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Influenza D virus (IDV) is considered a new agent involved in bovine respiratory disease (BRD). Based on seroprevalence studies or isolation from clinical samples, this virus has been detected on several continents and in several animal species, including cattle, pigs, camel, horses, and goats. We used an indirect in-house ELISA to detect anti-IDV antibodies in 165 serum samples from bulls on 116 farms in the province of La Pampa, Argentina. Eighty-five of 116 (73%) farms had at least 1 positive animal, and 112 of 165 (68%) of the analyzed samples were positive. There were no significant differences in the proportion of seropositive samples depending on the geographic region in which the samples were taken. Our results suggest that IDV infection is endemic in La Pampa; the clinical importance of IDV in Argentina remains to be investigated.

Las pérdidas reproductivas constituyen una causa importante de pérdida económica en el ganado bovino, aunque en más del 50% de los casos la etiología es desconocida. Las especies de la familia Chlamydiaceae han sido asociadas con abortos en bovinos y otras espeChlamydia abortus; cies animales, pero no existen datos al respecto en la República Argentina. El objetivo de este trabajo fue determinar la presencia de Chlamydia spp. y de Chlamydia abortus en pérdidas reproductivas de ganado bovino en La Pampa, Argentina. Se estudiaron 251 muestras provenientes de abortos y mortinatos. Se realizó PCR en tiempo real para la detección de la familia Chlamydiaceae y ArrayTube para la identificación de las especies presentes. Se detectó ADN de la familia Chlamydiaceae en 12 muestras (4,78%); el 83,33% (10/12) correspondió a abortos y el 16,66% (2/12) a mortinatos. El análisis por ArrayTube detectó C. abortus en 5 muestras (1,99% del total, 41,67% de las muestras con detección de Chlamydiaceae). Este trabajo presenta la primera confirmación de la presencia de ADN de diversas especies de Chlamydiaceae (incluida C. abortus) en muestras de pérdidas reproductivas de ganado bovino en Argentina. El valor de prevalencia hallado (4,78%) debe ser tomado como un valor basal, debido al tipo de muestras estudiadas. Se halló material genético de Chlamydiaceae que no coincidió con ninguna de las especies conocidas; esto podría deberse a variantes intraespecie o a especies autóctonas aún no descriptas. Es necesario avanzar en el estudio de la infección por estas bacterias en el ganado bovino de Argentina para conocer su dimensión y analizar su impacto económico y zoonótico, y también para planear medidas de prevención y control. Reproductive losses linked to an infectious etiology in bovine cattle are a major economic concern worldwide. In Argentina, more than 50% of abortion cases have unknown causes. Species belonging to Chlamydiaceae family are frequent etiologic agents of abortion around the world; however, there is yet no information on their prevalence in Argentina. The objective of this work was to identify Chlamydia spp., and particularly C. abortus in reproductive losses from bovine cattle in La Pampa, Argentina. Real time PCR targeting Chlamydiaceae-specific DNA fragments was performed on 251 samples obtained from bovine abortions and stillborns, and ArrayTube was used for species identification on positive samples. Chlamydiaceae DNA was detected in 12 samples of aborted fetuses (4.78%), 83.33% (10/12) accounting for abortions and 16.66% (2/12) for stillborns. C. abortus was detected by ArrayTube in 5 cases (1.99% of all samples, and 41.67% of Chlamydiaceae positive samples). This study shows the first detection of Chlamydiaceae and C. abortus DNA on reproductive losses of bovine cattle in Argentina, and the described prevalence value (4.78%) should be taken as baseline value due to the type of samples analyzed. Detection of genetic material from Chlamydiaceae not matching any of the studied species could be due to intraspecies variants or local species not yet described. Further research on Chlamydia infections in bovine cattle in Argentina is imperative to describe their range, to analyze their economic and zoonotic implications and to make recommendations about prevention and control measures.

