The aim of current study was to assess the seroprevalence of Neospora (N.) caninum and Toxoplasma (T.) gondii infections in aborted cattle and subsequent estimation of role of these parasites in cattle abortion in Hamedan province, Iran. Blood samples (n=85) of aborted cattle were collected from different locations of Hamedan province for a period of two years from January, 2010 to December, 2012. All the samples were evaluated for the presence of IgG-antibodies against N. caninum and T. gondii using Enzyme Linked Immuno Sorbent Assay (ELISA). The results showed that 61.2% (n=52/85), 5.9% (n=5/85) and 3.5% (n=3/85) animals were positive for N. caninum, T. gondii and co-infection of these two, respectively. There was no significant correlation between seroprevalence rates, age groups and breeding (p>0.05). However, a significant difference was found between T. gondii infection and the type of cattle (p=0.05), unlike to N. caninum (p=0.52). This is the first report of N. caninum and T. gondii co-infection in cattle in Iran. Although the rate of T. gondii infection was lower but this parasite might partly be responsible to be transmitted to humans. Further investigations and appropriate control strategies in management of cattle farms are highly recommended.

Neospora caninum is an Apicomplexan parasite which may cause abortion in cattle. This study investigated occurrences of antibodies against N. caninum in Iranian native cattle. From September 2010 to September 2011, blood samples (n=768) of native cows were collected randomly from different rural regions of Hamedan (n=400) and Kurdistan provinces (n=368) located to the western part of Iran. All the samples were evaluated for IgG antibodies against N. caninum using Enzyme Linked Immunosorbent Assay. The IgG antibodies to N. caninum were found in 14.2% (n=109/768) of serum samples (95% CI: 11.74 - 16.66). There was a significant difference between seropositivity and abortion history (p<0.0001, OR=2.9), unlike to age groups (p=0.105). This is the first report of N. caninum infection in Iranian native cattle. In conclusion, N. caninum is an important factor in abortion in Iranian native cattle. Further comprehensive studies and designing control strategies for improving management in cattle farms are highly recommended.

Infection with Neospora caninum in a dog is described. The predominant clinical findings were proprioception deficit, involuntary and continuous contracture of the pelvic extremity muscles. N. caninum is structurally similar to Toxoplasma gondii, but it is immunologically distinct. Therefore, infections had been misdiagnosed as toxoplasmosis. The diagnosis was confirmed by an immunofluorescence antibody test. The parasite is sensitive to clindamycin and trimethoprim/sulphonamide, however the prognosis is poor especially if the muscle contractures have occurred.

Objective--To evaluate the immunologic response of a killed tachyzoite vaccine against Neospora caninum and its effectiveness in preventing fetal loss associated with experimentally induced neosporosis in sheep. Animals--30 Dorset ewes. Procedure--Ewes were randomly allocated to receive vaccination on days 1 and 60 of the study with a killed N caninum tachyzoite preparation in a commercially available adjuvant or a saline-adjuvant mixture. A ram was placed on pasture with the ewes from days 15 to 60. Blood was collected from ewes before primary and booster vaccinations and prior to experimental challenge with N caninum tachyzoite performed on day 90; sera were assessed via Neospora agglutination (NA) and immunofluorescence antibody (IFA) assays. Blood was collected from lambs before they suckled, and sera were tested for antibodies against N caninum. Results--Of the 14 vaccinated ewes that became pregnant, 12 gave birth to live-born lambs; in contrast, 5 of 11 pregnant control ewes gave birth to live-born lambs. Whereas vaccination improved fetal survival in pregnant ewes challenged with N caninum tachyzoites, it did not appear to have any appreciable effect on transmission of N caninum to offspring, as indicated by results of NA and IFA assays. Conclusions and Clinical Relevance--The N caninum tachyzoite vaccine used in this study appeared to provide protection against fetal loss associated with experimentally induced neosporosis in a high proportion of pregnant ewes.

