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ABSTRACT: This study aimed to determine the frequency of anti-Neospora caninum antibody detection in three samples and sampling methods: A prevalence study, routine diagnostic laboratory samples, and fetal bovine serum samples. These samples were collected from cattle in Rio Grande do Sul (RS), southern Brazil, and analyzed using the indirect immunofluorescence reaction technique. For each sampling method, a historical study was used as a reference for comparison. In the prevalence study, 1,248 serum samples were collected from 2020 to 2022. The prevalence of N. caninum in the RS state was 22.8% (285/1248). This figure was statistically different compared to previous studies conducted in 2002, which reported a prevalence of 11.2% (p

IntroductionNeospora caninum is an important cause of abortion in cattle worldwide. Infection in cattle occurs horizontally by ingestion of oocysts shed by canids or vertically, from an infected dam to the fetus, and may result in abortion, stillbirth, or birth of seropositive offspring. The control of bovine neosporosis is difficult and costly. The objectives of this study were to estimate the current nationwide seroprevalence of N. caninum infections in Swiss cattle and to assess risk factors for infection with this parasite.MethodsWe conducted a cross-sectional study with cattle farms randomly selected and stratified according to population size, resulting in a sample of 780 female cattle. The cattle originated from 161 farms distributed over all Switzerland. The serum samples were tested for antibodies against N. caninum using a commercial ELISA and if inconclusive, retested using an in-house immunoblot technique. To collect farm parameters relevant to N. caninum transmission and prevention, farm owners were mailed a questionnaire which addressed topics putatively related to N. caninum infection such as husbandry, history of abortion, and presence of dogs on farm. Univariate analysis by generalized linear mixed model (with animal seropositivity as outcome variable) and logistic regression modeling (with farm seropositivity as outcome variable) was conducted on farm parameters investigated in the questionnaire.ResultsBy ELISA and immunoblot, 4.2% (33/780) of cattle sera yielded positive results. At the farm level, 16.2% (26/161) of the sampled farms had at least one seropositive animal. The return rate of the valid questionnaires was 54.0%. At the animal level, odds for farm seropositivity were 3.8 times higher when rodents had been recorded by the farmer as a problem on the farm. At the farm-level, two protective factors were identified: rearing of replacement heifers and feeding of concentrated feed.ConclusionWe recorded a low seroprevalence of N. caninum in a random sample of Swiss cattle representative for the years 2017–2018. Based on a questionnaire survey, we could identify risk and protective factors for infection with N. caninum, however their biological relevance needs to be confirmed in further studies.

Protozoan parasite Neospora caninum causes abortion in infected cattle while others remain asymptomatic. Host immunity plays a critical role in the outcome of bovine neosporosis. Despite extensive research, there is a critical gap in therapeutic and preventive measures, and no effective vaccines are available. Both beef and dairy cattle can suffer from N. caninum -induced abortions, but cumulative evidence suggests a breed susceptibility being higher in dairy compared with beef breeds. It has been established that the response to N. caninum infection primarily involves a cell-mediated immune response (CMIR) regulated by T-helper type 1 (Th1) cells and specific cytokines. The delayed-type hypersensitivity (DTH) skin test has been used to measure the ability of livestock to generate CMIR, in the context of breeding for disease resistance and as a method for diagnosis of several diseases. In this study, we evaluated the immune response triggered by an N. caninum -induced DTH skin test between Holstein – a dairy breed intensively selected- and Argentinean Creole heifers – a beef breed with minimal genetic selection- to assess differences in CMIR following experimental N. caninum infection. The immune response, measured through skinfold thickness and histological and immune molecular analysis, revealed variations between the breeds. Our study found an increased CMIR in Argentinean Creole heifers compared to Holstein heifers. Differential gene expression of key cytokines was observed at the DTH skin test site. Argentinean Creole heifers exhibited elevated IFN-γ, IL-12, IL-10, and IL-4, while Holstein heifers only showed higher expression of IL-17. This finding could underscore genetic diversity in response to neosporosis, which could be used in breeding cattle strategies for disease resistance in cattle populations. • The cellular immune response against N. caninum , differed among the cattle breeds. • N. caninum- sensitized Argentinean Creole heifers exhibited a higher immune response at the skin test site. • The cytokine gene expression at the skin test site revealed distinct expression patterns between different breeds. • Macrophages, T and B cells were labeled by immunohistochemistry at the N. caninum DTH bovine skin test site. [ABSTRACT FROM AUTHOR]

