Experimental Neospora caninum Infection in Pregnant Cattle: Different Outcomes Between Inoculation With Tachyzoites and Oocysts.

Neospora caninum is a globally distributed abortifacient protozoan of cattle. Experimental infections with N. caninum in cattle have provided valuable information on host-parasite interaction and immunopathogenesis. Experimental infection of pregnant cows has been reported in about 20 articles, with most studies using cultured parasite tachyzoites as the inoculum. Only three experimental studies have been conducted in pregnant cows using the parasite's oocysts which are shed by dogs, in large part because transmission experiments using oocysts take more time and are more complex and expensive than experiments using tachyzoites. In this minireview, we discuss differences between N. caninum tachyzoites and oocysts as inocula for experimental infection of pregnant cows, as well as the route animals are inoculated.

Congenital Neurological Disease Associated With HoBi-like Pestivirus Infection in a Newborn Dairy Calf From Brazil.

HoBi-like pestivirus (HoBiPeV) has been reported in several biological samples from cattle worldwide, but there are no descriptions of this virus associated with neurological symptoms. This report described the first occurrence of neurological disease associated with HoBiPeV in a newborn dairy calf. A mixed-breed Holstein calf had severe neurological symptoms at birth and died at 21 days old. The tissue fragments (central nervous system (CNS), myocardium, liver, kidney, lung, intestine, and spleen) were submitted to reverse transcription (RT)-PCR assay for the partial 5'-untranslated region (5'UTR) and N-terminal autoprotease (Npro) gene of the pestivirus genome, and the CNS tissue fragments were submitted to histopathological and immunohistochemical evaluation. The RT-PCR assay indicated that the kidney, CNS, and intestinal tissue fragments were positive for the pestivirus 5'UTR, and the CNS and intestinal tissue fragments were positive for the pestivirus Npro gene. Amplicons with high DNA quantification in the 5'UTR (CNS-cerebral cortex) and Npro (CNS-cerebral cortex and intestine) RT-PCR assays were sequenced. The nucleotide (nt) sequence and phylogenetic analysis of the 5'UTR strain exhibited 93.6 to 99.4%, 85%, 89.4 to 89.9%, 85.1%, and 90.5 to 91.5% nt identity with HoBiPeV strains from clades a, b, c, d, and e, respectively. The Npro amplicons showed 99.7% nt identity to each other and 90.4 to 96.5%, 85.1 to 85.3%, 79.2 to 79.7%, and 85.8 to 86.5% nt identity with HoBiPeV strains from clades a, c, d, and e, respectively. A histopathology revealed neuronal necrosis at the cerebrum, cerebellum, and brain stem. An immunohistochemical assay designed to identify antigens of bovine viral diarrhea virus revealed positive intracytoplasmic immunoreactivity within neurons at the cerebral cortex, cerebrum, cerebellum, and spinal cord. Thus, this report provides information about the first identification of HoBiPeV in tissues of the CNS in a newborn dairy calf with neurological symptoms.

Saccharomyces boulardii reduces the vertical transmission of Toxocara canis larvae in mice.

Probiotics have been shown to reduce the intensity of Toxocara canis infection in mice. However, larval transmission of this nematode also occurs via transplacental and transmammary routes. Thus, the aim of this study was to evaluate the effect of the Saccharomyces boulardii probiotic on the vertical transmission of T. canis in Swiss mice. The mice received 107S. boulardii colony-forming units per gram of food. The supplementation began 15 days before mating and was maintained throughout pregnancy and lactation. The animals were inoculated with 300 T. canis embryonated eggs on the 14th day of pregnancy. The presence of larvae was examined in the organs of the females and their offspring. The examined organs included the following: brain, liver, lungs, heart, kidneys, spleen, eye, skeletal muscle (carcass) and mammary glands of lactating females. There was a 42% (P = 0.041) reduction in the number of larvae transmitted to offspring in the group that received probiotic-supplemented food (GI). Additionally, there was a 50% reduction (P = 0.023) in the number of larvae found in the brains of lactating offspring in the GI group. These results reveal the potential of S. boulardii probiotic use as an auxiliary method of controlling visceral toxocariasis.

