Factors affecting plasma prolactin concentrations throughout gestation in high producing dairy cows

The aim of the present study was to investigate possible relationships between plasma concentrations of prolactin and the following factors throughout gestation in lactating dairy cows: photoperiod, season, milk production, Neospora caninum-seropositivity, twin pregnancy, and plasma concentrations of pregnancy associated glycoproteins-1 (PAG-1), progesterone and cortisol. Pregnancy was diagnosed by trans-rectal ultrasonography on Day 40 post-insemination and by palpation per rectum on Days 90, 120, 150, 180 and 210 or until abortion in aborting cows. Blood samples were collected from each animal immediately before each pregnancy diagnosis. The study population was comprised of 73 non-aborting (54 Neospora-seropositive cows) and 20 aborting cows (all Neospora-seropositive) cows. In non-aborting cows, GLM repeated measures of analysis of variance revealed that lactation number and days in milk had no effect on plasma prolactin concentrations throughout gestation, whereas high plasma prolactin concentrations were significantly associated with high plasma levels of cortisol and PAG-1, with Neospora-seropositivity, positive photoperiod (increasing day length), twin pregnancy, and with low plasma progesterone concentrations. An interaction among plasma prolactin, plasma cortisol and milk production was observed in that plasma prolactin concentrations differed significantly throughout gestation and were highest in high-producing cows with high cortisol levels. In Neospora-seropositive non-aborting versus aborting cows, mean prolactin concentrations failed to differ on Days 40, 90, 120, 150 and 180 of pregnancy, whereas prolactin values were significantly lower (P = 0.03) in aborting animals on Day 210 of pregnancy. Our results indicate that a positive photoperiod and Neospora-infection lead to increased plasma prolactin concentrations throughout gestation. Reduced prolactin concentrations detected in Neospora-seropositive aborting cows compared to non-aborting animals suggests a protective effect of prolactin in N. caninum infection.     

Gamma Interferon Production and Plasma Concentrations of Pregnancy‐Associated Glycoproteins 1 and 2 in Gestating Dairy Cows Naturally Infected With Neospora caninum

Gamma interferon (IFN‐γ) production and cross‐breed pregnancy have been attributed a role in protecting dairy cows infected with Neospora caninum against abortion. Plasma levels of pregnancy‐associated glycoproteins‐1 (PAG‐1) are a marker of placental/foetal well‐being and of PAG‐2 is an abortion risk indicator in chronically N. caninum‐infected animals. The present study examines, in cross‐breed pregnancies, interactions between IFN‐γ production and levels of PAG‐1 and PAG‐2 in non‐aborting naturally Neospora‐infected dairy cows. Data were obtained from 60 pregnant Holstein‐Friesian cows: 44 Neospora‐seropositive and 16 Neospora‐seronegative; 12 became pregnant using Holstein‐Friesian semen and 48 using Limousin semen. Blood samples were collected on Days 40, 90, 120, 150, 180 and 210 of gestation. Gamma interferon was only detected in the plasma of nine of the 44 Neospora‐seropositive cows, all of them became pregnant using Limousin semen. Through GLM procedures, in cows inseminated with Limousin semen and Neospora‐seropositive cows showing no IFN‐γ production, PAG‐1 concentrations were high and increased throughout gestation compared to the levels detected in cows inseminated with Holstein‐Friesian semen and Neospora‐seropositive cows producing IFN‐γ, respectively. In Neospora‐seronegative cows and in Neospora‐seropositive cows showing no IFN‐γ production, significantly increased PAG‐2 concentrations were observed on gestation Day 120. Our findings indicate that IFN‐γ production correlates negatively and the production of antibodies against N. caninum is uncorrelated with plasma PAG concentrations during gestation in Neospora‐infected dairy cows. Accordingly, IFN‐γ production could be linked to the transplacental migration of tachyzoites, which may cause a reduction in PAG levels.     

