Background and Aim: Extensive cattle rearing is a major source of economy for the inhabitants of the Amazon region of Peru. Milk and meat production is generally affected by the prevalence of various parasites, including hepatic and gastrointestinal parasites, as these products provide ideal conditions for parasitic growth. This poses a serious public health threat. This study aimed to estimate the prevalence, coinfection, and risk factors associated with the liver fluke (Fasciola hepatica) and other gastrointestinal parasites in cattle from the Amazon region of Peru. Materials and Methods: Fecal samples obtained from 1450 bovine specimens were analyzed using flotation and sedimentation methods to identify parasites, including Eimeria spp., strongyle-type eggs (STEs), and F. hepatica. We collected information about the specimens, including age, sex, origin, breed, category, frequency of deworming, farm size, herd size, water sources, and rearing system by conducting simple inspections and interviewing owners. The data obtained were statistically evaluated using the Chi-square test (p < 0.05) to determine the association between the qualitative variables. We also calculated the odds ratio at a 95% confidence interval to identify the risk factors. Results: We observed that F. hepatica, Eimeria spp., and STEs were 45.6%, 39.8%, and 35.3% prevalent, respectively. We found risk factors related to distomatosis in the animals from Huambo, where the drinking water sources are mainly streams, ditches, and rivers, while the specimens from Valle Chico were predisposed to coccidiosis. Further, the risk factors related to the presence of STEs in feces were age (61–90 months), origin (Valle Chico), herd size (<50 animals), and type of extensive rearing. Furthermore, significant coinfection was observed between Eimeria spp. and STEs. Conclusion: The high percentages of parasites in cattle observed were related to epidemiological factors, such as the origin of the sample, water sources, age, herd size, and extensive breeding. Similarly, the presence of STEs was a risk factor for contracting coccidiosis. Our future goals include investigating these parasites using a larger sample size and identifying more risk factors using more sensitive and specific diagnostic tests. Keywords: cattle, Eimeria spp., Fasciola hepatica, gastrointestinal parasites, prevalence, risk factors.
A major obstacle of widespread commercial application of bovine somatic cell nuclear transfer is the low overall efficiency, that is, healthy calf-late pregnancy per transferred embryo rate. In this study, we report a series of experiments with a limited number of embryos created with handmade cloning (HMC) and transferred without or after open pulled straw vitrification. Embryo reconstruction was performed by using in vitro matured oocytes and adult ear skin fibroblasts. In two experiments, a total of 53 D7 blastocysts were developed from 188 reconstructed embryos. Fresh transfer of seven blastocysts into six recipients has resulted in three early pregnancies, two of them developed over 90 days and eventually resulted in healthy calves (33% offspring/transfer rate). In another two experiments, a total of 11 D7 blastocysts were obtained from 36 reconstructed embryos. Out of these, eight have reexpanded 18 hours after vitrification and warming. Transfer of these blastocysts into eight recipients has resulted in four early pregnancies and two live births; 25% offspring/transfer rate. These results indicate that low overall efficiency may not be an intrinsic feature of cattle cloning, and selection of the right procedures may help to overcome the actual limitations.
RESUMENEn este estudio se demuestra el uso de la transferencia nuclear de células somáticas para producir los primeros bovinos clonados en el Perú. Se obtuvieron fibroblastos de piel y células de cúmulos de donantes adultos para ser usados como carioplastos; asimismo, ovocitos obtenidos a partir de ovarios de camal fueron madurados in vitro por 24 h. Los ovocitos madurados se incubaron 2 h en demecolcina (2.5 µg/ml) para promover la formación del cono con el plato metafásico y para orientar la enucleación manual. Se eliminó la zona pelúcida en pronasa (2 mg/ml) por 3 min. La enucleación fue manual con una microcuchilla dividiendo el óvulo en dos mitades, donde las mitades carentes de núcleo fueron fusionadas por el método «sandwich» (citoplasto-fibroblasto-citoplasto) por electrofusión. Las estructuras reconstruidas se activaron químicamente mediante incubación por 5 min en 7% de etanol absoluto, seguido por 5 h de citocalacina B (5µg/ml) y cicloheximida (10 µg/ml). Las estructuras se cultivaron durante 7 d hasta la fase de incubación/eclosión de blastocisto. Siete blastocistos fueron transferidos a seis vacas receptoras sincronizadas siete días después de la ovulación. Se logró la permanencia de cuatro y tres vesículas embrionarias hasta los días 28 y 60, respectivamente. Dos terneras llegaron a nacer a partir de embriones reconstruidos con células de piel y con células de cúmulos. Mediante el análisis de genotipos, utilizando 15 marcadores (SSR) para bovinos, se confirmó que los terneros clonados fueron derivados de las líneas celulares de las donantes.Palabras clave: reproducción asistida; bovino; clonación; transferencia nuclear de cé-lulas somáticas; clonación hecha a mano
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