COVID-19 y embarazo: caso clínico de presentación crítica, inflamación placentaria y transmisión vertical fetal demostrada / COVID-19 and pregnancy: clinical case of critical presentation, placental inflammation and demonstrated fetal vertical transmission

ANTECEDENTES: La pandemia global de COVID-19 llega al continente americano en marzo del año 2020 y en menos de dos meses reúne a más de la mitad de los casos a nivel global. OBJETIVO: Caso clínico de una mujer embarazada con una presentación crítica de COVID-19 y embarazo a las 25 semanas de gestación, en el contexto del peak de la pandemia en Chile en el otoño del año 2020. CASO CLÍNICO: El 20 de junio de 2020, una mujer de 34 años, con 25 semanas de embarazo, es trasladada desde Hospital de San Bernardo a Clínica Las Condes en Santiago, Chile, con un cuadro de 10 días de evolución de COVID-19, que evoluciona a una situación crítica con insuficiencia respiratoria severa. Ingresa a unidad de cuidados intensivos para ventilación mecánica. Las imágenes de radiología simple y de tomografía axial computarizada de tórax demuestran una neumopatía bilateral con imágenes características opacidades en vidrio esmerilado, asociado a engrosamiento intersticial, imágenes descritas previamente como características para COVID-19. La paciente permanece en unidad de cuidados intensivos en ventilación mecánica por siete días, con evolución favorable posterior, mejoría del cuadro séptico y alta después de 22 días de hospitalización. El parto ocurre en forma espontánea a las 38 semanas, la madre y el recién nacido evolucionan en buen estado general. El examen histopatológico placentario demuestra compromiso inflamatorio vellositario y los exámenes de anticuerpos en sangre del recién nacido demuestran la presencia de anticuerpos del tipo IgG e IgM. Se trata de uno de los pocos casos demostrados reportados de transmisión transplacentaria vía sanguínea de SARS-CoV-2 de la madre al recién nacido.

BACKGROUND: The global COVID-19 pandemic reaches the American continent in March 2020 and in less than two months it brings together more than half of the cases globally.OBJECTIVE: The clinical case of a 25-week pregnant woman with a critical presentation of COVID-19 and pregnancy at 25 weeks of gestation, is presented in the context of the peak of the pandemic in Chile in the fall of 2020. CLINICAL CASE: On June 20, 2020, a 34-year-old woman, 25 weeks pregnant, is transferred from Hospital de San Bernardo to Clinica Las Condes in Santiago, Chile, with a ten-day evolution of a COVID-19 that evolves to critical with severe respiratory failure. She is admitted to the intensive care unit for mechanical ventilation. Chest computerized axial tomography images demonstrate bilateral pneumopathy with characteristic images of ground-glass opacities, associated with interstitial thickening, images previously described as characteristics for COVID-19. The patient remains in the intensive care unit on mechanical ventilation for seven days, with subsequent favorable evolution, improvement of the septic condition, and discharge after 22 days of hospitalization. Delivery occurs at 38 weeks, the mother and the newborn evolve in good general condition. The placental histopathological examination demonstrates villous inflammatory involvement, and the newborn's blood tests show the presence of IgG and IgM antibodies. It is one of the few reported cases of transplacental transmission of SARS-CoV-2 from the mother to the newborn.

Anti-Sarcocystis Antibodies in Lambs Deprived of Colostrum.

INTRODUCTION: The objective of this study was to evaluate the presence of anti-Sarcocystis spp. specific IgG antibodies in serum samples from precolostral lambs to determine the occurrence of transplacental transmission of Sarcocystis spp. in sheep. METHODS: Blood samples were collected from 80 ewes and their respective lambs, immediately after lambing and before colostrum ingestion, respectively. The presence of anti-Sarcocystis spp. IgG was evaluated in serum samples using the indirect fluorescent antibody test (IFAT). Positive samples of the lambs were submitted to titration and IFAT to detect anti-T. gondii and anti-N. caninum specific IgG. RESULTS: Anti-Sarcocystis spp. IgG was detected in 62.5% of the ewes (50/80) and in 4% of the lambs of the seropositive ewes (2/50). None of the lambs from seronegative ewes were positive. The final titers of the positive lambs were 80. No cross reaction was detected among the positive samples to anti-Sarcocystis spp., anti-N. caninum, and anti-T. gondii IgG. The detection of anti-Sarcocystis spp. antibodies in serum samples of lambs deprived of colostrum suggests transplacental transmission of infection. Thus, the vertical transmission may be an alternative route of infection of Sarcocystis spp. also in sheep. Further studies are warranted to confirm transplacental transmission in sheep and to explain the importance of this infection pathway.

