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Toxoplasma gondii (T. gondii) is a zoonotic apicomplexan protozoan that can cause reproductive losses in ruminants across the world. Therefore, a meta-analysis was conducted to estimate the worldwide prevalence of T. gondii infection in the aborted fetuses and stillbirths of sheep, goat, and cattle. Moreover, it attempted to evaluate the prevalence rate of T. gondii infection in ruminants that had abortions using serological methods. Based on the keywords, a systematic search of six databases was conducted to retrieve cross-sectional articles in English-language. Data were synthesized to calculate the overall prevalence of T. gondii infection worldwide using the random-effects model with a 95 % confidence interval (CI). Moreover, the present study includes sensitivity analysis, publication bias test, and quality assessment of the studies. The final analyses included 37, 19, and 8 studies conducted on sheep (4383 aborted fetuses and stillbirths as well as 1940 abortive sheep), goat (248 aborted fetuses and stillbirths and 533 abortive goats), and cattle (460 aborted fetuses and stillbirths and 165 abortive cattle). The overall prevalence rates of T. gondii infection in aborted fetuses and stillbirths globally were 42 % (95 % CI: 17 %-67 %), and 31 % (95 % CI: 11 %-51 %) using molecular methods for sheep and goats, as well as 16 % (95 % CI: 11 %-22 %) and 27 % (95 % CI: 1 %-54 %) using serological tests for sheep and goats, respectively. In addition, the overall prevalence rates of T. gondii infection in sheep and goats that had abortions were 56 % (95 % CI: 35 %-76 %) and 50 % (95 % CI: 6 %-94 %), respectively. These analyses could not be performed in cattle due to the small number of studies and the small number of animals under study. According to the high prevalence of T. gondii infection in sheep and goats that had an abortion, T. gondii could be a potential factor of causing reproductive failures in ruminants worldwide. So, effective control measures and strategies are needed to reduce the rate of abortion in sheep and goats as well as reduce the economic damage to the livestock industry., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Purpose: The aim of this study was to describe for the first time a natural case of ovine abortion associated with Neospora caninum in a flock with reproductive losses in Argentina., Materials and Methods: The analyzed flock consisted of 256 Texel sheep, of which 134 had been mated. A single blood sample was obtained by jugular vein puncture from 220 ewes (116 adult ewes, 104 yearling ewes) and 93 lambs. Serum samples and fetal fluid were tested using the indirect fluorescence antibody test to detect antibodies against N. caninum and T. gondii. Fetal and placental tissues from aborted fetus were examined by standard gross pathology procedures and were tested using histopathology and immunohistochemistry. Moreover, DNA from fetal and placental tissues was isolated and a PCR assay to detect N. caninum, T. gondii and Chlamydia spp. was used., Results and Discussion: The pregnancy rate was 89% (119/134), the abortion rate was 8.4% (10/119) whereas the perinatal mortality rate was 15% (16/109). Out of 116 adult ewes sampled, 34.48% presented anti-N. caninum antibodies. Ten ewes had aborted, and one fetus was directly submitted to the diagnostic laboratory for further processing. Antibodies against N. caninum were detected in fetal fluid and in the aborted dam. Histopathological analysis in fetal tissues showed multifocal lymphohistiocytic glossitis, diffuse mild lymphohistiocytic endocarditis, pericarditis and focally extensive myocarditis. Severe multifocal necrotizing placentitis and diffuse mild lymphohistiocytic placentitis with the presence of lymphohistiocytic vasculitis were observed in placenta. N. caninum was immunolabeled in the placenta and fetal tongue. In addition, N. caninum DNA was detected in placenta, central neural system, lung and heart of the aborted fetus. There was no evidence of other infectious abortifacients in the aborted fetus., Conclusion: The present study described for the first time an ovine abortion caused by N. caninum in Argentina. Further investigations at a larger scale are required to establish the role of N. caninum as an important cause of reproductive losses in sheep flocks from the region.

Neospora caninum is an apicomplexan protozoan parasite that is a leading cause of abortion in cattle. Detection of parasite-specific DNA by PCR is a highly sensitive method for identifying the presence of N. caninum in a variety of tissues. We developed and validated a probe-based real-time PCR assay targeting the conserved Nc5 gene of N. caninum. Using N. caninum strain Nc-1 genomic DNA and a synthetic gene fragment as amplification standards, we determined the PCR amplification efficiency and the limit of detection to be 95.60% and 3 copies, respectively. Five pathogens frequently associated with bovine abortions, namely bovine viral diarrhea virus types I and II, bovine alphaherpesvirus-1, Chlamydia, and Leptospira, were tested to ensure analytical exclusivity. A total of 103 clinical samples from aborted fetuses were tested concurrently with a standard conventional PCR and the new probe-based real-time PCR assay. All tested samples showed 100% agreement between these two assays. In conclusion, the probe-based real-time PCR assay facilitates accurate and rapid detection of N. caninum from abortions in cattle., (Copyright © 2019 Elsevier B.V. All rights reserved.)

A captive 17-year old female Malayan tapir (Tapirus indicus) aborted a fetus with a crown rump length of 19cm in early pregnancy. The fetus showed an early state of mummification. Histologically, a multifocal mononuclear encephalitis, myocarditis and periportal hepatitis was present indicating a possible protozoal cause of abortion. Although immunohistologically, Neospora (N.) caninum antigen could not be demonstrated, N. caninum DNA was detected by Polymerase Chain Reaction (PCR) in brain, heart, liver and lung of the fetus. N. caninum DNA was extracted from the aborted fetus and the microsatellite marker MS10 was amplified by PCR and sequenced. The obtained MS10 microsatellite pattern has not been described in Germany yet. Nevertheless, the MS10 pattern was very similar to those reported for N. caninum isolated from dogs and cattle in Germany. Because of the histological pattern and extent of the lesions, neosporosis was suspected as the cause of fetal death and abortion. This case report describes for the first time transplacental transmission of N. caninum and abortion due to neosporosis in a tapir., (Copyright © 2017 Elsevier B.V. All rights reserved.)

N. caninum could cause abortion in small ruminants. The aim of the study was to detect N. caninum infection in ovine aborted foetuses in the Mashhad area by PCR examination. During the period 2009 to 2013, 71 ovine aborted foetuses were collected and their brain samples examined by PCR. Of the 71 brains of the aborted foetuses, N. caninum DNA was detected in seven (9.8%) samples. In conclusion, it seems that N. caninum may act as a causative agent of abortion in sheep in the Mashhad area.