Effects of crossbreed pregnancies on the abortion risk of Neospora caninum-infected dairy cows

Previous studies have shown that the use of beef bull semen significantly reduces the rate of abortions due to Neospora caninum in artificially inseminated (AI) seropositive dairy cows. In addition, certain beef breeds could be more resistant to N. caninum infection and abortion than others. The aim of the present study was to determine whether different crossbreed pregnancies, those derived from Limousin, Charolais, Piedmontese or Belgian Blue semen, carry different risks of abortion in Neospora-infected dairy cows. The effects of possible interactions between maternal levels of N. caninum antibodies and the different breed crosses were also evaluated. The study was performed on five commercial Holstein–Friesian dairy herds in Northeast Spain with previously confirmed diagnoses of N. caninum infection in aborted foetuses. The study population was comprised of 1115 pregnancies: 633 pregnancies recorded after AI using Holstein–Friesian semen from 18 bulls and 482 after AI using beef semen from 27 bulls (304 inseminations using semen from Limousin bulls, 191 from Belgian Blue bulls, 89 from Piedmontese bulls and 49 from Charolais bulls). Abortion rates were 32.2% (155/482) and 15.2% (96/633) for seropositive cows inseminated with Holstein–Friesian and beef breed semen, respectively. Logistic regression analysis revealed the herd and the interaction between maternal N. caninum antibody titre and the different crossbreeds as significant factors affecting the abortion rate. Lowest abortion rates, similar to that shown by seronegative animals in the analysed herds (3.2%, 239/7432), were observed in dams AI using Limousin semen that had low (<30 relative index (RI) units) N. caninum antibody titres (2.1% abortion, 3/145) and these cows were used as reference. Compared to the cows used as reference, cows with low N. caninum antibody titres (<30 RI units) showed a similar risk of abortion when inseminated with Piedmontese or Charolais bull semen, but higher risk of abortion when inseminated with Holstein (17.9 times) or Belgian Blue (7.2 times) bull semen. All cows with high N. caninum antibody titres (≥30 RI units) had a higher risk of abortion, ranging from 8.9 times (cows inseminated with Limousine semen) to 37.8 times (cows inseminated with Piedmontese semen), compared to the cows used as reference. In conclusion, different crossbreed pregnancies carried different abortion risks in Neospora-infected dairy cows. The use of beef bull semen dramatically reduced the risk of abortion in dairy cows, especially if Limousin breed semen was used. Moreover, this reduction was found to be dependent on the N. caninum antibody titre such that the lowest incidence of abortions was recorded in Limousin semen inseminated cows with low antibody titres. Insemination of Neospora-seropositive cows with beef bull semen could both reduce the risk of abortion and avoid breeding replacements for infected cattle.     

Plasma Concentrations of Pregnancy‐Associated Glycoproteins Measured Using Anti‐Bovine PAG‐2 Antibodies on Day 120 of Gestation Predict Abortion in Dairy Cows Naturally Infected with Neospora caninum

The present study sought to determine: (i) the effects of Neospora caninum infection and twin pregnancy on plasma pregnancy‐associated glycoprotein‐2 (PAG‐2) concentrations throughout pregnancy and (ii) whether plasma PAG‐2 concentrations could predict abortion in N. caninum‐infected cows. The study was performed on a commercial Holstein‐Friesian dairy herd in northeastern Spain and the final data included those recorded in 53 non‐aborting and 19 aborting animals. Blood samples were collected immediately before pregnancy diagnosis (on Days 40, 90, 120, 150, 180 and 210 post‐insemination) in non‐aborting cows or until the time of abortion detection in aborting cows. General lineal models (GLM) repeated measures anova revealed the different behaviour of PAG‐1 and PAG‐2, and significant effects of Neospora seropositivity, cool season and twin pregnancy on plasma PAG‐2 concentrations throughout gestation (between‐subject effects). In addition, based on the odds ratios, the likelihood of abortion increased in Neospora‐seropositive cows (by a factor of 7.0) compared to seronegative animals and decreased in cows with a high plasma PAG‐2 concentration (>4.5 ng/ml) on Day 120 of pregnancy (by a factor of 0.24), compared to the remaining cows. In conclusion, there is a relationship between plasma PAG‐2 concentrations and the risk of abortion in Neospora‐infected dairy cows. Thus, plasma PAG concentrations measured using anti‐boPAG‐2 antiserum on Day 120 of gestation could serve as an indicator of the abortion risk in N. caninum infected animals; values <4.5 ng/ml indicating a high risk of abortion in chronically infected animals.     

