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is a major cause of reproductive failure in cattle, responsible for epidemic abortion outbreaks that inflict annual billion-dollar losses on the global livestock industry. In water buffaloes ( ), however, a phylogenetically close relative often raised in the same environments, the same parasite typically establishes a subclinical persistent infection with markedly lower rates of clinical abortion. This review inverts the traditional narrative by arguing that the key to next-generation control strategies lies in understanding the tolerant host (buffalo) rather than solely the susceptible host (cattle). By dissecting this \" paradox\", we explore the molecular and immunological crosstalk that dictates pregnancy outcomes. We examine the parasite's invasion proteins, revealed by CRISPR-Cas9 screens, and the maternal-fetal interface, where the balance between immune tolerance and parasite control determines the fate of pregnancy. We also compare with the related zoonotic parasite to highlight how differential host immune recognition shapes infection outcomes. Finally, we propose that deciphering the buffalo's successful equilibrium with can illuminate novel pathways for vaccines and immunotherapeutic strategies, transforming the management of neosporosis worldwide.

Objectives: The study aimed to detect Neospora caninum by nested PCR (nPCR) in aborted fetuses of cattle, sheep, and goats in Bangladesh. Materials and Methods: The head portion of each aborted fetus (111) was dissected at each sampling site and transferred to the laboratory in an ice box. Data on risk factors associated with N. caninum infection were simultaneously collected. Deoxyribonucleic acid was extracted from brain tissue to perform nPCR targeting the internal transcribed spacer 1 (ITS1) ribosomal DNA (rDNA) gene of N. caninum and sequencing was performed from the representative positive samples. Results: By nPCR, N. caninum was found in 16.0% of aborted fetuses of cattle, followed by sheep (14.81%) and goats (11.78%). The highest prevalence was found in aborted fetuses of animals during the second trimester (27.78%) of pregnancy aged 2 to 4 years (18.75%). Obtained sequences showed they were completely matched with N. caninum ITS1 rDNA gene deposited in GenBank. Univariate analysis demonstrated that pregnancy stages (trimesters), abortion history of the animals, and access to dogs in animal farms were significantly (p ≤ 0.05) correlated with N. caninum infection. Conclusion: This study represents the first investigation into the molecular detection, phylogenetic characterization, and analysis of risk factors associated with N. caninum in livestock in Bangladesh. According to the research findings, N. caninum infection may have a role in abortion cases and the ensuing financial losses in the nation’s livestock industry.

PurposeThe aim of this study was to estimate the seroprevalence and molecular prevalence and perform a molecular identification of Neospora caninum in semen of Tunisian rams.MethodsA total of 92 blood samples were collected from four farms located in four Tunisian governorates (Jendouba, Kairouan, Zaghouan and Ben Arous) and samples were screened with a commercial ELISA kit for N. caninum antibodies. For the same rams, semen samples were collected and tested for the presence of N. caninum ITS1 gene using PCR. Five amplicons were randomly selected for sequencing. A phylogenetic tree was constructed to compare the partial sequences of the ITS1 gene with sequences deposited in GenBank.ResultsThe seroprevalence of N. caninum infection was 25% (23/92) and PCR revealed that the molecular infection prevalence in semen was 11.95% (11/92). Kappa test showed an average agreement between seroprevalence and parasite prevalence in semen (κ = 0.44). The highest molecular prevalence was for rams that accomplished more than two mating seasons (21.0 ± 12.1%) compared to those performed less than two mating seasons and yearling individuals (4.0 ± 5.5%) (P = 0.01). There were no differences in N. caninum molecular prevalence according to either breed or locality. Comparison of the partial sequences of the ITS1 gene revealed 99–100% similarity with those deposited in GenBank.ConclusionTo the best of our knowledge, this is the first detection and molecular identification of N. caninum in semen from rams in North Africa. Our findings indicate that N. caninum infection rate was high in rams.