The cyst-forming protozoan parasite Neospora caninum is one of the main causes of bovine abortion worldwide and is of great economic importance in the cattle industry. Recent studies have revealed extensive genetic variation among N. caninum isolates based on multilocus microsatellite genotyping. Currently, the most extensive study reported is based on the N. caninum genotyping of 96 samples from four countries on two continents (Spain, Argentina, Germany and Scotland) that demonstrate different clusters of multilocus genotypes (MLGs) implicated in cattle abortions as well as the population sub-structuring of N. caninum, which is partially associated with the geographical origin. The aim of this study was to genotype N. caninum from aborted bovine foetuses that originated from Mexico within the region of Aguascalientes and to investigate their genetic diversity. Parasite DNA was detected in 27 out of the 63 analysed foetuses recovered from 10 different herds. Complete or nearly complete profiles based on 9 microsatellite markers were obtained from 11 samples. Diverse N. caninum MLGs were implicated in the occurrence of abortion in each herd. All of the Mexican MLGs differed from the MLGs previously determined for the Argentinean, Spanish, German and Scottish N. caninum populations. The Mexican MLGs failed to cluster by eBURST analyses. The MLG relationships using PCoA showed a close genetic relationship between the Spanish population and a portion of the Mexican population, but a more distant genetic relationship with the Argentinean genotypes. These results demonstrate the genetic diversity of N. caninum in the studied areas that differed from other populations of N. caninum around the world., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Neospora caninum is a major cause of abortion in cattle. However, it is not known why not all infected animals abort. In this study, Th1 (IFN-γ), Th2 (IL4) and T reg (IL-10) cytokine gene expression was examined by real time PCR using the TaqMan approach in all of these dams and their foetuses after experimental infection with the isolate Nc-Spain7 at 110 days of pregnancy and euthanasia 6 weeks after infection. In prior published work, foetal death was observed in three of six infected dams and transplacental infection in all the 6 infected foetuses. In the spleen of the dams, IL-4 expression was down-regulated in dams with aborted/non viable foetuses compared to both uninfected dams (controls, n=3) and infected dams with live fetuses at euthanasia. In the lymph nodes draining the placenta, up-regulated expression of IL-4 was observed in infected dams with live foetuses compared to control dams. In the placenta, infected dams with live foetuses had significantly up-regulated IFN-γ in both caruncle and cotyledon and up-regulated IL-10 in cotyledon compared to control dams. Infected live foetuses showed up-regulated expression of IFN-γ and IL-10 in foetal spleen, and showed downregulated expression of IL-4 in the thymus compared to control uninfected foetuses. Expression of any cytokine in the thymus was significantly lower compared to the levels observed in foetal spleen. The results indicate an up-regulated expression of Th1, Th2 and Treg in infected dams with live foetuses and in their foetuses. On the other hand, down-regulation of Th2 immune responses and Treg cytokines were observed in infected dams which had aborted or had non-viable foetuses at euthanasia, suggesting an immunological recovery of cytokine gene expression levels in dams a few weeks after an abortion occurred., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The impact of spontaneous Neospora caninum infection on pregnancy loss and subsequent pregnancy in grazing lactating dairy cows was evaluated. Data from 1273 females (878 multiparous and 395 first-calving cows) from six preselected dairy herds were analyzed. Cows were classified as seropositive (SP) (prevalence, 24%; range, 11%-33%) or seronegative (SN) by indirect immunofluorescence detection of antibodies against N caninum. Seropositive cows (prevalence, 40.0%) presented higher (P < 0.001) incidence of abortion compared with SN cows (prevalence, 4.1%). Neospora caninum DNA was detected by real-time polymerase chain reaction in 44.4% of intact aborted fetuses from SP cows, whereas none was found in those aborted from SN cows. The average daily milk production adjusted to 305 days was lower (P < 0.001) in SP (22.5 ± 0.3 L/day) than in SN cows (24.8 ± 0.2 L/day). Furthermore, SP cows presented greater occurrence of retained placenta (17.1% vs. 6.0%; P < 0.001) and acute postpartum metritis (9.8% vs. 2.4%; P < 0.001). Despite similar pregnancy rates after first postpartum artificial insemination (27.6% vs. 31.8%; P = 0.40), cumulative pregnancy rates during 300 days in milk (94.7% vs. 98.5%; P = 0.005) were greater in SN cows. A reduced (P = 0.0001) Cox proportional hazard of pregnancy rate at 300 days in milk and a longer interval from parturition or abortion to conception (median, 111 vs. 101 days) were observed in SP compared with SN cows. Spontaneous N caninum infection is a significant contributing factor of pregnancy loss and occurrence of uterine disease (i.e., retained placenta and metritis), negatively affecting subsequent pregnancy in grazing lactating dairy cows., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Bovine neosporosis has emerged as a main cause of abortion in cattle worldwide. An important question to understand the disease is why not all infected cows abort. In the present review we summarize the knowledge on markers related to the diagnosis and more importantly to the risk of abortion in the infected cow. Markers considered herein include those based on specific antibodies, antibody titers and antibody subtypes, cellular immunological markers, hormones and other proteins related to gestation. The identification of parasite molecules that are specifically identified in the aborting cows might help to understand the mechanism of parasite-associated abortion and control the disease., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The water buffalo (Bubalus bubalis) is an important species in several countries for its milk and meat production, as well as for transport and other agricultural activities. It is, in general, considered more resistant than cattle to different parasitic diseases, also less demanding for forage quality. It has been postulated that buffalo may be resistant to abortion caused by neosporosis, because of high serological prevalences found in buffalo herds from different localities, with no description of Neospora caninum-related abortion. Recent studies have demonstrated the potential impact of neosporosis in pregnant water buffalo cows. In this work, three pregnant buffalo cows were experimentally infected with Nc-1 strain of N. caninum, and abortion was detected 35 days post-infection. Molecular and histopathological results found in post-mortem tissues are described and discussed, confirming the susceptibility of water buffalos to abortion caused by N. caninum.

The aims of this study were to identify the occurrence of Toxoplasma gondii and Neospora caninum abortions in goats from Argentina by serological, macroscopical and microscopical examination and bioassay, and to characterize the obtained isolates by molecular techniques. For this purpose, 25 caprine fetal fluids, 18 caprine fetal brains and 10 caprine placentas from 8 dairy/meat goat farms from Argentina were analyzed. Gestational age of the aborted fetuses was determined in 18 cases. Protozoal infections were detected by at least one of the applied diagnostic techniques in 44% (11/25) of examined fetuses; specifically, 24% (6/25) were positive to T. gondii, 8% (2/25) were positive to N. caninum and 12% (3/25) were positive to both parasites. In this study IFAT titers were similarly distributed in younger and older fetuses. Macroscopical and microscopical examination of one placenta revealed chalky nodules in the fetal cotyledons and normal intercotyledonary areas, as well as necrosis and calcification of mesenchymal cells in villi. Tachyzoites were observed in peritoneal wash from 2 mice inoculated with brain and a pool of brain and placenta of two fetuses. Cell culture growth of tachyzoites was achieved from one inoculated mouse, and confirmed as T. gondii by PCR. The T. gondii isolate was identified as atypical or non-canonical by nested-PCR-RFLP. This is the first study that investigated the involvement of N. caninum and T. gondii in cases of goat abortion in Argentina., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Serum samples from 528 aborted ewes from different sheep flocks in Kuwait were tested for Toxoplasma gondii. The indirect hemagglutination test (IHAT) was used with a titer of 1:80 considered positive. Antibodies to T. gondii were found in 94 (17.8%). The highest number of positive ewes (23; 24.5%) had a titer of 1:640. Only one aborted ewe had a titer of 1:2560. This study is the first to report Toxoplasma infection in sheep which may pose risk to humans who consume under cooked mutton. The detection of Toxoplasma antibodies in aborted ewes refers to the implication of toxoplasmosis in abortion among sheep in Kuwait.

