Effect of the racial group and body condition score at calving on production performance and metabolic profile of buffaloes during the transition period.

This study evaluated the body condition score (BCS) at calving and breed (B) effects on milk composition, yield, performance, physiological parameters, hemogram, blood metabolites, and urinary metabolites in the transition and early lactation periods of Mediterranean (MED) and Murrah (MUR) buffaloes. Twenty MED and fifteen MUR buffaloes were distributed into four experimental treatments, in a completely randomized design, considering their racial groups and BCS (LBCS = low; HBCS = high): LBCS MED (N = 9); HBCS MED (N = 11); LBCS MUR (N = 8); HBCS MUR (N = 7). Animals were monitored during the last 21 days of gestation and first 56 days postpartum and kept under the same management and feeding conditions. During data collection, milk composition, yield, performance, physiological parameters, hemogram, blood metabolites, and urinary metabolites were evaluated. Higher milk production and fat-corrected milk were observed in MED than MUR buffaloes. Breed effects were observed on body weight, rectal temperature, glucose, urea, calcium (Ca) concentrations, and BCS effects on total protein, albumin, urea, and Ca. There were BCS effects on hematocrit, neutrophils, eosinophils, and interactions between B × BCS for lymphocytes and platelets. There were breed effects on urinary concentrations of chlorine, uric acid, and interactions between weight (W) × B on chlorine and urea. The MED buffaloes can be considered the most prepared to undergo physiological changes, including the BCS value at calving, indicating higher physiological health. Besides, this study demonstrates more considerable preparation for the calving, regardless of the body condition score at calving.

Ruminal biohydrogenation and abomasal flow of fatty acids in lactating cows fed diets supplemented with soybean oil, whole soybeans, or calcium salts of fatty acids.

Ruminants have a unique metabolism and digestion of unsaturated fatty acids (UFA). Unlike monogastric animals, the fatty acid (FA) profile ingested by ruminants is not the same as that reaching the small intestine. The objective of this study was to evaluate whole raw soybeans (WS) in diets as a replacer for calcium salts of fatty acids (CSFA) in terms of UFA profile in the abomasal digesta of early- to mid-lactation cows. Eight Holstein cows (80 ± 20 d in milk, 22.9 ± 0.69 kg/d of milk yield, and 580 ± 20 kg of body weight; mean ± standard deviation) with ruminal and abomasal cannulas were used in a 4 × 4 Latin square experiment with 22-d periods. The experiment evaluated different fat sources rich in linoleic acid on ruminal kinetics, ruminal fermentation, FA abomasal flow, and milk FA profile of cows assigned to treatment sequences containing a control (CON), with no fat source; soybean oil, added at 2.68% of diet dry matter (DM); WS, addition of WS at 14.3% of diet DM; and CSFA, addition of CSFA at 2.68% of diet DM. Dietary fat supplementation had no effect on nutrient intake and digestibility, with the exception of ether extract. Cows fed fat sources tended to have lower milk fat concentration than those fed CON. In general, diets containing fat sources tended to decrease ruminal neutral detergent fiber digestibility in relation to CON. Cows fed WS had lower ruminal digestibility of DM and higher abomasal flow of DM in comparison to cows fed CSFA. As expected, diets containing fat supplements increased FA abomasal flow of C18:0 and total FA. Cows fed WS tended to present a higher concentration of UFA in milk when compared with those fed CSFA. This study suggests that under some circumstances, abomasal flow of UFA in early lactation cows can be increased by supplementing their diet with fat supplements rich in linoleic acid, regardless of rumen protection, with small effects on ruminal DM digestibility.

Effect of replacing ground corn and soybean meal with licuri cake on the performance, digestibility, nitrogen metabolism and ingestive behavior in lactating dairy cows.

