Histopathological and Serological Analysis of Aborted Ewes and Neonatal Death with Toxoplasma gondii in Duhok City, Kurdistan-Iraq.

This study was carried out on seven flocks of ewes suffered from late abortion and neonatal mortality with the prevalence rate of infection reported as 13.95%. The blood and tissue samples were collected from the aborted ewes in several flocks of Duhok province, Kurdistan Region, Iraq. Serological analysis indicated that all the aborted ewes were confirmed positive for agglutination to Toxoplasma gondii (T. gondii)antibody. The investigation of the aborted fetuses showed the blood-stained fluid in the thoracic and abdominal cavity. Most of the aborted fetuses had also enlarged, congested, and friable livers and lungs. The placenta was swollen, reddish, and friable, and its cotyledons also spotted with whitish foci. T. gondii tachyzoites were also demonstrated in the placental sections of some aborted ewes. Severe congestion, necrosis, and infiltration of multinucleated cells were the most predominant histopathological changes of the aborted fetuses, as well as presented tissue cysts, tachyzoites, and bradyzoites in the liver, brain, heart, and lung. There were also several clusters of dark purple banana-shaped T. gondii tachyzoites within the brain and heart tissues in most of the examined aborted fetuses in different flocks. T. gondii tachyzoites were also detected from the peritoneal ascites of mice inoculated experimentally 12 days following the infection. Moreover, T. gondii tissue cysts were detected from the impression smears of the mice brains 32 days after the infection. Accordingly, the demonstration of T. gondii in Giemsa-stained impression smears associated with characteristic histopathological changes of different organs is a great fundamental method for the diagnosis of T. gondii in aborted cases.

Macrophages and T Lymphocytes in the Ovine Placenta After Experimental Infection With Toxoplasma gondii.

Early abortion in ovine toxoplasmosis has had limited investigation. This study evaluated the immune response in the placenta of sheep orally infected with Toxoplasma gondii and euthanized between 2 and 4 weeks postinfection. Toxoplasma infection of the placenta was only found at 4 weeks after infection. Parasitic debris in foci of necrosis were immunolabeled in the maternal caruncle, whereas well-preserved intracellular parasitic vacuole-like structures were found in trophoblasts of fetal cotyledon. Early abortions had increased macrophages in caruncular septa, whereas in later abortions the placentas containing the parasite had an increase of T lymphocytes and macrophages mainly in the fetal cotyledons. This study suggests that the immune response in both the fetal and maternal compartments of the placenta may contribute to the pathogenesis of ovine toxoplasmosis and that these responses differ between early and late presentations of the disease.

Genetic characterization of Neospora caninum from Northern Italian cattle reveals high diversity in European N. caninum populations.

Recent studies have revealed extensive genetic variations among Neospora caninum, a cyst-forming protozoan parasite that is one of the main causes of bovine abortion in the cattle industry worldwide. Previous genetic studies based on multilocus microsatellite genotyping (MLGs) of different Ibero-American populations showed a high genetic diversity. These studies provided clear clues of a predominant clonal propagation in cattle and population sub-structuring partially associated with geographical origin. Although, these reports were limited to a reduced number of countries. In this study, the N. caninum isolates from aborted bovine fetuses and stillbirths and a goat abortion from Northern Italy were investigated genetically using 9 microsatellite markers. Complete or nearly complete isolate profiles were obtained from 30 fetuses and stillbirths. An extensive genetic diversity was also found in this Italian N. caninum population. The study of genetic relationships among Italian MLGs using network (eBURST) and principal component analyses based on the allele-sharing coefficient (PCoA) showed different clonal subpopulations disseminated throughout Northern Italy without apparent segregation depending on the geographic origin, cattle breed, or time of collection. The presence of linkage disequilibrium supports a predominant clonal propagation of Italian N. caninum. In addition, most of Italian MLGs segregated from other global populations including Spain, Argentina, Mexico, Brazil, Germany, and Scotland, suggesting the existence of specific N. caninum subpopulations in the Northern Italy and different subpopulations of N. caninum circulating in Europe.

