Seroprevalence of Neospora caninum-specific antibodies in German breeding bitches

Background: Neospora caninum is an intracellular obligate apicomplexan parasite responsible for multisystemic lesions in dogs. Being definitive hosts and reservoirs, dogs excrete environmentally resistant oocysts. Breeding bitches represent a susceptible dog group and infected bitches may spread this parasite through transplacental transmission. Results: A total of 218 serum samples of German breeding bitches were collected to determine the presence of N. caninum. Antibodies were detected in 16 (7.33%) bitches using a commercial indirect enzyme-linked immunosorbent assay (ELISA). Immunoblotting analysis confirmed all seropositive samples detected by ELISA, proving that the animals were infected with N. caninum. The owners were interviewed regarding breed, age, environment, type, vaccine status, feeding habits and the presence of reproductive disorders. Seropositive animals were between the ages of two to seven years; three of them were kept in kennels while the others were household dogs, one of which was additionally a hunting dog. Owners of four seropositive bitches reported one gestation, while multiple pregnancies had been recorded for the other twelve bitches. Fourteen bitches were regularly vaccinated and six were fed with fresh raw meat. Conclusions: Although the results confirmed a low incidence of N. caninum seropositive German breeding bitches, further epidemiological and surveillance studies are required to complement our findings regarding the current situation of neosporosis in this specific canine population of Germany

Bottlenose dolphins (Tursiops truncatus) do also cast neutrophil extracellular traps against the apicomplexan parasite Neospora caninum

Neutrophil extracellular traps (NETs) are web-like structures composed of nuclear DNA decorated with histones and cytoplasmic peptides which antiparasitic properties have not previously been investigated in cetaceans. Polymorphonuclear neutrophils (PMN) were isolated from healthy bottlenose dolphins (Tursiops truncatus), and stimulated with Neospora caninum tachyzoites and the NETs-agonist zymosan. In vitro interactions of PMN with the tachyzoites resulted in rapid extrusion of NETs. For the demonstration and quantification of cetacean NETs, extracellular DNA was stained by using either Sytox Orange® or Pico Green®. Scanning electron microscopy (SEM) and fluorescence analyses demonstrated PMN-derived release of NETs upon exposure to tachyzoites of N. caninum. Co-localization studies of N. caninum induced cetacean NETs proved the presence of DNA adorned with histones (H1, H2A/H2B, H3, H4), neutrophil elastase (NE), myeloperoxidase (MPO) and pentraxin (PTX) confirming the molecular properties of mammalian NETosis. Dolphin-derived N. caninum-NETosis were efficiently suppressed by DNase I and diphenyleneiodonium (DPI) treatments. Our results indicate that cetacean-derived NETs represent an ancient, conserved and relevant defense effector mechanism of the host innate immune system against N. caninum and probably other related neozoan parasites circulating in the marine environment