Neospora caninum is one of the main infectious agents that cause reproductive disorders in cattle. However, knowledge about the prevalence and causal factors of bovine neosporosis is needed in order to establish control measures. The purpose of this study was to estimate the pooled prevalence and determine the risk factors of bovine neosporosis using meta-analytic methods. Searches for data on N. caninum seroprevalence and potential risk factors were conducted in the PubMed, LILACS, and SciElo databases. The random effects model was used for the meta-analysis of the included studies. The pooled prevalence of N. caninum in cattle was 24% (95% confidence interval (CI), 19–29) in North and Central America, 24% (95% CI, 20–28) in South America, 18% (95% CI, 14–21) in Asia, 15% (95% CI, 12–18) in Europe, 13% (95% CI, 11–16) in Africa, and 8% (95% CI, 4–14) in Oceania. A significant correlation was found between N. caninum infection and abortion in cows (odds ratio (OR) = 2.66, 95% CI, 1.97–3.59). The following risk factors were signicantly associated with N. caninum seroprevalence: presence of dogs on the farm (OR = 2.84; 95% CI, 1.40–5.80) and the type of cattle production system, with dairy cattle being more susceptible to bovine neosporosis (OR = 1.60; 95% CI, 1.16–2.19) than beef cattle. The adoption of measures to prevent the dissemination of N. caninum in cattle herds is suggested, aiming to reduce the economic losses incurred to the dairy and beef industry. [ABSTRACT FROM AUTHOR]

Neospora caninum is a significant cause of abortion and economic losses in cattle worldwide. The main aim of the present work was to detect the prevalence of N. caninum infection in bulls in Hamedan (Iran) using serology and molecular techniques. All blood samples (n = 792) were screened for detecting the antibodies to N. caninum using enzyme-linked immunosorbent assay (ELISA). Then seropositive animals were rechecked using the immunofluorescent antibody test (IFAT). Also, blood, epididymis, and spinal cord samples were collected from animals for molecular analysis using nested PCR. In serology, using ELISA, 3.91% of animals were seropositive for N. caninum. Additionally, true prevalence based on the sensitivity and specificity of the ELISA was calculated 1.25% (95% CI: 0.48-2.02%). Neospora-infection in animals, calculated as the number of bulls seropositive and/or one sample positive to nested PCR, was 3.40%; and 19 bulls tested positive by both serology and molecular diagnostic methods. The overlaps between ELISA and molecular results were observed in 74.19% of whole blood samples, 80.64% of the epididymis, and 87.09% of the spinal cord. Using ELISA, the seroprevalence of N. caninum was detected 1.8% in ≤2 and 5.45% in >2 years old group of animals (p = 0.009, PR = 3.1). In addition, the seropositivity in Holstein and native breed animals was calculated 6.57% and 2.93%, respectively (p = 0.019, PR = 2.3). Seven sequences with 94.9-99.3% similarity were detected in multiple alignments of positive PCR products. Our work was the first comprehensive evaluation of Neospora-infection/neosporosis in Iranian bulls. We detected a low prevalence of infection in animals compared to previous reports. The ELISA is a sensitive serological technique for detecting the highest number of positive bulls in the present investigation and, the nested PCR is a reliable technique to identify Neospora-DNA., Competing Interests: Declaration of Competing Interest None., (Copyright © 2023 Elsevier B.V. All rights reserved.)

