Comparison of Meat Juice Serology and Bacteriology for Surveillance of Salmonella in the Brazilian Pork Production Chain.

This study assessed an enzyme-linked immunosorbent assay (ELISA) based assay to detect Salmonella in swine as a potential tool to predict the presence of Salmonella in swine carcasses. The following samples were collected from 10 swine batches: blood (n = 100); environment (barn floor, n = 10, and lairage floor, n = 10); meat juice (n = 100, obtained after defrosting of diaphragm); tonsils (n = 100); mesenteric lymph nodes (MLNs) (n = 100); and carcasses after bleeding (n = 100), after singeing (n = 100), after evisceration (n = 100), and after final rinsing (n = 100). Blood and meat juice were subjected to ELISA to detect antibodies against Salmonella, and other samples were subjected to Salmonella detection by ISO 6579. Salmonella was detected in 3 samples from barn floors, 7 lairage floors, 45 tonsils, 43 MLNs and in 3 carcasses. Based on ELISA, Salmonella positive samples were: 86 and 46 blood serum (20% and 40% cut-offs) and 68 and 46 meat juice (20% and 40% cut-offs). Optical density readings from blood serum and meat juice presented a high and significant correlation (r = 0.93, p < 0.001), and a substantial agreement for Salmonella detection (K = 0.69, ELISA 40% cut-off). The agreement between ELISA and microbiological analysis for Salmonella detection in pig carcasses were absent or poor, with the exception of results obtained by ELISA 40% cut-off from blood serum and meat juice with MLNs (K = 0.49 and 0.50, respectively) and tonsils (K = 0.29 and 0.30, respectively). Based on the obtained results, meat juice can be considered an alternative to blood serum as a matrix for ELISA for preliminary detection of Salmonella, allowing the identification of potential sources of contamination during slaughtering.

Calcium-dependent phospholipase A2 modulates infection-induced diaphragm dysfunction.

Calpain activation contributes to the development of infection-induced diaphragm weakness, but the mechanisms by which infections activate calpain are poorly understood. We postulated that skeletal muscle calcium-dependent phospholipase A2 (cPLA2) is activated by cytokines and has downstream effects that induce calpain activation and muscle weakness. We determined whether cPLA2 activation mediates cytokine-induced calpain activation in isolated skeletal muscle (C2C12) cells and infection-induced diaphragm weakness in mice. C2C12 cells were treated with the following: 1) vehicle; 2) cytomix (TNF-α 20 ng/ml, IL-1ß 50 U/ml, IFN-γ 100 U/ml, LPS 10 µg/ml); 3) cytomix + AACOCF3, a cPLA2 inhibitor (10 µM); or 4) AACOCF3 alone. At 24 h, we assessed cell cPLA2 activity, mitochondrial superoxide generation, calpain activity, and calpastatin activity. We also determined if SS31 (10 µg/ml), a mitochondrial superoxide scavenger, reduced cytomix-mediated calpain activation. Finally, we determined if CDIBA (10 µM), a cPLA2 inhibitor, reduced diaphragm dysfunction due to cecal ligation puncture in mice. Cytomix increased C2C12 cell cPLA2 activity (P < 0.001) and superoxide generation; AACOCF3 and SS31 blocked increases in superoxide generation (P < 0.001). Cytomix also activated calpain (P < 0.001) and inactivated calpastatin (P < 0.01); both AACOCF3 and SS31 prevented these changes. Cecal ligation puncture reduced diaphragm force in mice, and CDIBA prevented this reduction (P < 0.001). cPLA2 modulates cytokine-induced calpain activation in cells and infection-induced diaphragm weakness in animals. We speculate that therapies that inhibit cPLA2 may prevent diaphragm weakness in infected, critically ill patients.

Association between microbiological and serological prevalence of human pathogenic Yersinia spp. in pigs and pig batches.

