Histologic, infectious, and molecular correlates of idiopathic spontaneous abortion and perinatal mortality.

The purpose of this study was to analyze the placental and neonatal tissues in fatal cases for a wide variety of infectious agents and cytokine expression. Placentas and corresponding neonatal tissues in 21 consecutive cases of idiopathic spontaneous abortion or perinatal death, before or within 2 days of birth, were tested for an infectious agent. The controls included 10 consecutive cases of fetal and placental tissues from therapeutic abortions, 5 placentas from unremarkable childbirths, and 11 placentas from cases of spontaneous abortion or perinatal death of known cause (ruptured uterus, placenta abruption, prolapsed cord). An intrauterine infection was noted in 16 of 21 (76%) of the placentas associated with neonatal mortality; in each case, the same infectious agent was found in the neonatal tissues, primarily the spleen. The most common infectious agent was enterovirus/coxsackie virus (10 cases); the histologic findings in the placenta were nonspecific. There was strong expression of TNF-alpha in the placenta and spleen of each of the cases of intrauterine infection and in none of the 26 controls. It is concluded that in utero infection and the associated cytokine up-regulation are responsible for many cases of unexplained fetal and neonatal loss.     

Genetic structure of the tick Ornithodoros coriaceus (Acari: Argasidae) in California, Nevada, and Oregon

The argasid tick Ornithodoros coriaceus (Koch) is the only confirmed vector of epizootic bovine abortion (EBA) in the United States. The disease and its tick vector have historically been reported in the foothills of the Sierra Nevada Mountains and coast ranges of California. In the past two decades, the range of EBA has apparently expanded into southern Oregon and northern Nevada. Possible explanations for this expansion include 1) increased recognition and reporting of EBA in these regions; 2) widespread movement of tick-infested and EBA-infected hosts with subsequent colonization of these regions by infected ticks; and 3) widespread movement of the EBA agent, independent of tick movements, into extant tick populations in these new regions. The current study was performed to evaluate these hypotheses by examining patterns of variability in a 420-bp segment of the 16S mitochondrial rDNA gene sequence among 210 O. coriaceus individuals from 14 sites in California, Oregon, and Nevada. Sixty-three unique haplotypes were identified in the ticks tested, with 84% of the sequence variation attributable to among-population variation and 16% to within-population variation. A majority of the haplotypes were unique to their particular collection site, whereas only four collection sites shared haplotypes. Overall, very little evidence of gene flow among tick populations was detected, making it unlikely that widespread tick movement had introduced O. coriaceus and the EBA agent into new regions.     

A survey of Neospora caninum-associated bovine abortion in large dairy farms of Mashhad, Iran

Neospora caninum is an apicomplexan protozoon causing abortion in cattle worldwide. The present study was designed to assess the importance of this parasite for causing abortion in dairy farms in the Mashhad area of Iran. Of the aborted bovine fetuses, 151 were collected from dairy farms between 2006 and 2008. First, brain samples were examined by polymerase chain reaction (PCR) for the presence of N. caninum DNA, diagnosis was complemented with immunohistochemistry (IHC) and fetal serology (ELISA). Twenty-two (14.5%) of bovine fetuses were considered to be infected with N. caninum with at least one diagnostic technique being positive. PCR yielded 18 (11.9%) positive out of 151 brain samples. Only 52 brain samples were suitable for IHC examination, and N. caninum organism was detected in six (11.5%) of these 52 brain samples. Fetal fluids (n = 151) were assessed with a N. caninum-ELISA, resulting in 15 (9.9%) seropositive fetal fluids samples. In the present study, a good agreement was observed between PCR and ELISA, and a fair agreement between PCR and IHC. The results indicated that abortion due to N. caninum infection is prevalent among large-size dairy farms in the Mashhad area of Iran, and that different complementary diagnostic techniques should be used to increase the chance to detect N. caninum.     

