Macrophage migration inhibitory factor favors Neospora caninum infection in mice.

Neospora caninum is a protozoan parasite with worldwide incidence, acting as a major cause of reproductive failures in ruminants and neuromuscular symptoms in dogs. Macrophage Migration Inhibitory Factor (MIF) is produced by several cell types and exhibits a central role in immune responses against intracellular pathogens. The present study aimed to comprehend the role of MIF in the relationship between N. caninum and its host. We used in vivo, in vitro and ex vivo experiments in a model of infection based on genetically deficient mice to analyze the infection kinetics and inflammatory markers. MIF production was measured in response to N. caninum during the acute and chronic phases of the infection. While Mif-/- mice survived lethal doses of NcLiv tachyzoites, sublethal infections in these mice showed that parasite burden was controlled in target tissues, alongside with reduced inflammatory infiltrates detected in lung and brain sections. TNF was increased at the initial site of the infection in genetically deficient mice and the MIF-dependent reduction was confirmed in vitro with macrophages and ex vivo with primed spleen cells. In sum, MIF negatively regulated host immunity against N. caninum, favoring disease progression.

Herd-level prevalence of bovine leukemia virus, Salmonella Dublin and Neospora caninum in Alberta, Canada, dairy herds using ELISA on bulk tank milk samples.

Endemic infectious diseases remain a major challenge for dairy producers worldwide. For effective disease control programs, up-to-date prevalence estimates are of utmost importance. The objective of this study was to estimate the herd-level prevalence of bovine leukemia virus (BLV), Salmonella Dublin, and Neospora caninum in dairy herds in Alberta, Canada using a serial cross-sectional study design. Bulk tank milk samples from all Alberta dairy farms were collected 4 times, in December 2021 (n = 489), April 2022 (n = 487), July 2022 (n = 487), and October 2022 (n = 480), and tested for antibodies against BLV, S. Dublin, and N. caninum using ELISAs. Herd-level apparent prevalence was calculated as positive samples divided by total tested samples at each time point. A mixed effect modified Poisson regression model was employed to assess the association of prevalence with region, herd size, herd type, and type of milking system. Apparent prevalence of BLV was 89.4, 88.7, 86.9 and 86.9% in December, April, July, and October, respectively, whereas for S. Dublin apparent prevalence was 11.2, 6.6, 8.6, and 8.5%, and for N. caninum apparent prevalence was 18.2, 7.4, 7.8, and 15.0%. For BLV, S. Dublin and N. caninum, a total of 91.7, 15.6, and 28.1% of herds, respectively, were positive at least once, whereas 82.5, 3.6, and 3.0% of herds were ELISA-positive at all 4 times. Compared with the north region, central Alberta had a high prevalence (prevalence ratio (PR) = 1.13) of BLV-antibody positive herds, whereas south Alberta had a high prevalence (PR = 2.56) of herds positive for S. Dublin antibodies. Furthermore, central (PR = 0.52) and south regions (PR = 0.46) had low prevalence of N. caninum-positive herds compared with the north. Hutterite colony herds were more frequently BLV-positive (PR = 1.13) but less frequently N. caninum-positive (PR = 0.47). Large herds (>7,200 L/day milk delivered ∼ > 250 cows) were 1.1 times more often BLV-positive, whereas small herds (≤3,600 L/day milk delivered ∼ ≤ 125 cows) were 3.2 times more often N. caninum-positive. For S. Dublin, Hutterite-colony herds were less frequently (PR = 0.07) positive than non-colony herds only in medium and large stratum but not in small stratum. Moreover, larger herds were more frequently (PR = 2.20) S. Dublin-positive than smaller herds only in non-colony stratum but not in colony stratum. Moreover, N. caninum prevalence was 1.6 times higher on farms with conventional milking systems compared with farms with an automated milking system. These results provide up-to-date information of the prevalence of these infections that will inform investigations of within-herd prevalence of these infections and help in devising evidence-based disease control strategies.