We carried out an inter-laboratory trial to compare the serological tests commonly used for the detection of specific Neospora caninum antibodies in cattle in Ibero-American countries. A total of eight laboratories participated from the following countries: Argentina ( n = 4), Brazil ( n = 1), Peru ( n = 1), Mexico ( n = 1), and Spain ( n = 1). A blind panel of well-characterized cattle sera ( n = 143) and sera representative of the target population ( n = 351) was tested by seven in-house indirect fluorescent antibody tests (IFATs 1-7) and three enzyme-linked immunosorbent assays (ELISAs 1-3; two in-house and one commercial). Diagnostic performance of the serological tests was calculated and compared according to the following criteria: (1) the 'Pre-test information,' which uses previous epidemiological and serological data; (2) the 'Majority of tests,' which classifies a serum as positive or negative according to the results obtained by most tests evaluated. Unexpectedly, six tests showed either sensitivity (Se) or specificity (Sp) values lower than 90%. In contrast, the best tests in terms of Se, Sp, and area under the ROC curve (AUC) values were IFAT 1 and optimized ELISA 1 and ELISA 2. We evaluated a high number of IFATs, which are the most widely used tests in Ibero-America. The significant discordances observed among the tests regardless of the criteria employed hinder control programs and urge the use of a common test or with similar performances to either the optimized IFAT 1 and ELISAs 1 and 2. [ABSTRACT FROM AUTHOR]

Summary Neospora caninum , a tissue cyst-forming parasite, is the causative agent of bovine neosporosis. It is considered to be one of the most important transmissible causes of reproductive failure in cattle; abortion and neonatal mortality result in significant economic losses within the cattle industry worldwide. The balance between acute (mediated by the tachyzoite stage) and persistent (mediated by the bradyzoite stage) phases of the infection is influenced by the immune status of the animal, and for pregnant cows (the intermediate host) immune status is critical for transplacental (i.e. vertical) transmission of the parasite and associated disease outcomes. The horizontal route of transmission from the definitive host, the dog, occurs via ingestion of oocysts containing sporozoites, and plays a minor but important role in transmission of the infection to cattle. Despite the importance of this disease, there is no vaccine or treatment available currently, and at the present the only control measure to reduce the impact of disease is informed management on the farm. The development of vaccines, targeting key biological processes such as invasion and persistent infection, is needed urgently for the control of this widespread parasite. [ABSTRACT FROM AUTHOR]

Objetivou-se, no presente estudo, pesquisar a prevalência de anticorpos anti-Neospora caninum em 812 amostras de soros sangüíneos de bovinos leiteiros procedentes de propriedades rurais de sete municípios das microrregiões de Itapecuru-Mirim, Médio Mearim e Presidente Dutra, estado do Maranhão, Brasil. Para o cálculo do tamanho da amostra, considerou-se um soroprevalência de 34,7% para N. caninum, com erro máximo de 9,5% e intervalo de confiança de 95%. Para a detecção da presença de anticorpos da classe IgG, utilizou-se a técnica de Imunofluorescência Indireta (IFI), com ponto de corte 1:200, usando como antígeno, taquizoítos da cepa NC-1, mantida em cultura celular no Laboratório de Diagnóstico das Parasitoses dos Animais da Escola de Medicina Veterinária da UFBA. Do total de amostras analisadas, encontrou-se uma prevalência de 50,74%. Os títulos variaram de 1:200 a 1:6400, assim distribuídos: 108 (26,21%) amostras de soro apresentaram título de 1:200; 132 (32,04%) 1:400; 94 (22,81%) 1:800; 46 (11,16%) 1:1600; 23 (5,58%) 1:3200 e nove (2,18%) com títulos de 1:6400. Dentre as microrregiões, a Itapecuru-Mirim apresentou o menor percentual de animais soropositivos (20,69%) e Presidente Dutra o maior (47,66%). Com relação à variável sexo, observou-se maior prevalência de sororreagentes nas fêmeas (46,80%) do que nos machos (52,46%). Não se verificou diferença significativa (P>0,05) para as variáveis microrregiões, sexo e idade. Conclui-se que os bovinos leiteiros das regiões estudadas estão expostos à infecção por N. caninum.The objective in the present study was to research the prevalence of anti-Neospora caninum in 812 samples of blood serum of dairy cattle from farms of seven municipalities of microrregions of Itapecuru-Mirim, Middle Mearim and President Dutra, state of Maranhão, Brazil. For the calculation of sample size, it was considered a seroprevalence of 34.7% for N. caninum, with a maximum error of 9.5% and a confidence interval of 95%. To detect antibodies, it was used the technique of Indirect Immunofluorescence (IFI), with the cut-off of 1:200, using as antigen, tachyzoites strain NC-1, maintained in cell culture in the Laboratory of Diagnosis of Parasitism of the Animals, School of Veterinary Medicine of the Federal University of Bahia, Brazil. Of the total samples, it was obtained a prevalence of 50.74%. The titles ranged from 1:200 to 1:6400, distributed as follows: 108 (26.21%) serum samples showed title of 1:200; 132 (32.04%) 1:400; 94 (22.81%) 1:800; 46 (11.16%) of 1:1600; 23 (5.58%) of 1:3200 and nine (2.18%) with titers of 1:6400. Among the microrregiões the Itapecuru-Mirim showed the lowest percentage of animals seropositive (20.69%) and President Dutra the largest (47.66%). It was observed higher prevalence of seropositives in females (46.80%) than in males (52.46%). There was no significant difference (P> 0.05) for the microrregions variables, sex and age. Concluded that the dairy cattle of the regions studied are exposed to infection by N. caninum.