Presencia de Maedi-Visna en hatos ovejeros de Costa Rica

A pesar de que el virus de Maedi Visna (VMV) está ampliamente distribuido en el mundo, la presencia de este lentivirus de los pequeños rumiantes en Costa Rica era desconocida. Un total de 359 sueros procedentes de 15 fincas ovinas de Costa Rica fueron analizados mediante inmunoensayo enzimático para determinar la presencia de anticuerpos contra el VMV. También se aplicó una encuesta a los productores ovinos participantes, para determinar medidas de manejo en cada hato y la presencia de síntomas compatibles con Maedi Visna. Se detectó siete animales seropositivos, procedentes de seis fincas. Los porcentajes de positividad intrahato se establecieron entre 0 a 7.1%. El virus se presentó ampliamente distribuido dentro de las diversas regiones del país, ya que los ovinos positivos fueron encontrados en cuatro (Central, Huetar Atlántico, Huetar Norte y Pacífico Central) de las cinco regiones analizadas. La seropositividad en dichas regiones fue baja (entre 0 y 0.84%) al igual que la prevalencia global (1,95%). Por otra parte, la encuesta reveló que el 52% de las fincas visitadas habían introducido animales (machos y hembras), embriones o semen procedentes de otros hatos o de otros países, sin ningún tipo de certificación sanitaria que garantizara el estatus seronegativo para VMV. Además, solamente cuatro (26%) de los 15 productores encuestados habían escuchado previamente sobre el VMV. Ninguno de ellos había implementado aún un banco de calostro ni pasteurización de la leche ovina. La leche de cabra fue el remplazador más utilizado para crianza de los corderos, sin embargo, solamente una de las fincas empleaba leche de cabra de animales seronegativos al virus de la Artritis Encefalitis Caprina. Se recomienda confirmar la presencia de VMV, por medio de aislamiento celular o técnicas moleculares.Although Maedi-Visna Virus (VMV) was reported in most countries of the world; the presence of this small ruminant lentivirus in Costa Rica was unknown. A total of 359 sheep samples from 15 farms were analyzed for the presence of antibodies against VMV by immunoenzymatic assay. Also a survey was applied to the sheep owners to determine management measures and presence of clinical symptoms of Maedi-Visna in the flocks. Antibodies against VMV were detected in seven sheep from six different flocks, determining flock seropositivity percentages between 0 to 7.1%. The virus seemed to be distributed throughout the country, since the flocks with positive reactors were located in four (Central, Huetar Norte, Huetar Atlántico and Pacífico Central) out of five regions tested. The seropositivity detected in these regions was low, between 0% to 0,84%; as well as the global prevalence (1,95%). The survey revealed that 52% of the participating farms had introduced animals (males and females), embryos or semen from other farms or from abroad without any sanitary certification for VMV-free status. Only four (26%) out of 15 sheep farmers had heard before from VMV. None of the sheep farmers had implemented a bank with VMV free colostrum, and goat milk was the main milk substitute for the lambs, however, only one flock was aware to supplement milk from seronegative goats to Caprine Arthritis Encephalitis Virus. Confirmation of presence of VMV using isolation in cell culture or molecular methods is strongly recommended.Escuela de Medicina Veterinari

Seroprevalencia y factores de riesgo asociados al virus Lengua Azul (BTV) en hatos ovinos costarricenses

Detectar anticuerpos contra el virus Lengua Azul (BTV, Blue Tongue Virus) mediante técnica inmunoenzimática (ELISA) comercial, e identificar los factores de riesgo asociados a esta infección viral en hatos ovinos costarricenses.To detect antibodies against Blue Tongue Virus (BTV) by commercial enzyme-linked immunosorbent assay (ELISA), and to identify risk factors associated with this viral infection in Costa Rican sheep herds.Escuela de Medicina Veterinari

Seroprevalence of Bluetongue and associated risk factors in Costa Rican sheep flocks

A total of 359 sheep samples from 15 farms were analyzed for the presence of antibodies against bluetongue virus (BTV) by commercial enzyme-linked immunosorbent assay (ELISA). Antibodies were detected in 290 sheep from fourteen different flocks, distributed in all analyzed regions (Central, Chorotega, Atlantic Huetar, North Huetar, and Central Pacific) determining regional seropositivity between 63.5% and 100.0%, as well as an overall prevalence of 80.8 %. The within flock seropositivity percentages ranged between 28.6% and 100.0%. Two management practices were determined as risk factors for BTV seropositivity: buying animals, embryos, or semen from other farms without any sanitary control (59.1% of the participating farms, OR= 2.54; IC= 1.49 to 4.35), and the lack of quarantine areas or separated boxes for sick animals in each flock (70.47% of the studied flocks, OR= 6.47; IC= 3.68 to 11.40). All seronegative individuals (19.2%) came from flocks that used partial stabling, indicating this management practice as a protective factor (OR= 1.10; IC= 1.06 to 1.14) for BTV infection. Flocks with the highest seropositivity were found in low altitude regions close to the coast. The results of this study indicate that BTV is endemic in sheep herds from Costa Rica, and animals seem not show clinical signs. Further epidemiological studies are required to determine serotypes present in cattle, wild and small ruminants.Se analizaron muestras de sangre de un total de 359 ovejas de 15 explotaciones para detectar la presencia de anticuerpos contra el virus de la lengua azul (VLB ) mediante un ensayo comercial de inmunoabsorción ligado a enzimas (ELISA). Se detectaron anticuerpos en 290 ovejas de catorce rebaños diferentes, distribuidos en todas las regiones analizadas (Central, Chorotega, Huetar Atlántica, Huetar Norte y Pacífico Central) determinando una seropositividad regional entre el 63,5% y el 100,0%, así como una prevalencia global del 80,8%. Los porcentajes de seropositividad dentro de la bandada oscilaron entre el 0% y el 100,0%. Las bandadas con la seropositividad más alta se encontraron en las regiones de baja altitud cercanas a la costa. Los factores de riesgo y de protección determinados en el presente estudio no estaban en consonancia con esta enfermedad transmitida por los insectos. Los resultados de este estudio indican que el virus de la lengua azul es endémico en los rebaños de ovejas de Costa Rica, y los animales parecen no mostrar signos clínicos. Recomendamos que se lleven a cabo nuevos estudios para determinar la presencia del virus de la lengua azul en cabras y rumiantes salvajes, y para identificar los serotipos presentes en el país.Escuela de Medicina Veterinari

