Prevalence, Risk Factors, and Relationship between Reproductive Performance and the Presence of Antibodies against Coxiellosis in Dairy Farm Milk Tanks in the Northwest of Spain.

Q fever is a zoonotic disease that has been associated with reproductive problems in animals. As there is little epidemiological data regarding the distribution and risk factors of this disorder in cattle, the objective of this study was to evaluate the prevalence of Coxiella burnetii among dairy herds in the northwest of Spain, and to determine the on-farm risk factors associated with the disease and its effects on reproductive performance. Bulk tank milk (BTM) samples were collected from 262 commercial dairy herds from A Coruña, Lugo, and Pontevedra provinces. Data about location, mean age, and herd management features were obtained. A commercial indirect ELISA kit was used to determine the presence of antibodies against C. burnetii in BTM samples. The relationship between seropositivity to C. burnetii and the risk factors was checked using a Pearson's χ2 test and a classification tree analysis. In addition, a one-way ANOVA test and the Mann-Whitney U test were used to check the impact of seropositivity to C. burnetii on reproductive performance. A total of 60.1% of the farms tested positive for coxiellosis, the herd size, the external purchase of livestock, and the geographical area were identified as the main risk factors. Conception rate and first-service conception rate were significantly lower (p < 0.05) in positive farms (37.1 and 32.9%) compared to negative farms (39.8 and 36.1%). Similarly, positive farms had significant higher incidence of endometritis (13.7% vs. 11.2%, p < 0.05). Consequently, a high seropositivity and slightly negative effects of coxiellosis on reproductive performance were observed, which intensifies the need for further research, including the identification an active infection in positive herds and the characterization of the genotype.

Spatiotemporal monitoring of myxomatosis in European wild rabbit (Oryctolagus cuniculus) in Spanish Mediterranean ecosystems.

A long-term active epidemiological surveillance programme was conducted to determine seroprevalence to myxoma virus (MYXV), infection prevalence and spatiotemporal patterns and factors associated with MYXV circulation in wild rabbits (Oryctolagus cuniculus) in Spanish Mediterranean ecosystems. A total of 2376 animals were sampled over four study periods: 2009-2012 (P1), 2012-2015 (P2), 2015-2018 (P3) and 2018-2021 (P4). Antibodies against MYXV were detected by a commercial indirect ELISA in 59.9% (1424/2376; 95% CI: 58.0-61.9) of wild rabbits. At least one seropositive animal was detected on 131 (96.3%) of 136 game estates sampled. MYXV infection was confirmed by PCR in 94 of 1063 (8.8%; 95% CI: 7.3-10.7) wild rabbits. Circulation of the novel recombinant MYXV (ha-MYXV) was not found in wild rabbits analysed during P4. Five statistically significant spatiotemporal clusters of high MYXV seroprevalence were identified using a Bernoulli model: one in P2 and four in P3. A generalized linear mixed model (GLMM) analysis identified sampling season (autumn), age (adult and juvenile), outbreaks of myxomatosis in the month prior to sampling, mean annual temperature, humidity and seropositivity to rabbit haemorrhagic disease virus as factors potentially linked with MYXV seropositivity. GLMM analysis identified outbreaks of myxomatosis in the month prior to sampling, MYXV seropositivity and presence of lesions compatible with myxomatosis as factors associated with MYXV infection. The results indicate high exposure, widespread but non-homogeneous distribution, and endemic circulation of MYXV in wild rabbit populations in southern Spain during the last decade. Prevalence of antibodies against MYXV showed fluctuations both within the year and over the study periods, revealing variations in the immunity of wild rabbit populations in Mediterranean ecosystems that could increase the risk of MYXV re-emergence in immunologically naïve populations. The present study highlights the importance of long-term surveillance to better understand the epidemiology of MYXV in wild lagomorphs.

Epidemiological surveillance of Schmallenberg virus in small ruminants in southern Spain.