This study described and characterized clinical cases and outbreaks of neosporosis in cattle, sheep, and goats, considering the geographical distribution, clinical manifestations, lesions, and diagnostic methods reported in different regions of the world. An independent systematic review was conducted following the standard guidelines of the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA). Fifteen studies met the inclusion criteria: case reports and short communications published until December 2024, indicating the origin of the sampled animals; written in English, Portuguese, or Spanish; and published in indexed peer-reviewed journals. Case reports originated from four continents: America (North, Central and South), Europe, Asia, and Oceania. America (North, Central and South) had the highest number of case reports (60%; 9/15). Cattle were the most frequently reported species with neosporosis cases (46.7%; 7/15), while small ruminants accounted for eight cases (53.3%; 8/15). The most common clinical sign was abortion (86.6%; 13/15), and the most affected organs were the brain (93.3%; 14/15), heart (66.6%; 10/15), liver (40%; 6/15), and lungs (33.3%; 5/15). The most used diagnostic methods were histopathology (100%; 15/15) and immunohistochemistry (80%; 12/15). The most frequent microscopic findings included mononuclear cell infiltrates in the nervous tissues (66.6%; 10/15) and gliosis (44.6%; 7/15). Neosporosis showed wide geographical distribution, mainly affecting cattle in the Americas, with abortion as the main clinical sign and the brain as the most affected organ; diagnosis based on histopathology and immunohistochemistry, highlighting the importance of this disease in livestock production. RESUMO: O objetivo deste estudo foi descrever e caracterizar casos clínicos e surtos de neosporose em bovinos, ovinos e caprinos, considerando a distribuição geográfica, manifestações clínicas, lesões e métodos de diagnóstico em diferentes regiões do mundo. Realizou-se uma revisão sistemática independente seguindo as diretrizes padrão do Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA). E para realizar a descrição e a caracterização 15 estudos foram selecionados por atenderam aos critérios de inclusão: Relatos de casos e short communications, publicados até dezembro de 2024, com indicação da origem dos animais amostrados, redigidos em inglês, português ou espanhol, e publicados em periódicos indexados com revisão por pares. Esses relatos de caso foram provenientes de quatro continentes: América (Norte, Central e Sul), Europa, Ásia e Oceania. A América (Norte, Central e Sul) foi o continente que mais teve relatos de caso (60%; 9/15). A espécie bovina foi a mais relatada com casos de neosporose (46,7%; 7/15) e em pequenos ruminantes foram relatados oito casos de neosporose (53,3%; 8/15). Além disso, o sinal clínico mais comum foi o aborto (86,6%; 13/15) e os órgãos mais lesionados foram cérebro (93,3%; 14/15), coração (66,6%; 10/15), fígado (40%; 6/15) e pulmão (33,3%; 5/15). Os métodos de diagnóstico mais empregados foram o histopatológico (100%; 15/15) e a imuno-histoquímica (80%; 12/15). Os achados microscópicos mais frequentes foram infiltrados de células mononucleares no tecido nervoso (66,6%; 10/15) e gliose (44,6%; 7/15). A neosporose apresenta ampla distribuição, acomete principalmente bovinos nas Américas, com aborto como principal sinal clínico e cérebro como órgão mais afetado. O diagnóstico baseado em histopatologia e imuno-histoquímica, reforça o impacto da doença para a produção animal.

Recombinant NcPDI(recNcPDI), NcROP2(recNcROP2), and NcMAG1(recNcMAG1) were expressed in Escherichia coli and purified, and evaluated as potential vaccine candidates by employing the C57Bl/6 mouse cerebral infection model. Intraperitoneal application of these proteins suspended in saponin adjuvants lead to protection against disease in 50% and 70% of mice vaccinated with recNcMAG1 and recNcROP2, respectively, while only 20% of mice vaccinated with recNcPDI remained without clinical signs. In contrast, a 90% protection rate was achieved following intra-nasal vaccination with recNcPDI emulsified in cholera toxin. Only 1 mouse vaccinated intra-nasally with recNcMAG1 survived the challenge infection, and protection achieved with intra-nasally applied recNcROP2 was at 60%. Determination of cerebral parasite burdens by real-time PCR showed that these were significantly reduced only in recNcROP2-vaccinated animals (following intraperitoneal and intra-nasal application) and in recNcPDI-vaccinated mice (intra-nasal application only). Quantification of viable tachyzoites in brain tissue of intra-nasally vaccinated mice showed that immunization with recNcPDI resulted in significantly decreased numbers of live parasites. These data show that, besides the nature of the antigen, the protective effect of vaccination also depends largely on the route of antigen delivery. In the case of recNcPDI, the intra-nasal route provides a platform to generate a highly protective immune response.

