Hyperimmune egg yolk antibodies developed against Clostridium perfringens antigens protect against necrotic enteritis.

Necrotic enteritis (NE) is a widespread infectious disease caused by Clostridium perfringens that inflicts major economic losses on the global poultry industry. Due to regulations on antibiotic use in poultry production, there is an urgent need for alternative strategies to mitigate the negative effects of NE. This paper presents a passive immunization technology that utilizes hyperimmune egg yolk immunoglobulin Y (IgY) specific to the major immunodominant antigens of C. perfringens. Egg yolk IgYs were generated by immunizing hens with 4 different recombinant C. perfringens antigens, and their protective effects against NE were evaluated in commercial broilers. Six different spray-dried egg powders were produced using recombinant C. perfringens antigens: α-toxin, NE B-like toxin (NetB; EB), elongation factor-Tu (ET), pyruvate:ferredoxin oxidoreductase, a mixture of 4 antigens (EM-1), and a nonimmunized control (EC). The challenged groups were either provided with different egg powders at a 1% level or no egg powders (EN). The NE challenge model based on Eimeria maxima and C. perfringens dual infection was used. In Experiments 1 and 2, the EB and ET groups exhibited increased body weight gain (BWG; P < 0.01), decreased NE lesion scores (P < 0.001), and reduced serum NetB levels (P < 0.01) compared to the EN and EC groups. IgY against NetB significantly reduced Leghorn male hepatocellular cytotoxicity in an in vitro test (P < 0.01). In Experiment 3, the protective effect of the IgYs mixture (EM-2) against C. perfringens antigens (NetB and EFTu) and Eimeria antigens (elongation factor-1-alpha: EF1α and Eimeria profilin: 3-1E) was tested. The EM-2 group showed similar body weight, BWG, and feed intake from d 7 to 22 compared to the NC group (P < 0.05). On d 20, the EM-2 group showed comparable intestinal permeability, NE lesion scores, and jejunal NetB and collagen adhesion protein levels to the NC group (P < 0.05). In conclusion, dietary mixture containing antibodies to NetB and EFTu provides protection against experimental NE in chickens through passive immunization.

Collagen adhesin protein and necrotic enteritis B-like toxin as biomarkers for early diagnosis of necrotic enteritis in commercial broiler chickens.

Mouse monoclonal antibodies (mAbs) reactive with Clostridium perfringens collagen adhesin protein (CNA) and necrotic enteritis B-like toxin (NetB) were developed. The best capture/detection mAb pairs for CNA and NetB were selected based on their affinity and specificity to develop sandwich enzyme-linked immunosorbent assays (ELISAs) to detect CNA and NetB proteins, respectively, in jejunal digesta samples from commercial broiler farms in the United States. Prior to the analysis of samples from commercial broiler flocks, the specificity and sensitivity of the CNA and NetB ELISAs were validated using sera, jejunal digesta, and fecal samples from chickens coinfected with Eimeria maxima and CNA+/NetB+C. perfringens in an animal model of necrotic enteritis (NE). Subsequently, a total of 251 field samples were collected from 74 commercial poultry farms. Among these, 18 samples were from 6 broiler farms that used certified organics (CO), and 155 samples were from 42 farms with nonantibiotics (NA). In jejunal digesta samples, CNA levels ranged from 0.02 to 0.59 ng/mL and NetB levels ranged from 0.09 to 1.91 ng/mL. CNA and NetB levels showed a positive correlation with each other (Pearson correlation coefficient r = 0.772, P < 0.001). CNA and NetB levels in jejunal digesta were significantly decreased in CO farms compared with those from NA farms (P < 0.001). In conclusion, these new C. perfringens antigen-specific sandwich ELISAs offer a sensitive and specific means to detect C. perfringens CNA and NetB proteins as biomarkers of early NE occurrence in field samples from commercial broiler chickens.

Effect of stocking density and dietary tryptophan on growth performance and intestinal barrier function in broiler chickens.