Evidence of Australian wild deer exposure to N. caninum infection and potential implications for the maintenance of N. caninum sylvatic cycle.

Infections with the coccidian parasite Neospora caninum affect domestic and wild animals worldwide. In Australia, N. caninum infections cause considerable losses to the cattle industry with seroprevalence of 8.7% in beef and 10.9% in dairy cattle. Conversely, the role of wild animals, in maintaining the parasite cycle is also unclear. It is possible that native or introduced herbivorous species could be reservoir hosts of N. caninum in Australia, but to date, this has not been investigated. We report here the first large-scale screening of N. caninum antibodies in Australian wild deer, spanning three species (fallow, red and sambar deer). Consequently, we also assessed two commercial cELISA tests validated for detecting N. caninum in cattle for their ability to detect N. caninum antibodies in serum samples of wild deer. N. caninum antibodies were detected in 3.7% (7/189, 95% CI 1.8 - 7.45) of the wild deer serum samples collected in south-eastern Australia (n = 189), including 97 fallow deer (Dama dama), 14 red deer (Cervus elaphus), and 78 sambar deer (Rusa unicolor). Overall, our study provides the first detection of N. caninum antibodies in wild deer and quantifies deer's potential role in the sylvatic cycle of N. caninum.

Analytical sensitivity of a multiplex quantitative PCR for Toxoplasma gondii and Neospora caninum.

Cyst-forming coccidia, Toxoplasma gondii and Neospora caninum, are recognised as important causes of animal disease. Molecular diagnostics based on the presence of DNA in animal tissue are required to specifically detect T. gondii and N. caninum while achieving high levels of analytical sensitivity. We optimised available single-plex probe base qPCR assays into a multiplexed qPCR panel to detect cyst-forming coccidia, i.e. T. gondii and N. caninum. The T. gondii assay is based on a 529-bp repetitive (REP) element and the N. caninum assay on the NC5 repetitive region. Using target sequence synthetic DNA, the limit of detection (LOD) was determined to be 100 copies, that is less than a single tachyzoite of either T. gondii or N. caninum. The T. gondii and N. caninum multiplexed qPCR assay optimised in this study can be used to effectively detect parasite DNA for diagnostic purposes in animal tissue.

Molecular Detection of Toxoplasma gondii, Neospora caninum and Encephalitozoon spp. in Vespertilionid Bats from Central Europe.

Bats may carry various viruses and bacteria which can be harmful to humans, but little is known about their role as a parasitic source with zoonotic potential. The aim of this study was to test wild bats for the presence of selected parasites: Toxoplasma gondii, Neospora caninum and microsporidia Encephalitozoon spp. In total, brain and small intestine tissues of 100 bats (52 Myotis myotis, 43 Nyctalus noctula and 5 Vespertilio murinus) were used for the DNA isolation and PCR detection of the abovementioned agents. Toxoplasma gondii DNA was detected by real-time PCR in 1% of bats (in one male of M. myotis), while all bats were negative for N. caninum DNA. Encephalitozoon spp. DNA was detected by nested PCR in 25% of bats, including three species (twenty-two M. myotis, two N. noctula and one V. murinus). Positive samples were sequenced and showed homology with the genotypes Encephalitozoon cuniculi II and Encephalitozoon hellem 2C. This is the first study on wild vespertilionid bats from Central Europe and worldwide, with a relatively high positivity of Encephalitozoon spp. detected in bats.

Seroprevalence, risk factors, and serological cross-reactivity for diagnosis of Toxoplasma gondii and Neospora caninum infections in goats in India.