Licuri (Syagrus coronate) cake is a biodiesel by-product used in ruminant feed as a beneficial energy source for supplementation in managed pastures. The objective was to evaluate the performance, digestibility, nitrogen balance, blood metabolites, ingestive behavior and diet profitability of eight crossbred Holstein (3/4)×Gyr (5/8) multiparous cows (480±25 kg BW and 100 days milking) grazing and supplemented with licuri cake partially replacing ground corn and soybean meal in concentrate (0, 200, 400 and 600 g/kg in dry matter (DM)), distributed in an experimental duplicated 4×4 Latin square design. Licuri cake partially replacing ground corn and soybean meal increased (P<0.01) the intake and digestibility of ether extract and decreased the non-fiber carbohydrates; however, there were no influences on the intakes of DM, CP, NDF and total digestible nutrients (TDN). The digestibilities of DM, CP and NDF were not influenced by licuri cake addition. There was a decrease trend on TDN digestibility (P=0.08). Licuri cake replacing ground corn and soybean meal in concentrate did not affect the intake; fecal, urinary and mammary excretions; N balance; and triglycerides concentrations. However, the blood urea nitrogen (P=0.04) concentration decreased with the licuri cakes inclusion in cow supplementation. There was an increasing trend for serum creatinine (P=0.07). Licuri cake inclusion did not affect body condition score, production, yield, protein, lactose, total solids and solid non-fat contents of milk and Minas frescal cheese. There was a linear decrease in average daily weight gain (g/day). The milk fat concentration and cheese fat production (P<0.1) presented a linear increase with partial replacement of ground corn and soybean meal with licuri cakes. The addition of licuri cake did not alter the time spent feeding, ruminating or idling. There was an increasing trend in NDF feeding efficiency (P=0.09). The replacing of ground corn and soybean meal with licuri cake up to 600 g/kg decreased the concentrate cost by US$0.45/cow per day. Licuri cake replacing corn and soybeans (400 g/kg) in concentrate promoted a profit of US$0.07/animal per day. Licuri cake is indicated to concentrate the supplementation of dairy cows with average productions of 10 kg/day at levels up to 400 g/kg in the concentrate supplement because it provides an additional profit of US$0.07/animal per day and increased milk and Minas frescal cheese fat without negative effects on productive parameters.

Molecular identification of cetaceans from the West Atlantic using the E3-I5 region of COI.

Molecular identification is very useful in cases where morphology-based species identification is not possible. Examples for its application in cetaceans include the identification of carcasses of stranded animals in advanced state of decomposition and body parts that are illegally traded. One DNA region that is often used for molecular identification is the Folmer region of the mitochondrial gene cytochrome c oxidase subunit I (COI) (locus 48 to 705 bp). This locus has been used for the identification of several animal species, including whales and dolphins. The goal of the present study was to evaluate the usefulness of another region of COI, the E3-I5 (locus 685 to locus 1179; 495 bp) as a marker for identification of cetaceans from northeastern Canada and northeastern Brazil. The identification markers were successfully obtained for seven cetacean species after performing percent identity and Basic Local Alignment Search Tool analyses. The obtained markers are now publicly available and are useful for the identification of the endangered blue whale (Balaenoptera musculus), common minke whale (B. acutorostrata), vulnerable sperm whale (Physeter macrocephalus), harbor porpoise (Phocoena phocoena), common bottlenose dolphin (Tursiops truncatus), Guiana dolphin (Sotalia guianensis), and melon-headed whale (Peponocephala electra).

Influence of polymer-coated slow-release urea on total tract apparent digestibility, ruminal fermentation and performance of Nellore steers.

OBJECTIVE: Two experiments were performed to evaluate the effects of coated slow-release urea on nutrient digestion, ruminal fermentation, nitrogen utilization, blood glucose and urea concentration (Exp 1), and average daily gain (ADG; Exp 2) of steers. METHODS: Exp 1: Eight ruminally fistulated steers [503±28.5 kg body weight (BW)] were distributed into a d 4×4 Latin square design and assigned to treatments: control (CON), feed grade urea (U2), polymer-coated slow-release urea A (SRA2), and polymer-coated slow-release urea B (SRB2). Dietary urea sources were set at 20 g/kg DM. Exp 2: 84 steers (350.5±26.5 kg initial BW) were distributed to treatments: CON, FGU at 10 or 20 g/kg diet DM (U1 and U2, respectively), coated SRA2 at 10 or 20 g/kg diet DM (SRA1 and SRA2, respectively), and coated SRB at 10 or 20 g/kg diet DM (SRB1 and SRB2, respectively). RESULTS: Exp 1: Urea treatments (U2+SRA2+SRB2) decreased (7.4%, p = 0.03) the DM intake and increased (11.4%, p<0.01) crude protein digestibility. Coated slow-release urea (SRA2+SRB2) showed similar nutrient digestibility compwared to feed grade urea (FGU). However, steers fed SRB2 had higher (p = 0.02) DM digestibility compared to those fed SRA2. Urea sources did not affect ruminal fermentation when compared to CON. Although, coated slow-release urea showed lower (p = 0.01) concentration of NH3-N (-10.4%) and acetate to propionate ratio than U2. Coated slow-release urea showed lower (p = 0.02) urinary N and blood urea concentration compared to FGU. Exp 2: Urea sources decreased (p = 0.01) the ADG in relation to CON. Animals fed urea sources at 10 g/kg DM showed higher (12.33%, p = 0.01) ADG compared to those fed urea at 20 g/kg DM. CONCLUSION: Feeding urea decreased the nutrient intake without largely affected the nutrient digestibility. In addition, polymer-coated slow-release urea sources decreased ruminal ammonia concentration and increased ruminal propionate production. Urea at 20 g/kg DM, regardless of source, decreased ADG compared both to CON and diets with urea at 10 g/kg DM.

