PCR-based technology in veterinary parasitology.

DNA technology is having a major impact in many areas of veterinary parasitology. In particular, the polymerase chain reaction (PCR) has found broad applicability because its sensitivity permits enzymatic amplification of gene fragments from minute quantities of nucleic acids derived from limited amounts of parasite material. This paper discusses some recent applications of PCR-based methods to parasites and highlights their usefulness or potential for those of veterinary importance. The focus is on PCR tools for the accurate identification of parasites and their genetic characterisation, the diagnosis of infections, the isolation and characterisation of expressed genes, the detection of anthelmintic resistance, and mutation scanning approaches for the high resolution analysis of PCR products.     

Drug disposition and dosage determination of once daily administration of gentamicin sulfate in horses after abdominal surgery.

OBJECTIVE: To evaluate pharmacokinetics of once daily i.v. administration of gentamicin sulfate to adult horses that had abdominal surgery. DESIGN: Prospective study. ANIMALS: 28 adult horses that underwent abdominal surgery for colic. PROCEDURE: 14 horses were treated with each dosage of gentamicin (i.e., 6.6 or 4 mg/kg, i.v., q 24 h) and blood samples were collected for pharmacokinetic analysis. Plasma gentamicin concentrations were measured by use of a fluorescence polarization immunoassay. Pharmacokinetic analysis measured the elimination half-life, volume of distribution, and gentamicin total systemic clearance. Treatment outcome, CBC, and serum creatinine concentrations were recorded. RESULTS: 1 horse in the high-dosage group died. All other horses successfully recovered, and did not develop bacterial infection or have evidence of drug toxicosis resulting in renal injury. Mean pharmacokinetic variables for gentamicin administration at a high or low dosage (i.e., 6.6 or 4 mg/kg, i.v., q 24 h) were half-life of 1.47 and 1.61 hours, volume of distribution of 0.17 and 0.17 L/kg, and systemic clearance of 1.27 and 1.2 ml/kg/min, respectively. Mean serum creatinine concentration was 1.74 and 1.71 for the high and low dosages, respectively, and serum creatinine concentration was not correlated with gentamicin clearance. CONCLUSIONS AND CLINICAL RELEVANCE: Gentamicin administration at a dosage of 4 mg/kg, i.v., every 24 hours, will result in plasma concentrations that are adequate against susceptible bacteria with a minimum inhibitory concentration (MIC) of < or = 2.0 micrograms/ml. Gentamicin administration at a calculated dosage of 6.8 mg/kg, i.v., every 24 hours will result in optimum plasma concentrations against susceptible bacteria with a MIC of < or = 4.0 micrograms/ml.     

Influence of high ambient temperatures on performance of multiparous lactating sows.

Multiparous Large White sows (n = 63) were used to investigate the effects of five ambient temperatures (18, 22, 25, 27, and 29 degrees C) and two dietary protein contents on their lactation performance. At each temperature treatment, ambient temperature was maintained constant over the 21-d lactation period. Dietary protein content was either 14 or 17% with essential amino acids levels calculated not to be limiting. The animals had ad libitum access to feed between the seventh and the 19th day of lactation. Diet composition did not influence lactation performance. Over the 21-d lactation, feed intake decreased from 5.67 to 3.08 kg/d between 18 and 29 degrees C. Between d 7 and 19, the corresponding values were 7.16 and 3.48 kg/d, respectively. This decrease was curvilinear; an equation to predict voluntary feed intake (VFI) from temperature (T, degrees C) and body weight (BW, kg) is proposed: VFI = -49,052 + 1,213 T - 31.5 T2 + 330 BW - .61 BW2 (residual standard deviation: 1,018). Skin temperature increased regularly with increased ambient temperature (34.6 to 37.4 degrees C between 18 and 29 degrees C), whereas udder temperature reached a plateau at 25 degrees C (38.3 degrees C). The gradient of temperature between skin and rectum was minimal (2 degrees C ) at 27 degrees C and remained constant at 29 degrees C. This constancy coincides with the marked reduction of feed intake. The respiratory rate increased from 26 to 124 breaths/min between 18 and 29 degrees C, and this indicates that the evaporative critical temperature was below 22 degrees C. The BW loss increased from 23 to 35 kg between 18 and 29 degrees C, but its estimated chemical composition remained constant. Pig growth rate was almost constant between 18 and 25 degrees C (241 g/d) and was reduced above 25 degrees C (212 and 189 g/d at 27 and 29 degrees C, respectively). In conclusion, temperatures above 25 degrees C seem to be critical for lactating sows in order to maintain their performance.     