Anti-Neospora caninum IgG in fetal bovine serum / Anti-Neospora caninum IgG em soro fetal bovino

Neospora caninum is an obligate intracellular protozoan, recognized as an important cause of abortion in infection cattle worldwide, and is responsible for major economic losses in beef and dairy herds. There are scarce data on the presence of antibodies in fetal serum samples from beef cattle. The pathogen is straggling mainly by vertical transmission in ruminants. This study aimed to determine the frequency of anti-N. caninum antibodies in fetal bovine serum. Overall, 751 fetal serum samples from slaughterhouses in Uruguaiana and Santo Ângelo (Rio Grande do Sul state) were used. Serologic diagnosis was made using indirect immunofluorescence assay (IFA). Slides were prepared by culturing infected Vero cells using tachyzoites of the N. caninum NC-1 strain and a serum dilution of 1:25. Of the 751 samples analyzed, 11.7% (88/751) were positive in the IFA assay. The results indicate an occurrence of neosporosis in the studied region, as well as the possibility of transplacental transmission of the protozoon, characterizing an important mechanism of propagation for the disease.(AU)

O Neospora caninum é um protozoário intracelular obrigatório, reconhecido como uma causa importante de aborto no mundo todo, e é responsável pelas maiores perdas da economia de carne e leite. Há poucos dados da presença de anticorpos em amostras de soro fetal de bovinos de corte. Os patógenos são transmitidos principalmente por transmissão vertical em ruminantes. Este estudo pode ser usado para determinar a frequência de anti-N. caninum em soro fetal bovino. Ao todo, foram utilizadas 751 amostras de soro fetal bovino de matadouros de Uruguaiana e Santo Ângelo (RS). Os diagnósticos sorológicos foram realizados utilizando ensaios de imunufluorescência indireta (IFA). As lâminas foram preparadas por cultura de células Vero infectadas utilizando taquizoítos da estirpe N. caninum NC-1 e uma diluição de soro de 1:25. Das 751 amostras analisadas, 11,7% (88/751) foram positivas para o IFA. Os resultados indicam uma ocorrência de neosporose na região estudada, bem como a possibilidade de transmissão transplacentária do protozoário, caracterizando um importante mecanismo de propagação da doença.(AU)

Evidence of reduced vertical transmission of Neospora caninum associated with higher IgG1 than IgG2 serum levels and presence of IFN-γ in non-aborting chronically infected cattle under natural condition.

Neospora caninum infection of cattle can be vertically transmitted, resulting in abortion or birth of infected calves. Vertical transmission occurs both in acutely or chronically infected cattle. There is little information on the immune response needed to prevent endogenous transplacental transmission, particularly from chronically infected cattle to their offspring in a natural environment. In this study, N. caninum seropositive pregnant cattle from three different farms with high avidity antibodies and low IgM titers were selected and their newborn colostrum-deprived calves were tested for anti-N. caninum antibodies. Based on these results, dams were grouped according to their congenital transmission status. The analysis of the immune profile of the chronically-infected pregnant cattle revealed that higher ratio between IgG1 and IgG2 anti-N. caninum serum titers and higher levels of systemic IFN-γ were associated with diminished vertical transmission rates, compared to dams with the opposite profile. Our results evidenced an association between the immune profile and vertical transmission in non-aborting chronically infected dams, and confirm that vertical transmission, even when not leading to abortion, is related to a defined immune profile. This is important information to accomplish successful vaccine development efforts.

Congenital Zika syndrome: Pitfalls in the placental barrier.