Meat quality of lambs fed diets with peanut cake.

Replacement of soybean meal by peanut cake was evaluated on the meat quality of 45 Dorper × Santa Inês crossbred lambs. Animals were distributed in a completely randomized design, with five treatments and nine repetitions, and fed Tifton-85 hay and a concentrate mixed with 0.0%, 25.0%, 50.0%, 75.0% or 100.0% peanut cake based on the dry mass of the complete diet. The longissimus lumborum muscle was used to determine the proximate composition, physical-chemical characteristics and fatty acid profile. Significant differences (P<0.05) were found for the crude protein and ether extract levels, with average values of 23.38% and 2.15% in the sheep meat, respectively. The physical-chemical characteristics of the loin were not affected (P>0.05) by the diets. The fatty acid profile was affected by peanut cake supplementation for myristic, myristoleic, palmitoleic, linolenic and arachidonic fatty acids. Peanut cake can be added in the diet of lambs no effect on physical-chemical characteristics. However, the total replacement of the soybean meal altered the proximate composition and fatty acid profile of the meat.

Effects of whole flaxseed, raw soybeans, and calcium salts of fatty acids on measures of cellular immune function of transition dairy cows.

The objective of the current study was to evaluate the effects of supplemental n-3 and n-6 fatty acid (FA) sources on cellular immune function of transition dairy cows. Animals were randomly assigned to receive 1 of 4 diets: control (n=11); whole flaxseed (n-3 FA source; n=11), 60 and 80g/kg of whole flaxseed [diet dry matter (DM) basis] during pre- and postpartum, respectively; whole raw soybeans (n-6 FA source; n=10), 120 and 160g/kg of whole raw soybeans (diet DM basis) during pre- and postpartum, respectively; and calcium salts of unsaturated FA (Megalac-E, n-6 FA source; n=10), 24 and 32g/kg of calcium salts of unsaturated FA (diet DM basis) during pre- and postpartum, respectively. Supplemental FA did not alter DM intake and milk yield but increased energy balance during the postpartum period. Diets containing n-3 and n-6 FA sources increased phagocytosis capacity of leukocytes and monocytes and phagocytosis activity of monocytes. Furthermore, n-3 FA source increased phagocytic capacity of leukocytes and neutrophils and increased phagocytic activity in monocytes and neutrophils when compared with n-6 FA sources. Supplemental FA effects on adaptive immune system included increased percentage of T-helper cells, T-cytotoxic cells, cells that expressed IL-2 receptors, and CD62 adhesion molecules. The results of this study suggest that unsaturated FA can modulate innate and adaptive cellular immunity and trigger a proinflammatory response. The n-3 FA seems to have a greater effect on phagocytic capacity and activity of leukocytes when compared with n-6 FA.

Effects of flaxseed, raw soybeans and calcium salts of fatty acids on apparent total tract digestibility, energy balance and milk fatty acid profile of transition cows.