Some Factors Affecting the Abortion Rate in Dairy Herds with High Incidence of Neospora‐Associated Abortions are Different in Cows and Heifers

The aim of this study was to determine if the factors affecting the abortion rate in dairy herds with high incidence of Neospora‐associated abortions are different in pregnancies of cows and heifers chronically infected with Neospora caninum. In heifers (n = 229), an increase in the cumulative number of days with a mean relative humidity (RH) lower than 60% during the second trimester of gestation increases the risk of abortion. Yet, the likelihood of abortion was 7.6 times lower for pregnant heifers inseminated with Limousin bull semen, compared with those inseminated with Holstein‐Friesian bull semen. In pregnancies of parous cows (n = 521), an increase in rainfall and in the cumulative number of days with a mean RH lower than 60% during the second trimester of gestation increased the abortion rate. However, in contrast, an increase in the lactation number produced a decrease in the abortion rate, with a likelihood of abortion 4.8 times lower for pregnant cows inseminated with Limousin bull semen, and three times lower for those inseminated with Belgian Blue bull semen, compared with dairy cows inseminated with Holstein‐Friesian bull semen. Finally, the likelihood of abortion was 3.2 times lower for pregnancies of parous cows with low antibody titres against N. caninum (6–30 units) as compared to those with high antibody titres (≥30 units), whereas in heifers this variable had no effect. The practical recommendations of the present study include the control of the cow environment during the second trimester of gestation, the priority of culling for parous cows with higher antibody titres against N. caninum and the insemination of Neospora‐seropositive cows with semen from the Limousin breed.     

Neospora‐associated Abortion Episode over a 1‐Year Period in a Dairy Herd in North‐east Spain

This report describes a retrospective study based on serological screening, performed on a Neospora‐associated abortion episode over a 12‐month period in a dairy herd in north‐east Spain. During the 1‐year study period, the overall abortion rate for the herd was 23.2% (38 abortions of 164 diagnosed pregnancies). The data analysed were those derived from blood samples collected from the whole herd (n = 237) and from diagnosed pregnancies. Antibodies to Neospora caninum were found in 35.4% of the cattle with 44% of seropositive pregnant animals aborting over 1‐year period. Based on the odds ratio, the risk of abortion was 12.2 times higher (P < 0.0001) in the Neospora‐seropositive animals than in seronegative animals and significantly higher during the second term of gestation (P < 0.01) than during the first and third terms. Abortions were not found to be associated with parity status or season of pregnancy, and the common risk factors associated with pregnancy loss in the geographical area of the study. Age‐related differences in N. caninum seroprevalence were not statistically significant, indicating vertical transmission as the main route of infection. Indeed, a high percentage of congenitally infected offspring was observed (90.6%) and the farm had been free of dogs for the last 7 years. Our results suggest that, when a dairy herd shows an increased incidence of abortions due to Neosporosis, maternal serology can be a good indicator of the abortion risk in individual cows, and that the effects of factors normally related to abortion, such as parity and pregnancy season, may be masked.     

