (local isolate) sporozoites were adapted on the chorioallantoic membrane (CAM) of 10-12 days chicken embryos and completed its life cycle in 6~7 days at 39 degrees C and 70 per cent humidity. Only 23 embryos (4.6%) were found dead from 1~4 day post inoculation of sporozoites with mild lesions on CAM with no gametocytes but few sporozoites in chorioallantoic fluid (CAF). On 5~7 day post inoculation, 432 embryos (86.4%) were found dead with severe haemorrhages on CAM and CAF contained uncountable number of gametocytes. After seven days post inoculation, 45 embryos (9%) were found to be alive. Some oocysts were also detected in the CAF on 6~7 days post inoculation. In the histological sections of the CAM, there were abundant small dark colored rounded bodies of gametes; distributed extensively in tissues of CAM on 5~7 days post inoculation of sporozoites. In some cases, cluster of small mature and immature relatively large bodies were seen in increasing numbers on 5~6 days post inoculation.
Animals ; Chick Embryo ; *Chickens ; Chorioallantoic Membrane/*parasitology ; Coccidiosis/parasitology/*veterinary ; Eimeria tenella/*growth&development ; Histocytochemistry ; Poultry Diseases/*parasitology

The present study was attempted to compare the Neospora caninum (N. caninum) antigenic bands recognized by different bovine immunoglobulin classes. A total 10, 5, 2, and 6 antigenic bands were exhibited on immunoblot profiles against bovine IgM, IgE, IgA, and IgG, respectively. A 46 kDa band was probed as a common antigenic band except IgA; 69 kDa band was bovine IgM and IgE; 33, 37, 55, and 79 kDa bands were bovine IgM and IgG; 72 kDa band was found IgM and IgA profiles. Based on the analysis, it appeared that different immunoglobulin classes recognizing different antigenic molecules were cooperating to cope with neosporosis.
Animals ; Antigens, Protozoan/immunology ; Cattle ; Cattle Diseases/diagnosis/immunology/*parasitology ; Coccidiosis/diagnosis/immunology/parasitology/*veterinary ; Female ; Immunoblotting ; Immunoglobulin Idiotypes/diagnostic use/*immunology ; Neospora/*immunology

Coccidiosis is a ubiquitous intestinal protozoan infection of poultry seriously impairing the growth and feed utilization of infected animals. Conventional disease control strategies have relied on prophylactic medication. Due to the continual emergence of drug resistant parasites in the field and increasing incidence of broiler condemnations due to coccidia, novel approaches are urgently needed to reduce economic losses. Understanding the basic biology of host-parasite interactions and protective intestinal immune mechanisms, as well as characterization of host and parasite genes and proteins involved in eliciting protective host responses are crucial for the development of new control strategy. This review will highlight recent developments in coccidiosis research with special emphasis on the utilization of cutting edge techniques in molecular/cell biology, immunology, and functional genomics in coccidia vaccine development. The information will enhance our understanding of host-parasite biology, mucosal immunology, and host and parasite genomics in the development of a practical and effective control strategy against Eimeria and design of nutritional interventions to maximize growth under the stress caused by vaccination or infection. Furthermore, successful identification of quantitative economic traits associated with disease resistance to coccidiosis will provide poultry breeders with a novel selection strategy for development of genetically stable, coccidiosis-resistant chickens, thereby increasing the production efficiency.
Animals ; Biotechnology/methods ; Chickens ; Coccidiosis/prevention&control/*veterinary ; Eimeria/*immunology ; Poultry Diseases/parasitology/*prevention&control ; *Protozoan Vaccines