Real-time PCR as a surveillance tool for the detection of Trichinella infection in muscle samples from wildlife

Trichinella nematodes are the causative agent of trichinellosis, a meat-borne zoonosis acquired by consuming undercooked, infected meat. Although most human infections are sourced from the domestic environment, the majority of Trichinella parasites circulate in the natural environment in carnivorous and scavenging wildlife. Surveillance using reliable and accurate diagnostic tools to detect Trichinella parasites in wildlife hosts is necessary to evaluate the prevalence and risk of transmission from wildlife to humans. Real-time PCR assays have previously been developed for the detection of European Trichinella species in commercial pork and wild fox muscle samples. We have expanded on the use of real-time PCR in Trichinella detection by developing an improved extraction method and SYBR green assay that detects all known Trichinella species in muscle samples from a greater variety of wildlife. We simulated low-level Trichinella infections in wild pig, fox, saltwater crocodile, wild cat and a native Australian marsupial using Trichinella pseudospiralis or Trichinella papuae ethanol-fixed larvae. Trichinella-specific primers targeted a conserved region of the small subunit of the ribosomal RNA and were tested for specificity against host and other parasite genomic DNAs. The analytical sensitivity of the assay was at least 100 fg using pure genomic T. pseudospiralis DNA serially diluted in water. The diagnostic sensitivity of the assay was evaluated by spiking 10 g of each host muscle with T. pseudospiralis or T. papuae larvae at representative infections of 1.0, 0.5 and 0.1 larvae per gram, and shown to detect larvae at the lowest infection rate. A field sample evaluation on naturally infected muscle samples of wild pigs and Tasmanian devils showed complete agreement with the EU reference artificial digestion method (k-value=1.00). Positive amplification of mouse tissue experimentally infected with T. spiralis indicated the assay could also be used on encapsulated species in situ. This real-time PCR assay offers an alternative highly specific and sensitive diagnostic method for use in Trichinella wildlife surveillance and could be adapted to wildlife hosts of any region.     

Effects of norepinephrine and combined norepinephrine and fenoldopam infusion on systemic hemodynamics and indices of renal function in normotensive neonatal foals

Background: Norepinephrine increases arterial blood pressure but may have adverse effects on renal blood flow. Fenoldopam, a dopamine-1 receptor agonist, increases urine output in normotensive foals. The combination of norepinephrine and fenoldopam may lead to improved renal perfusion compared with an infusion of norepinephrine alone. The combined effects of these drugs have not been reported in the horse.
Hypothesis: Norepinephrine will alter the hemodynamic profile of foals without affecting renal function. Addition of fenoldopam will change the renal profile during the infusions without changing the hemodynamic profile.
Animals: Five conscious pony foals.
Methods: Each foal received norepinephrine (0.3 μg/kg/min), combined norepinephrine (0.3 μg/kg/min) and fenoldopam (0.04 μg/kg/min), and a control dose of saline in a masked, placebo-controlled study. Heart rate (HR), arterial blood pressure (direct), and cardiac output (lithium dilution) were measured, and systemic vascular resistance (SVR), stroke volume, cardiac index (CI), and stroke volume index were calculated. Urine output, creatinine clearance, and fractional excretion of electrolytes were measured.
Results: Norepinephrine and a combined norepinephrine and fenoldopam infusion increased arterial blood pressure, SVR, urine output, and creatinine clearance and decreased HR and CI compared with saline. The combination resulted in higher HR and lower arterial blood pressure than norepinephrine alone.
Conclusions and Clinical Importance: Norepinephrine might be useful for hypotensive foals, because in normal foals, this infusion rate increases SVR without negatively affecting renal function (creatinine clearance increased). Fenoldopam does not provide additional benefit to renal function. These findings warrant further investigation.     

Comparison of lithium dilution and thermodilution cardiac output measurements in anaesthetised neonatal foals

Knowledge of cardiac output is expected to help guide the treatment of hypotension associated with critical illness and/or anaesthesia in neonatal foals. However, a practical and safe method of measuring cardiac output has not been described for the foal. Lithum dilution, a new method of cardiac output determination not requiring cardiac catheterisation, has recently been reported in mature horses. We compared this method to thermodilution in isoflurane-anaesthetised foals age 30-42 h and found good agreement between the 2 methods in a range of cardiac outputs 5.4-20.4 l/min. The lithium dilution technique is a practical and reliable method of measuring cardiac output in anaesthetised neonatal foals, and warrants investigation in critically ill conscious foals.     

Use of lithium dilution and pulse contour analysis cardiac output determination in anaesthetized horses: a clinical evaluation

OBJECTIVE: To assess the suitability of a human algorithm for calculation of continuous cardiac output from the arterial pulse waveform, in anaesthetized horses. STUDY DESIGN: Prospective clinical study. ANIMALS: Twenty-four clinical cases undergoing anaesthesia for various conditions. MATERIALS AND METHODS: Cardiac output (Qt), measured by lithium dilution (QtLiDCO), was compared with a preceding, calibrated Qt measured from the pulse waveform (QtPulse). These comparisons were repeated every 20-30 minutes. Positive inotropes or vasopressors were administered when clinically indicated. Cardiac indices from 30.7 to 114.9 mL kg(-1) minute(-1) were recorded. Unusually shaped QtLiDCO curves were rejected and the measurement was repeated immediately. RESULTS: Eighty-nine comparisons were made between QtLiDCO and QtPulse. The bias between the mean (+/-SD) of the two methods (QtLiDCO - QtPulse) was -0.07 L minute(-1)(+/-3.08) (0.24 +/- 6.48 mL kg(-1) minute(-1)). The limits of agreement were -12.72 and 13.2 mL kg(-1) minute(-1) (Bland & Altman 1986; Mantha et al. 2000). Linear regression analysis demonstrated a correlation coefficient (r2) of 0.89. Cardiac output in individual patients varied from 49.1 to 183% of the initial measurement at the time of calibration. Linear regression of log-transformed Qt variation for each method found a mean difference of 9% with limits of agreement of -4.1 to 22.1%. CONCLUSIONS AND CLINICAL RELEVANCE: This method of pulse contour analysis is a relatively noninvasive and reliable way of monitoring continuous Qt in the horse under anaesthesia. The ability to easily monitor Qt might decrease morbidity and mortality in the anaesthetized horse.     

The physiological basis for genetic improvement of oil palm in Malaysia

Summary Yield of Deli dura oil palms after four generations of selection was 60% greater than that of the unselected base population. Total above-ground dry matter production was increased by selection, apparently through better utilisation of solar radiation. The dry matter requirement for vegetative growth was unchanged, so a greater surplus remained for fruit production in the selected palms. Crossing the dura and pisifera to give the thin-shelled tenera fruit type improved partitioning of dry matter within the fruit, giving a 30% increase in oil yield at the expense of shell, without changing total dry matter production.     

Boron: possible role in plant metabolism

Leaves of boron-deficient oil palm showed a total absence of the leucoanthocyanins usually present, well before onset of pathological symptoms. The association of boron with flavonoid synthesis is consistent with the otherwise anomalous situation that this element is essential for higher plants but not for animals or lower plants.     

IMPACT OF VIDEO IMAGES ON O.F.A. EVALUATION OF HIP STATUS

The Orthopedic Foundation for Animals (OFA), in an attempt to reduce reporting time and storage space, installed several remote video stations. However, these improvements would be of questionable benefit if the OFA consensus were to be effected. Therefore, the video image and corresponding radiographic image of OFA submissions were used to evaluate consistency in evaluations of the radiologists using the remote video stations. There was no apparent difference between the two methods.