Bovine neosporosis is a globally important disease, causing abortions and significant economic losses. In Brazil, studies on neosporosis in cattle are few and based on limited samples and/or from limited areas. We aimed to determine the herd and animal levels seroprevalence and associated factors for Neospora caninum infections in cattle from the state of Paraíba, Northeastern Brazil, using a planned sampling. Herds (n = 434) and cows aged ≥24 months (n = 1891) were randomly selected, and serum samples were tested with the immunofluorescence antibody test (IFAT) using as cut-off point the antibody titer 200. Herd-level and animal-level seroprevalences were 17.8% (95% CI = 14.3%-21.8%) and 18.1% (95% CI = 14.7%-22.1%), respectively. The factors associated with N. caninum infection were farm located in the Sertão mesoregion (Prevalence ratio [PR] = 2.37), mixed production (PR = 1.64), herd size of 34-111 animals (PR = 3.50) and herd size >111 animals (PR = 6.14). The results indicate high N. caninum circulation in the bovine population of the state of Paraíba, semiarid of Brazil, mainly in the Sertão mesoregion, where the highest apparent herd and animal-level prevalences of positive herds were identified. Control strategies should be adopted to mitigate the impact of disease on cattle production, as well as it's suggested the encouragement of conducting surveys in wildlife from Caatinga biome, mainly canids, to provide information on the importance of these animals on the epidemiology of bovine neosporosis., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Bovine neosporosis is a worldwide concern due to its global distribution and great economic impact. Reproductive failure in cattle due to abortion leads to major economic losses associated with the disease. Currently, there is no treatment or vaccine available against abortion or transmission caused by Neospora caninum infection in cattle. However, vaccination is considered the best measure of control against bovine neosporosis. Several host and parasite factors can influence the dynamics of the infection in bovines. Moreover, the availability of well-defined infection models is a key factor for the evaluation of vaccine candidates. However, working with cattle is not easy due to difficult handling, facilities and costs, and therefore, 'more affordable' models could be used for screening of promising vaccines to establish proof of concept. So far, live-attenuated vaccines have shown good efficacy against exogenous transplacental transmission; however, they have relevant disadvantages and associated risks, which render inactivated or subunit vaccines the best way forward. The identification of novel potential targets and vaccines, and the application of innovative vaccine technologies in harmonized experimental animal models, will accelerate the development of an effective vaccine against bovine neosporosis. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Infection with Neospora caninum is the leading cause of abortion in cattle. In cows naturally infected with N caninum , plasma concentrations of pregnancy-associated glycoproteins (PAG) 1 and 2 indicate fetal-placental well-being, whereas an excess of progesterone in the second trimester of gestation has been related to high abortion rate. The immunosuppressive action of progesterone on the uterus during gestation has been attributed in part to the uterine serpins (SERPINA14). This study examines expression patterns of the genes SERPINA14, PAG , and PAG2 at the fetal-maternal interface in dairy heifers experimentally infected with N caninum during the second trimester of pregnancy, when most abortions takes place in natural conditions. Irrespective of infection, expression of SERPINA14 was higher, and expression of PAG1 and PAG2 lower, for intercaruncular endometrium than for caruncles or cotyledons. Cotyledonary tissues showed the highest expression of both PAG genes but lowest expression of SERPINA14 . The expression of SERPINA14 was significantly higher in intercaruncular endometrium of control dams than for infected animals, pointing to potential disruption of modulation of maternal immune function during infection. Dramatically reduced SERPINA14 was particularly apparent in infected dams with aborted fetuses. There was also a negative association between N caninum antibody titers with SERPINA14 and PAG expression in infected animals, further suggesting that N caninum infection downregulates the uterine immunosuppressive function of SERPINA14. [ABSTRACT FROM AUTHOR]