Toxoplasma gondii and Neospora caninum are genetically related cyst-forming protozoan parasites that cause reproductive failures in ruminants. Given the limited information on the epidemiology of these infections in goats in India, the study aimed to estimate the seroprevalence, assess antibody cross-reactivity for diagnosis, and identify associated risk factors. A total of 695 sera were evaluated for antibodies to T. gondii and N. caninum infections using Modified Agglutination Test (MAT for Toxoplasma)/Neospora agglutination test (NAT), Enzyme-linked immunosorbent assay (ELISA), and Indirect Fluorescent Antibody Test (IFAT for tachyzoite and bradyzoite stages). The seroprevalence rate of T. gondii and N. caninum infections was 56.9% and 10.9%, respectively. Inter-rater agreement (kappa value - κ) was calculated to assess agreements between various diagnostic assays, using the IFAT as the gold standard (for detecting both infections), the agreements for MAT/NAT (κ = 0.97) and the ELISA (κ = 0.95) were excellent. The acute infection among seropositive goats were determined using serological (IgG avidity test - measures the binding strength between IgG antibodies and parasite antigens) and molecular diagnoses (PCR for repetitive DNA sequences - Toxoplasma B1 gene: 131 bp and Neospora NC5 gene: 328 bp). Among seropositive goats ≥80% had high IgG avidity and <10% of animals had low IgG avidity antibodies for both infections. Most low IgG avidity goats were PCR positive for the TgB1 gene (94.4%) or Nc5 gene (85.7%). In the serological assays, we used different dilutions of test serum to rule out the cross-reactivity owing to similar antigenic makeup between these two parasites. When the serological cross-reactivity was analyzed using invasion assay at a serum titer of ≥200, more than 90% T. gondii positive sera showed host cell invasion of N. caninum and vice versa. Largely, the serological results indicate that cut-off serum dilution of ≥1:200 for ELISA and IFAT and ≥1:25 for MAT/NAT avoids serological cross-reactivity between T. gondii and N. caninum. Further, the Univariate and multivariate analyses showed that adult animals (>2 years), reservoir hosts, and extensive rearing systems are common risk factors for these infections. However, the history of abortion was identified as a significant risk factor for T. gondii infection. This study revealed that T. gondii and N. caninum infections are highly prevalent in this region and the use of an appropriate cut-off serum dilution is necessary to avoid cross-reactivity between these closely related parasites.

Prevalence of Neospora caninum antibodies in fattening pigs and sows from intensive farms in northern Italy.

Neospora caninum (Apicomplexa, Sarcocystidae) is a major cause of reproductive failure in cattle. In pigs, only a few studies investigated the effects of this parasite on reproductive efficiency. Considering the relevance of swine farms in northern Italian regions, an epidemiological survey was designed to investigate the spread of N. caninum infection. Three hundred seventy fattening pigs and sows from 23 intensive farms in Lombardy were sampled. Sera were analyzed by a commercial immunofluorescence antibody test. Statistical analysis through univariate and multivariate generalized linear models was conducted to detect farm management practices enhancing the risk of infection. At the farm level, 52.1% (12/23) of the selected farms, 72.7% housing sows and 40% fattening pigs, scored positive. At the individual level, 25 animals (25/370, P = 6.7%) were positive to N. caninum antibodies: one fattening pig and two sows showed an antibody titer of 1:100, and in two sows, an antibody titer of 1:400 and 1:6400 was evidenced. A higher seroprevalence was detected in sows (17/151, P = 11.2%) if compared to fattening pigs (8/219, P = 3.6%) (OR = 1.19, P value = 0.000 in sows). Moreover, a higher seroprevalence was recorded in farms with low and moderate sanitary score (P = 100% and P = 64.2%, respectively) if compared to farms with high sanitary score (P = 22.2%) (OR = 1.24, P value = 0.007 in score = 1 and OR = 1.10, P value = 0.050 in score = 2). This study provides the first data on the circulation of N. caninum in intensive swine farms in Italy, demonstrating the spread of the parasite in fattening pigs and sows in Lombardy region.

Epidemic abortions due to Neospora caninum infection in farmed red deer (Cervus elaphus).