Neosporosis, which is caused by Neospora caninum, is a well-known disease in the veterinary field. Infections in pregnant cattle lead to abortion via transplacental (congenitally from mother to fetus) transmission. In this study, a N. caninum profilin (NcPROF), was expressed in silkworm larvae by recombinant Bombyx mori nucleopolyhedrovirus (BmNPV) bacmid and was purified from the hemolymph. Three NcPROF constructs were investigated, native NcPROF fused with an N-terminal PA tag (PA-NcPROF), PA-NcPROF fused with the signal sequence of bombyxin from B. mori (bx-PA-NcPROF), and bx-PA-NcPROF with additional C-terminal transmembrane and cytoplasmic domains of GP64 from BmNPV (bx-PA-NcPROF-GP64TM). All recombinant proteins were observed extra- and intracellularly in cultured Bm5 cells and silkworm larvae. The bx-PA-NcPROF-GP64TM was partly abnormally secreted, even though it has the transmembrane domain, and only it was pelleted by ultracentrifugation, but PA-NcPROF and bx-PA-NcPROF were not. Additionally, bx-PA-NcPROF-GP64TM was successfully purified from silkworm hemolymph by anti-PA agarose beads while PA-NcPROF and bx-PA-NcPROF were not. The purified bx-PA-NcPROF-GP64TM protein bound to its receptor, mouse Toll-like receptor 11 (TLR-11), and formed unique nanoparticles. These results suggest that profilin fused with GP64TM was secreted as a nanoparticle with binding affinity to its receptor and this nanoparticle formation is advantageous for the development of vaccines to N. caninum.

Microneme proteins (MICs) are important for Apicomplexan parasite invasion due to their adhesion to host cells. Several studies have indicated that Neospora caninum MIC3 and MIC8 are important adhesion factors and potential vaccine candidates against neosporosis. In this study, we evaluated the protective efficacy of recombinant proteins and DNA vaccines of NcMIC3 and NcMIC8. BALB/c mice were immunized with rNcMIC3, rNcMIC8, pcDNA3.1-NcMIC3 and pcDNA3.1-NcMIC8 respectively, and challenged with N. caninum tachyzoites. The immune responses were evaluated through cytokine, antibody measurements and the parasite burden in the mice brain tissues. Serological analysis showed that recombinant protein vaccines induced higher levels of immunoglobulin G (IgG) than other groups. The percentage of IgG1 and IgG2a in the recombinant protein groups was higher than the other groups, and with a predominance of IgG1 over IgG2a, suggesting that recombinant protein vaccines elicited a Th2-type immune response, while DNA vaccines mainly produce a Th1-type immune response. In addition, mice immunized with rNcMIC3 and rNcMIC8 a had lower parasite burden in brain tissue compared with the other groups. These results demonstrate that rNcMIC3 and rNcMIC8 could induce humoral and Th2-type immune response, leading to a considerable level of resistance against neosporosis.