The objective of the present experiment was to investigate the effect of stocking density (SD) and dietary supplementation of crystalline tryptophan (Trp) on growth performance and intestinal barrier function in broiler chickens raised in a floor pen. The experiment was conducted using a completely randomized design with a 2 × 2 factorial arrangement consisting of 2 different SD and 2 supplemental levels of dietary Trp. A total of 1,140 Ross 308 broiler chickens at 21 d of age were allotted to 1 of 4 treatments with 5 replicates. Low SD (9 birds/m2) and high SD (18 birds/m2) were achieved by raising different number of birds per identical floor pen (2.0 m × 2.4 m). The basal diet was formulated with no supplemental Trp in diets to meet or exceed nutrient recommendation of the Ross 308 manual. The calculated concentrations of total Trp and digestible Trp in the basal diet were 0.19 and 0.16%, respectively. The other diet was prepared by adding 0.16% crystalline Trp to the basal diet. Diets were fed to birds for 21 d. At the end of the experiment, 2 birds per replicate were euthanized to collect tissue samples for further analyses. Results indicated that there were no interactions between SD and dietary Trp for all measurements. For the main effects, birds raised at a low SD had greater (P < 0.01) body weight gain, feed intake, and feed efficiency than those raised at a high SD. However, supplementation of dietary Trp had no effect on broiler performance. Furthermore, there were no main effects of SD and dietary Trp on intestinal barrier functions. In conclusion, broiler chickens raised in a floor pen with a high SD (18 birds/m2) have decreased growth performance with little changes in intestinal barrier functions. Supplementation of dietary Trp at 0.16% has no positive effect on broiler chickens raised in a floor pen with either a low or high SD.

Effect of vitamin E sources and inclusion levels in diets on growth performance, meat quality, alpha-tocopherol retention, and intestinal inflammatory cytokine expression in broiler chickens.

Water-miscible vitamin E (WVE) has been known to be more bioavailable than conventional lipid-soluble vitamin E (LVE) in human foods because of its greater solubility in the gastrointestinal tract. However, no data regarding the comparison of the efficacy of WVE to LVE were available in poultry diets. The objective of the current experiment was to investigate the effect of vitamin E (VE) sources and inclusion levels in diets on growth performance, meat quality, alpha-tocopherol retention, and intestinal inflammatory cytokine expression in broiler chickens. A total of 420 6-d-old broiler chicks were allotted to 1 of 7 dietary treatments with 6 replicates in a completely randomized design. The corn-soybean meal-based basal diet was formulated without inclusion of supplemental VE. Additional 6 diets were prepared by supplementing the basal diet with either WVE or LVE at the inclusion levels of 33, 65, or 100 IU/kg in diets. Diets were fed to birds for 26 d. Results indicated that different VE sources and inclusion levels from 33 to 100 IU/kg in diets had no effects on growth performance in broilers. The VE sources did not affect lipid oxidation in both breast and thigh meat. However, increasing inclusion levels of VE decreased (linear and quadratic, P < 0.05) lipid oxidation in breast meat. Regardless of VE sources, increasing inclusion levels of VE increased (linear, P < 0.01) alpha-tocopherol concentrations in breast meat. Dietary WVE showed a greater reduction in the expression of both pro- and anti-inflammatory cytokine genes in the jejunum than dietary LVE. Increasing inclusion levels of both VE sources decreased (linear and quadratic, P < 0.05) the expression of both pro- and anti-inflammatory cytokine genes in the jejunum. The efficacy of dietary WVE for growth performance, meat quality, lipid oxidation, and alpha-tocopherol retention in broilers chickens is similar to that of dietary LVE. However, it is likely that dietary WVE is more effective in decreasing intestinal inflammatory responses than dietary LVE in broilers.

Effect of dietary melamine concentrations on growth performance, excreta characteristics, plasma measurements, and melamine residue in the tissue of male and female broiler chickens.