A near-full-term alpaca (Vicugna pacos) was stillborn 2 days before the expected date of delivery; necropsy examination was conducted within 6 hr of delivery. Gross lesions were enlarged liver and hydrocephalus. On histologic examination, mild inflammatory lesions were identified in the placenta, liver, and lungs, although no etiology was recognized. Within the brain, there was a mild nonsuppurative meningoencephalitis, hydrocephalus, and hydromyelia. Both kidneys had inflammatory foci in cortex and medulla associated with protozoal tissue cysts. Protozoa in brain and kidneys were identified as Toxoplasma gondii based on immunoreactivity to T. gondii polyclonal antibodies that stain both tachyzoites and tissue cysts and BAG1 T. gondii antibodies that are bradyzoite specific. The tissue cysts exhibited intense positivity to T. gondii and mild immunoreactivity to Neospora caninum antibodies. The dam had a high antibody titer (1 : 12,800) to T. gondii and low titer (1 : 100) to N. caninum using their respective agglutination tests. This is the first report of toxoplasmosis-associated abortion in alpaca.

This work studies the influence of Neospora caninum intra-species diversity on abortion outcome, infection dynamics in terms of parasite dissemination and peripheral-local immune responses in pregnant cattle. Animals were intravenously inoculated at day 70 of pregnancy with 10⁷ tachyzoites of two isolates showing marked differences in virulence in vitro and in pregnant mouse models: Nc-Spain7, a high virulence isolate, and Nc-Spain8, a low-to-moderate virulence isolate. After inoculation, pregnancy was monitored, and dams were culled when foetal death was detected. Foetal mortality occurred in all infected heifers between days 24 and 49 post-infection (pi), however, it was detected sooner in Nc-Spain7-infected animals (median day = 34) than those inoculated with Nc-Spain8 (median day = 41) with a trend towards significance (P < 0.11). Similar histological lesions were observed in placentomes and in most of the foetuses from the two infected groups. However, parasites were more frequently detected in the placenta and foetuses by PCR and in the foetal brain by immunohistochemistry in Nc-Spain7-infected animals. Specific antibodies were detected starting at day 13 post-infection in all infected cattle, with higher IgG levels in Nc-Spain7-infected group. IFN-γ and IL-4 profiles also varied between infected groups in PBMC stimulation assays. Infected animals showed significant increases in their cytokine mRNA levels (IFN-γ, IL-4, IL-10, IL-12p40 and TNF-α) in the caruncle at time of foetal death. Differences between the infected groups were also observed for cytokine profiles. These results demonstrate the influence of the N. caninum isolate on foetal death outcome, infection dynamics and immune responses in cattle.

The efficacy of sulfadimidine (4 doses of 33 mg/kg body weight, IM, q48h) against Toxoplasma abortion was assessed in 3 dairy goat herds suffering from Toxoplasma abortions during the 4th month of gestation. This protocol was very effective for the control of Toxoplasma abortions (P < 0.01).

The present study sought to determine: (i) the effects of Neospora caninum infection and twin pregnancy on plasma pregnancy-associated glycoprotein-2 (PAG-2) concentrations throughout pregnancy and (ii) whether plasma PAG-2 concentrations could predict abortion in N. caninum-infected cows. The study was performed on a commercial Holstein-Friesian dairy herd in northeastern Spain and the final data included those recorded in 53 non-aborting and 19 aborting animals. Blood samples were collected immediately before pregnancy diagnosis (on Days 40, 90, 120, 150, 180 and 210 post-insemination) in non-aborting cows or until the time of abortion detection in aborting cows. General lineal models (GLM) repeated measures anova revealed the different behaviour of PAG-1 and PAG-2, and significant effects of Neospora seropositivity, cool season and twin pregnancy on plasma PAG-2 concentrations throughout gestation (between-subject effects). In addition, based on the odds ratios, the likelihood of abortion increased in Neospora-seropositive cows (by a factor of 7.0) compared to seronegative animals and decreased in cows with a high plasma PAG-2 concentration (>4.5 ng/ml) on Day 120 of pregnancy (by a factor of 0.24), compared to the remaining cows. In conclusion, there is a relationship between plasma PAG-2 concentrations and the risk of abortion in Neospora-infected dairy cows. Thus, plasma PAG concentrations measured using anti-boPAG-2 antiserum on Day 120 of gestation could serve as an indicator of the abortion risk in N. caninum infected animals; values <4.5 ng/ml indicating a high risk of abortion in chronically infected animals., (© 2012 Blackwell Verlag GmbH.)

This work estimates the economic losses due to Neospora abortions in the humid pampa region of Argentina.The total dairy and beef cattle population at risk of abortion is 1,771,326 and 9,726,684 head, respectively. In dairy cattle, there was an 8 % risk of experiencing abortion due to a variety of causes, but 16.5 % of them were due to Neospora caninum. The economic losses were estimated at US$1,415 (1,400-1,431) per abortion, which equates to a total loss of US$33,097,221 (15,622,600-119,349,693) for the dairy industry at the humid pampa region of Argentina. In beef cattle, the overall risk of abortion was estimated to be 4.5 % for all pregnancies,whereas 6.7 % are specifically due to N. caninum, with an economic loss of US$440 (range, 150-730) per abortion.This amounts to an annual loss to the beef industry of US$12,903,440 (range, 1,130,700-42,070,630) in the same area. The results of this study show that Neospora infections and thus abortions cause severe economic impacts in the dairy and beef industries in the humid pampa region of Argentina, which is one the most important areas of cattle production in the world [corrected].