Much progress with respect to congenital Zika virus (ZIKV) pathogenesis has been achieved after the 2015 outbreak in Brazil. It is now accepted that ZIKV is vertically transmitted, infects cells of the developing central nervous system and the placenta, yet it is unclear to what extent placental affection contributes to the development of congenital ZIKV. The association between fulminant villitis and severe fetal involvement emerges as a possibility. ZIKV is unique among the Flaviviruses in its ability to be sexually transmitted, possibly responsible for its teratogenicity. Furthermore, there is controversy over the participation of antibody dependent enhancement (ADE) in patients with non-neutralizing anti-Flavivirus antibodies, a phenomenon previously recognized in serious DENV infections. Our aim was to analyze information regarding the contribution of the placental barrier as an actual player in neonatal ZIKV. Therefore, we underwent a systematic review with keywords "Zika virus" and "ZIKV". Articles were screened for relevance concerning the topics of microcephaly, transplacental transmission, sexual transmission, and ADE. We identified variables that affect the severity of congenital Zika syndrome: age of gestation at maternal infection, the extent of placental disruption (villitis), sexual transmission, initial viral replication at the uterine wall, anti-DENV antibodies, and the possibility of antibody-mediated transcytosis of ZIKV through the placenta. These questions may not seem relevant when Zika becomes endemic, and we are no longer witness to the extreme clinical sequelae seen when the virus moves through an immunologically naïve population; however, characterizing the pathogenesis of congenital Zika syndrome will continue to further our understanding.

Transplacental transmission of Neospora caninum in naturally infected small ruminants from northeastern Brazil / Transmissão transplacentária de Neospora caninum em pequenos ruminantes infectados naturalmente do Nordeste do Brasil

Toxoplasma gondii and Neospora caninum are causative agents of abortion in sheep and goats. Thus, the present study aimed to describe the transplacental transmission of these protozoans in small ruminants of northeastern Brazil. Seventeen fetuses (6 goats and 11 sheep) from farms with history of abortion were necropsied and samples were collected from different tissues (brain, liver, lung, kidney and heart). The samples were analyzed by PCR, histopathology (HP) and immunohistochemistry (IHC) to evaluate whether T. gondii and/or N. caninum infection were the cause of abortion. None of the samples was positive for T. gondii according to PCR and IHC results. Some brain, liver, lung, kidney and heart samples of goat fetuses were positive for N. caninum by PCR. In the histopathology, mild mononuclear infiltration and necrosis with calcification were observed in the liver and brain of one goat fetus, respectively, that also was positive for N. caninum by PCR and IHC. The results confirmed vertical transmission of N. caninum in naturally infected goats of northeastern, Brazil.(AU)

Toxoplasma gondii e Neospora caninum são reconhecidos como protozoário causadores de aborto em ovinos e caprinos. Desta forma, objetivou-se descrever a transmissão transplacentária desses agentes em pequenos ruminantes na região Nordeste do Brasil. Foram examinados seis fetos caprinos e onze fetos ovinos, totalizando 78 amostras de diferentes tecidos (cérebro, fígado, pulmão, rim e coração) provenientes de propriedades rurais com histórico de aborto. As amostras foram analisadas por Reação em Cadeia da Polimerase (PCR), Histopatologia (HP) e Imunohistoquímica (IHQ), com a finalidade de associar o abortamento a T. gondii e/ou N. caninum. Nenhuma amostra foi positiva na PCR e IHQ para T. gondii. Algumas amostras de cérebro, fígado, pulmão, rim e coração de fetos de caprinos e ovinos foram positivas na PCR para N. caninum. Na histopatologia foi observado leve infiltrado mononuclear no fígado e necrose com calcificação no SNC de um caprino, associada à imunomarcação positiva para N. caninum na IHQ e PCR positiva. Os resultados confirmam a transmissão vertical de N. caninum em caprinos naturalmente infectados na região nordeste do Brasil.(AU)

Transplacental transmission of Neospora caninum in naturally infected small ruminants from northeastern Brazil / Transmissão transplacentária de Neospora caninum em pequenos ruminantes infectados naturalmente do Nordeste do Brasil

Toxoplasma gondii and Neospora caninum are causative agents of abortion in sheep and goats. Thus, the present study aimed to describe the transplacental transmission of these protozoans in small ruminants of northeastern Brazil. Seventeen fetuses (6 goats and 11 sheep) from farms with history of abortion were necropsied and samples were collected from different tissues (brain, liver, lung, kidney and heart). The samples were analyzed by PCR, histopathology (HP) and immunohistochemistry (IHC) to evaluate whether T. gondii and/or N. caninum infection were the cause of abortion. None of the samples was positive for T. gondii according to PCR and IHC results. Some brain, liver, lung, kidney and heart samples of goat fetuses were positive for N. caninum by PCR. In the histopathology, mild mononuclear infiltration and necrosis with calcification were observed in the liver and brain of one goat fetus, respectively, that also was positive for N. caninum by PCR and IHC. The results confirmed vertical transmission of N. caninum in naturally infected goats of northeastern, Brazil.(AU)