Evaluation of abortions spontaneously induced by Neospora caninum and risk factors in dairy cattle from Lima, Peru / Avaliação de abortos induzidos espontaneamente por Neospora caninum e fatores de risco em bovinos leiteiros de Lima, Peru

Abstract Our objective was to identify the direct and indirect presence of Neospora caninum in dairy cattle and their aborted fetuses from Lima, Peru. A total 219 blood samples obtained from dairy cattle with records of spontaneous abortion were collected to detect antibodies against N. caninum in serum with indirect ELISA and search for risk-factor associations. 68 fetal aborted tissue samples of these cows were analyzed by PCR, indirect ELISA and histopathology assay to detect N. caninum presence. The prevalence ratio (PR) and 95% confidence intervals (CI) were estimated. Univariate analysis was performed using the chi-squared test. Among the 68 aborted fetuses collected, 10 (15%) were positive in at least two diagnostic tests. Among 219 serum samples, 46.6% (95% CI: 40.0%-53.3%) were positive. Cows with 4 years or older (PR: 7.10; 95% CI: 4.89-10.67) and multiparous (PR: 1.76; 95% CI: 1.11-2.80) were found to be more likely to possess N. caninum antibodies. This study detects presence of N. caninum in dairy cattle and their aborted fetus from Lima valley, suggesting biosecurity management improve to neosporosis control.

Resumo O objetivo do trabalho foi diagnosticar a presença direta e indireta de Neospora caninum em fetos abortados e de soros de bovinos leiteiros de Lima, Peru. Um total de 219 amostras de sangue obtido de vacas leiteiras, com registros de abortos espontâneos, foi coletado para detectar anticorpos contra N. caninum no soro (pelo ELISA indireto), e para estudar associações com fatores de risco. Foram examinadas 68 amostras de tecido fetal abortado das vacas soropositivas pela PCR, ELISA indireto e histopatologia para determinar a presença de N. caninum. A taxa de prevalência (PR) e o intervalo de confiança (CI) de 95% foram estimados. Análise univariada foi realizada usando o teste de Qui-quadrado. Entre os 68 fetos abortados, 10 (15%) foram positivos em pelo menos dois testes diagnósticos. Entre as 219 amostras de soro, 46,6% (95% IC: 40,0%-53,3%) foram positivas. Vacas com 4 anos ou mais de idade (RP: 7,10; 95% IC: 4,89-10,67) e multíparas (RP: 1,76; 95% IC: 1,11-2,80) apresentaram maior frequência de anticorpos contra N. caninum. Este estudo demonstrou a presença de N. caninum em bovinos leiteiros e em fetos abortados oriundos do vale de Lima, sugerindo a implementação de medidas de biossegurança, para o controle da neosporose.

Abortion in association with transplacental Theileria equi infection in a mare from the State of Espírito Santo, southeast Brazil: case report / Aborto equino associado à infecção transplacentária de Theileria equi, Espírito Santo, Brasil: relato de caso

Equine piroplasmosis is a tick-borne disease caused by the protozoan parasites Babesia caballi and Theileria equi. We report a case of abortion in association with transplacental Theileria equi infection in a mare from the State of Espírito Santo, southeast Brazil. An apparently healthy mare aborted at the tenth month of gestation. At necropsy, the subcutaneous tissue, skeletal muscles, and visible mucosae of the aborted fetus were pale, and there was moderate hydrothorax and marked splenomegaly. Microscopic findings included splenic lymphoid hyperplasia and nephrosis. Merozoite-infected erythrocytes were found within blood vessels of all organs examined and were most numerous in the brain. DNA extracted from the spleen, liver, kidney, and thymus was used as a template for PCR. Generic primers were employed for the detection of piroplasm 18S ribosomal gene. All samples were positive for piroplasm DNA by PCR. Amplicons were purified and then sequenced. Sequencing analysis of these amplicons revealed 98% identity to T. equi sequences. Based on our findings, we suggest that abortion in this case resulted from transplacental Theileria infection.(AU)