Pigs are the main reservoir of human pathogenic Y. enterocolitica, and the microbiological and serological prevalence of this pathogen differs between pig farms. The infection status of pig batches at moment of slaughter is unknown while it is a possibility to classify batches. A relation between the presence of human pathogenic Yersinia spp. and the presence of antibodies could help to predict the infection of the pigs prior to slaughter. Pigs from 100 different batches were sampled. Tonsils and pieces of diaphragm were collected from 7047 pigs (on average 70 pigs per batch). The tonsils were analyzed using a direct plating method and the meat juice collected from the pieces of diaphragm was analyzed by Enzyme Linked ImmunoSorbent Assay. The microbiological and serological results were compared using a mixed-effects logistic regression at pig and batch level. Yersinia spp. were found in 2031 (28.8%) pigs, antibodies were present in 4692 (66.6%) pigs. According to the logistic regression, there was no relation at pig level between the presence of Yersinia spp. in tonsils and the presence of antibodies. Contrarily, at batch level, a mean activity value of 37 Optical Density (OD)% indicated a Yersinia spp. positive farm and the microbiological prevalence in pig batches could be estimated before shipment to the slaughterhouse. This offers the opportunity to classify batches based on their potential risk to contaminate carcasses with human pathogenic Yersinia spp.

Mycobacterium avium subsp. avium in lymph nodes and diaphragms of pigs from one infected herd in the Czech Republic.

This study was performed on 40 finished pigs from one herd naturally infected with Mycobacterium avium subsp. avium. The aim was to investigate the presence and amount of M. a. avium in samples of lymph nodes and diaphragm tissues collected during routine postmortem inspection using the triplex quantitative real time PCR (qPCR) method. We collected, in total, 107 samples: various lymph nodes affected by gross tuberculosis (TB)-like lesions from 17 pig carcasses, as well as samples of head and mesenteric lymph nodes from 23 carcasses without TB-like lesions. Samples of diaphragm tissues were collected from all carcasses. M. a. avium was detected in one or more tissue samples collected from half of the slaughtered pigs tested. Samples of diaphragm tissues of three pigs with detected TB-like lesions contained M. a. avium (10(2) to 10(3) cells per g of sample); the organism was not detected in diaphragm tissues from pigs without TB-like lesions. The qPCR method may be useful for quantification of M. a. avium in pigs for the purposes of foodborne risk assessment.

[Concurrent interventions for pulmonary tuberculosis complicated by chronic pleural tuberculosis].

Basing on the literature data analysis the authors suppose the insufficient studying of issues, concerning simultant interventions for pulmonary tuberculosis, coinciding with chronic phthysic pleurytis. Surgical treatment of such processes was performed in 181 patients, total pleurectomy--in 172 (95%) of them, partial--in 9 (5%). Various kinds of pulmonary resection as the additional intervention were performed in 172 (95%) patients, extrapleural thoracoplasty--in 15 (8.3%), diaphragmatic plasty--in 15 (8.3%). In early postoperative period the tuberculosis micobacteria have had disappeared in all the patients, permitting to interpret surgical intervention as a sanation one for every patient, and as a socially directed antiepidemic measure.

The effects of pneumoperitoneum and controlled ventilation on peritoneal lymphatic bacterial clearance: experimental results in rats.

OBJECTIVE: To evaluate the effect of pneumoperitoneum, both alone and in combination with controlled ventilation, on peritoneal lymphatic bacterial clearance using a rat bacterial peritonitis model. METHOD: A total of 69 male Wistar rats were intraperitoneally inoculated with an Escherichia coli solution (10(9) colony-forming units (cfu)/mL) and divided into three groups of 23 animals each: A (control group), B (pneumoperitoneum under 5 mmHg of constant pressure), and C (endotracheal intubation, controlled ventilation, and pneumoperitoneum as in Group B). The animals were sacrificed after 30 min under these conditions, and blood, mediastinal ganglia, lungs, peritoneum, liver, and spleen cultures were performed. RESULTS: Statistical analyses comparing the number of cfu/sample in each of the cultures showed that no differences existed between the three groups. CONCLUSION: Based on our results, we concluded that pneumoperitoneum, either alone or in association with mechanical ventilation, did not modify the bacterial clearance through the diaphragmatic lymphatic system of the peritoneal cavity.