The objectives of the current experiment were to investigate the effect of dietary melamine concentrations on growth performance, excreta characteristics, plasma measurements, and melamine residue in the tissue of male and female broiler chickens. Finally, the safe levels of melamine in broiler diets were determined based on BW gain (BWG) and melamine residue in the breast meat. A total of 1,008 1-d-old Ross 308 male and female broiler chickens were allotted to 1 of 7 dietary treatments within each sex in a completely randomized design. There were 6 replicates per treatment and each replicate consisted of 12 birds. Dietary melamine concentrations were set to 0; 250; 500; 750; 1,000; 5,000; or 10,000 mg/kg by adding a purified form of melamine. Diets were provided to birds on ad libitum basis for 35 d. Results indicated that no significant interaction between sex and dietary melamine concentrations was observed for all measurements. The BW, BWG, and feed intake for birds fed diets containing 10,000 mg/kg melamine were less (P < 0.05) than for those fed other diets. Melamine residues in the kidney and breast for birds fed diets containing 10,000 mg/kg melamine were greater (P < 0.05) than for birds fed other diets. The toxic level of dietary melamine based on BWG was determined by the one-slope broken-line analysis. The resulting equation was Y = 1,851 - 0.0404 × (X - 4,292), which indicated that a greater than 4,292 mg/kg melamine in diets was toxic to broiler chickens. The safe level of dietary melamine to limit melamine residue in the broiler breast was analyzed using the linear regression, which indicated that the safe level of melamine in broiler diets was 814 mg/kg. In conclusion, less than 814 mg/kg melamine in broiler diets should be maintained to satisfy human food safety regulations for melamine residue in the breast meat of broiler chickens.

Effect of stocking density and sex on growth performance, meat quality, and intestinal barrier function in broiler chickens.

The objective of the current experiment was to investigate the effect of stocking density and sex on growth performance, meat quality, and intestinal barrier function in broiler chickens. The experiment was conducted in a completely randomized design with a 2 × 4 factorial arrangement consisting of sex and four different stocking densities in battery cages. A total of 540 1-d-old Ross 308 broiler chickens were allotted to one of eight treatments with five replicates. Within each sex, birds were raised at four different stocking densities of 15.2, 20.2, 25.3, or 30.4 birds/m2 from 1 to 28 d of age. Different stocking densities were achieved by raising a different number of birds per battery cage with identical floor size (0.76 m × 0.78 m). At the end of the experiment, two birds per replicate were euthanized by CO2 asphyxiation to collect tissue samples for further analyses. Results indicated that no interactions between sex and stocking density were observed for all measurements except for serum lipopolysaccharide (LPS) concentrations. Increasing stocking density decreased (linear, P < 0.01) body weight gain and feed intake, but had no negative effects on meat quality. Trans-epithelial electrical resistance values, a measure of intestinal permeability, were decreased (linear, P < 0.01) with increasing stocking density, regardless of sex. Accordingly, serum LPS concentrations were increased (linear, P < 0.01) with increasing stocking density. However, increasing stocking density increased serum LPS concentrations in male broiler chickens, but had no effects on female broiler chickens, showing an interaction (P < 0.01). The expression of zonula occludens-1 (ZO-1) and junctional adhesion molecule B (JAM-2) was decreased (linear, P < 0.05) with increasing stocking density. In conclusion, increasing stocking density decreases broiler performance regardless of sex and this negative effect is likely associated with decreased intestinal barrier function.

Arterial Embolism Occurring During Percutaneous Thrombectomy of Dialysis Graft.

PURPOSE: To evaluate the incidence, management methods and follow-up results of arterial embolism during percutaneous thrombectomy of hemodialysis grafts. MATERIALS AND METHODS: After Institutional Review Board approval, the radiologic database of our department for percutaneous thrombectomy procedure in hemodialysis access was retrospectively reviewed. Between 1998 and June 2014, 2975 percutaneous thrombectomy procedures using thromboaspiration technique were performed in 1524 patients with thrombosed hemodialysis grafts. After thrombectomy, angioplasty was performed for significant stenoses. The incidence of arterial embolism was analyzed according to the location/shape of the arteriovenous graft. Percutaneous management methods of arterial embolism and long-term follow-up results by fistulography were also evaluated. RESULTS: Arterial embolism was documented by angiography in 117 cases (3.9%). Of these, three were symptomatic and subsided after embolectomy. The incidence was significantly correlated with the location/shape of the graft (p = 0.001). Arterial emboli were retrieved using occlusion balloon/Fogarty balloon (n = 58), guiding catheter-assisted aspiration (n = 36), sheath-assisted aspiration (n = 2) and back-bleeding technique (n = 3). Others were observed without intervention (n = 17) or surgically removed (n = 1). Arterial emboli were completely retrieved in 86 cases and partially retrieved in 13 cases. Ulnar artery rupture occurred in one case due to over-inflation of the occlusion balloon. Follow-up fistulography performed in 60 patients among whom 99 percutaneous embolectomies were done revealed arterial stenosis/occlusion in 7 and residual emboli in one patient. In observed patients without intervention, follow-up documented complete resolution of the emboli without arterial stenosis in 9 patients. CONCLUSION: Radiologically perceivable arterial embolism is uncommon during percutaneous thrombectomy of thrombosed dialysis grafts. The majority of the emboli can be retrieved by percutaneous techniques, but may induce arterial damage in some patients. Clinical observation can be another option for patients without ischemic symptoms.

