The Frequency of Porcine Cysticercosis and Factors Associated with Taenia solium Infection in the Municipality of Tuchín-Córdoba, Colombia.

Taeniasis and cysticercosis are parasitic infections that affect humans and pigs. Their global distribution constitutes a serious public health issue with significant implications for pork production. The purpose of this study was to evaluate the presence of porcine cysticercosis in backyard swine from 42 indigenous communities throughout Tuchín-Córdoba, Colombia. Between December 2020 and March 2021, free-range pigs (n = 442) were assessed using the ELISA cysticercosis Ag test; 85 pigs were examined through sublingual visual evaluation, and 4 slaughtered pig carcasses were subjected to standard operation inspection. The collected cysticercus underwent histological and PCR analysis. Furthermore, 192 surveys of knowledge, attitudes, and practices (KAP) were used to identify the factors that facilitate infection transmission. Serological investigation revealed that 9.7% (46/472) of the animals were positive for cysticerci Ag. Sublingual inspection identified cysticercus in 28.7% (25/87) of the animals, while PCR analysis indicated that cysticercus corresponded to the T. solium American/African genotype. The factors associated with T. solium infection in the pigs in the surveyed areas numbered 14. The majority are associated with factors that promote the active persistence of Taenia solium's life cycle in an area, such as lack of environmental sanitation, a lack of coverage or care for drinking water and wastewater treatment services, and no solid waste disposal.

Glycolysis, monocarboxylate transport, and purinergic signaling are key events in Eimeria bovis-induced NETosis.

The protozoan parasite Eimeria bovis is the causative agent of bovine coccidiosis, an enteric disease of global importance that significantly affects cattle productivity. Previous studies showed that bovine NETosis-an important early host innate effector mechanism of polymorphonuclear neutrophil (PMN)-is elicited by E. bovis stages. So far, the metabolic requirements of E. bovis-triggered NET formation are unknown. We here studied early glycolytic and mitochondrial responses of PMN as well as the role of pH, distinct metabolic pathways, P2 receptor-mediated purinergic signaling, and monocarboxylate transporters 1 and 2 (MCT1, MCT2) in E. bovis sporozoite-induced NET formation. Seahorse-based experiments revealed a rapid induction of both neutrophil oxygen consumption rate (OCR) and early glycolytic responses, thereby reflecting immediate PMN activation and metabolic changes upon confrontation with sporozoites. The impact of these metabolic changes on NET formation was studied via chemical inhibition experiments targeting glycolysis and energy generation by the use of 2-fluor-2-deoxy-D-glucose (FDG), 6-diazo-5-oxo-L-norleucin (DON), sodium dichloroacetate (DCA), oxythiamine (OT), sodium oxamate (OXA), and oligomycin A (OmA) to block glycolysis, glutaminolysis, pyruvate dehydrogenase kinase, pyruvate dehydrogenase, lactate dehydrogenase, and mitochondrial ATP-synthase, respectively. Overall, sporozoite-induced NET formation was significantly diminished via PMN pretreatments with OmA and OXA, thereby indicating a key role of ATP- and lactate-mediated metabolic pathways. Consequently, we additionally studied the effects of extracellular pH, MCT1, MCT2, and purinergic receptor inhibitors (AR-C141900, AR-C155858, theobromine, and NF449, respectively). Pretreatment with the latter inhibitors led to blockage of sporozoite-triggered DNA release from exposed bovine PMN. This report provides first evidence on the pivotal role of carbohydrate-related metabolic pathways and purinergic receptors being involved in E. bovis sporozoite-induced NETosis.

Optimized excystation protocol for ruminant Eimeria bovis- and Eimeria arloingi-sporulated oocysts and first 3D holotomographic microscopy analysis of differing sporozoite egress.