A piroplasmose equina é transmitida por carrapatos, sendo causada pelos protozoários Babesia caballi e Theileria equi. O objetivo deste trabalho foi relatar um caso de aborto equino associado à piroplasmose fetal. Uma égua gestante, aparentemente saudável, abortou no 10º mês de gestação. Na necropsia do feto, foi observada palidez de subcutâneo, da musculatura esquelética e das mucosas visíveis, moderado hidrotórax e acentuada esplenomegalia. Histologicamente, havia hiperplasia linfoide esplênica e nefrose. Muitos merozoítos foram observados em eritrócitos nos vasos sanguíneos de todos os órgãos examinados, com maior intensidade no encéfalo. Amostras de DNA do baço, fígado, rim e timo foram utilizadas para PCR com primers genéricos para detecção de sequências do gene 18S ribossomal de piroplasmas, com resultado positivo para todas as amostras. Em seguida, os amplicons das amostras de baço foram purificados, sequenciados e, após análise, mostraram 98% de identidade com a sequência de T. equi. Portanto, as alterações microscópicas, os resultados de PCR e sequenciamento confirmam aborto associado à T. equi por transmissão transplacentária.(AU)

First case report of Toxoplasma gondii-induced abortions and stillbirths in sheep in Argentina.

The aim of this study is to report an episode of reproductive losses due to toxoplasmosis in a sheep flock in Argentina. A total of 15 abortions and 9 stillbirths were recorded in a flock of 190 Texel ewes. The affected ewes were more likely to be seropositive for Toxoplasma gondii (15/24) than ewes that delivered normal lambs (5/34, OR=9.6, 95%CI=2.7-34.0, p=0.0004). A pair of aborted twins was recovered for diagnostic investigation. One of these fetuses and its dam were seropositive for T. gondii. Histological examination of the two fetuses revealed non-suppurative myocarditis and epicarditis, portal hepatitis and multifocal necrotizing encephalitis with protozoal cysts in the brain. T. gondii was detected intralesionally by immunohistochemistry in one fetus and by PCR in both. Further investigations are necessary to evaluate the economic losses due to T. gondii in the Argentinean ovine industry.

Occurrence of abortions induced by Neospora caninum in dairy cattle from Santa Catarina, southern Brazil / Ocorrência de abortamentos induzidos por Neospora caninum em vacas de leite de Santa Catarina, sul do Brasil

Abstract The aim of the present study was to investigate the occurrence of N. caninum associated with abortions of dairy cattle from Santa Catarina state, southern Brazil by using enzyme-linked immunosorbent assay (ELISA), immunohistochemistry (IHC), and polymerase chain reaction (PCR). Blood from dairy cows that aborted along with intrathoracic fluid and tissue samples (brain, heart, liver, and lung) from their fetuses were collected and used for serology; PCR, histopathological, and immunohistochemistry (IHC) evaluations were also conducted. Twenty-one cows (51.2%) out of 41, and eight fetuses (26.7%) out of 30 were ELISA (HerdCheck, IDEXX) positive for N. caninum. Dams > 36 months of age had a higher risk of being serum positive than younger animals. PCR and IHC revealed that 38.8% (14/36) and 25.0% (9/36) of the fetuses were positive for N. caninum, respectively for each of the tests. Seropositive cows had a higher frequency of fetuses that were also positive by either intrathoracic fluid, PCR, or IHC. In summary, the present study observed a high frequency of N. caninum in abortions from dairy cows from southern Brazil, with a higher N. caninum prevalence found in cows that were older than 36 months. In addition, serology, PCR, and IHC should be used all together for better diagnosis of neosporosis in cattle.

Resumo O objetivo deste estudo foi avaliar a ocorrência de N. caninum associado a abortamentos em vacas de leite do estado de Santa Catarina, sul do Brasil pelo uso das técnicas de ELISA (HerdCheck, IDEXX), reação em cadeia pela polimerase (PCR) e imunohistoquímica (IHC). O sangue das vacas leiteiras que abortaram, bem como, o líquido intratorácico e amostras de tecidos (cérebro, coração, fígado e pulmão) de seus fetos foram coletados e usados para sorologia, PCR (Np21+ e Np6+), e IHC. Vinte e uma vacas (51,2%) de um total de 41, bem como, oito fetos (26,7%) de um total de 30 foram positivos no ELISA (IDEXX) para N. caninum. As vacas > 36 meses de idade tiveram um maior risco de serem soropositivas do que os animais mais jovens. PCR e IHC revelaram que 38,8% e 25,0% dos fetos foram positivos para N. caninum, respectivamente para cada um dos testes. As vacas soropositivas tiveram uma maior frequência de fetos que também foram positivos no fluído intratorácico, na PCR ou na IHC. Em resumo, o presente estudo observou uma alta frequência de N. caninum em abortos de vacas leiteiras na região estudada, com maior prevalência de N. caninum em vacas com mais de 36 meses de idade. Além disso, sorologia, PCR e IHC deveriam ser utilizadas ​​conjuntamente para melhor diagnóstico de neosporose em bovinos.