Oilseeds offer some protection to the access of ruminal microorganisms and may be an alternative to calcium salts of fatty acids (FA), which are not fully inert in the ruminal environment. This study aimed to evaluate the effects of different sources of FA supplementation on apparent total tract nutrient digestibility, milk yield and composition, and energy balance (EB) of cows during the transition period and early lactation. We compared diets rich in C18:2 and C18:3 FA. Multiparous Holstein cows were randomly assigned to receive one of the four diets: control (n=11); whole flaxseed (WF, n=10), 60 and 80 g/kg (diet dry matter (DM) basis) of WF during the prepartum and postpartum periods, respectively; whole raw soybeans (WS, n=10), 120 and 160 g/kg (diet DM basis) of WS during the prepartum and postpartum periods, respectively; and calcium salts of unsaturated fatty acids (CSFA, n=11), 24 and 32 g/kg (diet DM basis) of CSFA during the prepartum and postpartum periods, respectively. Dry cows fed WF had higher DM and net energy of lactation (NEL) intake than those fed WS or CSFA. The FA supplementation did not alter DM and NDF apparent total tract digestibility, dry cows fed WF exhibited greater NDF total tract digestion than cows fed WS or CSFA. Feeding WS instead of CSFA did not alter NEL intake and total tract digestion of nutrients, but increased milk fat yield and concentration. Calculated efficiency of milk yield was not altered by diets. FA supplementation increased EB during the postpartum period. Experimental diets increased long-chain FA (saturated and unsaturated FA) in milk. In addition, cows fed WS and CSFA had higher C18:1 trans-11 FA and C18:2 cis, and lower C18:3 FA in milk than those fed WF. Furthermore, cows fed CSFA had higher C18:1 trans-11 and cis-9, trans-11 FA than cows fed WS. Although supplemental C18:2 and C18:3 FA did not influence the milk yield of cows, they positively affected EB and increased unsaturated long-chain FA in milk fat.

Differences in maternal plane of nutrition and body condition during late gestation coupled with estrus synchronization at weaning do not result in differences in embryonic development at 4 days of gestation.

This study was conducted to assess the effects of dietary energy in late pregnancy and hormone therapy at weaning on plasma metabolite profile, litter performance, reproductive parameters, and embryo viability in the second pregnancy. A total of 23 first-parity sows at 75 d of pregnancy were randomly allocated to 4 treatments. Treatments were factorial (2 × 2) combinations of 2 nutritional strategies [standard-energy feed (SEF) and high-energy feed (HEF)] and 2 hormone therapies [600 IU eCG and 2.5 mg swine LH 72 h later (HO) and no hormone (WH)]. Sows were weighed weekly from 75 d of pregnancy until 3 d before farrowing; 1 d after farrowing; 7, 14, and 21 d into lactation; and at weaning. Back fat (BF) was measured at 75 d of pregnancy, 3 d before farrowing, and at weaning. Average daily gain and ADFI were also calculated. Plasma metabolites were analyzed after 82, 89, 96, and 103 d of pregnancy, at farrowing, and after 7, 14, and 21 d of lactation. Embryo viability was assessed after 4.55 d of second pregnancy. During pregnancy, HEF-treated sows displayed greater BW (P < 0.05) compared with SEF-treated females, but no differences were observed during lactation. There were no differences in BW of the piglets caused by the treatments. High-energy-treated females showed superior BF (P > 0.05) in all periods; however, significant differences were detected only at the prefarrowing measurement (P < 0.05). No differences in ADFI were observed during lactation. The SEF group showed positive ADG, whereas the HEF group showed negative ADG (0.216 vs. -0.266 kg/d for SEF and HEF, respectively; P < 0.05). High-energy-treated sows presented greater concentrations of total cholesterol after 89 and 103 d of pregnancy and greater concentrations of high-density lipid cholesterol (HDL) after 89 and 96 d. At farrowing and 14 and 21 d of lactation, NEFA concentrations were greater (P < 0.05) in the HEF group. After hormone treatment, no differences were observed on weaning-to-estrus intervals and estrus duration. Greater mobilization of body reserves observed in the HEF group during lactation did not affect reproductive performance negatively, suggesting that metabolic status was adequate for the first lactational catabolism.

Percolation on a multifractal.

We investigate percolation phenomena in multifractal objects that are built in a simple way. In these objects the multifractality comes directly from the geometric tiling. We identify some differences between percolation in the proposed multifractals and in a regular lattice. There are basically two sources of these differences. The first is related to the coordination number, which changes along the multifractal. The second comes from the way the weight of each cell in the multifractal affects the percolation cluster. We use many samples of finite size lattices and draw the histogram of percolating lattices against site occupation probability. Depending on a parameter characterizing the multifractal and the lattice size, the histogram can have two peaks. We observe that the percolation threshold for the multifractal is lower than that for the square lattice. We compute the fractal dimension of the percolating cluster and the critical exponent beta. Despite the topological differences, we find that the percolation in a multifractal support is in the same universality class as standard percolation.