Risk of spontaneous abortion in singleton and twin pregnancies after IVF/ICSI

BACKGROUND: The risk of spontaneous first trimester abortion is estimated to be between 10 and 20%. Although it is common knowledge that the incidence of abortion decreases as pregnancy progresses, exact data in relation to the duration of pregnancy are scarce. METHODS: We reviewed 1597 clinical IVF/ICSI pregnancies with known outcome and tabulated the number of miscarriages or fetal demise per intervals of 2 weeks. We furthermore compared the outcome in terms of fetal survival of 1200 singleton pregnancies with that of 397 twin pregnancies. RESULTS: The overall incidence of non-ongoing singleton pregnancies was 21.7%. Fetal death, after positive heart activity had been recorded, occurred in 12.2% of singleton pregnancies. The overall incidence of spontaneous abortion in twin pregnancies was 17.1% (12.1% vanishing twins and 5.0% complete miscarriages). The incidence of miscarriage in the twin pregnancies, expressed per gestational sac, was 11.1%. Once fetal heart activity was present, the risk of abortion (per gestational sac) was 7.3%, which is significantly lower than that in singleton pregnancies. CONCLUSIONS: Our data give an estimate of the probability of miscarriage or fetal demise at any given period of the first trimester both for singleton and twin pregnancies. Twin pregnancies after IVF have a better potential for survival than singleton pregnancies.     

复发性流产的免疫治疗效果分析

目的了解采用淋巴细胞对复发性流产进行主动免疫的疗效。方法采用丈夫外周血淋巴细胞治疗复发性流产,454例患者疗程结束并成功随访,妊娠至20周后者为治愈。结果116（25．55％）例足月分娩,治愈率70．48％（未包括已孕20周内）。其间1例孕20周诊断无脑儿,1例孕26周超声示胎儿先天性左心发育不良,畸形率0．44％。结论免疫治疗是安全、有效的治疗复发性流产的方法之一,能确实解决病患实际问题。     

自然流产组织应用FISH技术的染色体异常遗传因素分析

目的分析兰州地区自然流产组织中的染色体异常发生率,并探讨染色体异常与自然流产的年龄、孕周、次数、胚胎性别的关系。方法应用荧光原位杂交（fluorescent in situ hybridization,FISH）技术检测了299例自然流产的绒毛或胚胎组织。结果我们检测出了119例染色体异常病例,其中16-三体最为常见,占所有异常总数的27．73％,X单体次之,占18．49％。我们还发现自然流产与流产时的妊娠孕周和胚胎性别有关。结论染色体异常是自然流产的一个重要原因,尽早地遗传咨询能够有效的降低出生缺陷的发生率。     

Monoclonal antibodies against maternal major histocompatibility complex class I molecules induce rapid abortion in mice.

PROBLEM: The role of antibodies against fetal or maternal antigens in maintaining or losing pregnancy is not clear. METHOD OF STUDY: Term-pregnant mice were injected with monoclonal antibodies against only fetal or fetal and maternal major histocompatibility complex class I molecules. The development of pregnancy was then followed. RESULTS: Antibodies against maternal, but not fetal, major histocompatibility complex class I molecules induced abortion in mice. The abortion occurred 6-8 hr after the administration of autoreactive antibodies. The abortion could only be induced after the formation of placenta. Antibodies against tumor necrosis factor-alpha could not prevent or postpone the abortion. Extensive bleeding has been detected in the placenta of aborting mice 3 hr after the administration of the antibodies. CONCLUSIONS: This study indicates that autoreactive antibodies present risk for pregnancy and that the damage leading to abortion induced by such antibodies most likely occurs at the maternal side of placenta.     

Abnormal T-cell reactivity against paternal antigens in spontaneous abortion: adoptive transfer of pregnancy-induced CD4+CD25+ T regulatory cells prevents fetal rejection in a murine abortion model

Mammalian pregnancy is thought to be a state of immunological tolerance. The mechanisms underlying this phenomenon are still poorly understood. Here, we determined whether an inappropriate function of T regulatory (Treg) cells is involved in the pathogenesis of spontaneous abortion. We evaluated spleen and decidual lymphocytes from CBA/J mice undergoing immunological abortion (DBA/2J-mated) or having normal pregnancy (BALB/c-mated) on day 14 of gestation for ex vivo cytokine production after PMA or paternal antigen (alloantigen) stimulation. Treg activity was characterized by quantifying CD4(+)CD25(+) cells, foxp3 expression, and interleukin-10 secretion. Decidual lymphocytes from abortion CBA/J mice contained a significantly higher frequency of interferon-gamma-producing T cells specific for paternal antigens compared to those from normal pregnancy (7.8% versus 2.7%, P < 0.05). Compared to virgin CBA/J females, normal pregnant mice showed strongly elevated numbers of CD4(+)CD25(+) and interleukin-10(+) Treg cells in the thymus whereas significantly lower frequencies of Treg cells were observed in abortion mice. Very interestingly, CD4(+)CD25(+) Treg cells from normal pregnant and nonpregnant CBA/J mice could inhibit both proliferation and interferon-gamma secretion of lymphocytes from abortion mice in vitro whereas in vivo prevention of fetal rejection could only be achieved after adoptive transfer of Treg cells from normal pregnant mice. Our data suggest that pregnancy-induced Treg cells play a vital role in maternal tolerance to the allogeneic fetus.     