This study describes for the first time an abortion outbreak caused by Neospora caninum in farmed red deer. During a 5-year period, farmed hinds, naturally mated, were regularly ultrasound monitored to detect reproductive losses over their gestation. During the 4 years previous to the outbreak, abortion rates ranged from 4.7 to 8.6% (average 6.5%), and serology for indirect diagnosis of neosporosis and toxoplasmosis was performed. At the fifth year, the abortion rate increased to 25.3%. During this outbreak, three aborted foetuses and their placentas were recovered and submitted to laboratory for etiological diagnosis. Blood samples were collected from the 81 hinds at the end of the gestational period and the seropositivity rate for N. caninum, Toxoplasma gondii, Brucella abortus, bovine viral diarrhoea virus and bovine alphaherpesvirus type 1 was 66.7%, 67.9%, 0.0%, 8.6% and 0.0%, respectively. Neospora caninum-seropositive hinds (OR = 5.7, P = 0.0271) and hinds with high antibody titres to N. caninum (OR = 7.4, P = 0.0130) were more likely to abort than seronegative hinds. In addition, N. caninum seropositivity rate in the aborted hinds was higher (OR = 5.4, P = 0.033) than the non-aborted hinds. No association was found between T. gondii nor BVDV-seropositivity and abortions. Typical protozoal histopathologic findings (necrotizing non suppurative encephalitis, meningitis, myocarditis, hepatitis, among others) were observed in all foetuses. Neospora caninum was immunolabelled by immunohistochemistry in several tissues from two foetuses, and infection was also confirmed in the three foetuses by serology and/or DNA detection. No other abortifacient agent was detected in the foetuses. Their dams showed high N. caninum antibody titres (≥ 6400). Serologic evidence and epidemiological data recorded suggested a point-source of N. caninum infection before the occurrence of the outbreak, probably related with contaminated feedstuff with oocysts. Moreover, the intensive production system with a high stocking rate could be also considered a factor which might have increased the risk of horizontal N. caninum infection in this herd.

Evaluation of knowledge, attitudes and practices regarding neosporosis and toxoplasmosis among farmers and animal health practitioners in Namibia.

This study assessed the knowledge, attitudes and practices of livestock farmers in Namibia's Khomas region and animal health practitioners (veterinarians and animal health technicians) in the whole country concerning neosporosis and toxoplasmosis. Structured questionnaires were used, and a total of 63 farmers and 51 animal health practitioners responded out of an estimated 560 farmers in the Khomas region and 300 veterinarians and veterinary technicians in the country. Only 15.9% of the livestock farmers (n = 63) had heard about neosporosis or toxoplasmosis or knew how animals get infected (p < 0.0001). Only 5% of the farmers knew the risks associated with keeping dogs and cats concerning neosporosis and toxoplasmosis, respectively (p < 0.0001). None of the 51 animal health practitioners routinely requested Neospora caninum or Toxoplasma gondii laboratory tests in cases of cattle, sheep or goat abortions. Although all animal health practitioners indicated they routinely interacted with livestock farmers, none regularly discussed neosporosis or toxoplasmosis. Only 3.9% of animal health practitioners (n = 51) indicated that they had ever discussed either neosporosis or toxoplasmosis at a farmers' gathering (p < 0.0001), and only 21.6% had talked to at least one cat owner about toxoplasmosis in the previous 12 months (p < 0.0001). The authors concluded that farmers in the Khomas region were generally unaware of neosporosis and toxoplasmosis but could change their attitudes and practices if educated. The animal health practitioners lacked a deeper understanding and appreciation of the two diseases, which is required to cultivate enough confidence to educate farmers. Sharing this research and other relevant information on the two diseases at farmers' meetings, veterinary congresses, journals and newsletters could help educate farmers and animal health practitioners. Such platforms are likely to succeed because both these groups use these forums to get new information.

Prevalence and risk factors of Neospora caninum and Toxoplasma gondii infection in breeding ewes from central western Mexico.