Correlation of Mycobacterium avium subsp. paratuberculosis counts in gastrointestinal tract, muscles of the diaphragm and the masseter of dairy cattle and potential risk for consumers.

Tissues of cattle intended for human consumption can be contaminated by Mycobacterium avium subsp. paratuberculosis (MAP). Although different studies attribute varying roles of MAP in Crohn's disease progression it is thought that the exposure of humans to this bacterium should in any case be minimised. In this study, we have collected samples of intestine, mesenteric lymph nodes, muscles of diaphragm (musculus diaphragma) and masseter muscles (musculus masseter) from twenty-five cows in a slaughterhouse. The infectious status of all animals was confirmed by culture of faeces. MAP was found in almost all the intestines and mesenteric lymph nodes examined, including three faecal culture-negative animals indicating intermittent shedding. As intestine is used for the traditional production of sausages, it is alarming that 84.2% of intestine samples were positive for MAP. F57 and IS900 real time PCR revealed MAP in 40 to 68% of diaphragms and 11.1 to 38.9% of masseters. A noticeable dependence of the probability of MAP positivity of faeces versus gastrointestinal tract (GIT) and of GIT and muscles was observed. Due to the changing behaviour of consumers, both of these muscles have started to be widely used in cuisine. Therefore, the results of this paper imply that the processing of cows with paratuberculosis in abattoirs without any precautions (restrictions) and the usage of meat for human consumption should be rethought.

The effects of pneumoperitoneum and controlled ventilation on peritoneal lymphatic bacterial clearance: experimental results in rats

OBJECTIVE: To evaluate the effect of pneumoperitoneum, both alone and in combination with controlled ventilation, on peritoneal lymphatic bacterial clearance using a rat bacterial peritonitis model. METHOD: A total of 69 male Wistar rats were intraperitoneally inoculated with an Escherichia coli solution (109 colony-forming units (cfu)/mL) and divided into three groups of 23 animals each: A (control group), B (pneumoperitoneum under 5 mmHg of constant pressure), and C (endotracheal intubation, controlled ventilation, and pneumoperitoneum as in Group B). The animals were sacrificed after 30 min under these conditions, and blood, mediastinal ganglia, lungs, peritoneum, liver, and spleen cultures were performed. RESULTS: Statistical analyses comparing the number of cfu/sample in each of the cultures showed that no differences existed between the three groups. CONCLUSION: Based on our results, we concluded that pneumoperitoneum, either alone or in association with mechanical ventilation, did not modify the bacterial clearance through the diaphragmatic lymphatic system of the peritoneal cavity.

Chemokine receptor and ligand upregulation in the diaphragm during endotoxemia and Pseudomonas lung infection.

Sepsis-induced diaphragmatic inflammation has been associated with respiratory failure, but the role of chemokines in this process has not been evaluated. Here we sought to study the local expression and molecular regulation of the chemokines, regulated upon activation normal T cell expressed and secreted (RANTES) and macrophage inflammatory protein (MIP)-1alpha, in the murine diaphragm during sepsis. Constitutive expression levels of RANTES and MIP-1alpha, as well as their receptors, CCR1 and CCR5, were significantly higher in diaphragm than limb muscle. Sepsis was induced by acute lipopolysaccharide (LPS) delivery or subacutely by intratracheal administration of live Pseudomonas aeruginosa bacteria. Both sepsis models triggered a marked upregulation of RANTES and MIP-1alpha in the diaphragm. In vitro, stimulation of diaphragmatic muscle cells with LPS also led to RANTES upregulation. Inhibition of the NF-kB pathway using pharmacologic or dominant negative genetic approaches blocked the LPS-induced RANTES upregulation, while free radical scavengers had no effect. We conclude that sepsis leads to greatly increased expression of RANTES, MIP-1alpha and their cognate receptors in the diaphragm. Manipulation of the NF-kB pathway and other regulators of chemokine expression in the diaphragm could represent a novel method for mitigating the skeletal muscle inflammatory response associated with sepsis-induced diaphragmatic dysfunction.

