Heat identification by 17beta-estradiol and progesterone quantification in individual raw milk samples by enzyme immunoassay

Background: There is a substantial decline in first-service-pregnancy-rate in dairy cows. In this regard, future prospects are to measure milk hormones on-farm and progesterone levels in milk are not enough to precise ovulation unless connected to other data. The objectives of this study were to investigate whether 17beta-estradiol could be measured from individual cow milk samples using a commercially available non-radiolabelled enzyme immunoassay kit (EIA) with no previously reported milk application, and whether those detections could precisely illustrate 17beta-estradiol pre-ovulation peak in spite of its limited concentration and short manifestation in milk. Results: Milk sample treatments for progesterone and 17beta-estradiol EIA measurements are described. Hormonal profiles from daily milk samples of six different cows were reported and 17beta-estradiol pre-ovulation peak was visualized in all cases. Heat detection was possible by EIA using one every 2 days milking samples in almost all studied cases. Only in one case, morning and afternoon milking samples were required to visualize the 17beta-estradiol pre-ovulation peak. Conclusions: 17beta-estradiol EIA quantification in raw milk is a reliable, rapid, economic and a precise method to describe cow heat along with EIA progesterone determination.

Comparison of commercially-available RNA extraction methods for effective bacterial RNA isolation from milk spiked samples

Nucleic-acid based methods for bacterial identification are extremely useful in diagnostic applications due to their specificity and sensitivity. However, they require an optimal purification of the target molecules. As part of the development of a new diagnostic method for the detection of bacterial RNA in cow milk, we have compared four commercially available RNA extraction kits for the isolation of bacterial RNA from spiked UHT milk samples. The kits were compared in terms of extraction efficiency and RNA purity using two bacterial species, the Gram negative Escherichia coli and the Gram positive Staphylococcus aureus. Two kits are based in silica-matrix extraction, and the other two in the guanidinium thiocyanate-phenol-chloroform extraction. In our hands, the RNeasy Protect Bacteria Mini kit from QIAGEN was the best performing in terms of RNA yield, quality, reproducibility and consumable needs, under the conditions here described.

Queratitis por Alteraria alternata post queratoplastia penetrante / Keratitis caused by alternaria alternata after penetrating keratoplasty

Objetivo. Informar el caso segundo caso de queratitis por Alternaria alternata después de transplante corneal. Material y método. Se trata de una mujer de 46 años de edad sometida a cirugía refractiva con la colocación de un lente intraocular fáquico, con una queratopatía bulosa secundaria tratada con transplante corneal. Posteriormente desarrolló queratitis por Alternaria alternata. Resultados. El laboratorio de microbiología reportó la presencia de un hongo con hifas septadas, que se identificó como Alternaria alternata. El estudio histopatológico reveló la presencia de hifas septadas y ramificadas en el estroma corneal. Conclusiones. Se sugiere el uso de antimicóticos a dosis bajas en el medio de preservación, para evitar que se conserven esporas.