Statistics of largest cluster growth through constant rate random filling of lattices.

In this paper we consider a percolation model where the probability p for a site to be occupied increases linearly in time, from 0 to 1. We analyze the way the largest cluster grows in time, and in particular, we study the statistics of the "jumps" in the mass of the largest cluster, and of the time delay between those events. Different critical behaviors are observed below and above the percolation threshold. We propose a theoretical analysis, and we check our results against direct numerical simulations.

Therapeutic options in the management of toxic and nontoxic nodular goiter.

Nodular goiter is present in 500 to 600 million people and is usually secondary to endemic iodine deficiency. Despite adequate iodine intake, 4% to 6% of American adults are goitrous. Sporadic nodular goiter ensues from the natural heterogeneity of thyroid follicular cells, which, when amplified by yet unidentified trophic stimuli, results in episodes of proliferating, rapidly dividing micronodules. The initial small diffuse goiter evolves into a multinodular goiter (MNG) with 1 or more dominant nodules that may or may not be autonomous. An autonomous functioning thyroid adenoma (AFTA) usually possesses a somatic gain-of-function mutation of the thyrotropin (TSH) receptor associated with rapid growth, hemorrhagic necrosis, and reparative fibrosis that accentuate goiter nodularity. Diagnostic evaluation consists of patient history and physical examination, serum TSH determination, free thyroxine and free triiodothyronine measurements, and imaging studies assessing goiter function, size, and anatomy. If treatment is required, L-thyroxine, thionamides, surgery, radioiodine (I-131), and percutaneous ethanol injection (PEI) are effective in selected patients. In euthyroid patients, L-thyroxine reduces goiter size in some patients, but continued therapy is required to prevent regrowth. Thionamides control the hyperthyroidism of toxic nodular goiter in preparation for more definitive therapy, but are rarely used long term. Surgery and I-131 are most commonly selected for definitive therapy for the toxic AFTA, and the toxic or euthyroid MNG, but PEI is effective in selected toxic AFTAs.

The role of nuclear medicine in the management of Graves' disease.

131I therapy is safe and effective for most Graves' disease patients. It is being used more frequently in previously restricted populations such as children, adolescents, and women of childbearing age. It affords prompt, consistent relief of hyperthyroidism, but permanent hypothyroidism ensues in almost all patients. This complication is readily diagnosed with modern in vitro assays and inexpensive, life-long replacement therapy renders the patient asymptomatic and able to resume a normal lifestyle. Recent changes in NRC requirements have further liberalized the use of 131I for Graves' disease in the USA, permitting more patients to be treated with effective outpatient therapy. The controversial role of 131I in exacerbation of GO has been further clarified and preventive measures are available.

Modified PIOPED criteria used in clinical practice.

UNLABELLED: To assess the use of modified PIOPED scintigraphic criteria for lung scan (V/Q) interpretation to detect pulmonary embolism (PE), we prospectively applied these criteria in suspected PE patients referred for V/Q from 9/1/92 to 2/7/94. PIOPED criteria were modified by placing a moderate segmental perfusion mismatch in the intermediate instead of low probability of PE category and using the "stripe sign." METHODS: Patients were studied by six-view V/Q imaging using 74 MBq (2 mCi) 99mTc-MAA followed by 148-370 MBq (4-10 mCi) 99mTc-DTPA aerosol, contrast pulmonary selective angiography and Doppler sonography with leg compression as needed. Patients underwent follow-up (mean 13.9 mo) to detect subsequent thromboembolic events. In this study group, 1000 patients were studied by V/Q followed by angiography in 133 patients. RESULTS: The distribution of V/Q-assigned PE probabilities was: high probability 5.7%, intermediate 17.4%, low 41.4% and normal 35.5%. Group A patients (133) underwent angiography, which resulted in the determination of a 27.1% PE prevalence. Group B patients (867) did not have angiograms; the clinical prevalence of PE was 7.5%. In the total study population, the positive predictive value of a high probability V/Q study for PE (10.1% prevalence) was 98.2%, intermediate probability V/Q study for PE was 24.1% and a low probability study for PE was only 0.5%. CONCLUSION: Modified PIOPED V/Q interpretation criteria afford better angioproven PE discrimination between intermediate (31.8% PE prevalence) and low (5.5% PE prevalence) probability V/Q results than reported for PIOPED intermediate (32.6% PE prevalence) and low (16.3% PE prevalence) probability V/Q interpretation criteria.