Pelvic congestion syndrome diagnosed using endoscopic ultrasonography.

Pelvic congestion syndrome (PCS) presents with a non-cyclic pelvic pain lasting more than six months in duration in premenopausal women. Pelvic ultrasonography or computed tomography is usually the first imaging modality used to evaluate patients with suspected PCS. PCS is confirmed by visible congestion of the pelvic veins on selective ovarian venography. To our knowledge, the role of endoscopic ultrasonography (EUS) has not been reported. EUS showed multiple dilated structures especially on left side around the uterus and ovaries, which are compatible with other radiological investigations of PCS Although PCS is not typical areas within the scope of practice of endosonographers, it is useful to be familiar with the findings. We report a case of PCS that was diagnosed with the aid of EUS.

Magnetic resonance imaging of metaplastic carcinoma of the breast: sonographic and pathologic correlation.

Metaplastic carcinoma of the breast is a rare disease. We describe the MRI findings with the correlative sonographic and pathologic features of two cases. On MRI, T2-weighted images demonstrate a relatively well-defined mass with high signal intensity cystic components. Dynamic enhancement subtraction images showed an early enhancing and delayed washout peripheral rim and non-enhancing internal components. A microlobulated, isoechogenic mass with cystic components was seen sonographically, and was histopathology related to necrosis and cystic degeneration. Although these features are not unique, metaplastic carcinoma should be included in the differential diagnosis for breast masses.

Renal perfusion abnormality. Coded harmonic angio US with contrast agent.

PURPOSE: Coded harmonic angio (CHA) US is a recently developed technique that can depict the effects of contrast agents. The purpose of this study was to determine the role of this technique in depicting the enhancement patterns of various renal perfusion abnormalities compared with dynamic CT. MATERIAL AND METHODS: During a 6-month period, various renal lesions including renal cell carcinoma (n=12), transitional cell carcinoma (n=5), acute pyelonephritis (n=5), and renal trauma (n=2) were evaluated with CHA US using a microbubble contrast agent. US images were obtained before contrast administration and with a bolus injection of 4 g of microbubble contrast agent (300 mg/ml) every 10 s for 1 min and every minute for 5 min. The contrast enhancement patterns of various renal masses were compared with dynamic CT. RESULTS: Of 12 renal cell carcinomas, 9 (75%) showed heterogeneous enhancement and the remaining 3 (25%) showed homogeneous enhancement. Enhancement of more than adjacent renal parenchyma was seen 16-252 s after injection. The duration of enhancement was 13-208 s (mean, 80 s). All transitional cell carcinomas showed peripheral enhancement. Enhancement was seen 22-270 s after injection. The duration of enhancement was 191-238 s (mean, 291 s). Five patients with acute pyelonephritis and 2 with renal trauma showed focal perfusion defects not shown on the pre-contrast examinations. CONCLUSION: CHA US with microbubble contrast agent is an effective US technique for the evaluation of both tumor vascularity and renal perfusion abnormality.

When patients become cyanotic: acquired methemoglobinemia.

BACKGROUND: The authors conducted literature review to create a heightened awareness of the potential for developing toxic methemoglobinemia from local anesthetics. Methemoglobin normally is present in the blood at levels less than 1 percent. Levels may become toxic as hemoglobin is oxidized to methemoglobin after local anesthetics such as benzocaine and prilocaine are administered. TYPES OF STUDIES REVIEWED: The authors searched the medical and pharmaceutical industry literature. They found and reviewed case studies of incidences of methemoglobinemia that resulted from local anesthetic overdoses. RESULTS: Cases of local anesthetic-induced methemoglobinemia in dental practice are under-recognized and rare. Reported cases of prilocaine-induced methemoglobinemia have resulted in recent changes in some prilocaine literature. These changes include maximum recommended doses for patients of various weights. CLINICAL IMPLICATIONS: Dentists should identify patients who are at increased risk of developing methemoglobinemia before administering local anesthetics. They also should follow new recommended dosing guidelines for prilocaine and be aware of symptoms of this adverse reaction.