Neospora caninum-induced abortion is a major production problem in the daily cattle industry in the United States and worldwide. Abortions attributable to naturally acquired N caninum infection also have been observed in pygmy goats. We studied experimentally induced infections with N caninum in pregnant pygmy does to determine whether abortions attributable to N caninum infection would occur after inoculation. Seven pregnant pygmy does (1 control doe and 6 inoculated with N caninum) were studied. The control doe remained clinically normal throughout the study and delivered 2 healthy kids. Abortion, fetal death, and stillbirths were observed in some pregnant does inoculated with N caninum. Two pregnant pygmy does inoculated with N caninum early in gestation (day 51) had fetuses that died and were aborted, or died and were reabsorbed. Neospora caninum tachyzoites and lesions were observed in the brain, spinal cord, and heart of aborted fetuses; parasites also were isolated from the placenta. Four additional pregnant pygmy does (2 inoculated at mid-gestation [day 85], and 2 at late gestation [day 127]) did not abort after inoculation. However, 1 doe inoculated during mid-gestation delivered a stillborn fetus that had died about 1 week prior to parturition. This kid was congenitally infected with N caninum. Neospora caninum was isolated from the placentas of all inoculated does examined. Neonatal neosporosis was not observed in live-born kids, nor were stages of N caninum isolated from any live-born kid. Does did not undergo abortion or have congenitally infected kids when they were rebred and evaluated for neosporosis.

Neospora caninum causes serious disease in dogs, and it, or a similar parasite, is a major cause of abortion in cattle. Little is known about the susceptibility of this protozoan to antimicrobial agents. We studied several antimicrobial agents to determine which classes might have activity against this parasite. We also determined whether activity of such agents was coccidiocidal or coccidiostatic. A 2-day of treatment, monoclonal antibody-based enzyme immunoassay and a 5-day of treatment, cell culture flask (CCF), lesion-based assay were developed to examine the ability of test agents to inhibit tachyzoite multiplication. Seven sulfonamides were examined, with the following activities observed: sulfathiazole greater than or equal to sulfamethoxazole > sulfadiazine > sulfaquinoxaline greater than or equal to sulfamethazine > sulfadimethoxine > sulfamerazine. Dapsone, a sulfone, had little activity. Six dihydrofolate reductase/thymidylate synthase inhibitors were examined, with the following activities observed: piritrexim > pyrimethamine > ormetoprim > trimethoprim = diaveridine > methotrexate. Six ionophorous antibiotics were examined; lasalocid, maduramicin, monensin, narasin, and salinomycin had equivalent activities, but alborixin was toxic for host cells at the lowest concentration examined. Three macrolide antibiotics--azithromycin, clarithromycin, and erythromycin--were examined and had equivalent activities. Two tetracycline antibiotics, doxycycline and minocycline, were examined and had equivalent activities. Three lincosamide antibiotics were examined, with the following activities observed: clindamycin hydrochloride > clindamycin phosphate > lincomycin hydrochloride. Pentamidine and 6 of its analogs were examined, and only hexamidine and 1,4-Di[4-(2-imidazolinyl)-2-methoxy-phenoxylbutane had activity. Eight miscellaneous antiprotozoal agents were examined for activity. Amprolium, metronidazole, paromomycin, and roxarsone had little activity. Arprinocid, diclazuril, nitrofurazone, and robenidine had good activity. Eleven agents were examined in both assays, whereas 32 agents were examined in the CCF assay only. The enzyme immunoassay and CCF assay provided similar results for agents that rapidly killed tachyzoites. However, agents that inhibited development, but were not rapidly fatal for tachyzoites, had better activity in the CCF assay. Of the classes of agents examined, the dihydrofolate reductase/thymidylate synthase inhibitors, 2 of the 6 pentamidine analogs, and the ionophores were coccidiocidal and the sulfonamides, macrolides, tetracyclines, and lincosamides were coccidiostatic. Of the miscellaneous agents examined, arprinocid, nitrofurazone, and robenidine were coccidiocidal and diclazuril was coccidiostatic.