Associations of Neospora caninum seropositivity with gestation number and pregnancy outcome in Danish dairy herds

The prevalence and distribution of seropositivity towards the protozoan parasite Neospora caninum were studied in single blood samples from 1561 cows from 31 Danish dairy herds. Blood samples were analysed by an indirect enzyme-linked immunoassay and an indirect fluorescent-antibody test. Seroprevalence in 15 herds with previous abortions assigned to neosporosis ranged from 1% to 58%, with a mean frequency of 22%. In eight out of 16 herds without a history of N. caninum related abortions, no seroreactors were found. In the remaining eight herds, the seroprevalence ranged from 6% to 59%. The prevalence and distribution of seropositivity, gestation number prior to sampling, and breed were related to abortions and perinatal deaths using a random-effects logistic-regression model. Abortion risk was significantly increased in seropositive animals (OR=3) and in 2nd-gestation cows (OR=3). Perinatal death was significantly influenced by gestation number and breed, but not by serostatus. Reproductive performance and culling risk of cows were not affected by serostatus. Seropositivity increased with “age” (i.e. gestation number) (P=0.02). In open cows, seropositivity tended to decrease with distance from calving (P=0.05). The proportion of seropositive pregnant cows increased with trimester (P=0.02).     

The Use of Beef Bull Semen Reduced the Risk of Abortion in Neospora‐seropositive Dairy Cows

There is an evidence that the epidemiology of neosporosis differs in dairy and beef cattle, such that beef cattle carry a lower risk of abortion. The aim of the present study was to establish whether artificial insemination using semen from beef bulls could reduce the risk of abortion in dairy cows seropositive for the Neospora caninum parasite. Our study was based on yearly serological screening for neosporosis and on the confirmation of Neospora infection in aborted fetuses in two high‐producing dairy herds with a mean 28% seroprevalence of N. caninum antibodies. The study population comprised of 273 pregnancies in seropositive animals: 156 pregnancies monitored after insemination using Holstein–Friesian semen and 117 after insemination using beef bull semen. Abortion rates for these animals were 28.2% (77 of 273), 34.6% (54 of 156) and 19.7% (23 of 117). Logistic regression analysis indicated no significant effects of lactation number and previous abortion on the abortion rate. Based on the odds ratio, a 1‐unit increase in the Neospora antibody titre yielded a 1.01‐fold increase in the abortion rate. The likelihood of abortion was two times higher for cows in one of the two herds and 2.8 times lower (one of 0.36) for pregnant cows inseminated with beef bull semen rather than Holstein–Friesian semen. Our results indicate that the use of beef bull semen can reduce the risk of abortion in dairy cows, and suggest that annual screening for neosporosis, specifically the antibody titre to the protozoan, could be an useful predictor of abortion risk in reproductive health programmes.     

Relationship between Rainfall and Neospora caninum‐associated Abortion in Two Dairy Herds in a Dry Environment

The possible direct relationship between climate variations and abortion in Neospora caninum‐infected cows has not been studied. The objective of this study was to determine whether climate changes could be a risk factor for abortion in N. caninum‐infected cows, and was based on yearly serological screening for neosporosis and on the confirmation of N. caninum infection on aborted fetuses in two high‐producing dairy herds with a mean 27% seroprevalence of N. caninum antibodies. The final population study was comprised of 357 pregnancies in seropositive animals. Logistic regression analysis indicated no significant effects of herd, N. caninum antibody titre, climate variables during the first and third trimesters of gestation, mean and maximum temperature–humidity index values during the second trimester of gestation, and previous abortion on the abortion rate. Based on the odds ratio, a 1‐unit increase in lactation number yielded a 0.85‐fold decrease in the abortion rate. The likelihood of abortion was 1.9 times (1/0.54) lower for pregnant cows inseminated with beef bull semen compared with Holstein‐Friesian bull semen. The likelihood of abortion decreased significant and progressively by factors of 0.5, 0.41 and 0.3 for the respective classes 40–49, 30–39 and <30 rainfall mm during the second trimester of gestation (using the class ≥60 rainfall mm as reference). As a general conclusion, it seems that increased rainfall in a dry environment can compromise the success of gestation in N. caninum‐infected cows. Attempts should therefore be made to reduce environment effects during the second trimester of gestation, a period in which the immune response of cows is diminished.     