The objective of the study was to analyse the prevalence of infection by N. caninum and T. gondii in breeding ewes from central western Mexico. For this purpose, 184 blood samples were collected from sheep in the reproductive stage, which were analysed by ELISA and PCR tests, respectively, and the association between seroprevalence and some management factors was estimated by means of a logistic regression analysis. For N. caninum, a seroprevalence of 15.22% was identified, while the DNA detection in blood was 14.13% (26/184); positive animals were found in 75% of the farms (6/8). The variables poor placental waste management (OR 8.4), abortion history (OR 1.5) and presence of rodents (OR 1.3) were identified as risk factors for N. caninum infection. In the case of T. gondii, a seroprevalence of 61.96% was found, and antibodies were detected in eight farms included in the study; no positive samples were identified for T. gondii by the PCR test. The main identified risk factors for T. gondii were as follows: poor hygiene conditions (OR 12.5), presence of cats (OR 9.5), presence of other domestic animals (OR 5.7), urban context of the farm (OR 9.5) and public water supply (OR 5.3). The seroprevalence of co-infection of T. gondii and N. caninum in breeding ewes was 7% (13/184; 95% CI 6.92-7.20). T. gondii has a high seroprevalence while N. caninum was of lower proportion, and both may be associated with reproductive losses for these herds studied.

Molecular detection of coccidian Apicomplexa Parasites isolated from wild crab-eating and pampas foxes through novel TaqMan™ probes: a contribution to their molecular epidemiology.

Neospora caninum, Toxoplasma gondii and Hammondia spp. are coccidian parasites similar in morphology. Molecular techniques are necessary to detect parasite DNA isolated from stool samples in wild canids because they were reported as definitive hosts of N. caninum life cycle. The objective of this study was to develop a highly sensitive and accurate molecular method for the identification of coccidian Apicomplexa parasites in crab-eating fox (Cerdocyon thous) and pampas fox (Lycalopex gymnocercus). Tissue samples from road-killed animals (pampas fox = 46, crab-eating fox = 55) and feces (pampas fox = 84, crab-eating fox = 2) were collected, and species were diagnosed through molecular assay. PCR was used for the amplification of a fragment of the coccidian Apicomplexa nss-rRNA gene. Additionally, we developed a novel real-time PCR TaqMan™ probe approach to detect T. gondii- Hammondia spp. and N. caninum. This is the first report of N. caninum DNA in pampas fox feces (n = 1), thus it was also detected from pampas fox tissues (n = 1). Meanwhile, T. gondii was found in tissues of pampas (n = 1) and crab-eating (n = 1) foxes and H. triffittae in one crab-eating fox tissue. Despite the low percentage (2.5%) of positive samples, the molecular method developed in this study proved to be highly sensitive and accurate allowing to conduct an extensive monitoring analysis for these parasites in wildlife.

Experimental infection of tachyzoites of the NC1 strain of Neosporacaninum in female swine.

Neospora caninum is a protozoan that can cause reproductive problems in several animal species. Although N. caninum infection has been reported in swine, the pathogenesis and clinical signs are not fully known in this species. The objective of this work was to evaluate the effect of experimental infection with tachyzoites of the N. caninum strain Nc1 in swine matrices at different stages of gestation. For that purpose, 12 gilts, seronegative for N. caninum and T. gondii, were selected and allocated into four groups of three animals each. Animals in group A were not inoculated (control) and animals in groups B, C, and D were inoculated intravenously with of 2.9 × 107 tachyzoites, 30 days before conception, and at 45 and 90 days of gestation, respectively. Temperature, heart rate, blood, saliva, and vaginal mucus samples from the animals were collected periodically until the time of delivery for the investigation of IgG and IgM antibodies against N. caninum using IFAT and PCR to detect the parasite DNA. All gilts sero-converted from 5 and 7 DPI (days postinoculation) to IgM and IgG, respectively. Two gilts showed hypothermia on the 5th and 7th DPI, and five inoculated animals had leukocytosis on the 7th DPI. It was possible to detect DNA of N. caninum in samples of saliva (33/84), vaginal mucus (17/84), and blood (2/84). Based on serology (IgM) and PCR, three animals in group B showed evidence of reappearance of the infection during pregnancy. It is concluded that N. caninum can cause clinical signs in infected swine females, in addition to indicating saliva as a suitable diagnostic biological material for the detection of N. caninum DNA in this animal species.