Toxoplasma gondii e Neospora caninum são reconhecidos como protozoário causadores de aborto em ovinos e caprinos. Desta forma, objetivou-se descrever a transmissão transplacentária desses agentes em pequenos ruminantes na região Nordeste do Brasil. Foram examinados seis fetos caprinos e onze fetos ovinos, totalizando 78 amostras de diferentes tecidos (cérebro, fígado, pulmão, rim e coração) provenientes de propriedades rurais com histórico de aborto. As amostras foram analisadas por Reação em Cadeia da Polimerase (PCR), Histopatologia (HP) e Imunohistoquímica (IHQ), com a finalidade de associar o abortamento a T. gondii e/ou N. caninum. Nenhuma amostra foi positiva na PCR e IHQ para T. gondii. Algumas amostras de cérebro, fígado, pulmão, rim e coração de fetos de caprinos e ovinos foram positivas na PCR para N. caninum. Na histopatologia foi observado leve infiltrado mononuclear no fígado e necrose com calcificação no SNC de um caprino, associada à imunomarcação positiva para N. caninum na IHQ e PCR positiva. Os resultados confirmam a transmissão vertical de N. caninum em caprinos naturalmente infectados na região nordeste do Brasil.(AU)

Theileria equi infection causing abortion in a mare in Brazil.

The aim of this report was to confirm the cause of abortion to be Theileria equi acquired transplacentally in a mare in Federal District, Brazil. The aborted fetus showed a high parasitemia of erythrocytes (80%) by the oval and Maltese cross forms of T. equi. Necropsy of the fetus demonstrated congestion of organs, jaundice, anemia and pronounced spleno- and hepatomegaly. Twelve breeding mares, including the one that aborted and her fetus, tested positive for T. equi infection by universal and multiplex PCR assay. The absence of histopathologic changes such as inflammation and necrosis in the placenta and fetal tissues as well as the negative microbiological results and negative serology for Leptospira spp. in the mare that aborted were important to exclude other infectious agents that may be involved in equine abortion. This report highlights the transplacental transmission of T. equi infection in horses in endemic areas and reinforces the importance of investigating equine theileriosis as a possible cause of abortion in mares.

Transplacental transmission of Neospora caninum in naturally infected small ruminants from northeastern Brazil

ABSTRACT: Toxoplasma gondii and Neospora caninum are causative agents of abortion in sheep and goats. Thus, the present study aimed to describe the transplacental transmission of these protozoans in small ruminants of northeastern Brazil. Seventeen fetuses (6 goats and 11 sheep) from farms with history of abortion were necropsied and samples were collected from different tissues (brain, liver, lung, kidney and heart). The samples were analyzed by PCR, histopathology (HP) and immunohistochemistry (IHC) to evaluate whether T. gondii and/or N. caninum infection were the cause of abortion. None of the samples was positive for T. gondii according to PCR and IHC results. Some brain, liver, lung, kidney and heart samples of goat fetuses were positive for N. caninum by PCR. In the histopathology, mild mononuclear infiltration and necrosis with calcification were observed in the liver and brain of one goat fetus, respectively, that also was positive for N. caninum by PCR and IHC. The results confirmed vertical transmission of N. caninum in naturally infected goats of northeastern, Brazil.

RESUMO: Toxoplasma gondii e Neospora caninum são reconhecidos como protozoário causadores de aborto em ovinos e caprinos. Desta forma, objetivou-se descrever a transmissão transplacentária desses agentes em pequenos ruminantes na região Nordeste do Brasil. Foram examinados seis fetos caprinos e onze fetos ovinos, totalizando 78 amostras de diferentes tecidos (cérebro, fígado, pulmão, rim e coração) provenientes de propriedades rurais com histórico de aborto. As amostras foram analisadas por Reação em Cadeia da Polimerase (PCR), Histopatologia (HP) e Imunohistoquímica (IHQ), com a finalidade de associar o abortamento a T. gondii e/ou N. caninum. Nenhuma amostra foi positiva na PCR e IHQ para T. gondii. Algumas amostras de cérebro, fígado, pulmão, rim e coração de fetos de caprinos e ovinos foram positivas na PCR para N. caninum. Na histopatologia foi observado leve infiltrado mononuclear no fígado e necrose com calcificação no SNC de um caprino, associada à imunomarcação positiva para N. caninum na IHQ e PCR positiva. Os resultados confirmam a transmissão vertical de N. caninum em caprinos naturalmente infectados na região nordeste do Brasil.