Ultrasonographic detection of a live fetus in recurrent spontaneous abortion during the first trimester

To examine whether recurrent spontaneous abortion (RSA) canbe distinguished from repeated sporadic spontaneous abortion,the clinical course of 38 cases with three or more consecutiveand unexplained first trimester RSAs were retrospectively investigatedin this study. For comparison with controls, the clinical coursewas examined of 38 fertile females, who had had sporadic abortions.In 19 (50%) RSAs and 6 (16%) controls, fetal cardiac activitywas demonstrated by ultrasound during the course of pregnancy.The rate of detection of live fetus during pregnancy or at 8weeks ± 7 days gestation, was significantly greater inthe RSA group compared to the control. The rate of vaginal bleedingbefore spontaneous abortion was significantly less in the RSAgroup than in the control group. There was no difference betweenthe two groups in age or gestational age at spontaneous abortion.The patients with RSA were all examined for antiphospholipidantibodies in their sera and these were detected in eight ofthem. However, there was no difference in the rate of positivefetal cardiac activity between the RSA patients who tested positiveor negative for antibody. These results reveal that the clinicalcourse of RSA is very different from the course of sporadicabortion. Although sporadic abortion is a common complicationof pregnancy, RSA is not a random repeated abortion, but rathera separate disease from sporadic abortion in normal fertilefemales.     

Detection by PCR of Neospora caninum in fetal tissues from spontaneous bovine abortions

The routine diagnosis of Neospora caninum abortion is based upon histopathologic changes in fetal tissues and identification of tissue parasites by immunohistochemistry. Confirmation of N. caninum infection by immunohistochemistry has low sensitivity. In the present study, we examined the utility of PCR in detecting N. caninum infection in fetal tissues from spontaneous bovine abortion. DNA was obtained from fresh and formalin-fixed tissues from 61 bovine fetuses submitted for abortion diagnosis. Histopathology and immunohistochemistry determined the true status of N. caninum infection in each fetus. In formalin-fixed paraffin-embedded tissues, PCR detected N. caninum DNA in 13 of 13 true-positive fetuses (100%) and in 1 of 16 true-negative fetuses (6%). In fresh or frozen tissues, PCR detected N. caninum DNA in 10 of 13 true-positive fetuses (77%) and 0 of 11 true-negative fetuses (0%). PCR also detected N. caninum DNA in 6 of 8 fetuses that had typical lesions of N. caninum but were immunohistochemistry negative, indicating a higher sensitivity of PCR in comparison to that of immunohistochemistry. N. caninum DNA was amplified most consistently from brain tissue. PCR detection of N. caninum DNA in formalin-fixed, paraffin-embedded tissues was superior to that in fresh tissues, presumably because of the increased accuracy of sample selection inherent in histologic specimens.     

Predictable ultrasonographic findings of early abortion.

Early fetal growth delay and early oligohydramnios have been suspected as signs of embryonal jeopardy. However, little information is available for the prediction of early abortion. Sonographic examination of 111 early pregnancies between the sixth and ninth gestational week with regular, 28 day menstrual cycles was performed to investigate predictable sonographic findings of early abortion. Sonographic measurements of the gestational sac (G-SAC), crown-rump length (CRL) and fetal heart rate (FHR) were performed using a linear array real time transducer with Doppler. All measurements of 17 early abortions were compared to those of 94 normal pregnancies to investigate the objective rules for the screening of early abortion. Most of the early aborted pregnancies were classified correctly by discriminant analysis with G-SAC and CRL (G-SAC = 0.5222 CRL + 14.6673 = 0.5 CRL + 15, sensitivity 76.5% specificity 96.8%). With the addition of FHR, 94.1% of early abortions could be predicted. In conclusion, sonographic findings of early intrauterine growth retardation, early oligohydromnios and bradycardia can be predictable signs for the poor prognosis of early pregnancies.     