Neosporosis is a parasitic disease caused by the protozoan Neospora caninum which results in major economic losses for cattle breeding due to abortion and other reproductive disorders. Gerbils (Meriones unguiculatus) are commonly used as experimental models for neosporosis due to their high susceptibility to N. caninum infection, both by oocysts ingestion as by tachyzoites/bradyzoites parenteral inoculation. However, the risk of transmission by tachyzoites ingestion is not fully elucidated. In this study, infection of neonate gerbils by N. caninum (NC-1 strain) tachyzoites inoculated by the oral route and the parasite distribution in gerbils' tissues were evaluated by protozoan DNA detection. Seventeen neonate gerbils, aged 4-5 days, were inoculated with 4x1 05 tachyzoites by the oral route and one gerbil was kept as uninfected control. N. caninum DNA was detected in 100% of the inoculated gerbils, showing that the oral route is effective as a potential route of infection of neonates by N. caninum tachyzoites. N. caninum DNA was reported in all organs evaluated (heart, lungs, kidneys, liver, spleen and brain), with different frequencies. These results showed systemically distributed infection of neonate gerbils after oral inoculation of tachyzoites [ABSTRACT FROM AUTHOR]

The protozoan pathogen Neospora caninum is recognized as a leading cause of infectious abortions in cattle worldwide. To evaluate the impact of neosporosis on dairy and beef herd production, a retrospective, longitudinal study was performed to identify the impact of neosporosis alongside other causes of fetal abortion in British Columbia, Canada. Retrospective analysis of pathology records of bovine fetal submissions submitted to the Animal Health Centre, Abbotsford, British Columbia, a provincial veterinary diagnostic laboratory, from January 2007 to July 2013 identified 182 abortion cases (passive surveillance). From July 2013 to May 2014, an active surveillance program identified a further 54 abortion cases from dairy farmers in the Upper Fraser Valley, British Columbia. Of the total 236 fetal submissions analyzed, N . caninum was diagnosed in 18.2% of cases, making it the most commonly identified infectious agent associated with fetal loss. During active surveillance, N . caninum was associated with 41% of fetuses submitted compared to 13.3% during passive surveillance ( p < 0.001). Breed of dam was significantly associated with N . caninum diagnosis, with a higher prevalence in dairy versus beef breeds, and fetuses of 3–6 months gestational age had the highest prevalence of N . caninum. There was no significant association with dam parity. N . caninum was diagnosed in every year except 2009 and cases were geographically widespread throughout the province. Furthermore, the active surveillance program demonstrates that N . caninum is highly prevalent in the Upper Fraser Valley and is a major causal agent of production losses in this dairy intensive region. [ABSTRACT FROM AUTHOR]

Contents: This study examines gene expression patterns in dairy heifers experimentally infected with N. caninum during on Day 110 of pregnancy with live foetuses at euthanasia, 42 days later. The study population was constituted of four non‐infected controls and three infected dams. Gene expression was determined on gamma interferon (IFNγ), (Th1 pro‐inflammatory cytokine), interleukin‐4 (IL4) (Th2 pro‐gestation cytokine) or interleukin‐10 (IL10) (T regulatory cytokine) and the serine peptidase inhibitor SERPINA14 in intercaruncular, placental, uterine lymph node (UTLN) and luteal tissue samples. Intercaruncular SERPINA14 expression was negatively correlated with IFNγ expression in cotyledon samples and with IL4 expression in UTLN. No relationships were detected between cytokine gene expression at the foetal–maternal interface and SERPINA14 expression in the luteal samples. Our findings suggest that gene expression of the uterine serpin SERPINA14 correlates negatively with the expression of Th1 and Th2 cytokines at the foetal–maternal interface but not in the corpus luteum. [ABSTRACT FROM AUTHOR]