Genetic characterization of Neospora caninum from aborted bovine foetuses in Aguascalientes, Mexico.

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.

Toxoplasmosis-associated abortion in an alpaca (Vicugna pacos) fetus.

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.

Study on outbreak of Neospora caninum-associated abortion in dairy cows in Tabriz (Northwest Iran) by serological, molecular and histopathologic methods.

OBJECTIVE: To determine Neospora caninum (N. caninum) as a cause of bovine abortion in dairy cows by ELISA, PCR and Pathological methods in Tabriz, Northwest of Iran. METHODS: For study of outbreak of neosporosis, blood samples were collected from 76 Holstein aborted dairy cows in Tabriz (Northwest Iran). Antibodies to N. caninum were assayed by using a commercially ELISA kit (IDEXX, USA). IgG against N. caninum were found in 14 (18.4%) cases. Aborted fetuses of these seropositive dams were proposed for histopathological and molecular investigations. Brains, spinal cords and placentas of the fetuses were fixed in 10% buffered formalin for histopathology. Also 5-10 g of brain tissue was sampled for DNA extraction. In 6 out of 14 (42.8%) fetuses, brain tissue was positive in PCR. All dams of these fetuses were serologically seropositive in ELISA test. RESULTS: Histopathologically, the lesions consistent with N. caninum were observed in brains, spinal cords and placenta of all fetuses. Lesions in CNS included severe congestion, perivascular and perineuronal edema, status spongiosis, perivascular cuffing, focal gliosis, nourophagy and focal necrosis. There were some Neospora-like cysts in brain. In placentas, severe congestion, perivascular infiltration of mononuclear cells, vascular thrombosis, focal placentitis and necrotic foci in cotyledons were noticed. CONCLUSIONS: The results of present study agreed the results of similar studies about serological, histopathological and molecular results of other studies about neosporosis and it seems to support the outbreak of N. caninum-associated abortion in dairy cows in Tabriz (Northwest Iran).

Combination of monoclonal antibodies improves immunohistochemical diagnosis of Neospora caninum.

Histological analysis is commonly used for a conclusive diagnosis of neosporosis. Immunohistochemistry (IHC) using monoclonal (mAb) and polyclonal (pAb) antibodies can improve diagnosis; however, the use of pAb may induce cross-reactivity with other related parasites. The aims of this study were to compare the performance of mAbs and their combinations with that of pAb in IHC and evaluate the usefulness of mAb to identify Neospora caninum infection in aborted bovine fetal tissues. For this purpose, mAbs targeting NcSRS2 (4.15.15) or NcGRA7 (4.11.5 and 1/24-12) and one pAb collected from a rabbit inoculated with N. caninum tachyzoites were tested by IHC. Artificial standardized tissue sections were prepared as positive controls using homogenized bovine brain spiked with cultured tachyzoites of N. caninum. The numbers of labeled parasites were counted in each positive control section. In addition, four equal proportional combinations of the mAbs were also analyzed in the IHC. Finally, the pAb and the best combination of mAbs obtained in the positive control experiments were tested with tissue sections of naturally-infected cattle. To confirm analytical specificity, mAbs and a pAb were tested with Toxoplasma gondii and Besnoitia besnoiti positive control slides and tissues sections from naturally infected cattle containing Sarcocystis spp. and B. besnoiti antigens. The mAb 4.15.15 detected 57% of the total parasites in sections while 4.11.5 and 1/24-12 were able to detect 49% and 41%, respectively. For the mAb combinations (I: 1/24-12+4.11.5, II: 1/24-12+4.15.15, III: 4.15.15+4.11.5, IV: 1/24-12+4.11.5+4.15.15), the detection capacity was 32.4%, 79.4%, 66.6% and 60.7% for each combination, respectively. The best mAb combination (1/24-12 and 4.15.15) and the pAb serum detected 100% (18/18) of naturally-infected animals. Sarcocystis spp. or B. besnoiti were not detected by mAb combinations in IHC, however the pAb cross-reacted with Sarcocystis spp. cysts. These results confirm the usefulness of mAb application in IHC to N. caninum.