Prospective study investigating transplacental transmission of equine piroplasmosis in thoroughbred foals in Trinidad.

Equine piroplasmosis caused by Theileria equi and Babesia caballi is endemic in Trinidad and Tobago. Transmission occurs by ticks of the family Ixodidae. T. equi can also be transmitted transplacentally; however transplacental transmission of B. caballi is unknown. This study aims to investigate transplacental transmission of equine piroplasmosis from thoroughbred mares naturally infected via the tick vector. Whole blood and serum samples were collected from 117 mares in the fifth month of pregnancy. Blood samples were also collected from each of their foals (89 in total) within the first 36h of birth. Additionally, all foals were observed for clinical signs within 30days post - partum. All samples were examined microscopically for intra-erythrocytic piroplasms. Serum ELISA tests and PCR analysis on whole blood were performed to determine the presence of T. equi and B. caballi. Thirty-four (30.6%) mares and 14 (15.7%) of their foals were seropositive for T. equi. Twenty-seven (24.3%) mares were positive for T. equi by conventional (c) PCR. Real time (q) PCR analysis based on the ema - 1 gene revealed that seven (8%) foals were positive for T. equi. Eighty-nine (76.1%) mares and 38 (42.7%) foals were seropositive for B. caballi. Four (3.4%) mares were positive for B. caballi by cPCR. Three out of the four cPCR positive mares either had resorptions, or stillbirths for that pregnancy. From this study, there is strong evidence that transplacental transmission of B. caballi can occur leading to foetal losses. Six foals (7%) were positive for B. caballi by qPCR. Of these six, four were born to B. caballi seropositive mares. In this study a foal born of a T. equi seropositive mare was 55.7 times more likely to be serologically positive for T. equi than a foal born to a T. equi seronegative mare. Similarly a foal born of a B. caballi seropositive mare was 39.4 times more likely to be serologically positive for B. caballi than a foal born to a mare that was serologically negative for B. caballi at the fifth month of pregnancy. This is as a result of the ingestion of colostrum containing antibodies to these pathogens. Mares should be screened during pregnancy and their foals closely monitored at parturition for evidence of equine piroplasmosis so that treatment can be implemented earlier for a better prognosis.

Zika virus damages the human placental barrier and presents marked fetal neurotropism

An unusually high incidence of microcephaly in newborns has recently been observed in Brazil. There is a temporal association between the increase in cases of microcephaly and the Zika virus (ZIKV) epidemic. Viral RNA has been detected in amniotic fluid samples, placental tissues and newborn and fetal brain tissues. However, much remains to be determined concerning the association between ZIKV infection and fetal malformations. In this study, we provide evidence of the transplacental transmission of ZIKV through the detection of viral proteins and viral RNA in placental tissue samples from expectant mothers infected at different stages of gestation. We observed chronic placentitis (TORCH type) with viral protein detection by immunohistochemistry in Hofbauer cells and some histiocytes in the intervillous spaces. We also demonstrated the neurotropism of the virus via the detection of viral proteins in glial cells and in some endothelial cells and the observation of scattered foci of microcalcifications in the brain tissues. Lesions were mainly located in the white matter. ZIKV RNA was also detected in these tissues by real-time-polymerase chain reaction. We believe that these findings will contribute to the body of knowledge of the mechanisms of ZIKV transmission, interactions between the virus and host cells and viral tropism.

Transplacental transmission of bovine tick-borne pathogens: Frequency, co-infections and fatal neonatal anaplasmosis in a region of enzootic stability in the northeast of Brazil.