Tritrichomonas foetus isolates from feline and bovine origin has been previously shown to carry a certain degree of genetic heterogeneity. Here, novel candidate molecular markers were developed by means of multilocus sequence typing of the gap2 gene (encoding for T. foetus glyceraldehyde-3-phosphate dehydrogenase), ITS region, the TR7/TR8 variable-length repeat and microsatellite genotyping. These markers were used to characterize T. foetus field isolates from bulls and domestic cats and to compare phylogenetically with the following ATCC isolates: T. foetus isolated from cattle and pig (syn. Tritrichomonas suis), Tritrichomonas mobilensis, Tetratrichomonas gallinarum and Pentatrichomonas hominis. Among them, TFMS10 and TFMS7 were found to be the most polymorphic markers. Moreover, an 809 bp fragment of the gap2 gene was successfully amplified from all the trichomonads included in this study and the sequence analysis revealed differences between T. foetus porcine and feline genotypes and T. mobilensis in comparison to the bovine T. foetus ATCC isolate. The TR7/TR8 repeat pattern was not reproducible, being only consistent the fragments of approximately 110 and 217 bp. Sequence analysis of the latter revealed the existence of 3 SNPs resulting in 98.6 % homology between bovine and feline isolates. A search for similar sequences was carried out to develop a Restriction Length Fragment Polymorphism analysis. A 503 bp region, named TF1, revealed the existence of two BbvI restriction enzyme sites that were able to generate different length fragments for T. foetus feline and bovine isolates. Finally, the neighbour-joining analyses showed that T. foetus porcine genotype clusters together with bovine genotype, whereas T. mobilensis and the feline genotype form a separate cluster.

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

Brucella suis biovar 1 is the causative agent of brucellosis in several domestic and wild animals and it is a common agent of human brucellosis. European hares (Lepus europaeus) have been shown to be infected by B. suis biovar 1 and the transmission to other animals has been suggested. In this work, experimental rabbits (Cuniculus orictolagus) were infected with B. suis biovar 1 isolated from wild hares. Infected rabbits showed high serological response in 2 weeks after discharge and typical granulomatous lesions (2. mm diameter) were found in liver, spleen and kidneys after 50 days. B. suis biovar 1 was cultured from the lesion of the organs mentioned above as well as from urine, placenta and fetuses. These data suggest that hares are a potential source for horizontal transmission of B. suis biovar 1 to other mammalians. Fil: Fort, Marcelo. Instituto Nacional de Tecnología Agropecuaria. Centro Regional La Pampa-San Luis. Estación Experimental Agropecuaria Anguil; Argentina Fil: Baldone, Valeria Natalia. Instituto Nacional de Tecnología Agropecuaria. Centro Regional La Pampa-San Luis. Estación Experimental Agropecuaria Anguil; Argentina Fil: Fuchs, Lumila Ivana. Instituto Nacional de Tecnología Agropecuaria. Centro Regional La Pampa-San Luis. Estación Experimental Agropecuaria Anguil; Argentina Fil: Giménez, Hugo. Instituto Nacional de Tecnología Agropecuaria. Centro Regional La Pampa-San Luis. Estación Experimental Agropecuaria Anguil; Argentina Fil: Rojas, María. Instituto Nacional de Tecnología Agropecuaria. Centro Regional La Pampa-San Luis. Estación Experimental Agropecuaria Anguil; Argentina Fil: Breccia, Javier Dario. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Ciencias de la Tierra y Ambientales de La Pampa. Universidad Nacional de La Pampa. Facultad de Ciencias Exactas y Naturales. Instituto de Ciencias de la Tierra y Ambientales de La Pampa; Argentina Fil: Oyhenart, Jorge. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Ciencias de la Tierra y Ambientales de La Pampa. Universidad Nacional de La Pampa. Facultad de Ciencias Exactas y Naturales. Instituto de Ciencias de la Tierra y Ambientales de La Pampa; Argentina