Successful excystation of sporulated Eimeria spp. oocysts is an important step to acquire large numbers of viable sporozoites for molecular, biochemical, immunological and in vitro experiments for detailed studies on complex host cell-parasite interactions. An improved method for excystation of sporulated oocysts and collection of infective E. bovis- and E. arloingi-sporozoites is here described. Eimeria spp. oocysts were treated for at least 20 h with sterile 0.02 M L-cysteine HCl/0.2 M NaHCO3 solution at 37 °C in 100% CO2 atmosphere. The last oocyst treatment was performed with a 0.4% trypsin 8% sterile bovine bile excystation solution, which disrupted oocyst walls with consequent activation of sporozoites within oocyst circumplasm, thereby releasing up to 90% of sporozoites in approximately 2 h of incubation (37 °C) with a 1:3 (oocysts:sporozoites) ratio. Free-released sporozoites were filtered in order to remove rests of oocysts, sporocysts and non-sporulated oocysts. Furthermore, live cell imaging 3D holotomographic microscopy (Nanolive®) analysis allowed visualization of differing sporozoite egress strategies. Sporozoites of both species were up to 99% viable, highly motile, capable of active host cell invasion and further development into trophozoite- as well as macroment-development in primary bovine umbilical vein endothelial cells (BUVEC). Sporozoites obtained by this new excystation protocol were cleaner at the time point of exposure of BUVEC monolayers and thus benefiting from the non-activation status of these highly immunocompetent cells through debris. Alongside, this protocol improved former described methods by being is less expensive, faster, accessible for all labs with minimum equipment, and without requirement of neither expensive buffer solutions nor sophisticated instruments such as ultracentrifuges.

Haemonchosis en una ternera raza Brahman en el trópico alto del Nordeste Antioqueño / Haemonchosis in a Brahman calf in the high tropics of the Antioquian Northeast / Hemoncose em uma bezerra raça Brahman no trópico alto do nordeste da Antioquia

Resumen La haemonchosis es una parasitosis que, en bovinos, a diferencia de los pequeños rumiantes, raramente causa mortalidad. No obstante, cargas parasitarias que normalmente son subclínicas en terneros bien alimentados, pueden convertirse en clínicas causando la muerte, cuando la condición corporal es pobre. Este reporte describe un caso de haemonchosis fatal en un ternero Brahman de 7 meses y el resultado del tratamiento con albendazol (10 mg/kg PO) en 17 terneros congéneres. La finca es de cría y levante de ganado de carne y está ubicada en el trópico alto, en el municipio de Yarumal, Antioquia, a una altura de 2.353 m.s.n.m y una temperatura promedio de 14 ºC. En la producción no reportan plan sanitario adicional a la vacunación contra aftosa. El diagnóstico se sospechó inicialmente con base en observaciones clínico-patológicas de una anemia severa con hipoproteinemia de etiología no inflamatoria, y se confirmó en el examen post-mortem por la presencia de cientos de nematodos adultos de Haemonchus spp. en el abomaso, y un recuento de 19.200 huevos por gramo (HPG) de heces de trichostrongylidos. El coprocultivo de heces mostró que el 70, 20, y 10% de dichos huevos pertenecían a especies de Haemonchus, Coperia, y otros géneros de nematodos, respectivamente. El tratamiento con albendazol fue eficaz reduciendo las cargas parasitarias en los 17 terneros tratados desde niveles moderados (200-700 HPG) y altos (>700 HPG), a niveles bajos (16-100 HPG). Se redujo por completo la infección en la mitad de los animales tratados y solo estuvo por debajo del 90% en 3 animales. Debido a que la media del porcentaje de reducción de huevos fue del 95% se deduce que no existe resistencia al albendazol. En conclusión, este caso revela que la haemonchosis en terneros debe incluirse dentro de los diagnósticos diferenciales de mortalidad en el trópico colombiano y la realización de necropsias y exámenes coprológicos periódicos son imprescindibles para poder instaurar medidas preventivas y de control de dichas parasitosis.

Abstract Haemonchosis is a parasitic disease that in cattle, unlike small ruminants, rarely causes mortality. However, parasitic burdens that are usually subclinical in well-fed calves may become clinical causing death when body condition is poor. This report describes a case of fatal haemonchosis in a 7-month-old Brahman calf and the results of treatment with albendazole (10 mg/kg PO) in 17 cohort calves. The farm, dedicated to breeding and rearing beef calves, was located in the high tropics of the Andes, municipality of Yarumal (Antioquia), at 2,353 meters of altitude with an annual mean temperature of 14 °C. Apart from vaccination for foot and mouth disease, the animals are not subjected to any other treatments. The diagnosis was initially suspected based on clinico-pathological observations of a severe anemia with hypoproteinemia of non-inflammatory etiology, and was confirmed in the postmortem examination by the presence of hundreds of adult nematodes of Haemonchus spp. in the abomasum and a count of 19,200 eggs per gram of feces (EPG) of trichostrongylids. Stool culture showed that 70, 20, and 10% of these eggs belonged to Haemonchus, Coperia and other nematode genera, respectively. Albendazole treatment was effective in reducing parasitic loads in 17 treated calves from moderate (200-700 OPG) and high levels (> 700 EPG) to low levels (16-100 EPG). The infection was completely eliminated in half of the treated animals and was reduced to below 90% in only 3 animals. Because the mean egg reduction percentage was 95%, it can be inferred that there is no resistance to albendazole. In conclusion, this case reveals that haemonchosis should be included in the differential diagnosis of mortality in calves in the Colombian tropics, and that necropsies and periodic coprological examinations are necessary to establish preventive and control measures of this parasitosis.