Neospora caninum and Toxoplasma gondii are closely related cyst-forming apicomplexan parasites identified as important causes of reproductive failure in cattle and small ruminants, respectively. Protozoan abortion in small ruminants is traditionally associated with T. gondii, but the importance of N. caninum remains uncertain. The aim of this study was to investigate the presence of N. caninum and T. gondii infections in abortion cases in small ruminants submitted for diagnosis. For this purpose, 74 ovine and 26 caprine aborted foetuses were recovered from different areas in Spain. Foetal histopathology was used to detect the presence of protozoal-associated lesions in brain. The presence of N. caninum and T. gondii was confirmed by PCR. Protozoal infection was detected in 17 out of 100 (17%) foetuses examined by at least one of the diagnostic techniques used. Lesions suggestive of protozoal infection were observed in 10.8% (8/74) and 15.4% (4/26) of the ovine and caprine abortions respectively. N. caninum and T. gondii infection was detected by PCR in 6.8% (5/74) and 5.4% (4/74) of sheep foetuses, respectively, of which five showed protozoal-associated lesions. N. caninum DNA was detected in 11.5% (3/26) of goat foetuses, of which two showed protozoal-associated lesions, whereas T. gondii DNA was detected in one goat foetus with no lesions. The simultaneous presence of N. caninum and T. gondii DNA was detected in one sheep foetus with severe lesions. This study demonstrates that N. caninum plays a significant role in abortion in small ruminants in the studied population. In addition, our results highlight the importance of differentiating between protozoa whenever characteristic lesions are observed., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Two hundred and fifty-five biological samples (106 aborted foetal tissue samples and 149 blood samples from aborted sheep and goats) were collected from 188 animals during the lambing season from September 2009 to April 2010 from the Mafraq region of Jordan. The sampled animals belonged to 93 goat and sheep flocks that had cases of abortion. A total of 169 (66.3%) biological samples were collected from sheep and 86 (33.7%) from goats. Seventy-six (29.8%) biological samples (45 blood and 31 tissue samples) were positive for Toxoplasma gondii by PCR assay. The positive samples were obtained from 43 sheep and 23 goats. The overall toxoplasma-specific prevalence rate was 35.1% (66/188). Forty flocks (43%) had at least one T. gondii PCR-positive animal. The risk factors related to flock health status and farm management that are hypothesized to be associated with T. gondii PCR positivity were also assessed using multiple logistic regressions. The presence of cats (OR = 4.74), a large flock size (OR = 2.76) and the method of disposing the aborted foetuses (OR = 3.77) were all statistically significant (P<0.05) risk factors that were positively associated with toxoplasma positivity in goat and sheep flocks.

Neospora caninum is a major cause of abortion in cattle worldwide. However, little information is available for Algeria. Accordingly, 799 cattle from 87 farms in the north and northeast of Algeria were enrolled in a seroepidemiological survey. An indirect fluorescence antibody test (IFAT) revealed a seroprevalence of 19.6%. The animals were divided into 3 groups according to their breed: imported European cattle, local breeds, and crossed animals (European × local). Seroprevalences were 16.0%, 34.3%, and 18.6% in groups 1, 2, and 3, respectively. A case control study was performed to investigate the link between global seropositivity to N. caninum and abortion risk in those cattle farms. There was a significant (P < 0.01) association between the seroprevalence against N. caninum and the occurrence of abortion in those farms (odds ratio [OR]  =  12.03). This was also observed at the individual level (OR  =  2.79). The analysis of results according to the breed revealed a significant association between seroprevalence and abortion in groups 1 and 3, but not for group 2, despite the fact that the highest seroprevalence was observed in group 2. Cerebral tissues from 5 aborted fetuses were available for histology and polymerase chain reaction (PCR). One sample was found positive both by histology and by PCR, 2 samples were positive by PCR only, and 2 samples were negative in both tests.

Neospora caninum is a major cause of abortion in cattle, but the reasons why only some animals abort remain unclear. The immunological control of the parasite in the placenta or by the foetus could be the key to determining the mechanism of abortion and/or transplacental transmission to the foetus. In this study, cytokine gene expression, analysed by real-time RT-PCR, at the maternal (caruncle) and foetal placenta (cotyledon) of heifers infected at 110 days of gestation by intravenous inoculation of N. caninum tachyzoites was compared with the responses in uninfected heifers. Animals were euthanized 3 weeks after infection. Upregulated Th1, Th2 and T-regulatory (Treg) cytokine gene expression was observed in both the maternal and the foetal placenta in the infected group. In the caruncle of infected animals, the main changes included upregulation of IFN-γ, IL-12p40, IL-6 and IL-10. In the cotyledon, the main changes included upregulation of IFN-γ and downregulation of TGF-β, being the later the only cytokine downregulated in the infected group. The observed cytokine expression pattern was associated with alive but transplacentally infected foetuses, suggesting that such cytokine pattern is beneficial to foetal survival, but could have a role in the transplacental transmission of the parasite., (© 2011 Blackwell Publishing Ltd.)

In December 2008, a southern white rhinoceros (ãsimum simum) aborted a 7-mo gestation male fetus. On hematoxylin and eosin-stained sections of fetal tissues, foci of necrosis were noted in the hepatic parenchyma and were associated with low numbers of lymphocytes, plasma cells, and neutrophils. Protozoal zoites were identified within the hepatic lesions and within the cerebellum. Evaluations utilizing immunohistochemistry, polymerase chain reaction, and DNA sequencing identified the protozoan as Neospora caninum. A microsatellite analysis using MS10 marker showed a unique trinucletoide repeat pattern (ACT), (AGA)19 (TGA)8 distinct from all studied N. caninum to date. This is the first report of N. caninum-related abortion of a rhinoceros fetus of any species and the first report of polymerase chain reaction-confirmed N. caninum infection in any rhinoceros.

Neosporosis is considered to be one of the main causes of abortions in bovines. In this study we evaluated the congenital transmission and occurrence of abortions by Neospora caninum in an embryo transfer center in Neropolis, Goiás. Serological samples from 101 recipients, 61 donors, and 90 calves were analyzed. Among these animals, 32.67% of the recipients, 22.22% of the donors, and 6.66% of the calves were positive for N. caninum. The rate of vertical transmission was 24%. There was a statistically significant difference between antibody titer of the recipient cows and the serological status of the calves. There was no statistically significant relationship between N. caninum serological status and reproductive rates such as estrum repetition and embryo production. Eight abortions occurred, six (75%) in positive females and two (25%) in negative females. These results indicate that neosporosis may be an important cause of failing reproduction and that in embryo transfer centers recipients should be previously examined, and those that are seronegative for Neospora should be chosen to reduce abortion and the birth of seropositive calves., (Copyright © 2010. Published by Elsevier B.V.)