Schmallenberg virus (SBV) is an emerging Culicoides-borne Orthobunyavirus that affects ruminant species. Between 2011 and 2013, it was responsible for a large-scale epidemic in Europe. In the present study, we aimed to determine the seroprevalence, spatial distribution and risk factors associated with SBV exposure in sheep and goats in the region where the first Schmallenberg disease outbreak in Spain was reported. Blood samples from 1,796 small ruminants from 120 farms were collected in Andalusia (southern Spain) between 2015 and 2017. Antibodies against SBV were detected in 536 of 1,796 animals (29.8%; 95%CI: 27.7-32.0) using a commercial blocking ELISA. The individual seroprevalence according to species was 31.1% (280/900; 95%CI: 28.1-34.1) in sheep and 28.6% (256/896; 95%CI: 25.6-31.5) in goats. The farm prevalence was 76.7% (95%CI: 69.1-84.2). Seropositivity to SBV was confirmed in both sheep and goats in all provinces by virus neutralization test. Two significant (p < .001) spatial clusters of high seroprevalence were identified. The generalized estimating equation analysis showed that management system (extensive), temperature (>14ºC) and altitude (<400 metres above sea level) were risk factors associated with SBV exposure in small ruminants. Our results highlight widespread but not homogeneous circulation of SBV in small ruminant populations in Spain.

Epidemiological study of the association between bovine gammaherpesvirus type 4 and reproductive disease in dairy cattle from northwestern Spain.

Bovine gammaherpesvirus 4 (BoHV-4) has controversially been related with cattle reproductive disease. In the present study we analyze the relationship between exposure to BoHV-4 and reproductive performance in dairy cattle from northwestern Spain. A total of 2022 sera from 50 farms were examined to detect anti-BoHV-4 antibodies. Herd and individual reproductive records were collected to analyze association with exposure to BoHV-4. In addition, 52 abortion cases were examined to detect BoHV-4 DNA. An individual seroprevalence of 66.6 % and a herd prevalence of 98 % were found. Exposure to BoHV-4 increased with age, particularly in individuals between 26-36 months old (OR = 2.7; CI 95 %: 1.2-5.0, compared to animals < 26 months). Seroprevalence was not associated with herd fertility and herd abortion rate, but seropositive animals between 26-36 months presented prolonged calving to fertilizing insemination intervals (HR: 1.4; CI 95 %: 1.2-2.0) as well as higher odds of an unsuccessful 1st insemination (OR: 2.5; CI 95 %: 1.2-5.0). In abortion cases, BoHV-4 DNA was found in 12 vaginal swabs from 5 farms but not in any fetal tissue. Our results reveal an endemic, high and widespread exposure to BoHV-4 among dairy cattle from NW Spain with a limited impact in the reproductive performance of herds. The significantly worse reproductive performance of seropositive animals of 26-36 months of age may be the consequence of the establishment of primo-infections when moving heifers to lactation lots. Our findings may be useful to understand the potential population impact of BoHV-4.

Spatiotemporal monitoring of selected pathogens in Iberian ibex (Capra pyrenaica).

An epidemiological surveillance programme was carried out to assess exposure and spatiotemporal patterns of selected pathogens (Brucella spp., Mycobacterium avium subsp. paratuberculosis (MAP), Mycoplasma agalactiae, Pestivirus and bluetongue virus (BTV)) in Iberian ibex (Capra pyrenaica) from Andalusia (southern Spain), the region with the largest population of this species. A total of 602 animals in five distribution areas were sampled during 2010-2012 (P1) and 2013-2015 (P2). The Rose Bengal test (RBT) and complement fixation test (CFT) were used in parallel to detect anti-Brucella spp. antibodies. Commercial ELISAs were used to test for antibodies against the other selected pathogens. Sera positive for BTV and Pestivirus by ELISA were tested by serum neutralization test (SNT) to identify circulating serotypes/genotypes. The overall seroprevalences were as follows: 0.4% for Brucella spp. (2/549; CI 95%: 0.1-1.3) (14/555 positive by RBT; 2/564 by CFT), 0.5% for MAP (3/564; CI 95%: 0.1-1.5), 5.7% for M. agalactiae (30/529; CI 95%: 3.9-8.0), 11.1% for Pestivirus (58/525; CI 95%: 8.5-14.1) and 3.3% for BTV (18/538; CI 95%: 2.0-5.2). Significantly higher seropositivity to both M. agalactiae and BTV was observed in P1 compared with P2. Spatiotemporal clusters of high seroprevalence were also found for M. agalactiae in four of the five sampling areas in 2010, and for BTV in one of five areas in 2012. Specific antibodies against BTV-4, BDV-4 and BVDV-1 were confirmed by SNT. Our results indicate that the Iberian ibex may be considered spillover hosts of Brucella spp. and MAP rather than true reservoirs. The prevalence of antibodies against M. agalactiae and BTV suggests spatiotemporal variation in the circulation of these pathogens, while Pestivirus has a moderately endemic circulation in Iberian ibex populations. Our study highlights the importance of long-term surveillance for a better understanding of the spatiotemporal distribution of shared infectious diseases and providing valuable information to improve control measures at the wildlife-livestock interface.

