DNA diagnosis in veterinary medicine: II. Polymerase chain reaction (PCR)

DNA diagnostic has become very important for genetic and infectious diseases. With a new method, called the polymerase chain reaction (PCR), specific pieces of genomes from viruses, bacteria, parasites or even cells can be amplified more than hundred million fold in a very short time period. For the following reasons this opens a new dimension in diagnostics: (1) The technique is so sensitive that a single virus, bacterium, parasite or cell is sufficient to be detected provided part of its nucleic acid sequence is known. (2) The material does neither need to be fresh nor be stored under special conditions since the nucleic acids are much more stable than e.g. enzymes or other proteins. (3) The method is quicker than the usual immunodetection methods. (4) The polymerase chain reaction has been standardized recently so that it can be carried out in diagnostic laboratories. The purpose of this minireview is to give an introduction into the basic technique and to mention possible and already existing applications in veterinary medicine.     

Effects of long- or short-term exposure to a calf identified as persistently infected with bovine viral diarrhea virus on feedlot performance of freshly weaned, transport-stressed beef heifers

A single experiment with a completely randomized design was conducted to evaluate the effects of long- or short-term exposure to a calf identified as persistently infected with bovine viral diarrhea virus (PI-BVD) on feedlot performance and carcass characteristics of freshly weaned, transport-stressed beef heifers. Two hundred eighty-eight heifers that had been vaccinated for BVD before weaning and transport were processed and given a metaphylactic antibiotic treatment at arrival and were fed common receiving, growing, and finishing diets for a 215-d period. Treatments were designed to directly or adjacently expose the cattle to a PI-BVD heifer. Directly exposed treatments were 1) negative control with no PI-BVD calf exposure (control), 2) PI-BVD calf commingled in the pen for 60 h and then removed (short-term exposure), and 3) PI-BVD calf commingled in the pen for the duration of the study (long-term exposure); and spatially exposed treatments were 1) negative control with no PI-BVD calf exposure (adjacent pen control), 2) PI-BVD calf commingled in the adjacent pen for 60 h and then removed (adjacent pen short-term exposure), and 3) PI-BVD calf commingled in the adjacent pen for the duration of the study (adjacent pen long-term exposure). Exposure to a PI calf transiently (60 h) or for the duration of the feeding period (215 d) did not affect (P > or = 0.25) final BW compared with heifers that were not exposed. Neither period nor overall DMI was affected (P > or = 0.37) by PI-BVD calf exposure, and no differences (P > or = 0.44) were observed between short- and long-term exposed heifers in the direct or spatially exposed groups. Likewise, total trial ADG was not affected (P > or = 0.36) and overall efficiency of gain (P > or = 0.19) was unaffected by PI-BVD calf exposure in the direct or spatially exposed groups. The results from this study suggest that exposing previously vaccinated, freshly weaned, transport- stressed beef calves to a calf that is persistently infected with bovine viral diarrhea virus has little, if any, marked effects on health, performance, or carcass characteristics.     

Locking compression plate osteosynthesis of sacral fractures in three heifers

Objective: To describe a technique used for reduction and internal fixation of sacral fractures in cattle. Study Design: Case series. Animals: Three heifers aged 19–32 months with deformation of the tail head and tail paralysis attributable to a closed fracture of S5 or S4 and S5. Methods: Fractures were reduced and repaired under general anesthesia. A narrow, 6–10‐hole, locking compression plate (LCP) was placed on the spinous processes of S5 and the first coccygeal (Co1) using 5.0 mm locking head screws. Follow‐up inquiries were made by telephone (3/3), and cows were examined clinically and radiographically 2 (1 cow) to 9 (1cow) months postoperatively. Results: No complications were encountered. The range of movement of the tail improved so that soiling of the tail during defecation was no longer a problem. In all heifers, the normal top line was restored and there was no narrowing of the pelvic canal. Unassisted calving occurred in all heifers, and follow‐up radiographs for 2 cows showed complete fracture healing. Conclusion: LCP‐osteosynthesis of the spinous processes provided sufficient stability for internal fixation of sacral fractures. Marked improvement in tail function occurred postoperatively without decompression by laminectomy.     

Growth of the digestive organs in ducks with considerations on their growth in birds in general.

1. Growth of the oesophagus, proventriculus, gizzard, intestine, liver and pancreas weight was investigated in Mallards, White Pekins, Muscovies and a Muscovy x White Pekin cross. The birds varied in age between hatching and 154 d. The data were analysed by fitting both the Janoschek growth curve and the allometric formula. 2. The growth rate of all organs, except the oesophagus, peaked earlier (30 d for Muscovies and 14 d for the other breeds) than body weight and they grew faster to any given percentage of their final weight. In contrast, oesophagus weight showed growth curve characteristics similar to body weight. 3. Oesophagus weight showed simple, slightly negative allometry. The remaining organs followed complex allometry that can approximately be described by 2 allometric stages. The 1st phase was isometric to positive allometric. The 2nd showed marked negative allometry. 4. These growth patterns are assumed to be generally present in birds.     