Adrenal cortical and medullary imaging.

Adrenal disease can be manifested by endocrine dysfunction or anatomic abnormalities detected by cross-sectional imaging modalities. With the advent of newer and more reliable in vitro assays and a better understanding of the spectrum of adrenal pathology, the physician can now adopt a more accurate and cost-effective approach to the diagnosis of adrenal disease. Both functional and anatomic imaging modalities can play an important role in the evaluation of the incidental adrenal mass, the early detection of adrenal metastases, differentiation of the various causes of Cushings's syndrome, selection of patients for potentially curative surgery in primary aldosteronism and adrenal hyperandrogenism, and localization of pheochromocytomas and neuroblastomas. The usefulness of the adrenal cortical radiopharmaceutical, 131I-6-beta-iodomethylnorcholesterol (NP-59), and the adrenal medullary radiopharmaceuticals, 131I and 123I-metaiodobenzylguanidine (MIBG), is detailed for these various clinical settings and the role of NP-59 and MIBG is contrasted to that of the cross-sectional modalities, computed tomography and magnetic resonance imaging (MRI). Incidental adrenal masses are common, but malignancies are few. Imaging studies select those patients who require a further evaluation by biopsy examination or adrenalectomy. In the hyperfunctioning endocrine states, such as Cushing's syndrome, primary aldosteronism, adrenal androgenism, and pheochromocytoma, correlation of biochemical findings with both functional and anatomic imaging is necessary to avoid inappropriate and ineffective surgical intervention, yet not miss an opportunity for curative resection. Lastly, MIBG and MRI are complementary in the detection and staging of neuroblastoma.

Thyroid and parathyroid imaging.

With the advent of better thyroid function tests, a tumor marker, and fine-needle aspiration, the role of thyroid imaging studies in the evaluation of the patients with thyroid disease has diminished. Although multimodality thyroid imaging had improved our understanding of thyroid disease, current indications for thyroid imaging are the solitary or dominant thyroid nodule, an upper mediastinal mass, differentiation of hyperthyroidism, detection and staging of postoperative thyroid cancer, neonatal hypothyroidism, thyroid developmental anomalies, and the thyroid mass post-thyroidectomy for benign disease. To provide optimal, cost-effective, care for the thyroid patient, the physician must understand the advantages and disadvantages of each imaging modality--scintigraphy, real-time sonography (RTS), computed tomography, and magnetic resonance--in specific clinical settings. Similarly, preoperative noninvasive localization of hyperfunctioning parathyroid tissue in patients with primary hyperparathyroidism undergoing their initial neck exploration usually is not warranted. In this situation, the best localization procedure is to enlist the services of an experienced parathyroid surgeon. However, if this is not feasible because of local constraints, both sestamibi methoxy isobutyl isonitrile (MIBI) scintigraphy and magnetic resonance imaging (MRI) provide excellent localization (< 90%) of juxta-thyroidal and ectopic parathyroid adenomas. Hyperplastic glands are more difficult to detect because of their smaller size, and tandem studies (MIBI and MRI) should provide higher sensitivity before initial exploration, especially in patients with ectopic glands. In patients with persistent or recurrent disease, multimodality imaging with MIBI, MR, computed tomography and RTS in a sequential fashion is warranted to optimize two-test, site-specific localization.

Interference by spironolactone on adrenocortical scintigraphy and other pitfalls in the location of adrenal abnormalities in primary aldosteronism.

A case of primary aldosteronism is presented in which the CT scan was initially misleading, adrenocortical scintigraphy was rendered inaccurate by pharmacological interference of spironolactone, and selective adrenal venous sampling of aldosterone was technically difficult. When dexamethasone suppression adrenocortical scintigraphy was performed with attention to technical detail and exclusion of interference by spironolactone, the causative lesion was scintigraphically demonstrated. This finding was confirmed by the results of venous sampling and the correctly located tumor removed.