Toxoplasma gondii abortion storm in sheep on a Texas farm and isolation of mouse virulent atypical genotype T. gondii from an aborted lamb from a chronically infected ewe.

Sheep are commonly infected with the protozoan parasite, Toxoplasma gondii. Infection may cause early embryonic death and resorption, fetal death and mummification, abortion, stillbirth, and neonatal death. Most sheep acquire T. gondii infection after birth. Recent studies reported that congenital ovine transmission of T. gondii may be more common than previously believed, but these findings are solely based on PCR data and require confirmation using other techniques to verify the findings. In the present study, during the lambing season of 2005 a toxoplasmosis abortion storm occurred in a flock of purebred Suffolk ewes on a farm in Texas. Only 14 healthy lambs were born, and 38 abortuses, mummies and weak or stillborn lambs were delivered. Another 15 fetuses identified by ultrasound were presumably resorbed or were aborted undetected. Antibodies to T. gondii were found in 37 (94.8%) of the 39 ewes and 30 of them had high titers (1:3200 or higher) when tested in the modified agglutination test (MAT). In the 2006 lambing season, two (both with MAT titers of ≥ 3200 in 2005) of 26 ewes delivered T. gondii infected lambs. T. gondii tissue cysts were found histologically in lesions of encephalitis in a lamb from one ewe and viable T. gondii (designated TgShUs55) was isolated from the brain and heart of a lamb from the second ewe. TheTgShUs55 had an atypical genotype using 10 PCR-RFLP markers, and was 100% lethal for Swiss Webster mice, irrespective of the dose or the stage of the parasite inoculated. In subsequent seasons, the ewes lambed normally. The results of the present study support the hypothesis that most sheep that have aborted due to T. gondii develop protection against future toxoplasmosis induced abortion, but the protection is not absolute.

Factors associated with seroprevalence of Neospora caninum in dairy cattle in southeastern Brazil.

Neosporosis is an infectious disease caused by Neospora caninum, an obligate intracellular cyst-forming protozoan considered a major cause of miscarriage in dairy cattle in many parts of the world. This cross-sectional study evaluated the relationship between reproductive abnormalities and seropositivity to N. caninum in 1,204 dairy cows from 40 farms located in the state of Minas Gerais, southeastern Brazil. Producers were interviewed, and blood samples were collected to perform indirect immunofluorescence tests (IFAT 1:200). Association between reproductive abnormalities and seropositivity in cattle was evaluated with generalized estimating equations. The true herd-level seroprevalence of N. caninum was 95 % (83.3-99.1), while the individual-level true seroprevalence was 21.6 % (19.2-24.2). Several reproductive abnormalities were significantly associated with seropositivity to N. caninum: occurrence of repeated estrus (p=0.02; OR=3.84; 95 % CI=1.239-11.893), repeated miscarriages (p=0.001; OR=2.54; 95 % CI=1.423-5.402), and temporary anestrus (p=0.001; OR=3.44; 95 % CI=1.976-5.994). Furthermore, loose dogs (p=0.041; OR=2.20; 95 % CI=1.033-4.672) when fed raw meat (p=0.001; OR=1.91; 95 % CI=1.443-2.519) are risk factors for N. caninum infection. We observed that seropositivity to N. caninum in cattle increases risk of miscarriage by almost twice throughout the reproductive life of cows (p=0.004; OR=1.978; 95 % CI=1.249-3.131). Serologic evidence in this study indicates that N. caninum infection is widely distributed among dairy herds and significantly associated with reproductive disorders, especially miscarriage, repeated estrus, and temporary anestrus.

Absence of link between abortion and seropositivity of cystic hydatid disease in ewes and female goats in Turkey.

This study was conducted to test whether there is a statistically significant association between seropositivity to cystic echinococcosis and abortion in ewes and female goats from the Turkish provinces of Elazig (east Anatolia), Sanliurfa (south-east Anatolia) and Kayseri (Inner Anatolia) using the enzyme-linked immunosorbent assay (ELISA). A total of 20 of 133 sera (15.1%) from ewes and 5 of 101 sera (4.9%) from goats with a history of abortion gave seropositive results that were not significantly different (p>0.05) from these, 9.9% (10/101) were reported for ewes and 1.6% (2/122) for female goats without a history of abortion. Serological prevalence rates among those animals with a history of abortion were not significantly different from the control group. No positive association was established between seropositivity for cystic echinococcosis and abortion in ewes and female goats.