Neospora caninum is a protozoan parasite that causes abortion in cattle around the world. Although the clinical signs of disease in both dogs and cattle have now been recognised for over 20years, treatment and control options are still limited, despite the availability of a commercial vaccine in some countries of the world. The case for an efficacious vaccine has not been convincingly waged by farmers, veterinarians and other members of the agricultural and rural communities. In recent times, however, economic modelling has been used to estimate the industry losses due to Neospora-associated abortion, providing, in turn, the business case for forms of control for this parasite, including the development of vaccines. In this review, we document progress in all areas of the vaccine development pipeline, including live, killed and recombinant forms and the animal models available for vaccine evaluation. In addition, we summarise the main outcomes on the economics of Neospora control and suggest that the current boom in the global dairy industry increases the specific need for a vaccine against N. caninum-associated abortion.

Previous studies have shown that the use of beef bull semen significantly reduces the rate of abortions due to Neospora caninum in artificially inseminated (AI) seropositive dairy cows. In addition, certain beef breeds could be more resistant to N. caninum infection and abortion than others. The aim of the present study was to determine whether different crossbreed pregnancies, those derived from Limousin, Charolais, Piedmontese or Belgian Blue semen, carry different risks of abortion in Neospora-infected dairy cows. The effects of possible interactions between maternal levels of N. caninum antibodies and the different breed crosses were also evaluated. The study was performed on five commercial Holstein-Friesian dairy herds in Northeast Spain with previously confirmed diagnoses of N. caninum infection in aborted foetuses. The study population was comprised of 1115 pregnancies: 633 pregnancies recorded after AI using Holstein-Friesian semen from 18 bulls and 482 after AI using beef semen from 27 bulls (304 inseminations using semen from Limousin bulls, 191 from Belgian Blue bulls, 89 from Piedmontese bulls and 49 from Charolais bulls). Abortion rates were 32.2% (155/482) and 15.2% (96/633) for seropositive cows inseminated with Holstein-Friesian and beef breed semen, respectively. Logistic regression analysis revealed the herd and the interaction between maternal N. caninum antibody titre and the different crossbreeds as significant factors affecting the abortion rate. Lowest abortion rates, similar to that shown by seronegative animals in the analysed herds (3.2%, 239/7432), were observed in dams AI using Limousin semen that had low (<30 relative index (RI) units) N. caninum antibody titres (2.1% abortion, 3/145) and these cows were used as reference. Compared to the cows used as reference, cows with low N. caninum antibody titres (<30 RI units) showed a similar risk of abortion when inseminated with Piedmontese or Charolais bull semen, but higher risk of abortion when inseminated with Holstein (17.9 times) or Belgian Blue (7.2 times) bull semen. All cows with high N. caninum antibody titres (> or =30 RI units) had a higher risk of abortion, ranging from 8.9 times (cows inseminated with Limousine semen) to 37.8 times (cows inseminated with Piedmontese semen), compared to the cows used as reference. In conclusion, different crossbreed pregnancies carried different abortion risks in Neospora-infected dairy cows. The use of beef bull semen dramatically reduced the risk of abortion in dairy cows, especially if Limousin breed semen was used. Moreover, this reduction was found to be dependent on the N. caninum antibody titre such that the lowest incidence of abortions was recorded in Limousin semen inseminated cows with low antibody titres. Insemination of Neospora-seropositive cows with beef bull semen could both reduce the risk of abortion and avoid breeding replacements for infected cattle.

Gamma interferon (IFN-gamma) production has been attributed a protective role against abortion in Neospora-infected dairy cows. This study investigate possible relationships between plasma levels of the N. caninum-specific immunoglobulin (Ig) isotypes IgG1 and IgG2 and IFN-gamma production throughout gestation in dairy cows naturally infected with the parasite. Data were obtained from 31 pregnant Neospora-seropositive animals. Blood samples were collected on gestation days 40, 90, 120, 150, 180 and 210 or until abortion. Ten pregnancies ended in abortion. One of the 11 cows producing IFN-gamma suffered abortion (9.1% of the cows producing IFN-gamma). Non-aborting cows producing IFN-gamma (n=10) showed a significantly higher IgG2/IgG1 antibody ratio throughout the gestation period than cows not producing IFN-gamma (n=11). A significant negative effect of IFN-gamma production on IgG1 antibodies was also observed, while IFN-gamma production did not affect IgG2 antibody levels. In contrast, higher levels of IgG2 antibodies compared to IgG1 antibodies were observed over the whole of gestation in aborting animals, both in those not producing IFN-gamma and in the single aborting cow that produced the cytokine. Our findings indicate that a Th1 immune response, in which IgG2 antibodies prevail, could be protective against N. caninum abortion, but only in the presence of IFN-gamma production. On their own, elevated IgG2 antibody titres appear to be insufficient to protect dams chronically infected with N. caninum against abortion.

Neospora caninum, a protozoan parasite, has been considered as one of the most important etiological agents responsible for abortion in dairy cattle throughout the world since it was first identified in dogs in 1988. In this report, characteristics of neosporosis, detected in a dairy cow ranch having epidemic abortions as high as 18.4%, were described. Blood samples were collected from 25 infertile or aborted dairy cattle, 6 calves born in 2006 and 40 heifers that were born in 2005 and raised in the same ranch. Necropsy was conducted in a 20-day-old Simmental calf that exhibited neurological signs including incoordination, head shaking, hyperextension in forelimbs and hindlimbs and tremor. The seroprevalance in aborted or infertile dairy cattle, heifers, and calves was 60%, 40%, and 33.3%, respectively. The mothers of seropositive two calves including clinically affected calf and its dam were N. caninum seropositive. In immunoperoxidase examinations, N. caninum antigen immunopositivity was observed in the degenerative and necrotic neurons in the brain, cerebellum as well as neurons in dorsal root ganglia of the cervical and thoracic regions of the spinal cord. In the heart, myocytes and Purkinje cells exhibited granular and linear patterns of immunoreactivity. Striated myofibers around the eyeball also showed immunolocalization for N. caninum antigen. Ultrastructurally, tachyzoites with typical apical complex, rhoptries and double-layered parasitic membrane were detected in the brain and heart sections. In conclusion, this report described clinical neosporosis for the first time in Turkey with tissue localization of the causative agents. This scientific communication also discusses the possible impact of cattle neosporosis by clinical, serologic and pathologic evidences collected from the survey of calves born in two successive generations in a ranch.