Polymorphism of human leukocyte antigen-E gene in the Japanese population with or without recurrent abortion

PROBLEM: The aim of this study was to investigate the gene frequencies and shared alleles of human leukocyte antigen (HLA)-E gene in Japanese couples with or without recurrent abortion. METHOD OF STUDY: Polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP) analysis was carried out to detect polymorphism in exon 3 of the HLA-E gene in 30 Japanese couples with recurrent abortion and 38 normal Japanese couples with proven fertility. RESULTS: No point mutation was detected in exon 3 of HLA-E in both recurrent aborters and normal controls. HLA-EG and HLA-ER alleles were detected with frequencies of 66.7% and 33.3% in couples with recurrent abortion and 69.2% and 30.8 in normal couples, respectively. The gene frequency of HLA-EG was higher than that of HLA-ER, which is contrary to that found in Caucasian, African-American and Hispanic people but similar to Chinese people. The frequency of each allele was not significantly different between recurrent aborters and normal controls. The number of shared alleles between each couple with recurrent abortion is not significantly different from that with normal controls. CONCLUSION: Allele frequencies of HLA-E were suggested to be different in Asian people from those in other ethnic people. In light of no specific distribution pattern in recurrent aborters, HLA-E polymorphism does not seem to play a role in the pathogenesis of recurrent abortion.     

Attitudes of Mexican geneticists towards prenatal diagnosis and selective abortion

Prenatal diagnosis (PD) provides the physician information on whether the unborn fetus has a genetic or chromosomal disorder, and offers patients a new option: selective abortion. In the present study, we analyzed the answers Mexican geneticists provided to a few selected questions from a multi-national survey designed by Wertz and Fletcher [1988: Am J Hum Genet 42:592–600]. The selected questions were related to the use of PD, the acceptance of selective abortion, and the self-reported directiveness of counselling following the diagnosis of a fetal anomaly. Our results show that the great majority of Mexican geneticists participating in the study agree with PD when medically indicated, but not on free demand. Specific cases stimulated the group on thinking more than the general statements provided in the survey. Although the majority agreed that PD should be available to all women, when faced with cases of nonmorbid maternal anxiety, paternity testing, and sex selection, the proportion of geneticists willing to perform the test decreased substantially. When counselling patients on a fetal anomaly, the minority would be as unbiased as possible, and this seems to be the tendency in developing countries where counselling, as stated in the respondents' comments, reflects the belief that the goal of genetics is the prevention of or opposition to abortion. Counselling was influenced by the severity of the disorder. The geneticists' personal attitude toward abortion in the same situations was stronger than when counselling others. Analysis of directiveness in counselling for fetal anomaly showed that older geneticists, with more years of experience in medical genetics, were more likely to be neutral. When counselling directively, the group showed an overall direction toward continuing affected pregnancies. However, older geneticists and those with more than 10 years of practice were more likely than their younger counterparts to counsel towards terminating affected pregnancies. In personal situations of fetal disorder, the general tendency was to abort; however, geneticists seeing more than 5 patients per week, and those who believe that religion is important, were more likely to reject abortion. The sample is representative of Mexican geneticists, and the main limitation of this study is that the geneticists have very little experience in PD, and that their responses were mostly based on theory. However, their opinions may influence the demand and the availability of PD and abortion, as well as the possibility of legalization of abortion on the basis of a fetal defect. Am. J. Med. Genet. 75:426-431, 1998. © 1998 Wiley-Liss, Inc.     

Ovine chlamydial abortion: characterization of the inflammatory immune response in placental tissues