Neospora caninum is an obligate intracellular protozoan that causes abortion in cattle worldwide. Plasma concentrations of pregnancy-associated glycoprotein 1 (PAG-1), produced by binucleate trophoblast cells, are used for pregnancy diagnosis and as a marker of foetal-placental well-being, while PAG-2, produced by both mono- and binucleate trophoblast cells, may serve as an indicator of abortion risk. In prior work, natural N. caninum infection was found to modify plasma PAG-1 and PAG-2 patterns. The present study examines PAG-1 and PAG-2 dynamics and trophoblast cell populations following experimental infection with N. caninum. The study population was comprised of 17 N. caninum seronegative Holstein-Friesian heifers. On day 110 of gestation, 6 heifers were inoculated intravenously with 10 7 taquyzoites of N. canimum. Plasma samples for PAG-1 and PAG-2 determinations were collected on days 0, 7, 14, 21 and 42 post infection. During the study course, pregnancy was normally expressed in all controls while three infected heifers lost their foetuses. All heifers were euthanised on day 42 post infection and placentome samples from the 14 non-aborting heifers were collected to examine trophoblast cell populations. Plasma PAG-1 and PAG-2 concentrations in non-infected heifers increased until the day of euthanasia while non-aborting infected heifers showed a temporary fall in PAG-1 ( P < 0.004) and PAG-2 ( P < 0.002) concentrations from 7 to 14 days post infection (dpi). The two dams aborting at 14 and 21 dpi and a third dam with a mummified foetus upon euthanasia showed dramatic PAG-1 and PAG-2 reductions from 14 dpi to undetectable levels upon euthanasia. A stereological study of placentomes revealed significantly higher relative proportions of mono- ( P = 0.035) and binucleate ( P = 0.029) trophoblast cells at 42 dpi in non-infected heifers than infected non-aborting heifers. According to our findings, following experimental N. caninum infection on day 110 of gestation, non-aborting heifers showed a brief reversible drop in plasma PAG-1 and 2 concentrations two weeks later and reduced proportions of bi- and mono-nucleate trophoblast cells 42 days after infection. In aborting dams, dramatically reduced PAG levels were related to severe placental damage and a non-viable pregnancy. [ABSTRACT FROM AUTHOR]

SUMMARY: At present, bovine neosporosis is an important worldwide concern because of its wide geographic distribution and economic impact. Abortion is the main clinical sign of bovine neosporosis in both dairy and beef cattle. Ruminant challenge models are critical to evaluate potential vaccine candidates to help tackle bovine neosporosis and to study pathogenesis and host responses to infection. Several research groups have developed ruminant models of Neospora caninum infection independently of others, resulting in a high degree of variability due to the use of different species of animals, breeds, strains/isolates of N. caninum , doses, routes and times of inoculation. Standardization is greatly needed to advance research in a more collaborative, timely and efficient manner. In the absence of widely accepted international guidelines, this manuscript serves to summarize and discuss the different models and parameters currently in use. Parameters essential for the development of non-pregnant and pregnant ruminant models are outlined and the main knowledge gaps are identified. This information could act as the basis to develop a consensus for international standard guidelines for ruminant models of neosporosis that would be helpful for researchers in this field worldwide. [ABSTRACT FROM AUTHOR]