Neosporosis is considered to be a contributing risk factor for abortions in dairy cows and other farm animals and has negative economic impact on their breeding. In respect of the rapid spread of neosporosis in herds throughout the world, our aim was to detect the prevalence of anti-Neospora antibodies in cows post-abortion (PA) (PA Group n=716) and in cows without any reproduction problems (Control Group n=247) on large breeding farms from Eastern Slovakia. The overall mean seropositivity in PA Group (20.1%) was significantly higher (p<0.0001) in comparison with the Control Group (2.3%), and this demonstrates the causal dependency of abortions on neosporosis. On farms from south-east region, 15.6% of cows were seropositive on average ranging from 7.8% to 25.8% in different districts. In the northern region, significantly (p=0.0002) higher 26.2% mean seropositivity was detected. The seroprevalence in different districts ranged from 2.9% to 39.4%. In the PA Group a high seropositivity to infectious bovine rhinotracheitis (41.5%) and sporadic occurrence of Q-fever, leptospirosis and chlamydiosis was also detected. From 11 Toxoplasma-seropositive cows, only one animal was simultaneously positive to Neospora. The relatively high prevalence rate in cows warrants the attention and a need of surveillance in Slovak herds. A better knowledge of epidemiology of this etiological agent and the mechanisms of its transmission may help in the introduction of more effective preventive and control measures.

Several bacterial species have been implicated in canine and feline pregnancy loss. Brucella canis is one of the more important bacterial infectious agents that cause pregnancy loss in the bitch. Brucella has been documented in the queen but in general infectious abortion from bacteria and protozoal agents is uncommon in the species. Protozoal causes of pregnancy loss in the bitch and queen are less common than in other species. Etiology, clinical signs, diagnosis and treatment of bacterial and protozoal causes of pregnancy loss in the bitch and queen are reviewed. Veterinary practitioners should be aware that many of these organisms have zoonotic potential.

In the Philippines, insufficient consideration has been given to the implementation of systematic control measures against major abortifacient infectious agents in livestock. To elucidate the epidemiology of abortifacient infectious agents in livestock, the prevalence of four abortifacient agents was assessed. Initially, a total of 96 cattle including 17 cows with history of abortion were examined in a herd in Luzon at the request of the farm owner. Six (35.3%) of the 17 aborting cows were found to be serologically positive for Neospora caninum (N. caninum), whereas the seroprevalence in non-aborting cows was 15.9% (10/63). Four of the 6 serologically positive aborting cows were also RT-PCR-positive for bovine viral diarrhea virus (BVDV). Two (12.5%) of the 16 bulls examined were also found to be infected with BVDV, suggesting a putative risk factor of transmission via semen. Based on sequence analysis, the isolates detected belong to BVDV type 1b group. Furthermore, an epidemiological survey of abortifacient infectious agents was conducted with various species of livestock from herds located in Luzon. Out of the 105 water buffalo samples collected, 4 (3.8%) were indicated positive to N. caninum, 2 (1.9%) to Toxoplasma gondii (T. gondii) and 2 (1.9%) to Trypanosoma evansi (T. evansi). The overall seroprevalence of N. caninum in goat and sheep were 23.6% (21/89) and 26.3% (10/38), respectively. BVDV was not detected in these herds. The findings of this exploratory study indicate a relationship between infection and bovine abortion and that a lager study is required to statistically confirm this relationship.

A study carried out on a sheep farm examined whether Toxoplasma gondii foetal infection and associated abortion occur in successive lambings. We identified 29 ewes that gave birth to lambs on at least 2 successive years over our study period, 2000-2003. Tissue samples from the progeny of these ewes were analysed by PCR to determine infection status with T. gondii. T. gondii-infected lambs were born in 31% of successive pregnancies. T. gondii-positive lambs were aborted in successive pregnancies in 21% of lambings during study period, 2000-2003. The frequency of successive abortions within this flock over the period 1992-2003 was 18%. If a lamb was congenitally infected there was a high risk (69%) that the successive lamb from that ewe would also be congenitally infected. Similarly, if a lamb was aborted there was a high risk (55%) of abortion in the next lamb produced. These data suggest that life-long immunity to T. gondii infections may not always be acquired following an initial infection and raises the question as to whether the mechanisms of T. gondii transmission prior to and during ovine pregnancies are fully understood.

A 1-year-old, 5- to 6-week pregnant cat was presented with a history of aborting 3 kittens the previous night. During the examination, the cat began to seizure and died. At necropsy, formalin-fixed tissues and blood smears prepared from an EDTA blood sample collected via cardiac puncture were submitted to a diagnostic laboratory. Examination of the blood smears revealed numerous large (15-75 microm) clumped macrophages containing dark blue intracytoplasmic organisms consistent with Cytauxzoon merozoites. Scattered erythrophagocytic macrophages were also observed. Within several erythrocytes, 1-2 small, round, ring-like piroplasms consistent with Cytauxzoon were observed. Histologic examination revealed numerous large, schizont-laden macrophages filling the blood vessels of multiple organs. The cytologic and histologic findings were diagnostic for Cytaxuzoon felis infection. This case was of particular interest because of the unusually large number of organism-laden macrophages in the cardiac blood sample, an uncommon finding in peripheral blood. Although the cat was presented for complications of abortion, it remains uncertain whether C felis organisms crossed the placenta and infected the fetuses or lead to the abortions.

Progesterone has been reported to reduce Th1 responses during pregnancy. Thus, given that an excessive Th1 response could lead to abortion in Neospora-infected dairy cows, we hypothesized a reduction in abortion risk following progesterone supplementation during the mid-gestation period in these animals. Seropositive animals were assigned on day 120 of gestation to a Control (n=33) or Treatment (n=34) group. Treatment consisted of progesterone supplementation by fitting a progesterone releasing intravaginal device containing 1.55g of progesterone for 28 days. Mean Neospora caninum antibody titres tested by ELISA on days 40, 90 and 120 of gestation were used to classify the cows as showing high (> or =30 units) or low (<30 units) titration. Abortion rate showed a significant increase (P<0.01) in the Treatment group (35%) compared to the Control (9%). Using logistic regression procedures, there was a significant (P<0.0001) interaction between progesterone treatment and antibody titration. Progesterone-treated cows with a high antibody titre showed a likelihood of abortion 14.3 times higher than those not treated, while in cows with low antibody titres, exogenous progesterone had not effect. Our results do not support our starting hypothesis and suggest that progesterone supplementation in cows with high N. caninum antibody titres increases the risk of abortion possibly by affecting cell-mediated immune responses.

In a retrospective study, covering the period from 1986 to 1995, tissues of aborted fetuses were re-examined. A total of 347 cases were tested immunohistochemically, among them samples of 223 brains were examined for Neospora caninum, Toxoplasma gondii and Bovine Virus Diarrhoea Virus (BVDV), and 249 placentae for Chlamydiaceae. Two real-time PCR assays, one for N. caninum, and one for T. gondii, were developed. These potential abortion-inducing agents were detected - and confirmed by PCR, except for BVDV - in 16.1% (N. caninum), 0% (T. gondii), 9.9% (BVDV) and 0.8% (Chlamydiales) of the cases examined. Immunohistochemistry proved to be inadequate for the detection of the protozoal epitopes, whereas it was confirmed as a very useful tool for the detection of BVDV. In abortion material, PCR is considered to be more suitable for the detection of protozoa and Chlamydophila abortus, an adequate sampling presupposed.