Differential acute-phase protein responses in dogs seropositive or seronegative for Neospora caninum.

Neospora caninum is one of the most prevalent Apicomplexa parasites that causes abortion in cattle, as it infects dogs as its definitive host, causing subclinical disease or active neosporosis, marked by meningoencephalitis, and myopathies with muscle and neuromuscular signs of disease. This study aimed to evaluate the acute phase protein response in dogs seropositive and seronegative for N. caninum. Serum samples of 72 dogs were tested by an immunofluorescence antibody test using N. caninum NC-1 strain, and the study population was divided into four groups: symptomatic - muscular and/or neuromuscular signs - and seropositive (n = 16); symptomatic and seronegative (n = 9); asymptomatic and seropositive (n = 34); and asymptomatic and seronegative (n = 13). C-reactive protein (CRP) was measured via immunoturbidimetric assay and serum haptoglobin (Hp) via hemoglobin-binding capacity assay. In the symptomatic groups, seropositive dogs had higher levels of Hp, but not CRP, while seronegative dogs had higher CRP levels. There was no difference in CRP concentration in asymptomatic dogs. Dogs with neuromuscular signs had higher concentrations for Hp in the group seropositive. Hp concentration did not differ between dogs seropositive and seronegative dogs for each group. Serum Hp and CRP could not sufficiently alone flag subclinical infections. Measurement of CRP and Hp concentrations could be clinically valuable to the diagnosis of neurological diseases, and their relative change may indicate the stage of the infection, although their sole use is not able to support the diagnosis of canine neosporosis. Further studies are encouraged to evaluate the specific dynamics of acute phase proteins in canine neosporosis. Serum samples of 72 dogs were tested by an immunofluorescence antibody test using N. caninum NC-1 strain, and the study population was divided into four groups: (1) dogs with muscular and/or neuromuscular signs and seropositive for N. caninum; (2) dogs with muscular and/or neuromuscular signs and seronegative for N. caninum; (3) dogs seropositive for N. caninum with no neuromuscular signs; and (4) healthy dogs and seronegative for N. caninum. The study evaluated if N. caninum infection could have pathophysiological changes activating the acute phase response and an increase in the concentration of acute phase proteins in serum.

Neosporosis in bulls: potential for venereal transmission, and effect on semen quality and production.

Neospora caninum caninum is a protozoan parasite that can cause abortions, birth of congenitally infected calves, and reduced milk production in cattle. Neosporosis in cattle can be transmitted through vertical transplacental transmission from dam to calf, or through horizontal transmission between animals. While it is well established that horizontal transmission from dogs (the definitive host) to cattle (an intermediate host) occurs, it is not clear whether horizontal transmission may occur between cows and bulls during mating. Transmission via infected semen of seropositive bulls was suggested when N. caninum DNA was found in bull semen. The primary objective of this article is to critically evaluate the evidence for venereal transmission of N. caninum between cattle, to determine whether this transmission route is likely to occur. A secondary objective is to review and evaluate published evidence for whether neosporosis in bulls may affect semen quality and production. N. caninum DNA can be detected by PCR in occasional batches of semen straws from bulls that are seropositive for N. caninum. Insemination of heifers with semen experimentally infected with 5 × 104 N. caninum tachyzoites has caused persistent seroconversion in some animals. However, the highest concentration of parasites in the semen of naturally infected bulls was estimated, using quantitative PCR, as 10 tachyzoites. Thus it appears that, whether used for natural or artificial insemination, the semen of bulls naturally infected with N. caninum, does not contain the necessary intrauterine dose of tachyzoites to cause persistent seroconversion in a heifer or cow. Thus, based on the available evidence, the venereal transmission of N. caninum via bull semen is extremely unlikely. Nevertheless, neosporosis in bulls may influence semen quality and production. Epididymal semen concentration, viability, and motility appeared "to be" significantly decreased in N. caninum-seropositive bulls compared to seronegative bulls. Furthermore, semen production could be altered if neosporosis affects the thyroid gland. The thyroid hormones such as triiodothyronine and thyroxine (T4) may affect testicular development. Significantly lower concentrations of T4 were observed in the blood of N. caninum-seropositive, compared with those of seronegative bulls. Longitudinal studies are needed to confirm the long-term effect and significance of neosporosis on semen quality and thyroid hormone concentrations in bulls.