Resumo A hemoncose é uma doença parasitária que no gado, ao contrário dos pequenos ruminantes, raramente causa mortalidade. No entanto, cargas parasitas que geralmente são subclínicas em bezerros bem alimentados, podem tornar-se clinicamente causadores de morte quando a condição corporal é ruim. Este relato descreve um caso de hemoncose fatal em um bezerro Brahman de 7 meses de idade e os resultados do tratamento com albendazol (10 mg/kg PO) em 17 bezerros da coorte. A fazenda, dedicada à criação de bezerras, está localizada no trópico alto dos Andes, município de Yarumal (Antioquia), a 2.353 metros de altitude, com temperatura média anual de 14 ºC. Além da vacinação para a febre aftosa, os animais não são submetidos a outros tratamentos. O diagnóstico foi inicialmente suspeito com base em observações clínico-patológicas de uma anemia grave com hipoproteinemia de etiologia não inflamatória, e foi confirmado no exame post-mortem pela presença de centenas de nematóides adultos de Haemonchus spp. no abomaso e uma contagem de 19.200 ovos por grama de fezes (EPG) de tricostrongilídeos. A cultura de fezes mostrou que 70, 20 e 10% desses ovos pertenciam a Haemonchus, Coperia e outros gêneros de nematóides, respectivamente. O tratamento com albendazol foi eficaz na redução das cargas parasitas em 17 bezerros tratados de níveis moderados (200-700 OPG) e altos (> 700 EPG) para níveis baixos (16-100 EPG). A infecção foi completamente eliminada em metade dos animais tratados e foi reduzida para menos de 90% em apenas 3 animais. Como a porcentagem média de redução de ovos foi de 95%, pode-se inferir que não há resistência ao albendazol. Em conclusão, este caso revela que a hemoncose deve ser incluída no diagnóstico diferencial de mortalidade em bezerros nos trópicos colombianos, e que necropsias e exames coprológicos periódicos são necessários para estabelecer medidas preventivas e de controle desta parasitose.

Molecular characterization of Marek's disease virus in a poultry layer farm from Colombia.

Marek's disease (MD) is a lymphoproliferative disease caused by an Alphaherpesvirus, genus Mardivirus, serotype 1 (Gallid Herpesvirus 2, GaHV-2) that includes all known pathogenic strains. In addition to Marek's disease virus (MDV) serotype 1, the genus includes 2 distinct nonpathogenic serotypes: serotype 2 (GaHV-3) and serotype 3 (Meleagridis Herpesvirus 1, MeHV-1) which are used in commercially available vaccines against MD. As a result of vaccination, clinical signs are not commonly observed, and new cases are usually associated with emerging variant strains against which the vaccines are less effective. In this study, a commercial layer farm showing clinical signs compatible with MDV infection was evaluated. Histological lesions and positive immunohistochemistry in the sciatic nerve and thymus were compatible with cytolytic phase of MD. GaHV-2, GaHV-3 and MeHV-1 were identified by PCR and qPCR in blood samples from 17 birds with suspected MD. Analysis of the Meq gene of the Colombian GaHV-2 isolate revealed a 99% sequence identity with Asian strains, and in the phylogenetic analysis clustered with vv+ MDV. The analysis of amino acid alignments demonstrated an interruption of the proline rich region in P176A, P217A and P233L positions, which are generally associated with vv+ strains. Some of these changes, such as P233L and L258S positions have not been reported previously. In addition, primary cell cultures inoculated with lymphocytes isolated from the spleen showed typical cytopathic effect of GaHV-2 at 5 d post infection. Based on the molecular analysis, the results from this study indicate the presence of vv+ MDV infection in commercial birds for the first time in Colombia. It is recommended to perform further assays in order to demonstrate the pathotype characteristics in vivo.