Bovine tick-borne disease (TBD) constitutes a worldwide group of diseases that result in great losses for dairy and beef cattle. With regard to the epidemiological profile of the diseases, the importance of transplacental transmission is still not very well understood. The aim of this study was to determine the transplacental transmission of TBD agents (Anaplasma marginale, Babesia bovis and B. bigemina) in a herd of dairy cattle that had been naturally infected in an area of enzootic stability in northeastern Brazil. Blood for serology of the three agents was collected from cows within 120 days of gestation and serology, haemogram and nPCR assays were performed after birth. Blood was collected from the calves within 3h of birth, and haemogram and nPCR assays were performed in all animals. Pre-colostrum serology was achieved in 34 animals. The Student's t-test was used to compare the haemogram results between animals that were positive and negative for the haemoparasites. The cows were seropositive for all agents in at least one of the examinations. We detected 15 cases of vertical transmission of A. marginale, 4 of B. bovis and 2 of B. bigemina in the 60 cows. In infected animals, co-infection was detected for A. marginale and B. bovis in 1 of 60 calves, and a triple infection was detected in one other calf. Fatal neonatal anaplasmosis was observed in 1 of 15 calves, in which death occurred within 24h of birth. From the results, we concluded that transplacental transmission of TBD agents occurs, including in cases of co- and triple-infection. Such transplacental transmission can cause neonatal death, increasing the importance of this form of epidemiological transmission and suggesting its role as a cause of undiagnosed neonatal death.

Transplacental transmission of Anaplasma marginale in beef cattle chronically infected in southern Brazil / Transmissão transplacentária de Anaplasma marginale em bovinos de corte cronicamente infectados no sul do Brasil

In this study, we have investigated the incidence of transplacental transmission of Anaplasma marginale in chronically infected cows with no history of acute anaplasmosis during gestation. In addition, we evaluated various techniques for detection of transplacental transmission of A. marginale. Blood samples were collected from 30 cows at three different periods: at the time of artificial insemination, at gestational diagnosis, and after calving. Also, blood was collected from the newborn calves, including one sample before colostrum intake, and another three days after birth. A. marginale-specific antibodies were detected in 100% of the cows with an indirect fluorescent antibody test (IFAT), and in 97% of them, using an indirect enzyme-linked immunosorbent assay (ELISA). Also, we observed that all of the three-day-old newborn calves were seropositive by IFAT. According to polymerase chain reaction, 63.3% of the cows were carriers of A. marginale, as well as 6.7% of the newborn calves. This represented a transplacental transmission rate of 10.5%. Furthermore, a correlation of 93.3% was observed between the two serodiagnostic techniques, demonstrating that both ELISA and IFAT can be used in epidemiological surveys of A. marginale. These results confirm the occurrence of transplacental transmission of A. marginale in chronically infected cows and suggest the importance of this transmission route in areas of enzootic instability.

O objetivo do estudo foi avaliar a transmissão transplacentária de A. marginale em bovinos de corte cronicamente infectados utilizando diferentes testes de diagnóstico. Amostras de sangue foram colhidas de trinta matrizes durante o período de inseminação artificial, no diagnóstico de gestação e após o parto. Foi colhido sangue dos bezerros antes da ingestão do colostro e três dias após o nascimento. Anticorpos foram detectados em 100% das matrizes pela Reação de Imunofluorescência Indireta (RIFI) e 97% pelo Ensaio de Imunoadsorção Enzimático Indireto (ELISA). Aos três dias de idade 100% dos bezerros estavam soropositivos na RIFI. Conforme os resultados da PCR, 63.3% das matrizes e 6.7% dos bezerros eram portadores do agente, representando taxa de transmissão transplacentária de 10.5%. Correlação de 93.3% foi observada entre os resultados das técnicas sorológicas, demonstrando que tanto ELISA quanto RIFI podem ser utilizadas em levantamentos epidemiológicos do agente. Os resultados confirmaram a transmissão transplacentária do agente em vacas cronicamente infectadas. Isto demonstra a importãncia desta via de transmissão do A. marginale em áreas de instabilidade enzoótica.