► We demonstrate the immune enhancement of a tailored N. caninum vaccine in mice. ► Vaccine antigen was selected based on its capacity to activate dendritic cells. ► Vaccines with Providean-AVEC® adjuvant limited N. caninum infection. ► A tenth of the antigen payload was needed compared to other aqueous adjuvants. ► Protective responses against N. caninum entailed systemic IgG2a and IFN-γ. Efficient, cost-effective and safe Th1-immunity-inducing vaccine formulations are paramount for achieving protection against Neospora caninum. In this study, a new adjuvant (Providean-AVEC®) was used in the development of a N. caninum vaccine and evaluated in a mouse model. Soluble N. caninum tachyzoite native protein extract (sNcAg) was selected as vaccine antigen based on its capacity to activate production of pro-inflammatory cytokines on dendritic cells. Vaccines containing 4 and 0.4μg of sNcAg, and Providean-AVEC®, ISCOM-Matrix or aluminum hydroxide (Alum) were tested in BALB/c mice. While mice vaccinated with 4μg of sNcAg+Providean-AVEC® developed specific antibodies shortly after the first dose, the rest of the high antigen payload formulations only induced seroconversion after the booster. Mice immunized with the high payload ISCOM vaccine (4μg sNcAg) or with either low or high payload Providean-AVEC® formulations (0.4μg and 4μg sNcAg, respectively) elicited higher IgG2a than IgG1 serum levels, and IFN-γ anamnestic responses with a Th1-cytokine biased profile. These animals had no histological signs of cerebral lesions and parasite burden assessed by quantitative real-time PCR was not detected. Vaccine preparations including Providean-AVEC® as adjuvant limited N. canimum multiplication even with only a tenth of antigen payload compared to vaccines containing other adjuvants. Using adjuvants to specifically activate dendritic cells, combined with a careful antigen selection can enhance cellular responses to inert N. caninum vaccines.

A 3-mo-old red fox (Vulpes vulpes) developed generalized crusty plaques on its body during rehabilitation after an automobile accident requiring amputation of one leg. Histologic examination of skin lesion biopsy revealed granulomatous dermatitits with many intralesional protozoal tachyzoites. The protozoa stained positively with antibodies to Neospora caninum but not to Toxoplasma gondii. Treatment with clindamycin hydrochloride (10 mg/kg, twice daily, s.c.) for 1 mo completely resolved lesions, and protozoa were not demonstrable in biopsy of skin after treatment. The fox had agglutinating antibodies to T. gondii (modified agglutination test, titer 1 : 3200) and N. caninum (Neospora agglutination test, titer 1 : 25), and viable T. gondii (genotype III) was isolated from the skin biopsy after treatment. This is the first report of clinical neosporosis in a wild canid.

Neospora caninum, the causative agent of neosporosis, is a protozoan parasite responsible for high rate of abortion in cattle worldwide. In dogs, consequences of infection vary from severe neuromuscular disorders to asymptomatic infection and shedding of environmentally resistant oocysts. In this study, we determined the occurrence of N. caninum antibodies in dairy cattle and dogs in Vojvodina (Northern Province of Serbia) and possible risk factors. N. caninum antibodies were found in 15.4% (55/356, CI 95%:12.0–19.6) of cows and 17.2% (17/99, CI 95%: 10.8–26.2) of dogs. Cows from smallholdings showed significantly greater odds (OR = 5.28, CI 95%: 2.0–13.6, p = 0.0006) of being seropositive in comparison to the farm cows. Epidemiological importance of results is discussed. Neospora caninum, l’agent qui cause la néosporose, est un parasite protozoaire responsable d’un taux élevé d’avortements dans le bétail à travers le monde. Chez les chiens, les conséquences de l’infection varient entre un trouble neuromusculaire sévère et une excrétion asymptomatique d’oocystes résistants dans l’environnement. L’objectif de cette étude était d’examiner le statut sérologique des vaches laitières et des chiens en Voïvodine (province du nord de la Serbie) et les facteurs de risque possibles. Des anticorps contre N. caninum ont été détectés dans 15,4 % (55/356, CI 95 %:12,0–19,6) des vaches et dans 17,2 % (17/99, CI 95 %:10,8–26,2) des chiens. Les vaches des petits propriétaires ont manifesté une probabilité significativement plus importante (OR = 5,28, CI 95 %:2,0–13,6, p = 0,0006) d’être séropositives que les vaches de fermes. L’importance épidémiologique de ces résultats est discutée.