Development of an enzyme immuno assay for the determination of porcine haptoglobin in various body fluids: testing the significance of meat juice measurements for quality monitoring programs

Quantification of haptoglobin (Hp), an acute phase protein, in blood is presently discussed as being useful to monitor animal health. We developed an enzyme immuno assay (EIA) which is specific for porcine Hp, is not impaired by hemolytic samples and is sufficiently sensitive to be applied in meat juice. Hp was purified from porcine serum by affinity chromatography on hemoglobin Sepharose followed by gel filtration. A specific rabbit antiserum was obtained. In a competitive approach, biotinylated porcine Hp was used as tracer and incubated with Hp standard or sample in microtiter plates. The limit of detection was 0.02 mg/l, parallelism of sample dilutions was proven; recovery of Hp added to serum samples was 96.4 +/- 4.7%. The coefficients of intra and inter-assay variation were 3.3 (n=5) and 10.2% (n=16), respectively. Hp was reliably quantified in blood serum and plasma, whole blood, saliva and meat juice. For healthy pigs of different ages (4 weeks and 6 months), mean Hp concentrations of about 0.5-0.7 mg/ml were observed. To test the significance of Hp measurements in other matrices, samples were obtained from fattening pigs or from slaughter pigs. Blood serum or plasma was collected in parallel. In whole blood, Hp concentrations were about 40% lower than in plasma, but were closely related (n=24,r=0.85,P<0.001). Saliva Hp concentrations ranged between 0.3 and 3.0 microg/ml and were marginally related with blood plasma concentrations (n=93,r=0.35,P<0.001). From 106 hybrid slaughter pigs (100-110 kg) blood and muscle samples (diaphragmatic pillar, d.p.; m. brachiocephalicus, m.b.) were collected. Meat juice was obtained after freezing and thawing. Concentrations were 0.39+/-0.5 mg/ml in serum and 0.04+/-0.06 mg/ml in meat juice. Hp concentrations in blood were closely correlated with those in d.p. juice (P<0.001,r=0.750) and m.b. juice (P<0.001,r=0.776). In view of the many reports on Hp measurements being predictive for animal health even in the subclinical range, we conclude that Hp quantification in meat juice might be useful to assess meat quality at slaughter and further along the processing chain in terms of animal health.     

Pneumoperitoneum secondary to a perforated gastric ulcer in a cat

A 7-year-old domestic cat was examined because of a history of weight loss, intermittent diarrhea, and anorexia of 6 months' duration. Results of physical examination were normal except for marked abdominal tension. Results of a CBC and serum biochemistry profile were unremarkable. Severe pneumoperitoneum was noted radiographically, and abdominocentesis yielded 640 mL of air. Abdominal distension recurred 1 day after abdominocentesis. Exploratory laparotomy was performed 3 days after recurrence of abdominal distension and revealed a gastric perforation along the greater curvature of the stomach with omental and pancreatic adhesions at the site of perforation. Mild local peritoneal inflammation was also noted. A partial gastrectomy was performed to resect the lesion, and histologic examination confirmed a severe perforating ulcer. The gastric wall adjacent to the ulcer was histologically normal. Aerobic and anaerobic bacteriologic cultures of abdominal swab specimens were negative for bacterial growth. The cat was discharged 4 days after surgery and clinically normal at suture removal.     

Treatment of nonunions with nonglycosylated recombinant human bone morphogenetic protein-2 delivered from a fibrin matrix

OBJECTIVE: To report the results of the treatment of nonunions with nonglycosylated recombinant human bone morphogenetic protein-2 (nglBMP-2) delivered from a designed fibrin matrix. STUDY DESIGN: Experimental trial in rodents and prospective clinical study in dogs and cats with nonunion fractures. ANIMALS: Twenty adult female, albino, Sprague-Dawley rats; 8 client-owned cats and dogs. METHODS: After development of a fibrin matrix and evaluation of nglBMP-2 in a rodent femoral defect model, 8 consecutive long bone nonunion fractures (no progression in healing in > or = 3 months), were treated using 300 microg nglBMP-2 in a liquid fibrin precursor, injected into the defect gap after fracture revision and stabilization, or through a stab incision into the fracture site. The fibrin matrix was designed to clot in the wound after 60 seconds and to release the nglBMP-2 continuously over several days. RESULTS: Using only fibrin gel, 7% of the rat femoral defect was filled with new formed bone compared with 79% defect filling using 2 microg nglBMP-2 (P=.006). Five and 10 microg nglBMP in fibrin resulted in union of all femoral defects with complete filling of the gap with new bone. Bony bridging and clinical healing was achieved in 7 patients within 24 weeks of administration of nglBMP-2. CONCLUSIONS: Application of nglBMP-2 in a functional matrix can induce bone healing. Controlled release of nglBMP-2 from a fibrin matrix mimics the natural fracture hematoma. CLINICAL RELEVANCE: nglBMP-2/fibrin can successfully replace a cancellous bone autograft in fracture treatment with an associated reduction in graft donor site morbidity and surgical time.     