Transplacental transmission of Anaplasma marginale in beef cattle chronically infected in southern Brazil

In this study, we have investigated the incidence of transplacental transmission of Anaplasma marginale in chronically infected cows with no history of acute anaplasmosis during gestation. In addition, we evaluated various techniques for detection of transplacental transmission ofA. marginale. Blood samples were collected from 30 cows at three different periods: at the time of artificial insemination, at gestational diagnosis, and after calving. Also, blood was collected from the newborn calves, including one sample before colostrum intake, and another three days after birth. A. marginale-specific antibodies were detected in 100% of the cows with an indirect fluorescent antibody test (IFAT), and in 97% of them, using an indirect enzyme-linked immunosorbent assay (ELISA). Also, we observed that all of the three-day-old newborn calves were seropositive by IFAT. According to polymerase chain reaction, 63.3% of the cows were carriers of A. marginale, as well as 6.7% of the newborn calves. This represented a transplacental transmission rate of 10.5%. Furthermore, a correlation of 93.3% was observed between the two serodiagnostic techniques, demonstrating that both ELISA and IFAT can be used in epidemiological surveys of A. marginale. These results confirm the occurrence of transplacental transmission of A. marginale in chronically infected cows and suggest the importance of this transmission route in areas of enzootic instability.

O objetivo do estudo foi avaliar a transmissão transplacentária deA. marginale em bovinos de corte cronicamente infectados utilizando diferentes testes de diagnóstico. Amostras de sangue foram colhidas de trinta matrizes durante o período de inseminação artificial, no diagnóstico de gestação e após o parto. Foi colhido sangue dos bezerros antes da ingestão do colostro e três dias após o nascimento. Anticorpos foram detectados em 100% das matrizes pela Reação de Imunofluorescência Indireta (RIFI) e 97% pelo Ensaio de Imunoadsorção Enzimático Indireto (ELISA). Aos três dias de idade 100% dos bezerros estavam soropositivos na RIFI. Conforme os resultados da PCR, 63.3% das matrizes e 6.7% dos bezerros eram portadores do agente, representando taxa de transmissão transplacentária de 10.5%. Correlação de 93.3% foi observada entre os resultados das técnicas sorológicas, demonstrando que tanto ELISA quanto RIFI podem ser utilizadas em levantamentos epidemiológicos do agente. Os resultados confirmaram a transmissão transplacentária do agente em vacas cronicamente infectadas. Isto demonstra a importãncia desta via de transmissão do A. marginale em áreas de instabilidade enzoótica.

Experimental infection with Neospora caninum in pregnant bitches / Infecção experimental com Neospora caninum em cadelas prenhes

In this study, transplacental transmission of Neospora caninum in bitches at different stages of pregnancy was evaluated. Three bitches were inoculated in the 3rd week and three in the 6th week of gestation with 10(8) tachyzoites of N. caninum (Nc-1 strain). All the infected bitches and at least one of their offspring presented anti-N. caninum antibodies according to the indirect fluorescent antibody test (IFAT > 400). The pups and their mothers were sacrificed and tissues from the central nervous system (CNS), popliteal lymph nodes, skeletal muscle, brain, lungs, heart and liver were analyzed for the presence of N. caninum using the nested polymerase chain reaction (nested PCR), restriction fragment length polymorphism (RFLP) and immunohistochemistry (IHC). The parasite was found in the pups in lymph node, CNS, heart and liver tissues using nested PCR. There was no difference in perinatal mortality between the offspring from bitches infected in the 3rd week of gestation (60%) and in the 6th week (53.8%).

Neste estudo a transmissão transplacentária de Neospora caninum foi avaliada em fêmeas em diferentes estágios de gestação. Três cadelas foram inoculadas na 3ª semana e três na 6ª semana de gestação com 10(8) taquizoítos de N. caninum (cepa Nc-1). Todas as cadelas infectadas, e pelo menos um de seus filhotes, apresentaram anticorpos anti-N. caninum por imunofluorescência indireta (RIFI > 400). Os filhotes e suas mães foram sacrificados e tecidos de sistema nervoso central (SNC), linfonodo poplíteo, músculo esquelético, cérebro, pulmões, coração e fígado foram analisados para a presença de N. caninum pela reação em cadeia da polimerase (nested PCR), polimorfismo de comprimento de fragmentos de restrição (RFLP) e imunoistoquímica (IHQ). O parasita foi encontrado em filhotes em linfonodo, SNC, coração e fígado pela nested PCR. Mortalidade perinatal não apresentou diferença entre os filhotes das cadelas infectadas na 3ª semana (60%) ou na 6ª semana de gestação (53,8%).