Esophageal rupture during balloon dilation of strictures of benign or malignant causes: prevalence and clinical importance.

PURPOSE: To review the prevalence and clinical importance of esophageal rupture during balloon dilation for treatment of esophageal stricture. MATERIALS AND METHODS: Fluoroscopically guided esophageal balloon dilation was performed within 9 years in 96 consecutive patients with esophageal strictures. Esophageal rupture was classified into three types: type 1 was intramural; type 2, transmural; and type 3, transmural with mediastinal leakage. RESULTS: Each patient underwent one to seven procedures, for a total of 191 procedures. Esophageal rupture occurred in 20 patients (21%). Type 1 esophageal rupture occurred in eight patients, type 2 in 11, and type 3 in one. All esophageal ruptures were detected immediately after the procedure. Sixteen patients were treated with fasting, parenteral alimentation, and antibiotics; two were treated surgically; and two were treated with stent placement. No treatment-related deaths occurred. CONCLUSION: The overall prevalence of esophageal rupture was 21%. A substantial number of patients who developed type 1 rupture had associated clinical symptoms, such as pain and fever, but responded to conservative management and are thus included as having complications of esophageal balloon dilation.

Isolation and characterization of Saccharomyces cerevisiae SAB2, a suppressor gene for temperature-sensitive phenotype of ARS-binding factor 1 mutant.

A high-copy number suppressor of yeast abf1-5 mutant, a temperature-sensitive lethal mutant, was isolated and named SAB2 (suppressor of ABF1). Hybridization to a yeast chromoblot and to prime clone grid filters revealed that the SAB2 gene was located near the yeast SUP3 on chromosome XV. The suppressor activity was contained in a 2.5 Kbp DNA region of the SAB2 plasmid. The nucleotide sequence of the DNA region contained a long open reading frame, which turned out to encode for yeast tryptophan permease. Four putative ABF1 binding sites were found in the promoter and the structural regions of the tryptophan permease gene. Binding of ABF1 to two of the sites tested in this study was detected. Our results indicate that ABF1 may be involved in the transcriptional control of the yeast tryptophan permease gene.

Protracted anemia associated with chronic, relapsing systemic inflammation induced by arthropathic peptidoglycan-polysaccharide polymers in rats.

Mild hypoproliferative anemia with abnormal iron metabolism frequently accompanies chronic inflammation and infection in humans. To determine whether anemia is associated with chronic relapsing arthritis induced by bacterial cell wall polymers, serial determinations of the hematocrit were measured in rats injected intraperitoneally with sonicated peptidoglycan-polysaccharide fragments from group A streptococci. Acute anemia peaked 5 to 10 days after injection, and chronic, spontaneously relapsing anemia persisted for 309 days. 51Cr labeling demonstrated decreased erythrocyte survival, i.e., a half-life of 8.4 days in cell wall-injected rats versus 11.8 days in controls. Erythrocytes were mildly microcytic, and leukocyte counts were elevated during early spontaneous reactivation of arthritis, 15 days after injection of peptidoglycan-polysaccharide. Bone marrow myeloid/erythroid precursor ratios were elevated in arthritic rats (P less than 0.0001). Purified peptidoglycan produced an acute anemia lasting 10 days, while injection of group A streptococcal polysaccharide and mutanolysin-digested cell wall did not affect the hematocrit. The minimal effective dose of peptidoglycan-polysaccharide was 5 micrograms of rhamnose per g (body weight). Serum iron and transferrin levels were decreased in cell wall-injected rats (P less than 0.005) and were closely correlated with hematocrit values and joint inflammatory scores. Stainable iron was increased in the liver, spleen, and mesenteric lymph nodes and unchanged in the bone marrow of cell wall-injected rats. Anemia accompanying chronic, relapsing systemic inflammation induced by peptidoglycan-polysaccharide polymers appears to be an excellent animal model of the anemia of chronic disease.