Seroepidemiology of Toxoplasma gondii in extensively raised Iberian pigs in Spain.

Pigs reared under extensive farming conditions are currently in high commercial demand because they are associated with high-quality products. Nevertheless, the risk of contact with different pathogens of animal and public health concern is also higher in extensive production systems. Toxoplasma gondii is a widely prevalent zoonotic pathogen and transmission by contaminated pork is likely one of the main routes of human toxoplasmosis. The aim of this study was to determine the seroprevalence, risk factors and spatial distribution of T. gondii on extensive Iberian pig herds in Spain. Sera from 2245 Iberian pigs from 114 herds were collected between 2015 and 2017 and analyzed using a commercial ELISA. The apparent individual prevalence of antibodies against T. gondii was 24.1 % (542/2245) and the estimated true seroprevalence was 24.3 % (CI95 %: 22.5-26.1). Seropositivity was detected in 86.0 % (98/114; CI95 %: 77.4-91.1) of 114 herds analyzed. A multi-level logistic regression model showed that T. gondii infection was significantly more frequent in sows than in fattening pigs (OR: 2.6; CI95 %: 1.5-4.8) and in herds with more than three cats compared to no cats (OR: 2.9; CI95 %: 1.1-8.7). Our results indicate a widespread but heterogenous distribution of T. gondii in extensively reared Iberian pig herds, which may have important implications for public health through the consumption of undercooked or improperly cured pork products.

Prevalence and molecular characterization of Rickettsia spp. in questing ticks from north-western Spain.

Tick-borne rickettsioses, most of them belonging to the spotted fever group (SFG), have been recognized as important emerging vector-borne zoonotic diseases. In order to determine the presence of Rickettsia spp. in questing ticks from north-western Spain, 1056 Ixodes ricinus, 19 Dermacentor marginatus, 17 Dermacentor reticulatus and one Ixodes acuminatus were processed. Rickettsia DNA was detected by PCR targeting rOmpA and rOmpB genes. A total of 219 (20.7%) I. ricinus, 19 (100%) D. marginatus and four D. reticulatus (23.5%) were positive. The prevalence was significantly higher in I. ricinus from coastal areas and in winter. Five species were identified: Rickettsia felis, Rickettsia monacensis, Rickettsia raoultii, Rickettsia slovaca and "Candidatus Rickettsia rioja". Our results reveal a significant presence of some pathogenic Rickettsia species in questing tick populations from this area which involves a noticeable risk of rickettsiosis. As R. raoultii, R. slovaca and "Ca. R. rioja" DNA were identified in I. ricinus, considered an unusual vector for these Rickettsia species, further studies are needed to unravel the role of that tick species in the maintenance and transmission of these three Rickettsia species in north-western Spain.

Serological survey of Toxoplasma gondii in captive nonhuman primates in zoos in Spain.

Toxoplasma gondii is a widely distributed zoonotic protozoan parasite, which can affect most warm-blooded species. Some species of non-human primates (NHPs) are highly susceptible to T. gondii infection. The aim of the study was to determine the seroprevalence and risk factors associated with T. gondii infection in NHPs housed in zoos in Spain. Sera from 189 NHPs belonging to 33 species were collected in eight zoos. Additionally, 10 of the 189 animals were longitudinally sampled. Anti-T. gondii antibodies were detected in 48 NHPs (25.4%; confidence interval of 95% (CI95%): 19.2-31.6) using a modified agglutination test (MAT; cut-off = 25). Seropositive animals had titers of 25 (6.3%), 50 (8.3%), 100 (8.3%) and ≥500 (68.8%). Seropositivity was detected in 15 of the 33 species (45.5%). Of the 10 NHPs sampled more than once, two animals (one Barbary macaque [Macaca sylvanus] and one common chimpanzee [Pan toglodytes]) seroconverted along the study period, while one seropositive chimpanzee increased antibody titers over time. The Hominidae family (OR = 5.9; CI95%: 2.7-12.8) and sex (females) (OR = 2.1; CI95%: 1.1-4.1) were risk factors potentially associated with seropositivity to T. gondii. Our results evince a widespread circulation of T. gondii in NHPs in zoos in Spain, which may be of conservation concern. Control measures should be implemented to minimize the risk of exposure of these species to T. gondii.