Pinniped- and Cetacean-Derived ETosis Contributes to Combating Emerging Apicomplexan Parasites (Toxoplasma gondii, Neospora caninum) Circulating in Marine Environments

Leukocytes play a major role in combating infections either by phagocytosis, release of antimicrobial granules, or extracellular trap (ET) formation. ET formation is preceded by a certain leukocyte cell death form, known as ETosis, an evolutionarily conserved mechanism of the innate immune system also observed in marine mammals. Besides several biomolecules and microbial stimuli, marine mammal ETosis is also trigged by various terrestrial protozoa and metazoa, considered nowadays as neozoan parasites, which are circulating in oceans worldwide and causing critical emerging marine diseases. Recent studies demonstrated that pinniped- and cetacean-derived polymorphonuclear neutrophils (PMNs) and monocytes are able to form different phenotypes of ET structures composed of nuclear DNA, histones, and cytoplasmic peptides/proteases against terrestrial apicomplexan parasites, e.g., Toxoplasma gondii and Neospora caninum. Detailed molecular analyses and functional studies proved that marine mammal PMNs and monocytes cast ETs in a similar way as terrestrial mammals, entrapping and immobilizing T. gondii and N. caninum tachyzoites. Pinniped- and cetacean leukocytes induce vital and suicidal ETosis, with highly reliant actions of nicotinamide adenine dinucleotide phosphate oxidase (NOX), generation of reactive oxygen species (ROS), and combined mechanisms of myeloperoxidase (MPO), neutrophil elastase (NE), and DNA citrullination via peptidylarginine deiminase IV (PAD4).This scoping review intends to summarize the knowledge on emerging protozoans in the marine environment and secondly to review limited data about ETosis mechanisms in marine mammalian species

Seroprevalence of Toxoplasma gondii and Neospora caninum infections and associated factors in sheep from Costa Rica

The presence of antibodies against Toxoplasma gondii and Neospora caninum were analyzed in 392 sheep sera from ten Costa Rican ovine flocks using indirect immuno-enzymatic assays. Additionally, general information about sheep management, environment, and clinical reproductive disorders was assessed through a questionnaire to inquire factors related to these apicomplexan parasites. A total of 161 (41.1%) serum samples reacted positive to T. gondii, 43 (10.9%) to N. caninum and 26 (6.63%) to both parasites. Toxoplasma gondii serorreactors were detected in all the analyzed flocks (100.0%), meanwhile N. caninum antibodies were found in nine flocks (90%), from the six Costa Rican regions. Factors associated with T. gondii were the co-presence of cattle (OR = 5.06; C.I.95%; 2.08–12.30; p: <0.001), grey foxes (Urocyon cinereoargenteus) and opossums (Didelphis marsupialis) (OR = 2.44; C.I.95%; 1.50–3.95; p: <0.001) inside or around the farms, and the presence of peccaries (Tayassu sp.) (OR = 0.35; C.I.95%; 0.16–0.74; p: 0.0058) was a variable associated with N. caninum seropositivity. The obtained results of T. gondii and N. caninum infections in sheep flocks from Costa Rica should be considered for the proper prevention and control strategies against these apicomplexan abortive parasites. Keywords: Abortion, Sheep, Toxoplasmosis, Neosporosis, Costa Ric

Erster Bericht über die Seroprävalenz von Neospora caninum in hessischen Ziegenbeständen