Neospora caninum Infection Does Not Affect the Fertility of Dairy Cows in Herds with High Incidence of Neospora‐associated Abortions

This present study, was designed to establish whether Neospora caninum infection affects the fertility of high‐producing dairy cows, and was based on yearly serological screening for neosporosis and on the confirmation of N. caninum infection in aborted foetuses. Assessment was made of 7518 artificial inseminations (AI) (applied to parous cows) performed in three herds. Of these inseminations, 2540 (33.8%) resulted in pregnancy; 34% of which corresponded to seronegative cows (2226 of 6556 AI performed in seronegative cows) and 32.6% to seropositive animals (314 of 962). Abortion occurred in 97 (30.1%) of the 314 pregnancies recorded in dams seropositive for N. caninum, while only 93 (4.2%) of the 2226 pregnancies in seronegative animals ended in abortion. Logistic regression analysis indicated no significant effects of N. caninum antibody titre, days in milk and milk production at insemination on fertility. Factors found to affect fertility were herd, season of insemination, lactation and insemination number, semen‐providing bull and AI technician. Our results indicate that N. caninum infection does not affect the fertility of high‐producing dairy cows.     

Immune responses during the peripartum period in dairy cows with postpartum endometritis

Determining the immune responses to the development of endometritis during the peripartum period may assist in the development of more efficient reproductive management regimens for dairy herds. In this study, we compared the peripartum immune responses of dairy cows that develop endometritis by 4 weeks postpartum (n = 11) to cows that did not develop this disease (n = 19). Blood samples were collected 1 week before calving, just after or during calving, and then at weeks 1, 2, 3, and 4 postpartum. Cows that developed endometriris had significantly higher total leukocyte, neutrophil, lymphocyte, and monocyte counts than the control cows (P < 0.05) at all time points. The leukocytes from cows that developed endometritis were significantly less phagocytic than those from control cows at all sampling time points (P < 0.01). The serum TNFalpha concentrations of the control cows decreased linearly from the prepartum time point (P = 0.0029), but the endometritis cows showed a different profile (P > 0.05). As a result, the serum TNFalpha concentrations were greater in the endometritis group (P < 0.01) than in the control group during the third and fourth weeks postpartum. The greater total leukocyte numbers and neutrophil, lymphocyte and monocyte counts, and the maintenance of elevated serum TNFalpha levels in the cows with endometritis may be due to infection in the postpartum period. Furthermore, the decreased phagocytic capacity of leukocytes during the peripartum period, including at the prepartum time point, makes cows more susceptible to postpartum endometritis.     

Neospora caninum versus Brucella spp. exposure among dairy cattle in Ethiopia: A case control study

This case-control study aimed at assessing the relative association of Neospora caninum and Brucella species exposure with reproductive disorders. The study was carried out between October 2011 and June 2012 on 731 dairy cows sampled from 150 dairy farms in selected 17 conurbations of Ethiopia. Two hundred sixty-six of the cows were categorized as cases based on their history of abortion or stillbirth while the remaining 465 were controls. The presence of antibody to N. caninum was screened using indirect ELISA, while Brucella spp. exposure was assayed serially using Rose Bengal Plate Test and Complement Fixation Test. Exposure to N. caninum was more frequently observed among cases (23.8 %) than controls (12.7 %), while no significant difference (p?>?0.05) was noted for Brucella exposure between the two groups. Moreover, the proportion of cows with disorders like retention of fetal membrane, endometritis and increased inter-calving period were significantly higher (p?<?0.05) among Neospora seropositive cows. In conclusion, the finding discloses the strong association of N. caninum with reproductive disorders compared to Brucella spp. exposure. However, neither N. caninum nor Brucella spp. could explain the majority (73.2 %) of the reported abortions and stillbirths in cattle. Hence, this observation underscores the need for more intensive investigation on the identification of causes of the aforementioned disorders in dairy cattle of Ethiopia.     