OBJECTIVE To determine the extent of environmental exposure to heteroxenous coccidia from wild canid feces in southeastern Ohio. SAMPLE 285 presumed wild canid fecal samples collected across an ecological system in southeastern Ohio. PROCEDURES Morphological classification and molecular analysis were used to determine the canid genus for collected fecal samples. Microscopic and molecular analysis were used to detect coccidian oocysts and DNA. Several variables were analyzed for associations with coccidian DNA detection or prevalence. RESULTS Coccidian DNA was detected in 51 of 285 (17.9%) fecal samples. Of those positive samples, 1% (95% confidence interval, 0.4% to 3%) had positive results for Hammondia heydorni and none had positive results for Neospora caninum, for an estimated environmental N caninum prevalence of 0% (95% confidence interval, 0% to 7%)/1-km2 hexagonal area evaluated. Morphological classification revealed that 78.9% (225/285) of fecal samples were from coyotes and 17.2% (49/285) were from foxes. No difference in proportions of coccidian DNA-positive fecal samples was identified among canid species. Environmental temperature and fecal freshness were associated with coccidian DNA detection. Land use type, relative canid density, and cattle density were not associated with the prevalence of coccidian DNA-positive samples. CONCLUSIONS AND CLINICAL RELEVANCE The low prevalence of coccidia shed in wild canid feces in this study, including the estimated 0% environmental prevalence of N caninum, suggested that the role of the oocyst environmental phase in coccidia transmission to ruminants is likely minor in rural southeastern Ohio.

This survey was carried out to investigate the seroprevalence of antibodies to Neospora (N.) caninum in dairy cattle raised in Kangwon province in Korea. A total of 349 sera collected from dairy cattle were tested for N. caninum antibodies using an indirect immunofluorescent antibody test (IFAT). Ninety six (27.5%) dairy cattle were positive by IFAT. Seroprevalence of cows was 28.9% (91/315) and seroprevalence of boars was 14.7% (5/34). The seroprevalences of cows were increased according to the ages from 19.6% in cows less than 2 year-old to 50.0% in cows more than 7 year-old. Among the seroprevalences of cattle according to the raised areas, five counties or cities, Wheongsung was 34.6% (27/78), Wonju was 29.3% (46/157), Hongcheon was 28.9% (13/45), Chuncheon was 15.0% (3/20) and Cheolwon was 13.3% (2/15). It was proved that dairy cattle raised in Kangwon provinces exposed extensively and seriously to N. caninum.

This study was performed to detect Neospora caninum in blood of 61 Korean native cattle and 50 dairy cows in Chungnam province. All of them were healthy and did not show any clinical signs. DNA was isolated from blood samples and a 328 bp fragment was amplified by PCR using primer pair Np21 and Np6. The PCR positive rate was 14.8% in Korean native cattle and 0% in dairy cows. Cows with 15.6% were a little higher than bulls with 12.5% in gender. The detection rate of over 3-year-old Korean native cattle was 28.6% in age. The results demonstrate that N. caninum DNA can be detected in blood by PCR. PCR analysis in blood may be useful to annually screening test for N. caninum infection in clinically healthy cattle.

A murine monoclonal antibody (MAB) 6G7 generated against tachyzoites of Neospora caninum recognized 8 major and several minor antigens, as observed by western blot analysis. Relative rate of migration of the 8 major antigens ranged from 31 to 97.4 kd. In addition, MAB 6G7 recognized a Toxoplasma gondii tachyzoite antigen with a relative rate of migration of 107 kd. Immunogold labeling of N caninum tachyzoites grown in human foreskin fibroblast cells indicated that MAB 6G7 binds to micronemes, dense granules, basal portions of rhoptries, and intravacuolar tubules within the parasitophorous vacuole. Monoclonal antibody 6G7 also bound to micronemes and basal portions of rhoptries within tachyzoites of T gondii. Monoclonal antibody 6G7 did not significantly inhibit development of tachyzoites in vitro.