Effects of sire growth potential, growing-finishing strategy, and time on feed on performance, composition, and efficiency of steers.

Beef production systems that increase use of unharvested forages and use animals with greater potential for gain affect age and size of animals placed on a finishing regimen. This experiment was conducted to evaluate effects of genetic potential for gain, age at the start of a finishing period, and time on feed on composition, quantity, and quality of beef produced and efficiency of production during finishing. Crossbred cows were bred by AI to Charolais or Line 1 Hereford bulls that represented potentially high (HG) or moderate growth (MG) rates, respectively, to produce spring- or fall-born calves. Steer calves from these matings were placed on an individually fed finishing diet at three ages (A). Spring-born steers were started at 6 or 18 mo of age (A6 and A18), and fall-born steers were started at 12 mo of age (A12). Slaughter times (T) were at 0, 90, 180, and 270 d for A6; 68, 136, and 204 d for A12; and 0, 45, 90, and 135 d for A18. Data collected on each animal included feed intake, growth, chemical composition of the complete body and carcass, and quantitative and qualitative assessment of the meat produced. Four steers of each sire group were slaughtered in each of the 11 A-T treatment groups, and the experiment was repeated for 2 yr in the A12 groups and 3 yr in the A6 and A18 groups (n = 237). Steers sired by HG bulls were larger and produced larger carcasses and more carcass protein than MG-sired steers (S, P < .05 or .01). Steers sired by MG bulls were fatter, had higher quality grades, and accumulated fat at a faster rate than HG-sired steers, and this effect was greater in older steers (G and GA, P < .05 or .01). Sire growth potential did not affect gain, intake, live weight efficiency, tenderness, or taste panel scores (P > .2). Steers sired by HG bulls were more efficient at producing carcass weight and carcass protein at A12 and A18 than were MG-sired steers. At the end of the finishing period, older (A18), HG-sired steers were too large with insufficient fat by current industry standards, and younger (A6), MG-sired steers were too small. Our conclusions are that both HG- and MG-sired steers can produce acceptable carcasses for current market standards with comparable efficiencies of live-weight gain, but the growing and finishing strategy must be adapted to the genotype.     

Comparison of the kinetics of the specific cellular immune response to duck hepatitis B virus in infected and immune ducks.

The kinetics of the cell mediated immune response by ducks acutely and chronically infected with, or immune to infection by duck hepatitis B virus (DHBV) was determined. This was measured by an antigen specific blastogenesis assay to duck hepatitis B surface antigen (DHBsAg) and duck hepatitis B core antigen (DHBcAg) using peripheral blood mononuclear cells (PBMC). The three outcomes of acute infection by DHBV were either clearance from both serum and liver, clearance from serum but not liver, or the development of persistent viraemia. Acutely infected ducks that failed to clear the infection also failed to develop a significant cellular immune response to both antigens. Ducks with chronic infection acquired as neonates or as the result of the failure to clear acute infection had an increasing cellular immune response over time. Two groups of immune ducks were examined. These were either ducks that had become immune following infection or that had been vaccinated. Both groups of ducks demonstrated significant cellular responses following challenge with DHBV irrespective of the level of their responses before challenge. However, there was a reduction in the response of their PBMC over a 4-week-period postchallenge. The range of cellular immune responses to DHBV antigens observed in this study has a number of counterparts in hepatitis B infection of humans. Coupled with the defined clinical outcomes that can be established in the duck/DHBV model, further study of the cellular immune response to DHBV is warranted.     