Glucocorticoid receptor down-regulation in neutrophils of periparturient cows

OBJECTIVE: To determine effects of parturition on glucocorticoid receptor (GR) expression in neutrophils, serum cortisol concentration, and total blood leukocyte and neutrophil counts in periparturient dairy cows. ANIMALS: 23 Holstein cows. PROCEDURE: Blood samples were collected from 8 multiparous and 5 primiparous periparturient cows at various times from 28 days before parturition until 14 days after parturition. Glucocorticoid receptor expression in neutrophils, serum cortisol concentration, and total blood leukocyte and neutrophil counts were determined. Results were compared with results from control samples obtained from 5 multiparous and 5 primiparous Holstein cows in midpregnancy. RESULTS: Neutrophils from periparturient cows had 49% reduction in GR expression at calving, compared with GR expression 2 to 4 weeks before calving, and 39% reduction, compared with neutrophils from cows in midpregnancy. Reduction in neutrophil GR expression began 1 week before calving and was most severe at calving and 24 hours after calving; a significant difference in GR expression was detected between primiparous and multiparous cows. Serum cortisol concentrations and total leukocyte and neutrophil counts were significantly increased at calving and returned to baseline values by 24 hours after calving. Significant negative correlations were detected between neutrophil GR expression and serum cortisol concentration, total leukocyte count, and neutrophil count. CONCLUSIONS AND CLINICAL RELEVANCE: Reduced GR expression in blood neutrophils of periparturient dairy cows was associated with increased serum cortisol concentrations, leukocytosis, and neutrophilia. Thus, GR down-regulation in neutrophils may be involved in periparturient neutrophil dysregulation and may cause increased susceptibility to mastitis.     

Experimental Neospora Caninum Infection in Pregnant Dairy Heifers Raises Concentrations of Pregnancy‐Associated Glycoproteins 1 and 2 in Foetal Fluids

Plasma concentrations of PAG‐1 are used for pregnancy diagnosis and as a marker of placental/foetal well‐being, while those of PAG‐2 may be an indicator of abortion risk in Neospora caninum‐infected cows. Studies have shown that N. caninum infection modifies PAG‐1 and PAG‐2 patterns in maternal blood plasma. However, no prior work has examined the effects of N. caninum infection on concentrations of PAGs in foetal fluids. In this study, PAG‐1, PAG‐2 and pH levels were determined in the amniotic and allantoic fluids of foetuses collected at 152 days of gestation from control uninfected dams and from dams experimentally infected with N. caninum on Day 110 of gestation. Foetal fluids from infected foetuses had significantly higher PAG‐2 concentrations (p = 0.026) and pH values (p = 0.02) than fluids from non‐infected foetuses. In infected foetuses, significantly higher concentrations of PAG‐1 (p < 0.001) and PAG‐2 (p < 0.001) were detected in fluid samples showing antibodies against N. caninum than those without antibodies. Moreover, pH values were significantly higher (p = 0.011) in foetal fluid samples with antibodies than in samples from non‐infected foetuses. In conclusion, this is the first report on the effect of N. caninum infection on PAG levels in foetal fluids. Our results indicate that following the experimental infection of dams with N. caninum on Day 110 of gestation, foetal fluids collected from the infected foetuses of these dams featured higher PAG‐1 and PAG‐2 levels and pH values than fluids from non‐infected controls, provided that the samples tested showed the presence of antibodies. The clinical implications of these findings are that following infection with N. caninum, most cows will experience some level of placental damage and that this injury correlates with foetal fluid PAG levels and pH.     

Associations between pregnancy outcome and serological response to Neospora caninum among a group of dairy heifers

AIM: To monitor pregnancy in a group of rising 2-year-old dairy heifers on a farm on which abortion due to Neospora caninum was known to occur in previous years.

METHODS: A prospective cohort study group of 164 rising 2-year-old heifers was pregnancy-tested and blood-sampled at 4–5-week intervals throughout gestation. Sera were tested for antibodies to N. caninum at 3–4-month intervals, using an enzyme-linked immunosorbent assay (ELISA). When loss of pregnancy was detected, an N. caninum indirect fluorescent antibody test (IFAT) was conducted retrospectively on stored sera collected the month before abortion, the month abortion was detected, and for the following 2 months, from heifers that aborted. All fetal and placental material detected following abortion was subjected to gross post-mortem and histopathological examination.