Leptospirosis is a zoonosis of worldwide distribution. The aim of this study was to examine the presence of antibodies against 21 Leptospira reactive serovars in Chaetophractus villosus in La Pampa province, Argentina, using the microscopic agglutination test (MAT). Pathologic changes compatible with leptospirosis and in situ detection of the agent by immunohistochemistry were studied in 24 and 3 individuals respectively. Only 35/150 (23.3%) serum samples had antibodies against Leptospira sp. Six percent of the samples reacted with serovar Canicola, 4.7% with serovar Castellonis, 1.3% with serovar Icterohemorrhagieae and 0.7% with serovar Hardjo. Sixteen (10.6%) serum samples agglutinated with Castellonis-Icterohemorrhagiae and Canicola-Castellonis serovars, both with 4.7%, and Canicola-Hardjo and Castellonis-Canicola-Icterohemorrhagiae both with 0.6%. Fourteen animals had variable degrees of lesions, which were more severe in animals with higher serological titers (3200), and Leptospira sp. was detected in 3 animals by immunohistochemistry. These results represent the first record of the presence of Leptospira in C. villosus in La Pampa. Fil: Kin, Marta S.. Universidad Nacional de La Pampa. Facultad de Ciencias Exactas y Naturales; Argentina Fil: Brihuega, Bibiana. Instituto Nacional de Tecnología Agropecuaria; Argentina Fil: Fort, Marcelo. Instituto Nacional de Tecnologia Agropecuaria. Centro Regional La Pampa-San Luis; Argentina Fil: Delgado, Fernando. Instituto Nacional de Tecnología Agropecuaria; Argentina Fil: Bedotti, Daniel. Instituto Nacional de Tecnología Agropecuaria. Centro Regional la Pampa-San Luis. Estación Experimental Agropecuaria Anguil; Argentina Fil: Casanave, Emma Beatriz. Universidad Nacional del Sur; Argentina. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico Bahia Blanca. Instituto de Ciencias Biologicas y Biomedicas del Sur; Argentina

Toxoplasma gondii is an obligate intracellular apicomplexan parasite that causes abortion and reproductive disorder in domestic animals. T. gondii is a common worldwide disease in homeothermic animals, including birds and humans. The aim of the present study was to determine the presence of antibodies against T. gondii in the armadillo Chaetophractus villosus in the province of La Pampa, Argentina. Serum samples were collected from 150 individuals (70 males and 80 females). For serological detection of T. gondii, a latex agglutination test was first performed and then positive sera were confirmed with an indirect hemagglutination test, using 1:4 to 1:64 dilutions. Results showed that 27% (41) of the samples presented titers for antibodies against T. gondii. There were not significant differences between the presence of antibodies against T. gondii and age or sexes of the armadillos. Results show that presence of T. gondii antibodies in armadillos were associated with presence of pigs, and sheep, however there was not association with chickens and dairy cattle in capture site. T. gondii has an important presence in C. villosus population, suggesting a potential zoonotic risk for humans and wildlife animals when C. villosus meats are consumed raw or undercooked. This is the first record of the presence of antibodies against T. gondii in C. villosus. Fil: Kin, Marta S.. Universidad Nacional de la Pampa. Facultad de Ciencias Exactas y Naturales; Argentina Fil: Fort, Marcelo. Instituto Nacional de Tecnologí­a Agropecuaria. Centro Regional La Pampa-San Luis. Estación Experimental Agropecuaria Anguil; Argentina Fil: Gimenez, Hugo D.. Instituto Nacional de Tecnología Agropecuaria. Centro Regional La Pampa-San Luis. Estación Experimental Agropecuaria Anguil; Argentina Fil: Casanave, Emma Beatriz. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico Bahia Blanca. Instituto de Ciencias Biologicas y Biomedicas del Sur; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina

Neospora caninum is considered one of the major causes of abortion in cattle. The aim of this study was to examine and quantify the extent of the infection in cattle in a representative region of Argentina (La Pampa, province). An average sample size of 36 sera per herd was selected from 97 beef and 24 dairy herds. A total of 4334 serum samples were tested for specific anti- Neospora caninum IgG using an indirect-ELISA and 302 seropositive-ELISA sera were re-examined using an Avidity-ELISA procedure for N.caninum. The overall estimated seroprevalence for N.caninun was 9.6% (95%CI: 8.7%; 10.5%). Levels of seroprevalence were significantly different in beef 7.0% and dairy 20.3% cattle. Disease distribution seems to be associated with climatic conditions as well as the management system. Cows in the east and central regions were at a 4.5-fold and 2.0-fold higher risk, respectively, of being N. caninum seropositive compared with cows in west region. Levels of recent infection were evaluated through an avidity ELISA in seropositive animals, being registered a 0.56% and a 1.71% of recent infection in beef and dairy cattle respectively (p = 0.006). The results revealed that dairy cows had 3.1(95%CI: 1.4; 7.0) higher risk of contracting Neoporosis through horizontal transmission than beef cows. A relationship between Brucella abortus and N. caninum seroprevalence was also observed. The risk of being N. caninum seropositive was two times higher where Brucellosis seroprevalence was >3.5%. These results reveal the distribution of N. caninum infection in the cattle population in La Pampa, Argentina

Brucellosis is a zoonotic disease transmitted from an animal reservoir to humans. Both, wildlife and domestic animals, contribute to the spreading of these zoonosis. The surveillance of the animal health status is strictly regulated for domestic animals, whereas disease monitoring in wildlife does not exist. The aim of the present study was to provide data on the prevalence of anti-Brucella antibodies in Chaetophractus villosus from a region of La Pampa, Argentina to assess public health risks. The C. villosus is endemic to South America, and in Argentina it represents a food resource for human consumption. A total of 150 sera of armadillos bleeding between 2007 and 2010 were tested using buffered plate antigen test (BPAT), serum agglutination test (SAT), 2-mercaptoethanol (2-ME) and complement fixation test (CFT), for the detection of anti-Brucella antibodies. Antibodies to Brucella sp. were found in 16% (24:150) of the armadillos tested using the BPAT test. All 24 positive samples were confirmed by the SAT, 2-ME and CFT tests. Strain isolation was attempted from liver and spleen samples of two animals with positive serology. Isolates were characterized by conventional biotyping and identification of specific DNA using polymerase chain reaction (PCR). A total of 2 isolates were recovered from spleen and liver. Both of them were identified as Brucella suis biovar 1. This preliminary study provides the first report on the seroprevalence of brucellosis and describes the first isolate of B. suis biovar 1 in C. villosus in Argentina.

The protozoan parasite Tritrichomonas foetus is well known as an important causative agent of infertility and abortion in cattle (bovine trichomonosis). This World Organisation for Animal Health (O.I.E.) notifiable disease is thought to be under control in many countries including Switzerland. In recent studies, however, T. foetus has also been identified as an intestinal parasite that causes chronic large-bowel diarrhoea in cats. Since the feline isolates were considered indistinguishable from bovine isolates, the possibility and risk of parasite transmission from cats to cattle and vice versa has been intensively discussed in current literature. Therefore, we investigated if cat and cattle isolates are genetically distinct from each other or in fact represent identical genotypes. For this purpose, two independent genetic loci were selected that turned out to be well-suited for a PCR sequencing-based genotyping of trichomonad isolates: (i) previously published internal transcribed spacer region 2 (ITS-2) and (ii) a semi-conserved sequence stretch of the elongation factor-1 alpha (EF-1α) gene used for the first time in the present study. Respective comparative analyses revealed that both loci were sufficiently variable to allow unambiguous genetic discrimination between different trichomonad species. Comparison of both genetic loci confirmed that T. suis and T. mobilensis are phylogenetically very close to T. foetus. Moreover, these two genetic markers were suited to define host-specific genotypes of T. foetus. Both loci showed single base differences between cat and cattle isolates but showed full sequence identity within strains from either cat or cattle isolates. Furthermore, an additional PCR with a forward primer designed to specifically amplify the bovine sequence of EF-1α was able to discriminate bovine isolates of T. foetus from feline isolates and also from other trichomonads. The implications these minor genetic differences may have on the biological properties of the distinct isolates remain to be investigated.