Ovine chlamydial abortion is a serious cause of fetal mortality in several sheep-rearing countries. The causal agent, Chlamydophila abortus (Chlamydia psittaci), does not generally induce clinical signs in the ewe other than abortion; this is associated with macroscopically visible damage in the placenta, which may be inflamed and thickened. To investigate the nature of the placental inflammation, seven pregnant sheep were inoculated subcutaneously at 70 days' gestation with C. abortus (strain S 26/3). A further five pregnant sheep received control inoculum by the same route at the same stage of pregnancy. Three of the infected ewes produced stillborn lambs and four produced live lambs. Lesions characteristic of chlamydial infection were present in all placentas except for two from one ewe that gave birth to twins. Histopathological examination of placental tissues from aborted fetuses showed a mixed inflammatory cell infiltrate with vasculitis and thrombosis in the mesenchyme of the intercotyledonary membranes. Cells expressing the macrophage-associated molecule CD 14 were found to be numerous, as were cells expressing major histocompatibility complex class II (MHC II) molecules. Many cells expressing messenger RNA (mRNA) encoding for tumour necrosis factor-alpha (TNF-alpha) were demonstrated, but few cells expressing interferon gamma mRNA and none expressing interleukin-4 mRNA were detected. The fetal immune response included small numbers of CD4+ and CD8+ cells, gamma delta T cells and B cells. It is concluded that abortion is the result of several factors, including destruction of tissue by C. abortus, vascular thrombosis, and an inflammatory response by the fetus. Production of TNF-alpha by fetal macrophages expressing MHC II molecules may be of considerable significance in the pathogenesis of abortion.     

Roles of the surface layer proteins of Campylobacter fetus subsp. fetus in ovine abortion

The role of the surface (S)-layer proteins of Campylobacter fetus subsp. fetus has been investigated using an ovine model of abortion. Wild-type strain 23D induced abortion in up to 90% of pregnant ewes challenged subcutaneously. Isolates recovered from both dams and fetuses expressed S-layer proteins with variable molecular masses. The spontaneous S-layer-negative variant, strain 23B, neither colonized nor caused abortions in pregnant ewes. A series of isogenic sapA and recA mutants, derived from 23D, also were investigated in this model. A mutant (501 [sapA recA(+)]) caused abortion in one of five challenged animals and was recovered from the placenta of a second animal. Another mutant (502 [sapA recA]) with no S-layer protein expression caused no colonization or abortions in challenged animals but caused abortion when administered intraplacentally. Mutants 600(2) and 600(4), both recA, had fixed expression of 97- and 127-kDa S-layer proteins, respectively. Two of the six animals challenged with mutant 600(4) were colonized, but there were no abortions. As expected, all five strains recovered expressed a 127-kDa S-layer protein. In contrast, mutant 600(2) was recovered from the placentas of all five challenged animals and caused abortion in two. Unexpectedly, one of the 16 isolates expressed a 127-kDa rather than a 97-kDa S-layer protein. Thus, these studies indicate that S-layer proteins appear essential for colonization and/or translocation to the placenta but are not required to mediate fetal injury and that S-layer variation may occur in a recA strain.     

Risk factors for spontaneous abortion: a case-control study in France.

A case-control study was conducted in seven maternity hospitals in the Paris area in 1988 to evaluate the role of several risk factors in spontaneous abortion. A total of 279 cases and 279 controls were compared for socio-demographic characteristics, reproductive history and for conditions of conception. Prior fetal losses [odds ratio (OR) = 2.30 for n greater than or equal to 2; 95% confidence interval (CI) = 1.17-4.61] and maternal age at pregnancy (greater than 30 years) appeared to be major and independent risk factors of spontaneous abortion. Other factors associated with an increased risk of fetal loss were: geographical or ethnic origin (OR = 2.85 for North African women; 95% CI = 1.58-5.10); psychological problems at the time of conception, either related to the outcome of the pregnancy (OR = 3.08; 95% CI = 0.92-10.25) or unrelated to this outcome (OR = 3.35; 95% CI = 1.41-8.00). The following factors were not associated with spontaneous abortion: gravidity, parity, prior induced abortion, prior sexually transmitted diseases and Chlamydia trachomatis serology, menstrual cycle abnormalities, induced conception cycle and in-vitro fertilization, cigarette smoking, current or past use of combined oestrogen/progestagen pill or intrauterine device. These findings confirm the importance of two risk factors for fetal loss: maternal age and number of prior spontaneous abortions. Two risk factors, ethnic origin and psychological problems at the time of conception are also identified, which require further study.     

Virus-induced abortion. Studies of equine herpesvirus 1 (abortion virus) in hamsters.

A hamster-adapted strain of equine herpesvirus-1 (equine abortion virus) caused severe hepatic degeneration in both pregnant and nonpregnant hamsters and, in addition, regularly induced abortion in pregnant hamsters inoculated at midgestation. In nonpregnant hamsters, the only consistently affected organ was the liver despite a prolonged viremia. Newborn animals usually died 1 to 2 days after inoculation; adults died 5 to 9 days after inoculation. In pregnant hamsters, the virus had a tropism for the placenta as well as the liver. The placental infection was confined almost exclusively to one cell type in the fetal portion of the placenta: the trophoblast cells of the syncytiotrophoblast zone. Necrosis of this zone led to fetal death and abortion. Infection of the fetus did not occur.     

