Use of BACTEC radiometric culture method and polymerase chain reaction for the rapid screening of faeces and tissues for Mycobacterium paratuberculosis

SUMMARY The BACTEC radiometric culture method for detection of Mycobacterium paratuberculosis was evaluated on faeces from cattle on a farm in quarantine for Johne's disease. A multiplex polymerase chain reaction (PCR) based on the IS900 sequence specific for M paratuberculosis and a genus specific 16S rRNA region was developed and used to test cultures showing evidence of mycobacterial growth in the BACTEC liquid radiometric culture medium. Using the BACTEC - PCR combination, confirmation of M paratuberculosis from faeces and tissue of clinically affected animals was achieved within 2 to 4 weeks and 1 week, respectively, a substantial improvement on traditional culture and identification methods. The PCR provided rapid exclusion of M paratuberculosis when other Mycobacterium spp were grown. The radiometric culture medium proved to be very sensitive for culturing Mycobacterium spp.     

Responses of pregnant ewes and young lambs to cold exposure

The effects of cold stress were studied in pregnant ewes during the last three weeks of gestation and in their progeny during the first three days of life. In general, ewes were unaffected by treatment whereas changes were observed in the cold-stressed lambs. Cold-induced changes in lambs included physical weakness, depression, and poor nursing response. Serum concentrations of glucose and insulin were lowered whereas concentrations of blood urea nitrogen, alkaline phosphatase, aspartate aminotransferase, lactate dehydrogenase, triglycerides, and cortisol tended to be higher in cold exposed lambs. The mortality rate was higher (40%) in cold-stressed lambs than in lambs kept at warmer temperatures (10%). At necropsy, cold-exposed lambs had reduced amounts of adipose tissue in perirenal areas, and extensive subcutaneous hemorrhages and edema in the distal portions of the thoracic and pelvic limbs.     

CD4+ and CD8+ T- lymphocytopenia in a filly with Pneumocystis carinii pneumonia

Decreased proportion of CD4+ and CD8+ T lymphocytes in peripheral blood likely contributed to susceptibility to Pneumocystis carinii in a foal. Cytological evaluation of bronchoalveolar lavage was required for identification of the pathogen and serial flow-cytometric analysis of peripheral blood lymphocytes documented transient low expression of CD4+ and CD8+ T lymphocytes. Although immunodeficiency is uncommon, it must be included in the differential diagnosis for patients suffering from chronic or opportunistic infections and may provide an indication for immunostimulant therapy.     

Monitoring changes in back fat thickness and its effect on the restoration of ovarian activity and fertility in Bos indicus cows

With the objective of testing the hypothesis if animals with a stable layer of body fat (FAT) during the peripartum have a better chance of becoming pregnant after calving, fifty‐nine multiparous Brahman cows in their last trimester of pregnancy were used. Animals averaged four parturitions and were stocked at a rate of 1.25 animal units per hectare and divided into two groups depending on the time postpartum (dpp) that the intravaginal releasing device CIDR was inserted; Group 1 (<30 dpp; n = 30) received the implant at 25.2 ± 4.21 and withdrawn 9 days later. Group 2 (≥30 dpp; n = 29) received the CIDR at 38.41 ± 5.8. Animals were AI at detected oestrus until 170 dpp and calculated as pregnant at first service or requiring more than one service (1s and >1s), not pregnant but cycling (not pregnant) and those not cycling at all (anestrus). The FAT measurements were taken twice each month from the last trimester of gestation until 96 dpp. The onset of ovarian activity was monitored through blood levels of progesterone (P4) at days 14 and 9 prior to CIDR insertion and days 10, 13, 30 and 33 after CIDR withdrawal. Animals pregnant did not have any major changes in their fat thickness. In contrast, cows pregnant in the group ≥30 dpp had changes in their FAT homoeostasis, and pregnant animals in the 1s and >1s groups did not show differences in dorsal back fat in the last trimester of pregnancy and early postpartum. In contrast, animals not pregnant and in anestrus FAT values decreased considerably after parturition. Overall, fertility was 49%, but 18% of all the animals remained anestrus losing FAT. Thus, animals with adequate metabolic conditions will have a better chance of pregnancy regardless of the time postpartum when the reproductive programme starts.     

The relationship between intestinal and oral mucosa microcirculation in anaesthetized horses

Objective

To compare alteration in intestinal blood flow in anaesthetized horses with changes in oral mucosa blood flow.

24-Hour Secretion Patterns of Plasma Oestradiol 17β in Pony Mares in Late Gestation

The mare exhibits nocturnal uterine contractions in the last 6 days of gestation. It is hypothesized that estradiol 17β (O17β) may be associated with the nightly increase in uterine contractions. The 24‐h secretion pattern of plasma O17β was measured in 3 pony mares in late gestation to identify changes in release as the mare neared parturition. Blood was collected weekly at 08:00 hours beginning on day 240 and every third day from day 330 until delivery. Serial blood samples were collected from each mare every 30‐min for 24‐h beginning on gestation day 310 and every sixth day thereafter until parturition. Concentrations of O17β were elevated at night with lowest concentrations occurring directly before sunset (p < 0.01). The natural log of the variance was increased at sunset (p < 0.01) and was decreased during the 6‐h period immediately after sunrise. This pattern was especially evident in the 6 days that preceded parturition. The contrast between nocturnal and daytime concentrations of O17β in the last 6 days of gestation may contribute to night‐time delivery in the mare.     

Influence of xylazine on the function of the LiDCO sensor in isoflurane anaesthetized horses

ObjectivePrevious studies showed an influence of xylazine on the LiDCO sensor in vitro and in standing horses, but did not prove that this interaction caused error in LiDCO measurements. Therefore, agreement of cardiac output (CO) measurements by LiDCO and bolus-thermodilution (BTD) was determined in horses receiving xylazine infusions.Study designProspective, experimental study.AnimalsEight Warmblood horses.MethodsAll horses were premedicated with xylazine. Anaesthesia was induced with midazolam and ketamine and was maintained with isoflurane in oxygen. During six hours of anaesthesia CO measurements and blood samples were taken before, during and after a 60 minute period of xylazine infusion. Pairs of LiDCO and bolus thermo-dilution (BTD) measurements of CO were performed. Sensor voltages exposed to blood and saline were measured before, during and after xylazine infusion and compared using Bland-Altman method of agreement with corrections for repeated measures.ResultsThe CO values (mean ± SD) before xylazine were 34.8 ± 7.3 and 36.4 ± 8.1 L minute⁻¹ for BTD and LiDCO, respectively. After starting the xylazine infusion, the CO values for BTD decreased to 27.5 ± 6.1 L minute⁻¹ whereas CO values measured by LiDCO increased to 54.7 ± 18.4 L minute⁻¹. One hour after discontinuing xylazine infusion, CO values were 33 ± 6.7 and 36.5 ±11.9 L minute⁻¹ for BTD and LiDCO, respectively. The difference between saline and blood exposed sensor voltages decreased during xylazine infusion and these differences were positive numbers before but negative during the infusion. There were correlations between xylazine plasma concentrations, CO differences and sensor voltage differences (saline – blood).Conclusions and clinical relevanceThis study proved that xylazine infusion caused concentration dependent bias in LiDCO measurements leading to an overestimation of readings. Sensor voltage differences (saline – blood) may become valuable clinical tool to predict drug-sensor interactions.