A nested PCR that successfully detected Neospora caninum DNA in serum of cattle was used for investigation of selected abortion cases and in a study of healthy pregnant cows at an abattoir. N. caninum DNA was not detected in serum from antibody positive dams that aborted due to N. caninum, but was present in serum of some antibody negative dams that aborted due to other causes. N. caninum DNA was also found in the serum of about half of the animals that aborted of undetermined cause, but was not detected in cow sera from two beef cattle herds in Western Australia with no recent history of abortion. In the abattoir study of 79 dams and their foetuses N. caninum DNA was found in serum of 3 dams and in material from 11 foetuses. The majority of the cows and all foetuses were antibody negative. Our findings suggest that there is no obvious relationship between the presence or absence of N. caninum DNA in serum and the presence of antibodies to N. caninum in dams, the presence of N. caninum DNA in foetuses or abortion due to N. caninum. This is the first report of the detection of N. caninum DNA in serum of cattle rather than the white blood cell fraction. It indicates the presence of free tachyzoites and/or parasite DNA in circulation. The results suggest that persistent infection in the absence of antibodies is a possible outcome of N. caninum infection. Infection of foetuses in the absence of antibodies supports the possibility of persistent infection due to immunotolerance to an early in utero infection. It is therefore important to test for N. caninum DNA as well as antibodies for the detection of exposed and/or infected animals. However, the presence or absence of N. caninum antibodies or DNA did not support nor exclude N. caninum as the cause of abortion. Additional criteria are required for a positive diagnosis of abortion caused by N. caninum.

Neospora caninum-associated abortion was diagnosed in a 7-mo gestational age beef cow fetus from Andorra. The fetus had a multifocal necrotizing encephalitis and nonpurulent multifocal myocarditis. The diagnosis was confirmed by demonstration of N. caninum DNA by polymerase chain reaction and tachyzoites by specific staining with N. caninum polyclonal antibodies in the fetal brain. The dam of the aborted fetus had serum N. caninum antibodies at the time of abortion but not 2 mo before abortion took place. This is the first report of N. caninum abortion in Andorra and the first confirmed N. caninum abortion in an acutely infected cow.

It has been suggested that the abortion herd pattern could influence bovine foetal neosporosis. Here, a comparison of (i) Neospora caninum DNA-detectability by PCR, (ii) N. caninum-associated lesions and (iii) parasite loads in target organs was made between epidemic and endemic abortion cases. We observed that N. caninum DNA was predominantly detected in more than one organ in the foetuses from herds with epizootic rather than endemic abortion cases (P<0.05, Fisher F-test). The highest parasite burdens were found in the heart in foetuses from outbreaks of epidemic abortion and in the brain in endemic cases (P<0.05, Kruskal-Wallis H-test). Moreover, foetuses from epidemic outbreaks had significantly higher parasite burdens in heart (P<0.05, Mann-Whitney U-test) than endemic abortion cases. Epidemic abortion cases showed higher lesion frequencies in liver (P<0.05, Fisher F-test). This report confirms that the abortion herd pattern is an important factor that influences pathogenesis in natural N. caninum infections.

Recent work on Neospora caninum, a protozoan parasite that causes abortions in dairy cattle has focused on a number of different control options. Modelling has suggested the most effective options for control but the present paper argues that the most effective option might not necessarily be optimal from an economic point of view. Decision trees, using published quantitative data, were constructed to choose between four different control strategies. The costs of these interventions, such as 'test and cull', therapeutic treatment with a pharmaceutical, vaccination or "doing nothing" were compared, and modelled, in the first instance, on the New Zealand and Australian dairy situation. It is argued however, that the relative costs in other countries might be similar and that only the availability of a registered vaccine will change the decision tree outcomes, as does the within-herd prevalence of N. caninum infection. To "do nothing" emerged as the optimal economic choice for N. caninum infections/abortions up to a within-herd prevalence of 18%, when viewed over a 1-year horizon, or 21% when costs were calculated over a 5 years horizon. For a higher (>or=21%) within-herd prevalence of N. caninum infection vaccination provided the best (i.e. most economic) strategy. Despite being the most efficacious solutions, 'test and cull' or therapeutic treatment never provided a viable economic alternative to vaccination or "doing nothing". Decision tree analysis thus provided clear outcomes in terms of economically optimal strategies. The same approach is likely to be applicable to other countries and the beef industry, with only minor changes expected in the relationships of decisions versus within-herd prevalence of N. caninum infection.

This article presents the clinical and laboratorial findings in an outbreak of abortions and high neonatal mortality attributable to Trypanosoma evansi infection in camels. A total of 16 females were diagnosed, 2 of which showed moderate signs of chronic form, particularly hyporexia and intolerance to exercise. The main laboratorial findings were regenerative anemia (hemolytic anemia), lymphocytic and monocytic leukocytosis, hyperproteinemia, hyperglobulinemia, hypoglycemia, serum urea increased, and serum iron decreased. The most characteristic finding in the examined females would be the uremia, probably due to the higher protein metabolism.

We used a prospective seroepidemiological approach to investigate endemic abortion in a dairy herd in Arequipa, Peru, and its association with Neospora caninum and bovine viral-diarrhoea virus (BVDV) infections. Between January 2002 and March 2004, 1094 pregnancies were confirmed in 538 cows. Of these, 137 pregnancies (13%) in 121 cows ended in abortion. The serological status to N. caninum was assessed using a single serological screening, whereas BVDV status was assessed at the herd level through consecutive samplings of young stock. Cox proportional-hazards models were used to estimate the effect of N. caninum and BVDV on the hazard of early (between day 42 and day 100 in gestation), and late (after day 100) abortions, respectively. Serological status to N. caninum was included as a dichotomous variable, and the effect of BVDV estimated at the herd level, as a time-dependent seasonal effect. Because data from repeated pregnancies were included, we considered possible lack of independence between observations and included frailty effects into the models. Our models also considered the possible confounding by parity and animal origin. Only multiparity was associated with the hazard of early abortion (HR=2.8 compared to nulliparous heifers). N. caninum seropositivity significantly affected the hazard of late abortion, but interacted with parity. The HRs for Neospora-positive animals were 6.4, 3.7 and 1.9, respectively, for nulliparous heifers, first-lactation cows and multiparous cows. Evidence of BVDV circulating (or not) among the young stock was not associated with abortions, but few cows in this herd were susceptible to incident infection.