Dual-component video image analysis system (VIASCAN) as a predictor of beef carcass red meat yield percentage and for augmenting application of USDA yield grades

An improved ability to quantify differences in the fabrication yields of beef carcasses would facilitate the application of value-based marketing. This study was conducted to evaluate the ability of the Dual-Component Australian VIASCAN to 1) predict fabricated beef subprimal yields as a percentage of carcass weight at each of three fat-trim levels and 2) augment USDA yield grading, thereby improving accuracy of grade placement. Steer and heifer carcasses (n = 240) were evaluated using VIASCAN, as well as by USDA expert and online graders, before fabrication of carcasses to each of three fat-trim levels. Expert yield grade (YG), online YG, VIASCAN estimates, and VIASCAN estimated ribeye area used to augment actual and expert grader estimates of the remaining YG factors (adjusted fat thickness, percentage of kidney-pelvic-heart fat, and hot carcass weight), respectively, 1) accounted for 51, 37, 46, and 55% of the variation in fabricated yields of commodity-trimmed subprimals, 2) accounted for 74, 54, 66, and 75% of the variation in fabricated yields of closely trimmed subprimals, and 3) accounted for 74, 54, 71, and 75% of the variation in fabricated yields of very closely trimmed subprimals. The VIASCAN system predicted fabrication yields more accurately than current online yield grading and, when certain VIASCAN-measured traits were combined with some USDA yield grade factors in an augmentation system, the accuracy of cutability prediction was improved, at packing plant line speeds, to a level matching that of expert graders applying grades at a comfortable rate.     

Clinical examination of fish in private collections.

The rapid advancement of fish medicine over the past 10 years has led to a drastically improved ability to manage the individual pet fish successfully. Principle physiologic differences between fish and higher vertebrates include the lack of ability to regulate body temperature (this has a profound effect on the immune system), the use of gills for respiration, and the importance of osmoregulation in the aquatic environment. Components of the clinical history for a fish patient should include information on the environment, tank mates, recent medical history, and specific information on the affected individual. Fish can be manually restrained or safely anesthetized with methane tricaine sulfonate, depending on their size, condition, and circumstances. Diagnostic techniques useful for fish patients include gill, skin, and fin biopsy; hematology; cytology; radiology; and ultrasound. Preventive medicine programs should include water quality, nutritional management, sanitation, and quarantine.     

Clinical features of avian vacuolar myelinopathy in American coots

OBJECTIVE: To characterize clinical features of avian vacuolar myelinopathy (AVM) in American coots. DESIGN: Case-control study. ANIMALS: 26 AVM-affected American coots and 12 unaffected coots. PROCEDURES: Complete physical, neurologic, hematologic, and plasma biochemical evaluations were performed. Affected coots received supportive care. All coots died or were euthanatized, and AVM status was confirmed via histopathologic findings. RESULTS: 3 severely affected coots were euthanatized immediately after examination. Seventeen affected coots were found dead within 7 days of admission, but 5 affected coots survived > 21 days and had signs of clinical recovery. Abnormal physical examination findings appeared to be related to general debilitation. Ataxia (88%), decreased withdrawal reflexes (88%), proprioceptive deficits (81%), decreased vent responses (69%), beak or tongue weakness (42%), and head tremors (31%), as well as absent pupillary light responses (46%), anisocoria (15%), apparent blindness (4%), nystagmus (4%), and strabismus (4%) were detected. Few gross abnormalities were detected at necropsy, but histologically, all AVM-affected coots had severe vacuolation of white matter of the brain. None of the control coots had vacuolation. CONCLUSIONS AND CLINICAL RELEVANCE: Although there was considerable variability in form and severity of clinical neurologic abnormalities, clinical signs common in AVM-affected birds were identified. Clinical recovery of some AVM-affected coots can occur when supportive care is administered. Until the etiology is identified, caution should be exercised when rehabilitating and releasing coots thought to be affected by AVM.