A total of 415 goat serum samples from 26 flocks in Hesse, Central Germany, were analyzed for the presence of specific antibodies against the abortion-causing apicomplexan parasite Neospora (N.) caninum by immuno-enzyme assay (ELISA). In total, three serum samples were seropositive for N. caninum with two of them originating from a flock in Middle-Hesse and one sample coming from a flock in Northern Hesse. Western Blotting confirmed two of three ELISA-positive samples proving a low overall prevalence of 2/415 (0.48%) for caprine neosporosis. No clinical signs related to neosporosis were detected in any seropositive animal. Additionally, there was no familiar relationship between them and N. caninum-positive goats were purchased from different breeders. The low numbers of N. caninum-seropositive animals excluded risk factors assessment. Based on the current seroprevalence data, N. caninum infections appear of minor importance in German goat flocks. Nevertheless, taking into account that caprine neosporosis was detected in some European countries bordering Germany, further epide-miological and surveillance studies on caprine N. caninum infections are required to complement our findings regarding the current situation in goat populations from the other German Federal States.Insgesamt wurden 415 Ziegenserumproben von 26 Herden in Hessen, Deutsch-land, auf das Vorhandensein spezifischer Antikörper gegen den abortiven api-komplexen Parasiten Neospora (N.) caninum mittels Immunoenzymassay (ELISA) untersucht. Insgesamt konnten nur drei positive Proben ermittelt werden, wobei zwei aus einem Bestand in Mittelhessen und eine aus einem nordhessischen Bestand kamen. Ein anschließend durchgeführter Western Blot bestätigte zwei der drei ELISA-positiven Proben, die auf eine insgesamt niedrige Gesamtprävalenz von 2/415 (0,48 %) für Ziegenneosporose hinweisen. Keines der seropositiven Tiere wies klinische Symptome der Neosporose auf. Darüber hinaus lagen keine verwandtschaftlichen Beziehungen zwischen ihnen vor und die N. caninum-positiven Ziegen wurden von unterschiedlichen Züchtern erworben. Die geringe Seroprävalenz der vorliegenden Studie schloss die Möglichkeit, Risikofaktoren zu analysieren, aus. Die tatsächliche Bedeutung der N. caninum-Infektionen in deut-schen Ziegenbeständen scheint aufgrund der gemessenen geringen Seropräva-lenz vernachlässigbar. Da die Neosporose in Ziegen in einigen an Deutschland angrenzenden europäischen Ländern bereits nachgewiesen wurde, sind weitere epidemiologische Studien zu N. caninum-Infektionen von Ziegen erforderlich, um die aktuellen Erkenntnisse bezüglich anderer Bundesländer zu ergänzenSe analizaron 415 muestras de suero de cabra de 26 rebaños en Hesse, Alemania central, para detectar la presencia de anticuerpos específicos contra el parásito apicombustible causante del aborto, Neospora (N.) caninum, mediante un ensayo de inmunoenzimas (ELISA). En total, tres muestras de suero resultaron seropositivas para N. caninum, dos de ellas procedentes de un rebaño de Hesse Medio y una muestra procedente de un rebaño de Hesse del Norte. Western Blotting confirmó dos de las tres muestras positivas para ELISA, demostrando una baja prevalencia global de 2/415 (0,48%) para la neosporosis caprina. No se detectaron signos clínicos relacionados con la neosporosis en ningún animal seropositivo. Además, no había ninguna relación familiar entre ellos y las cabras N. caninum-positivas fueron compradas a diferentes criadores. El bajo número de animales seropositivos para N. caninum excluía la evaluación de los factores de riesgo. Basándose en los datos actuales de seroprevalencia, las infecciones por N. caninum parecen tener una importancia menor en los rebaños de cabras alemanas. No obstante, teniendo en cuenta que se detectó neosporosis caprina en algunos países europeos limítrofes con Alemania, es necesario realizar más estudios epidemiológicos y de vigilancia de las infecciones por N. caninum en caprinos para complementar nuestros hallazgos sobre la situación actual de las poblaciones caprinas de los demás Estados Federados alemanes.UNIVERSIDAD JUSTUS-LIEBIG, GIESSEN, ALEMANIAEscuela de Medicina Veterinari

Seroprevalence of Neospora caninum-specific antibodies in German breeding bitches