Abstract: Bovine neosporosis control programs are currently based on herd management and serodiagnosis because effective treatments and vaccines are unavailable. Although a wide variety of serological tools have been developed, enzyme-linked immunosorbent assays (ELISAs) are the most commonly commercialized tests. Partial comparative studies have been performed in the past, and the panel of available ELISAs has notably changed in the last few years. Therefore, diagnostic laboratories are requesting updated information about the performance of these tests. Accordingly, the aim of this study was to compare all of the commercially available ELISAs (n =10) by evaluating their performance and to re-standardize them based on TG-ROC analyses when necessary. For this purpose, a well-characterized serum panel from experimentally and naturally infected bovines and non-infected bovines (n =458) was used. Two different definitions of gold standard were considered: (i) the result of the majority of tests and (ii) pre-test information based on epidemiological, clinical and serological data. Most of the tests displayed high sensitivity (Se) and specificity (Sp) values when both gold standard criteria were considered. Furthermore, all the tests showed near perfect agreement, with the exception of the pair-wise comparisons that included the VMRD and SVANOVIR. The best-adjusted ELISAs were the HIPRA-CIVTEST, IDVET, BIOVET and IDEXX Rum (Se and Sp>95%). After the TG-ROC analyses, higher Se and Sp values were obtained for the BIO-X, LSI Bov, LSI Rum and IDEXX Bov, though the increases were more significant for the SVANOVIR and VMRD. The Kappa values also increased with the new adjusted cut-offs. This is the first study that offers updated performance evaluations of commercially available ELISAs. Such analyses are essential for diagnostic laboratories and are valuable to the companies that develop and distribute these tests. [Copyright &y& Elsevier]

Neospora caninum is a protozoan parasite with a wide host range but with a preference for cattle and dogs. Since the description of N. caninum as a new genus and species in 1988, bovine neosporosis has become a disease of international concern as it is among the main causes of abortion in cattle. At present there is no effective treatment or vaccine. This review focuses on the epidemiology of the disease and on prospects for its control in cattle. Finally, based on the implications of clinical findings reported to date, a set of recommendations is provided for veterinarians and cattle farmers. [ABSTRACT FROM AUTHOR]

Abstract: The Nc-Spain 7 isolate of Neospora caninum, which was newly obtained from an asymptomatic congenitally infected calf, demonstrated a similar virulence as Nc-1 strain in mouse models. The aim of this study was to characterize the pathogenesis of Nc-Spain 7 isolate in cattle after experimental infection at 65 days of gestation. For this purpose, thirteen pregnant heifers were divided into three groups as follows: group A: 7 heifers inoculated with 1×108 tachyzoites of Nc-Spain 7 isolate; group B: 4 heifers inoculated with 1×108 tachyzoites of Nc 1 strain; and group C: 2 heifers received PBS. Serum samples were collected weekly and heparinized blood samples were collected three times (0, 28 and 42 days after inoculation) by jugular venipuncture. Placenta and fetal tissue samples were collected at time of necropsy. Specific antibody response in the dams was tested by IFAT, indirect ELISA, and rNcGRA7 and rNcSAG4 based-ELISA. Specific antibody response in fetal fluids was tested by IFAT. IFN-γ production was measured after in vitro culture of PBMC and the supernatant was assessed using a commercial kit (BOVIGAM). A significant increase in N. caninum antibody responses was detected in groups A and B by IFAT and by i-ELISA from day 14 after inoculation onwards. Besides, antibody response against rNCGra7 protein was also detected in all inoculated heifers by rNcGra7-based ELISA. Four fetuses from group A and one from group B were aborted between 3 and 5 weeks after infection. In the recovered fetuses, only 3 out of 4 fetal fluids from fetuses of group A and 1 out of 3 of group B were seropositive by IFAT, but all of them were positive by PCR. Transplacental transmission could be determined in all fetuses from groups A and B by PCR and/or IHC. Heifers of group C and their fetuses remained negative by all techniques. The results of this study demonstrate that the NC-Spain 7 isolate could be transmitted transplacentally, and produced fetal death and abortion in cattle. [Copyright &y& Elsevier]