Caffeine intake and the risk of first-trimester spontaneous abortion

BACKGROUND: Some epidemiologic studies have suggested that the ingestion of caffeine increases the risk of spontaneous abortion, but the results have been inconsistent. METHODS: We performed a population-based, case-control study of early spontaneous abortion in Uppsala County, Sweden. The subjects were 562 women who had spontaneous abortion at 6 to 12 completed weeks of gestation (the case patients) and 953 women who did not have spontaneous abortion and were matched to the case patients according to the week of gestation (controls). Information on the ingestion of caffeine was obtained from in-person interviews. Plasma cotinine was measured as an indicator of cigarette smoking, and fetal karyotypes were determined from tissue samples. Multivariate analysis was used to estimate the relative risks associated with caffeine ingestion after adjustment for smoking and symptoms of pregnancy such as nausea, vomiting, and tiredness. RESULTS: Among nonsmokers, more spontaneous abortions occurred in women who ingested at least 100 mg of caffeine per day than in women who ingested less than 100 mg per day, with the increase in risk related to the amount ingested (100 to 299 mg per day: odds ratio, 1.3; 95 percent confidence interval, 0.9 to 1.8; 300 to 499 mg per day: odds ratio, 1.4; 95 percent confidence interval, 0.9 to 2.0; and 500 mg or more per day: odds ratio, 2.2; 95 percent confidence interval, 1.3 to 3.8). Among smokers, caffeine ingestion was not associated with an excess risk of spontaneous abortion. When the analyses were stratified according to the results of karyotyping, the ingestion of moderate or high levels of caffeine was found to be associated with an excess risk of spontaneous abortion when the fetus had a normal or unknown karyotype but not when the fetal karyotype was abnormal. CONCLUSIONS: The ingestion of caffeine may increase the risk of an early spontaneous abortion among non-smoking women carrying fetuses with normal karyotypes.     

Maternal T cells promote placental growth and prevent spontaneous abortion

Transplantation immunologists have long been intrigued by the natural allograft that results from normal mammalian pregnancy. Its general success contrasts with the rejection problems associated with most artifactual organ transplantation and raises intriguing questions concerning the nature of the mechanisms involved in that success. This area of research has recently taken on added momentum because it is now clear that immunological maneuvers can prevent recurrent spontaneous abortion in mice, horses and humans [1-3]. The purpose of this review is to discuss some of these recent developments, which lead to the surprising conclusion that maternal T cells, rather than being potentially detrimental to the fetal allograft, promote its growth and viability during normal pregnancy. This review will address these questions by considering: (a) the nature of the exposure of fetal alloantigens to the maternal circulation in the chimeric zone of the placenta; (b) the evidence for maternal immune recognition of the fetal alloantigens; and (c) the consequences of that recognition with respect to the prevention of spontaneous abortion. As in other areas of immunology the T cell emerges as the most important component of this immune recognition.     

Detection of Neospora from tissues of experimentally infected rhesus macaques by PCR and specific DNA probe hybridization.

Neospora is a newly recognized Toxoplasma-like cyst-forming coccidian parasite that causes abortion or congenital infections in naturally or experimentally infected animals. In this study, pregnant rhesus macaques were inoculated with culture-derived tachyzoites of a bovine Neospora isolate, and tissue samples from various major organs were collected from dams and fetuses for the detection of parasite DNA by using oligonucleotide primers COC-1 and COC-2 for PCR amplification of a conserved coccidial nuclear small-subunit rRNA gene sequence, and amplification products were confirmed by hybridization with a Neospora-specific DNA probe. PCR products were amplified from DNAs of different fetal monkey tissues, including brain, heart, lung, liver, spleen, skeletal muscle, skin, and placenta. In addition, Neospora DNA was amplified from the brain, heart, and lung tissues of infected rhesus macaque dams. The PCR and probe hybridization system may provide an effective method for the detection of Neospora infection in fetuses and dams from nonhuman primates and may be useful in determining the zoonotic potential of Neospora.