Reproductive losses in camelids are due to infertility, pregnancy loss, udder diseases and neonatal mortality caused by a variety of infectious diseases. Uterine infection and abortion represent the major complaint in camelid veterinary practice. The major infectious organisms in endometritis and metritis are E. coli and Streptococcus equi subspecies zooepidemicus. Abortion rates due to infectious diseases vary from 10% to more than 70% in some areas. Leptospirosis, toxoplasmosis and chlamydiosis have been diagnosed as the major causes of abortion in llamas and alpacas. In camels, brucellosis and trypanosomiasis represent the major causes of infectious abortion in the Middle East and Africa. Mastitis is rare in South American camelids. The prevalence of subclinical udder infection in camels can reach very high proportions in dairy camels. Udder infections are primarily due to Streptococcus agalactiae and Staphylococcus aureus. Neonatal mortality is primarily due to diarrhea following failure of passive transfer and exposure to E. coli, rotavirus, coronavirus, Coccidia and Salmonella. This paper reviews the etio-pathogenesis of these causes of reproductive losses, as well as the major risk factors and strategies to prevent their occurrence.

Our understanding of the epidemiology of bovine neosporosis is advancing rapidly with considerable research activity being facilitated by improving methods. The dynamics of the infection in the known definitive hosts, the dog and the coyote, are being described. Improved procedures for production of oocysts enables the horizontal transmission to intermediate hosts and the subsequent more natural infection process to be studied. Details of the sylvatic cycles, potentially involving other animals in the dairy environment, are also emerging.

No significant association existed between Neospora caninum titer and serostatus to Leptospira serovar hardjo, icterohaemorrhagiae, or pomona in cattle on 78 dairy herds in Ontario. Leptospira titer increased with parity. Amongst herds not vaccinated against Leptospira, the proportions of herds with > or = 1 animal seropositive to serovar hardjo, icterohaemorrhagiae, or pomona were 45%, 42%, and 58%, respectively.

We conducted a longitudinal study to follow-up the anti-Neospora caninum serologic status in 30 initially seropositive and 83 initially seronegative cows during their pregnancy. Study cows were blood-sampled every other month during pregnancy until parturition. Blood serum samples were screened for anti-N. caninum antibodies by ELISA. Cows that seroconverted were re-tested by immunoblot as a confirmation test. Among 30 seropositive cows, 28 cows remained seropositive during the whole pregnancy, whereas 2 cows transiently tested negative at least once during pregnancy. Among 83 seronegative cows, 82 cows remained seronegative and 1 cow tested positive three times during the sixth, eighth and last month of pregnancy. As only 2 out of 30 seropositive animals and 1 out of 83 animals changed their serologic status during pregnancy, the study results indicate that there is only a minor temporal instability of anti-N. caninum antibody reactivity in adult cattle.

The aim of this study was to determine by nested PCR probe the presence of Neospora caninum in fetal brain tissue from aborted dairy fetuses in Aguascalientes, a state in the central part of Mexico. Forty-four fetal brains from eight dairy farms were analyzed using single tube nested PCR probe with primers NF1, NS2, NR1 and SR1, with histopathology as the reference technique. From the 44 bovine fetuses studied, 35 (80%) were diagnosed as being infected by N. caninum using PCR probe and 20 (45%) were considered positive by histopathology, because they had lesions suggestive of bovine neosporosis. Fair agreement was observed (31%) between both probes. The age of aborting cows ranged from 1.6 to 2.9-year-old in 17 (49%) of the positive cases diagnosed by PCR. Gestational ages of aborted fetuses ranged from <3 to 9 months, with average of 5.6 months, and no statistical significant difference was observed when ages of infected and no infected fetuses were compared (P > 0.05). The frequency of N. caninum positive cases observed could be considered to be higher than other areas of Mexico and other countries.

In the present work we have studied in Neospra caninum aborted bovine foetuses the influence of foetal age (first, second and third gestational periods) on parasite distribution by nested PCR, parasite loads by real-time PCR and N. caninum associated lesions. For this purpose, a total of 220 aborted foetuses were analysed and detection of N. caninum infection was accomplished by nested-PCR in brain, heart and liver, detecting the presence of the parasite in 72 (32.7%) bovine foetuses. When the different age classes were compared, parasite DNA-detectability in heart and liver was reduced over time of gestation (P < 0.05, Fisher F-test). N. caninum distribution, parasite loads and lesions were studied on 34 out of 72 N. caninum-infected foetuses selected according to the stage of pregnancy and organs recovered. A higher number of positive-PCR tissue samples were observed in the foetuses corresponding to the first and second pregnancy periods. In the last trimester, the parasite could only be detected in the brain and, sporadically, in the diaphragm, heart and lymph nodes. The parasite loads decreased during pregnancy and the foetuses from the first period had higher parasite burdens in brain, heart, kidney and lung (P < 0.05, Kruskal-Wallis H-test) than in those corresponding to the other two trimesters of pregnancy. In addition, the observed lesions were more severe in foetuses from the first and second pregnancy periods than those from the third period (P > 0.05, Kruskal-Wallis H-test). Our results confirm the influence of N. caninum foetal age on pathogenesis in natural N. caninum infections.

A seroepidemiology study of Neospora caninum was conducted in Mashhad area, Khorasan province, Iran on 337 cattle from 30 dairy using ELISA. All herds had seropositive cattle and antibodies were formed in 46% of the cattle. Abortion was significantly associated with seropositivity of cattle. The highest risk of abortion was observed in the 1-2-year-old cows. Our results indicate that Neospora infection is widespread in Iran.

A 1.5-year-old Coonhound from Maryland aborted 7 fetuses. Placenta and internal tissues of 1 fetus were examined histologically. The predominant lesion was placentitis characterized by necrosis and infiltration of mixed leukocytes. Numerous Leishmania spp amastigotes were identified in placental trophoblasts, and the diagnosis was confirmed by use of immunohistochemical staining with Leishmania-specific antibodies. Protozoa were not found in the fetal tissues. An indirect fluorescent antibody test yielded a serum titer of 1:100, and a recombinant K39 immunoassay of serum yielded positive results for the K39 Leishmania antigen.

An immunochromatographic test (ICT) with recombinant surface antigen 1 of Neospora caninum (NcSAG1) was developed for the rapid detection of antibodies to N. caninum in cattle. The ICT was used to clearly discriminate between immunofluorescent-antibody test (IFAT)-positive bovine sera and IFAT-negative bovine sera. Serum samples collected from cattle in Yanbian, China, were examined by the ICT. Of the 96 serum samples, 23 (24.0%) were positive by the ICT, and 19 (19.8%) samples were positive by a previously developed enzyme-linked immunosorbent assay (ELISA). Eighteen of 19 ELISA-positive samples were positive according to the ICT. A good agreement was found between the results of the ICT and the ELISA. The results presented here suggest that the ICT with recombinant truncated NcSAG1 fused to glutathione S-transferase is a useful and reliable method for the detection of antibodies to N. caninum in cattle.