The aim of this study was to describe the prevalence of Neospora caninum based on the detection of specific antibodies in bulk tank milk (BTM) from dairy cattle farms in the Mar y Sierras Basin by using an enzyme-linked immunosorbent assay (ELISA). A total of 98 BTM samples from 49 dairy farms were collected during autumn and spring of 2019. Additionally, 147 paired individual milk and serum samples were collected from two dairy farms to assess the prevalence within-herd by ELISA and indirect fluorescent antibody test (IFAT, serum samples). Additionally, 12 individual serum samples were also assessed to test the agreement between IFAT and ELISA (total serum samples 159). Noteworthy, 100 and 91.84% of 49 dairy farms were positive in the BTM in autumn and spring, respectively. For the within-herd individual samples, a good agreement between serum and milk results was obtained for ELISA and IFAT (0.86-0.90). This is the first study in Argentina in which milk samples were tested to determine the N. caninum infection status at herd and within-herd levels in dairy farms, providing a base for further research., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Neosporosis is one of the leading causes of abortion in cattle worldwide, posing a great economic burden on cattle producers. The aim of this study was to determine the national seroprevalence and putative risk factors of Neospora caninum (N. caninum) in dairy cattle in Uruguay. A cross-sectional study was carried out. Herds were stratified by size (1-50, 51–250, and >250 cattle) and up to 60 dairy cows per herd were randomly selected. Four thousand two hundred twenty-three serum samples from 102 dairy herds were analyzed by indirect ELISA test, under the manufacturer's recommendations. In addition, the herdsman was surveyed and a population study was carried out. The in degree data, geographical coordinates, and seroprevalence of bovine viral diarrhea, enzootic bovine leukemia and infectious bovine rhinotracheitis were available for each herd. A sampling design was used to estimate population seroprevalence of N. caninum. In order to determine the factors associated with the disease, herds with an intra-herd seroprevalence over 20% were considered as case herds. Seroprevalence of N. caninum was 22.3% (95% CI: 18.7-25.9 %) and 96.0% (95% CI: 92.1-99.8%) at the animal and herd level, respectively. The number of dogs on the dairy farms were associated with infection levels (OR: 1.43, 95% CI: 1.02 to 2.03). It was concluded that N. caninum is endemic in the country, and is spreading over dairy herds. Although this study showed evidence that the number of dogs were associated with high levels of infection, more studies should be conducted, to better understand the epidemiology of the disease and thus develop efficient control measures. [ABSTRACT FROM AUTHOR]

The aim of this study was to describe bovine neosporosis in dairy cattle from the Sierra region, Ecuador. A case-control study was performed on 841 dairy cattle from 5 dairy herds. The overall seroprevalence was 23.4% having significant association between abortion and seropositivity (p < .05). Additionally, 46 fetuses were recovered from a local slaughterhouse to evaluate the frequency of vertical transmission. Seventeen and 3 fetuses were positive by PCR and had compatible histopathological lesions, respectively. N. caninum infection must be considered as a relevant cause of reproductive losses in Ecuador., Competing Interests: Declaration of Competing Interest The authors do not have any conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Neospora caninum is a protozoan parasite that causes abortion and important economic losses in cattle worldwide. The accurate diagnosis of neosporosis is essential for management and control measures. The aims of this study were: i) to evaluate the performance of an in-house enzyme-linked immunosorbent assay based on the 38kDa native antigen (p38-ELISA) to diagnose bovine neosporosis in Argentina using a well- characterized local sera panel from experimentally infected and naturally exposed cattle and ii) to compare the diagnostic performance and agreement of three N. caninum serological tests: p38-ELISA, indirect fluorescence antibody test (IFAT) and immunoblotting (IB) using the same sera panel. Serum samples testing either positive or negative by IFAT and IB were considered "Relative Standards of Comparison" (RSC) and used for p38-ELISA evaluation. Receiver operating characteristics analysis revealed that p38-ELISA was highly accurate (area under the curve= 0.982) according to RSC with a cut-off index of 0.0905. Relative sensitivity and specificity of p38-ELISA were 97.8% and 99.5%, respectively and agreement between RSC and p38-ELISA was almost perfect (k= 0.97). The evaluation and performance comparison of serological tests were performed according to the definition of gold standard based on the decision of the "majority of tests". All tests displayed high sensitivity and specificity values (greater than 95%); and excellent agreement. This study describes the accurate performance of p38-ELISA evaluated locally and the highly accurate diagnostic performance of the studied tests for the detection of anti-N. caninum antibodies in cattle from Argentina., (Copyright © 2015 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.)