[Advances in the development of vaccines for bovine neosporosis]. / Avances en el desarrollo de vacunas contra la neosporosis bovina.

Neosporosis, a disease caused by the obligate intracellular protozoan Neospora caninum, produces abortions in cattle. The severe economic losses in cattle industry justify the need to develop control measures for preventing bovine abortion. Apicomplexan parasitic resistance is associated with T helper 1 immune response mediated by CD4 cytotoxic T lymphocytes, the production of interferon-gamma, interleukin-12, tumor necrosis factor and immunoglobulin G2. The reduction of vertical transmission in subsequent pregnancies and the low levels of abortion repetition suggests the existence of protective immune mechanisms. Inoculation with live tachyzoites before mating protects against infection and abortion. Antecedents of the development of live vaccines against other protozoa stimulate research to develop a live vaccine against N. caninum. On the other hand, an inactivated vaccine with low efficacy against neosporosis is useful in the prevention of abortion in farms with epizootic disease. A neosporosis vaccine should avoid abortion, transplacental transmission and infection persistence. In the present work, advances in vaccine development including lysate of tachyzoites, live parasites, recombinant antigens and vaccine vectors are reviewed.

Neosporose bovina: avaliação da transmissão vertical e fração atribuível de aborto em uma população de bovinos no Estado do Rio Grande do Sul / Bovine neosporosis: evaluation of vertical transmission and population etiological fraction of abortion in a bovine population in the State of Rio Grande do Sul, Brazil

O protozoário Nespora caninum é um parasito que causa grandes perdas reprodutivas e econômicas em bovinos no mundo inteiro. Os objetivos deste estudo foram verificar tanto a associação entre o histórico de aborto e a presença de anticorpos contra N. caninum, quanto a transmissão vertical como forma de manutenção da infecção nos rebanhos bovinos em regiões do Rio Grande do Sul, através da sorologia pareada de mães e filhas. Foi realizada amostragem de 60 propriedades distribuídas em duas regiões do Rio Grande do Sul, das quais foi coletado sangue de 40% dos animais presentes para a detecção de anticorpos anti-N. caninum por imunofluorescência indireta (IFI). Para verificar a relação aborto e soropositividade foi utilizado o teste de regressão logística univariada, e para sorologia de mães e filhas o teste de qui-quadrado de McNemar para dados pareados. Foram confrontados os dados de sorologia e aborto, sendo encontrada a frequência de 58,5% (24/41) de soropositivos quando havia histórico de aborto, e 16,4% (199/1215) dentre os sem histórico de aborto. Os animais soropositivos apresentaram um risco 7,21 (IC 95%, 3,65-14,32) vezes maior de possuir histórico de abortamento (estatística de Wald χ2=44,93, P<0,001). A fração atribuível à neosporose como causa de aborto na população em risco nas duas regiões foi estimada em 9,73% (λpop). O resultado sorológico de cada mãe foi pareado com o de sua filha e, pelo teste de qui-quadrado de McNemar (χ2=59,84, P<0,001), houve associação significativa entre as sorologias de mães e filhas, sugerindo transmissão vertical. Ressalta-se ainda a importância do acompanhamento sorológico para N. caninum, evitando assim manutenção de animais portadores que sirvam como reservatório do protozoário nas propriedades.

The protozoa Nesporora caninum is a parasite that causes great economic and reproductive losses in cattle worldwide. The objective of this study was to verify the association between abortion and the presence of antibodies against N. caninum and the vertical transmission as a means of maintaining the infection in cattle herds in Rio Grande do Sul by matching the serology of mothers and daughters. Sampling was performed in 60 dairy farms of two regions of the state, where blood was collected from 40% of the herds for the detection of anti-N. caninum by indirect immunofluorescence (IFI). To verify the association between abortion and seropositivity we used the univariate logistic regression test, and for the serology of mothers and daughters the chi-square McNemar test for paired data. In comparing serology and abortion data, a prevalence of 58.5% (24/41) among cattle with history of abortion, and 16.4% (199/1215) prevalence between those with no history of abortion was found; seropositive animals were 7.21 times more likely (95% CI, 3.65-14.32) to have previously aborted (Wald statistic χ 2=44.93, P<0.001). The fraction affected by neosporosis in the population studied was estimated as 9.73% (λpop). The serologic result of each mother was paired with her daughter and showed by the McNemar chi-square (χ2=59.84, P<0.001) significant association between the serology of mothers and daughters, suggesting vertical transmission. It is worth to note the importance of serological monitoring for N. caninu to avoid maintenance of animals that may serve as carries of the parasite reservoir on the farms.