Antibody seropositivity for Neospora caninum in Blanco Orejinegro cattle in Colombia and factors associated with the infection.

Neosporosis is a parasitic disease that causes abortions and economic losses in bovine production systems, but no studies have been found concerning its effect on the Creole cattle breed, Blanco Orejinegro (BON). The aim of this research was to establish the serological status of Colombian BON cattle against Neospora caninum and to determine the factors associated with seropositivity. Blood samples were taken from 363 animals from 13 herds in six states of Colombia, and analyzed by indirect ELISA using a commercial test. Information on sex, herd, generation group, and state was recorded. A survey was carried out with 26 questions related to productive, reproductive, and health factors per herd. A logistic regression analysis was performed and the ORs for significantly associated variables were estimated using the R software. General seropositivity of 73.5% (95% CI 68.6-77.9%) was obtained, and sex, age group, and herd were the variables significantly associated with seropositivity (p < 0.05). For the sex variable, seroprevalence levels of 79.6% (95% CI 74.3-84.1%) were recorded for females and 54.5% (95% CI 43.6-65.1%) for males. Herd seroprevalence varied between 58.3 and 95.8%, and the last generation showed the lowest positivity (51.2%, 95% CI 42.1-60.2%). The inadequate disposal of fetuses was a risk factor, while carrying out serological tests to new animals that enter the herd, the use of new gloves and palpation utensils for each animal, supplementation, and stabling were stated as protective factors. No effect of positivity was found in the last calving interval. The implementation of bovine neosporosis control programs to support breeding and conservation programs of the BON breed in Colombia is recommended.

Experimental infection by Neospora caninum in gerbil reduces activity of enzymes involved in energy metabolism.

Neospora caninum is a protozoan that has tropism for the central nervous system. The aim of this study was to determine whether experimental infection of gerbils would interfere with activity of enzymes associated with energy metabolism. We randomized 20 gerbils into two groups (ten animals per group): the control group (healthy animals; uninfected) and the infected group (experimentally infected with dose 7.8 × 102 tachyzoites of N. caninum per gerbil). On day six and twelve post-infection (PI), brain and spleen tissues were collected for biochemical and histopathological analyses. No histopathological lesions were observed in the brains of infected animals; however, inflammatory infiltrates were found in the spleen. Significantly greater levels of reactive oxygen species (ROS) were observed in the brain and spleen of infected gerbils than in the control group at 12 days PI. Cytosolic creatine kinase (CK-CYT), mitochondrial creatine kinase (CK-MIT), and pyruvate kinase (PK) activities were lower in the brains of infected gerbils than in those of the control group on day 12 PI. There was significantly less CK-CYT activity in the spleens of infected gerbils on day 6 and 12 PI. Finally, there was significantly less sodium-potassium ion pump (Na+/K+ ATPase) activity in the brains and spleens of infected gerbils on day 12 PI. These data suggest that experimental infection with N. caninum interfered with energy metabolism associated with ATP homeostasis in the brain and spleen, directly or indirectly, apparently mediated by ROS overproduction, contributing to inhibition of Na+/K+ ATPase activity.

Urological detection of specific antibodies against Neospora caninum infection in mice: A prospect for novel diagnostic approach of Neospora.