Elephant herpes virus--a problem for breeding and housing of elephants]

Herpesvirus infections which take a fatal turn on African elephants as well as on Asian elephants seem to occur increasingly not only in the USA but also in European stocks. The endotheliotropic herpesvirus causes a rapidly progressing and severe disease which makes any therapeutical effort unsuccessful and finally results in death of the animal, especially in young Asian elephants. As all attempts to culture the virus failed up to now, molecular biological procedures have to be used more often for diagnostical purpose together with the common methods of pathology, virology, and electronmicroscopical evaluation. This is a report on the case of 'KIBA', an eleven year old male elephant at the Zoological Garden Berlin, infected with the endotheliotropic elephants herpesvirus. 'KIBA' was born at the Zoo in Houston, Texas, and raised within his herd. Upon arriving in Berlin in November 1997 he adapted to the new premises and climate and new social circumstances without any problems. In June 1998 he already serviced three females of his new herd several times. In August 1998 he died after passing a peracute progression of the disease after residenting in Berlin for only 9 months. The dissection of the animal revealed some evidence on an agent damaging the endothelium. Major signs indicating this agent were bleedings in several serous membranes, mucosa and on the the right atrium, as well as other parts of the myocardium. Furthermore there have been ulcerations at various localisations of the whole digestive tract. Slightly basophilic intranuclear inclusion bodies have been found histologically in endothelial cells of different organ samples. An examination of altered organ-material by electronmicroscopy made some herpesvirus-like particles visible. A virological investigation first revealed evidence of giant cell formations with solitary basophilic intranuclear inclusion bodies in different cell cultures, however, without any distinct cytopathogenic effect. Supported by molecular biological procedures the infection of 'KIBA' could be verified as the elephants herpesvirus. By means of PCR and subsequent sequence analysis a DNA-sequence typical for the elephants herpesvirus could be obtained which showed an identity of 97% with the terminase sequence of the elephant herpesvirus described by American authors. The deduced amino acid-sequences were 100% identical. To the terminase of the human cytomegalovirus, the elephant sequence had an identity of 53% (similarity: 74%). Based on the cooperation of ILAT, Institute of Veterinary-Pathology/Free University Berlin, Robert-Koch-Institut Berlin, and Zoological Garden Berlin, the cause of 'KIBA's' death could be discovered immediately. Possible implications of this case especially on breeding and keeping elephants are discussed briefly.     

Comparison of haemodynamic changes during two surgical methods for neutering female dogs

The aim was to investigate if intraoperative registrations of blood pressure and heart rate could be used to estimate surgical trauma. In a prospective clinical trial, registrations in eight dogs neutered by laparoscopic ovariectomy (LOE) were compared with eight dogs subjected to open ovariohysterectomy (OHE). For comparisons phases were used: phase zero = steady state after induction of anaesthesia; phase one = opening of abdomen; phase two = severing of ovarian pedicles; and, phase three = abdominal closure. During LOE, mean systolic blood pressure increased by 7 mm Hg (p = 0.05) and 15 mm Hg (p = 0.01) from phase zero to phase one and two, respectively, and during OHE by 3 mm Hg (ns) and 29 mm Hg (p < 0.0001), respectively. The increase from phase one to phase two differed between groups (p = 0.03). Heart rate did not change. Frequent intraoperative measurements of blood pressure appear a promising method for evaluating surgical trauma.     

Effects of supplementation of whole corn germ on reproductive performance, calf performance, and leptin concentration in primiparous and mature beef cows

A 2-yr study using primiparous and multiparous, spring-calving, crossbred beef cows was conducted to evaluate the effects of supplemental whole corn germ on reproductive performance, calf performance, and serum leptin concentrations. Each year, cows were blocked by age and BCS and assigned randomly to one of three treatments: PRE (n = 115) cows received 1.14 kg/d (DM basis) of whole corn germ for approximately 45 d before calving; POST (n = 109) cows were fed 1.14 kg/d of whole corn germ for approximately 45 d after calving; and control cows (n = 118) were fed similar energy and protein from dry-rolled corn (1.82 kg of DM/d) for 45 d before and after calving. Additionally, PRE cows were grouped with controls after calving, and POST cows were grouped with control cows before calving, so that corn germ-supplemented cows received the control supplement in the alternate feeding period. Cow BW (538 +/- 13 kg) and BCS (5.4 +/- 0.13) did not differ among treatments at any time during the experiment. Calf birth weight (39 +/- 2 kg), weaning weight (225 +/- 7 kg), and age-adjusted weaning weight (234 +/- 8 kg) did not differ because of dam supplementation regimen. Treatment did not affect the proportion of cows exhibiting ovarian luteal activity before the start of the breeding season (67%) or pregnancy rate (91%). The interval from exposure to bulls until subsequent calving did not differ (P = 0.16) among PRE (298 +/- 2.3 d), POST (303 +/- 2.6 d), and control (304 +/- 2.3 d) cows. Leptin concentrations did not differ among treatments and were 2.15 +/- 0.75, 1.88 +/- 0.76, and 1.91 +/- 0.75 ng/mL for control, POST, and PRE cows, respectively. Age and week relative to calving influenced leptin concentration. Primiparous cows had similar leptin concentrations to 3-yr-old and mature cows for wk -7 and -6 relative to calving, but lower (P < 0.10) concentrations than mature cows for wk -5, and lower (P < 0.05) concentrations than either 3-yr-old or mature cows for wk -4 to +7 relative to calving. Serum leptin was correlated with BCS (P < 0.0001; r = 0.35) at initiation of the feeding period and was correlated with BCS (P = 0.02; r = 0.12) and weight (P < 0.01; r = 0.14) at the completion of the supplement period, but it was not correlated with initial BW or interim BCS. Calving interval was not correlated (P > 0.12) with weekly measures of serum leptin concentration. Supplementing beef cows with whole corn germ had no effect on cow performance, calf performance, or serum leptin concentrations of cows.