RESULTS: Eleven of 18 (61%) heifers that were seropositive and 4/146 (3%) heifers that were seronegative to N. caninum by ELISA, aborted. The relative risk for abortion among ELISApositive heifers was 23.6. Abortion occurred predominantly between Days 120 and 152 gestation among the ELISA-positive heifers and throughout gestation among the ELISA-negative heifers. IFAT titres rose around the time of abortion in most of the heifers that were previously seropositive by ELISA, but dropped rapidly again in post-abortion samples. IFAT titres among 4/6 ELISA-positive heifers that did not abort increased, but later in gestation than the time other heifers aborted. IFAT titres remained negative in heifers that aborted that were ELISAnegative.

CONCLUSIONS: Heifers that were seropositive to N. caninum by ELISA had a much greater risk of abortion than seronegative heifers. Most seropositive heifers showed evidence of a reactivation of infection during pregnancy. High (≥1:2,000) N. caninum IFAT titres also occurred in non-aborting heifers.

CLINICAL RELEVANCE: Culling of replacement heifers seropositive to N. caninum may be a cost-effective strategy for minimising risk of abortion. Pregnancy testing heifers before 5 months gestation may overestimate the number that calve in N. caninum-infected herds, but would assist in documenting the occurrence of abortion. Reliance on a high (>1:2,000) IFAT titre to rule-in N. caninum as a cause of abortion is likely to produce false-positive results.     

Seroprevalence of Neospora caninum antibodies and its consequences for reproductive parameters in dairy cows from Dakar–Senegal,West Africa

The aim of this study was to determine the seroprevalence of Neospora caninum antibodies and its effects on reproductive parameters in cows in intensive dairy herds in Dakar. Randomised blood samples were taken for serology from 196 cows in four herds with a history of sporadic abortion. All of the sera were assayed for antibodies against N. caninum, Candida guillermondii, Coxiella burnetii, and Chlamydophila sp. The associations between serostatus and reproductive parameters (abortion, number of inseminations to conception, and calving to conception interval) were assessed over a period of 5 years (2004–2008). The seroprevalence of N. caninum antibodies in dairy cattle was 17.9%. The local Gobra breed and crossbreeds had higher levels of N. caninum antibodies than exotic breeds (p < 0.05). For the most recent pregnancies, seropositive cows required more inseminations to establish conception than seronegative cows (p < 0.05). The results indicate that dairy cattle from Dakar are exposed to N. caninum. Neosporosis should, therefore, be systematically considered as a cause when the calving to conception interval is prolonged.     

Changes in plasma concentrations of S100A7 and S100A8 in dairy cows during pregnancy

This study was carried out to examine the changes in plasma concentrations of the Ca‐binding antimicrobial proteins S100A7 and S100A8 during pregnancy in dairy cows. Holstein Friesian cows (n = 19) were inseminated with Holstein Friesian semen. Blood was collected at days 30, 60, 90, 120, 150, 180, 210, 240 and 270 after insemination. Plasma was used for measuring the concentrations of S100A7 and S100A8. Both S100A7 and S100A8 concentrations showed similar patterns during gestation; they increased during the midgestation, between days 90 and 180, and then declined before calving. The findings indicated that plasma concentrations of S100A7 and S100A8 did not change significantly during pregnancy in cows. Further studies are required to determine the roles of S100A7 and S100A8 in physiological function during pregnancy in dairy cows.     