Occurrence of Neospora caninum and Toxoplasma gondii infections in ovine and caprine abortions.

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.

Implications of wild dog ecology on the sylvatic and domestic life cycle of Neospora caninum in Australia.

Neospora caninum is transmitted either transplacentally or horizontally by ingestion of tissue cysts present in tissues or oocysts shed by dogs. Neosporosis is a significant disease, causing cattle abortion at 5-7 months of pregnancy. Infected cows may remain infective for life transmitting the infection in several consecutive or non-consecutive pregnancies. A great deal is known about the epidemiology of neosporosis, although only limited information is available on the main routes of horizontal transmission. In Australia, the presence of the dingo as the top-order predator suggests a potential sylvatic route of transmission between dingoes and as yet unknown native wildlife in addition to the domestic route via dogs with access to infected tissue on farms. This review article critically evaluates the overlap between the domestic and sylvatic routes, taking into account canine ecology, and summarises current understanding of the transmission of N. caninum to provide a foundation for epidemiologists, farmers and conservation biologists dealing with neosporosis and wild dog control programs.

Association of BoLA DRB3 and DQA1 alleles with susceptibility to Neospora caninum and reproductive outcome in Quebec Holstein cattle.

The BoLA DRB3 and DQA1 genes are part of the major histocompatibility complex (MHC) class II in cattle. These genes are highly polymorphic and have been associated with resistance to several diseases, such as mastitis, Bovine Leukemia Virus (BLV) and dermatophilis. Sequenced based typing of these genes has been carried out extensively from blood samples; however it is often impractical or expensive to obtain such samples. Repositories of well-characterized serum from cattle are readily available in many veterinary research facilities. This paper reports a retrospective analysis of BoLA class II genotypes of cattle obtained from stored serum samples from Holstein cattle from Québec dairy farms, which were obtained as part of a previous study on bovine neosporosis. It was possible to genotype 56 cattle with known infection status for Neospora caninum. We identified 14 different DRB3 and 10 different DQA1 alleles in this population. The allele frequency distribution was consistent with previously studied cattle populations, and alleles known to be associated with BLV and mastitis were present. No association was found between allele frequency distribution of DRB3 or DQA genes and infection with N. caninum. However, an association of allele DRB3*1001 and allele DRB3*2703 with resistance and susceptibility to pregnancy loss, irrespective of infection status, was identified.

[Ovine abortion associated with toxoplasmosis: serological, anatomo-pathological and immunohistochemistry characterization]. / Aborto em ovinos associado à toxoplasmose: caracterização sorológica, anátomo-patológica e imunoistoquímica.

Ovine toxoplasmosis is a major cause of abortion and placentitis. Lambs with congenital infection are born with encephalitis. Abortions occurred between May and August 2006 in an estate in Três Palmeiras, southern Brazil. This study aimed to assess the frequency of abortion associated with toxoplasmosis among ewes in southern Brazil using indirect hemagglutination and pathoanatomical and immunohistochemical analyses. Blood samples were collected from nine animals for indirect hemagglutination. One fetus underwent necropsy and histopathological and immunohistochemical analyses. Three samples, one of which belonged to the necropsied animals mother, were positive on the serological test. The necropsy revealed pale foci in the liver, lungs with a marbled appearance, heart with severe pallor and brain and cerebellar congestion. Fragments of all organs were collected, fixed in 10% buffered formalin, processed, and stained with hematoxylin-eosin. Microscopically, there were cysts and tachyzoites in the brain in areas of malacia with microgliosis and lymphoplasmacytic infiltration, suggesting toxoplasmic encephalitis. There was also interstitial pneumonia, centrolobular necrosis with structures compatible with tachyzoites, focal lymphocytic myocarditis and acute tubular nephrosis. The immunohistochemical test was positive for Toxoplasma gondii. The results allowed diagnosing the ovine toxoplasmosis.