Metastatic anal sac adenocarcinoma in a dog presenting for acute paralysis

A 4-year old, female spayed terrier was referred for hind end paresis that rapidly progressed to paralysis. Spinal radiographs revealed vertebral collapse and bony lysis. Myelography confirmed spinal cord compression and surgical exploration found an extradural soft tissue mass. Metastatic anal sac adenocarcinoma was diagnosed at postmortem examination.     

Pneumopericardium associated with a pulmonary-pericardial communication in a dog

A 6-year-old 18-kg (39-lb) spayed female Standard Poodle was referred for treatment of pneumopericardium. The dog did not have severe clinical signs relating to the pneumopericardium, and the diagnosis was made incidentally while investigating the cause of a cough. Computed tomography revealed an air-filled structure consistent with a bulla to the right of the heart base that appeared to communicate with the pericardial cavity Because spontaneous resolution of the pneumopericardium seemed unlikely and cardiac tamponade was a possibility, exploratory thoracotomy was performed. A lobulated bulla was found at the hilus of the right middle lung lobe adhered to the underlying pericardial sac, and a 4-cm-diameter communication from the pericardium to the pulmonary bulla was found. Right middle lung lobectomy was performed, and the pericardiotomy and pericardial opening were sutured. The dog recovered without complications. In previously reported cases of pneumopericardium involving a cat and a dog, the condition resolved spontaneously. In this dog, in contrast, surgical resection of the affected lung lobe with pericardial reconstruction was required for resolution of the pneumopericardium.     

Control of late blight in organic potato production: evaluation of copper-free preparations under field,growth chamber and laboratory conditions

In order to replace copper fungicides in organic potato production, 53 copper-free preparations (CFPs) based on natural compounds, including plant extracts and microorganisms, and five copper preparations were evaluated for their potential to control Phytophthora infestans, the pathogen that causes late blight of potatoes. In in vitro assays, 30% of the CFPs inhibited indirect germination of sporangia, 26% mycelial growth and in growth chamber experiments, 21% efficiently reduced foliar blight of tomato plants. In micro-plot field trials with applications twice a week, the copper preparations were the most effective and reduced foliar blight by 99%. Of the CFPs tested, Oekofluid P, Mycosin and other sulphuric clays, and C-2000 reduced late blight the most, from 63% to 37%. In small-plot trials in 2001, 2002 and 2004, 27 CFPs with different formulations and four copper preparations were examined. In 2004, copper preparations at full and reduced rates and sulphuric clays were applied either weekly or according to the decision support system Bio-PhytoPRE. With Bio-PhytoPRE, copper preparations reduced foliar blight of potatoes by 23–77% and increased tuber yield by 2–28%, depending on the copper rate applied and year. With CFPs, maximal efficacy was 17% and no effect on tuber yield was observed. In vitro and in vivo trials showed that the rainfastness and the persistence of CFPs was low compared with copper preparations. This indicates that the failure of CFPs under field conditions is probably due to a lack of stability under prevailing environmental conditions and not to a lack of efficacy. Until stable formulations for CFPs are developed, an optimised and restricted use of copper fungicides using a decision support system could help to control late blight in organic potato production and to reduce copper input into the environment.     

Modification of the effective mesh size for measuring landscape fragmentation to solve the boundary problem

Patch-based landscape metrics can be biased by the boundaries and the extent of a reporting unit if the boundaries fragment patches. We call this the “boundary problem”. The effective mesh size m _eff is a convenient method to quantify landscape fragmentation, that is based on the probability that two points chosen randomly in a region will be connected, e.g., not be separated by roads, railroads, or urban development. The cutting-out (CUT) procedure, used in the original computation of m _eff, suffers from the boundary problem because the boundaries of the reporting units are considered to be additional barriers. Therefore, m _eff will be underestimated, particularly if reporting units are embedded within the broader landscape. In this paper, we present a solution to overcome this limitation by a new method called “cross-boundary connections” (CBC) procedure. It attributes the connections between two points that are located in different reporting units to both reporting units. We systematically compare the CBC procedure to the CUT procedure and show that the boundary problem is intrinsic to the CUT procedure, while the CBC procedure is independent of the size and administrative boundaries of reporting units. In addition, we elucidate the superior performance of the new procedure in the case study of South Tyrol where m _eff is being used for sustainability reporting on the level of municipalities. The new CBC procedure eliminates the bias due to the boundaries and the size of reporting units in measuring landscape fragmentation through m _eff.