The intracellular protozoan parasite Neospora caninum is incriminated to induce drastic economic losses in both livestock and pet animal industries. Neosporosis is primarily characterized by abortion in cattle and paralytic symptoms in dogs. Because there are no effective treatments or vaccines, diagnosis is critical for Neospora control. Thus, diversification of laboratory tests and specimens used for diagnosis of N. caninum is an essential scientific endeavor to judge and select the most appropriate diagnostic tool. Herein, we provide the first evidence for the utility of urine samples for demonstration of specific antibodies against N. caninum employing an experimentally infected murine model. Specific antibodies to recombinant N. caninum dense granule 7, surface antigen 1, and lysate antigen were assayed using different antibodies-based ELISAs. Urine based IgG ELISA efficiently discriminated between infected mice (acute or chronic infection), and those of non-infected mice. This effect was also noticed for IgG1 and IgG2a suggesting the utility of urine for assessment of T-helper 2- and T-helper 1-mediated immunities, respectively. In addition, reactivity of specific antibody in urine was also confirmed against parasites when indirect fluorescent antibody test was employed. Usefulness of urine as an additional clinical sample for Neospora diagnosis was confirmed via comparison with the relevant control non-infected and infected mouse sera as reference samples. Because of minimum invasiveness and ease of urine collection, this approach might offer new diagnostic opportunities for N. caninum either for the field or research purposes. However, further studies are required to extrapolate this preliminary study and results in the animal species of interest particularly in dogs.

Absence of Neospora caninum DNA in Human Clinical Samples, Spain.

Low antibody titers to Neospora caninum have been reported in humans, but infection has not been confirmed. We used N. caninum-specific PCR to test 600 clinical samples from patients with toxoplasmosis signs but Toxoplasma gondii-negative PCR results. We did not detect N. caninum DNA, demonstrating it is an unlikely opportunistic zoonotic agent.

Toxoplasma gondii, Neospora caninum and Encephalitozoon cuniculi in Animals from Captivity (Zoo and Circus Animals).

Problems with parasitic infections are common in zoological gardens and circuses. In some animals it can lead to several disorders such as systemic disease, reproductive disorders (abortions and neonatal mortality), and even to death if severe illness is untreated. Thus, the aim of this study was to evaluate the prevalence of three common parasites in 74 animals from three zoos, and four circuses in Southern Italy. Antibodies to Toxoplasma gondii, Neospora caninum, and Encephalitozoon cuniculi were detected in 51%, 12%, and 20% of animals, respectively. Co-infections of T. gondii and N. caninum were reported in seven animals (9%) and co-infection of T. gondii and E. cuniculi in one animal. T. gondii, N. caninum and E. cuniculi seroprevalence differed in type of diet (P ≤ 0.0001; P ≤ 0.037 and P ≤ 0.004, respectively). T. gondii and E. cuniculi seroprevalence also differed in animal families (P ≤ 0.0001) and according to type of housing (P ≤ 0.003), respectively. Statistical differences were not found in other characteristics (gender, age, country of birth, origin, and contact with cats or dogs). This is the first serological study focusing on protozoan and microsporidian parasites in zoo and circus animals from Southern Italy and the first detection of antibodies to E. cuniculi in camels in Europe.

Serology for Neosporosis, Q fever and Brucellosis to assess the cause of abortion in two dairy cattle herds in Ecuador.

BACKGROUND: Determining the infectious cause of abortion in cattle is difficult. This case-control study was set up to investigate the infectious causes of abortion by determining the seroprevalence of three reproductive pathogens in dairy cattle in Ecuador and their association with abortion: Brucella abortus, Neospora caninum and Coxiella burnetii. RESULTS: Ninety-five blood samples were obtained from cows that had experienced a mid- or late gestation abortion of their first calf and seventy-seven samples from a control group of cows with the same age that did not experience abortion problems. No antibodies were detected for B. abortus in any of the serum samples, but a high seroprevalence for both C. burnetii (52.9%) and N. caninum infection (21.5%) was found in group of cows. The seroprevalence of N. caninum infection in cattle that had experienced abortions was significantly higher (p < 0.05) than the seroprevalence in the control cows on one of the cattle farms, but no association between abortion and seropositivity for C. burnetii was found. CONCLUSION: We conclude that Neosporosis plays an important role in the epidemiology of abortion on one cattle farm, but that Q fever is apparently not an important cause for abortion in this setting.