Applying a kinetic method to an indirect ELISA measuring Ostertagia ostertagi antibodies in milk

Indirect enzyme-linked immunosorbent assays (ELISAs) are frequently run as endpoint ELISAs (e-ELISAs). However, kinetic ELISAs (k-ELISAs) have certain advantages over e-ELISAs. The objective of this study was to understand the relationship between e-ELISA and k-ELISA results. Specifically, to determine whether it was possible to run both k-ELISA and e-ELISA on the same plate and establish an appropriate time interval for k-ELISA measurements. A normalization method for k-ELISA slopes (slope ratio) is proposed. Using an indirect e-ELISA test measuring antibodies against Ostertagia ostertagi in milk from dairy cattle, we found that running a k-ELISA had no effect on optical density ratio results of an e-ELISA on the same plate, and that agreement was very strong at 10, 15, and 28 min, allowing for a reduction in the total processing time for ELISA tests.     

Normal and aberrant tissue distribution of Loma salmonae (Microspora) within rainbow trout, Oncorhynchus mykiss (Walbaum), following experimental infection at water temperatures within and outside of the xenoma-expression temperature boundaries

Temperatures above 20 °C or below 9 °C interrupt the life cycle of the gill intracellular microsporidian parasite Loma salmonae (Microspora) prior to sporogony, inhibiting the production of xenomas. This study intended to characterize this life-cycle failure. Juvenile rainbow trout, Oncorhynchus mykiss (Walbaum), were experimentally infected with L. salmonae spores, and the effect of water temperature on the progress of infection, as determined by polymerase chain reaction, was compared for fish held at water temperatures of 5, 15 and 21 °C. At 15 °C, parasite DNA was first detected in the heart (3 days post-exposure [PE]), and then in the gills and spleen (2 weeks PE). Branchial xenomas developed by week 4 PE. In contrast, at 5 °C, the arrival of the parasite in the heart was delayed until 7 days PE. However, even though parasite DNA was detected in the gills at 7 days PE, xenomas failed to form in the gill, and by week 4 PE, parasite DNA was no longer detected. In fish held at 21 °C, parasite DNA was detected in the heart, gills and spleen by 3 days post-infection, and similar results were observed at 7 days PE. Xenomas also failed to form in these fish and parasite DNA was no longer detected by week 2 PE. Within the range of temperatures tested in this study, spore germination and delivery of their DNA into the host through the intestinal wall was not blocked by temperature. At 5 or 21 °C, migration to the heart and gills occurred, but at aberrant periods of time. The normal life cycle of L. salmonae may depend on the completion of relatively lengthy, but yet unknown, stages of development within the heart, prior to reaching the gill. This development may be adversely affected by temperature, and explain the temperature limits of this parasite.     

Serologic studies of bovine respiratory syncytial virus in Minnesota cattle

Serum antibody titers to bovine respiratory syncytial virus were determined for 559 cattle. Serum samples were obtained through the Minnesota State-Federal Brucellosis Laboratory and were collected over a 1-year period. Results of this study revealed an antibody prevalence of 65.5% to bovine respiratory syncytial virus. The distribution of antibody titers is presented, as well as analysis of titers based on breed, sex, and age of the cattle.     

Inhibition of phagosome-lysosome fusion in macrophages by soluble extracts of virulent Brucella abortus

Water extracts of virulent Brucella abortus were able to inhibit phagosome-lysosome fusion in unelicited murine peritoneal macrophages following ingestion of yeast. Extracts from an avirulent strain were unable to produce a similar effect. Lipopolysaccharide from B abortus did not appear to be involved with the ability of the extracts to inhibit fusion.     

Liver biopsy techniques for adult horses and neonatal foals to assess copper status

Objective To evaluate standing, percutaneous, ultrasound-guided, transthoracic liver biopsy in mares, and transabdominal, laparoscopically-guided, liver biopsy under general anaesthesia in foals, as techniques for obtaining tissue for assessment of copper status. The techniques were evaluated with respect to ease of use and effect on the animal.
Procedure Twenty of 24 Thoroughbred mares and 21 of their foals were biopsied. The animals were part of a larger study of the effect of copper supplementation on copper status and the prevalence of developmental orthopaedic disease. Livers were also collected from unrelated horses and sampled to investigate the variability in the distribution of copper in liver tissue.
Result The biopsy technique caused no lasting effect on the mares, but there was an increased risk of viscus penetration associated with taking multiple biopsy cores. The use of ultrasonography to scan the target area for the liver identified four cases that were not appropriate candidates for liver biopsy, because of large intestine being located in the biopsy area. In the foals there were no serious postoperative adverse effects, nor was there any evidence of problems caused by the procedure when the abdomen was examined post-mortem at 5 months of age. In livers collected to investigate the variability of copper concentration, copper appeared to be relatively evenly distributed through the liver.
Conclusion Standing, percutaneous, ultrasound-guided, transthoracic liver biopsy in mares, and transabdominal, laparoscopically-guided, liver biopsy under general anaesthesia in foals are convenient procedures for obtaining liver tissue for assessing copper status in horses. The use of ultrasound to identify liver tissue is recommended, especially in older mares.