Abstract Background Neospora caninum is an intracellular obligate apicomplexan parasite responsible for multisystemic lesions in dogs. Being definitive hosts and reservoirs, dogs excrete environmentally resistant oocysts. Breeding bitches represent a susceptible dog group and infected bitches may spread this parasite through transplacental transmission. Results A total of 218 serum samples of German breeding bitches were collected to determine the presence of N. caninum. Antibodies were detected in 16 (7.33%) bitches using a commercial indirect enzyme-linked immunosorbent assay (ELISA). Immunoblotting analysis confirmed all seropositive samples detected by ELISA, proving that the animals were infected with N. caninum. The owners were interviewed regarding breed, age, environment, type, vaccine status, feeding habits and the presence of reproductive disorders. Seropositive animals were between the ages of two to seven years; three of them were kept in kennels while the others were household dogs, one of which was additionally a hunting dog. Owners of four seropositive bitches reported one gestation, while multiple pregnancies had been recorded for the other twelve bitches. Fourteen bitches were regularly vaccinated and six were fed with fresh raw meat. Conclusions Although the results confirmed a low incidence of N. caninum seropositive German breeding bitches, further epidemiological and surveillance studies are required to complement our findings regarding the current situation of neosporosis in this specific canine population of Germany

Presence of Maedi-Visna in Costa Rican sheep flocks

A total of 359 sheep serum samples from 15 farms were analyzed for the presence of antibodies against Maedi Visna Virus (MVV) by Enzyme-linked Immunosorbent Assay (ELISA). Additionally, a survey was applied to the sheep owners in order to determine management measures and presence of clinical symptoms of Maedi-Visna in the flocks. Logistic regression with random effects was performed to assess risk factors to seropositivity. Seven serum samples were positive to MVV belonging to six flocks, determining low flock seropositivity (0 to 7.1%). Only one of the five regions studied was negative, determining low overall prevalence (1.95%) and low positivity by region (0.0% to 0.84%). One seropositive animal with symptoms related to MVV was sacrificed for necropsy. An enlargement of mesenteric and celiac lymph nodes was determined, histopathology of lungs revealed acute interstitial pneumonia. Analysis of organs, fluids and blood of this sheep by Polymerase Chain Reaction (PCR) to detect MVV yielded negative results. In order to confirm the serological results, serum and blood samples from two remaining seropositive animals were analyzed six months later, one resulted seronegative and the other seropositive in ELISA, but both negative in PCR. No risk factors were associated to seropositivity. Since MVV produces latency, we conclude, that the positive serological results were due to false positive reactions. Even though, 52.0% of the participating farms had introduced animals, embryos, or semen from other farms or from abroad without any sanitary certification, MVV seems to be present in Costa Rica probably in a very low prevalence. Further studies are required to implement control measures and prevent the spread of the disease.Se analizaron 359 muestras de suero de oveja de 15 explotaciones para detectar la presencia de anticuerpos contra el virus Maedi Visna (VMV) mediante el ensayo de inmunoabsorción ligado a las enzimas (ELISA). Además, se realizó una encuesta entre los propietarios de las ovejas para determinar las medidas de gestión y la presencia de síntomas clínicos del Maedi-Visna en los rebaños. Se realizó una regresión logística con efectos aleatorios para evaluar los factores de riesgo de seropositividad. Siete muestras de suero dieron positivo al MVV de seis rebaños, lo que determinó una baja seropositividad del rebaño (0 a 7,1%). Sólo una de las cinco regiones estudiadas fue negativa, lo que determinó una baja prevalencia general (1,95%) y una baja positividad por región (0,0% a 0,84%). Se sacrificó para la necropsia un animal seropositivo con síntomas relacionados con el VMV. Se determinó un agrandamiento de los ganglios linfáticos mesentéricos y celíacos, la histopatología de los pulmones reveló una neumonía intersticial aguda. El análisis de los órganos, fluidos y sangre de esta oveja por reacción en cadena de la polimerasa (PCR) para detectar el MVV dio resultados negativos. Para confirmar los resultados serológicos, seis meses después se analizaron muestras de suero y sangre de los dos animales seropositivos restantes, uno resultó seronegativo y el otro seropositivo en la prueba ELISA, pero ambos negativos en la PCR. No se asociaron factores de riesgo a la seropositividad. Dado que el MVV produce latencia, concluimos que los resultados serológicos positivos se debieron a reacciones positivas falsas. Aunque el 52,0% de las granjas participantes habían introducido animales, embriones o semen de otras granjas o del extranjero sin ninguna certificación sanitaria, el VMV parece estar presente en Costa Rica probablemente en una prevalencia muy baja. Es necesario realizar más estudios para aplicar medidas de control y prevenir la propagación de la enfermedad.Universidad Nacional, Costa RicaEscuela de Medicina Veterinari