Neospora caninum tachyzoites inoculated by the conjunctival route are not vertically transmitted in pregnant cattle: A descriptive study

The aim of this study was to evaluate whether Neospora caninum tachyzoites (Nc-1) inoculated by the conjunctival route in pregnant cows were able to generate infection in their fetuses. Group 1 contained 2 naturally infected cows; group 2 contained two cows inoculated intravenously with 2.5 × 10⁸ tachyzoites, group 3 contained two cows inoculated with 2.5 × 10⁸ tachyzoites by the conjunctival route, and group 4 contained two uninfected control cows. The four inoculated cows from groups 2 and 3 were challenged at 23 weeks of gestation. An indirect fluorescent antibody test (IFAT), recombinant NcGRA7-based ELISA, ELISA for IgG subisotypes and Western blot analysis were assessed to characterize the humoral immune response in dams. Sera from their fetuses were tested also using Western blot analysis. Routine microscopic evaluation of H&E stained fetal tissues was made and any fetal tissues and placentas with lesions compatible with Neospora-infection were processed by immunohistochemistry (IHC). DNA extraction from fresh and formalin-fixed, paraffin-embedded fetal tissues were tested by nested PCR. All dams from groups 1, 2 and 3 were seropositive by IFAT, rNcGRA7-based-ELISA and Western blot. IgG₁/IgG₂ ratios were ≤1 at weeks 27 and 29 of gestation. Only fetuses from groups 1 and 2 developed N. caninum specific antibodies by Western blot. Histopathological lesions compatible with those caused by N. caninum were observed in fetuses from groups 1 and 2. N. caninum cysts and tachyzoites were observed by IHC on fetal tissues from groups 1 and 2. Only fetal samples from group 2 were positive by PCR. Further work is needed not only to characterize the cellular immune response but also to clarify the consequences on the dam after conjunctival inoculation of N. caninum tachyzoites. This study shows that N. caninum tachyzoites inoculated by the conjunctival route were not vertically transmitted in pregnant cows.     

Cholinesterase as an inflammatory marker of subclinical infection of dairy cows infected by Neospora caninum and risk factors for disease

The aim of this study was to evaluate the seroprevalence of Neospora caninum in dairy cows, as well as to verify the risk factors for N. caninum infection and inflammatory response in dairy cows. Using the indirect immunofluorescence reaction, the seroprevalence of neosporosis was determined to be 32.82%. Based on regression analysis, the presence of dogs at a farm increased the probability of a cow testing positive for N. caninum (OR = 20.01 [5.21–123.12]). These data suggest that N. caninum has a relevant prevalence in dairy cows of the Microregion of Rio do Sul (Brazil), with elevated frequencies of anti-N. caninum IgG. The data also suggest that the parasite is widely distributed in dairy herds of the micro-region, because 94.4% of properties screened had at least one seropositive animal. The principal risk factors for disease maintenance in herds may be considered the presence of dogs and absence of a diagnostic test when introducing new animals. Blood from these cows was used to measure variables related to the inflammatory response. Serum cholinesterase activity, as well as serum levels of globulins and C-reactive protein were higher in seropositive to N. caninum than in seronegative cows. Furthermore, the infection by parasite causes an intense inflammatory process, contributing to disease pathophysiology.     

Phenotypic characterisation of the cellular immune infiltrate in placentas of cattle following experimental inoculation with Neospora caninum in late gestation

Despite Neospora caninum being a major cause of bovine abortion worldwide, its pathogenesis is not completely understood. Neospora infection stimulates host cell-mediated immune responses, which may be responsible for the placental damage leading to abortion. The aim of the current study was to characterize the placental immune response following an experimental inoculation of pregnant cattle with N. caninum tachyzoites at day 210 of gestation. Cows were culled at 14, 28, 42 and 56 days post inoculation (dpi). Placentomes were examined by immunohistochemistry using antibodies against macrophages, T-cell subsets (CD4, CD8 and γδ), NK cells and B cells. Macrophages were detected mainly at 14 days post inoculation. Inflammation was generally mild and mainly characterized by CD3⁺, CD4⁺ and γδ T-cells; whereas CD8⁺ and NK cells were less numerous. The immune cell repertoire observed in this study was similar to those seen in pregnant cattle challenged with N. caninum at early gestation. However, cellular infiltrates were less severe than those seen during first trimester Neospora infections. This may explain the milder clinical outcome observed when animals are infected late in gestation.