Impact of IL-10 on diaphragmatic cytokine expression and contractility during Pseudomonas Infection.

Severe weakness of the respiratory muscles, with attendant respiratory failure and death, has been documented in sepsis. In this study, we show that during murine pulmonary infection with Pseudomonas aeruginosa, multiple proinflammatory genes are up-regulated not only within the lungs, but also within the diaphragm. Significant induction of TNF-alpha, IL-1alpha, IL-1beta, IL-6, and IL-18 gene expression occurred within the diaphragm in a bacterial dose-dependent manner. We determined whether the anti-inflammatory cytokine IL-10 could blunt proinflammatory gene expression within the diaphragm under these conditions. The IL-10 receptor was found to be expressed by the diaphragm in vivo as well as in primary diaphragmatic muscle cell cultures. Transduction of myoblasts with an adenoviral vector (Ad-IL-10) induced strong IL-10 expression, and intramuscular injection of the same vector in vivo produced significant increases in IL-10 serum levels. Ad-IL-10 treatment of mice infected with P. aeruginosa significantly inhibited the induction of proinflammatory cytokines within the diaphragm, but not in the infected lungs. Ad-IL-10 treatment also led to greatly improved diaphragmatic force production in infected mice. These results suggest that pulmonary infection triggers proinflammatory gene expression by the diaphragm along with diaphragmatic weakness. Shifting the balance between pro- and anti-inflammatory mediators in favor of the latter by IL-10 gene delivery was able to restore normal diaphragmatic force-generating capacity under these conditions, suggesting a possible avenue for therapeutic intervention.

Primary pulmonary botryomycosis with multiple adjacent organ involvement mimicking mucosa-associated lymphoid tissue lymphoma.

Botryomycosis is an unusual chronic suppurative disease characterized by the histological finding of sulphur granules containing the causative bacteria in an abscess. We describe a previously healthy 14-year-old girl who presented with chronic productive cough. Chest computed tomography (CT) revealed a right lower mass with involvement of the parietal pleura, chest wall, diaphragm, liver and costovertebral junction. Thoracostomy biopsy was performed due to suspicion of low-grade mucosa-associated lymphoid tissue lymphoma. However, histological analysis revealed a pulmonary botryomycosis. The right lower lung mass completely resolved after 3 months of antibiotics treatment. This case emphasizes that primary pulmonary botryomycosis with extensive contiguous involvement may not be distinguishable from lung cancer, lymphoma or fungal infection on chest CT.

Protein carbonyl formation in the diaphragm.

Although protein carbonyl formation is an index of oxidative stress in skeletal muscles, the exact proteins, which undergo oxidation in these muscles, remain unknown. We used 2D electrophoresis, immunoblotting, and mass spectrometry to identify carbonylated proteins in the diaphragm in septic animals. Rats were injected with saline (control) or Escherichia coli lipopolysaccharides (LPS) and killed after various intervals. Diaphragm protein carbonylation increased significantly and peaked 12 h after LPS injection, and it was localized both inside muscle fibers and in blood vessels supplying muscle fibers. Aldolase A, glyceraldehyde 3-phosphate dehydrogenase, enolase 3beta, mitochondrial and cytosolic creatine kinases, alpha-actin, carbonic anyhdrase III, and ubiquinol-cytochrome c reductase were all carbonylated in septic rat diaphragms. In addition, we found significant negative correlations between the intensity of carbonylation and creatine kinase and aldolase activities. We conclude that glycolysis, ATP production, CO2 hydration, and contractile proteins are targeted by oxygen radicals inside the diaphragm during sepsis.

Prevalence and antimicrobial resistance pattern of Salmonella isolates from apparently healthy slaughtered cattle in Ethiopia.

The prevalence and antimicrobial resistance pattern of Salmonella isolates was determined from apparently healthy slaughtered cattle at Debre Zeit (Ethiopia). A total of 323 cattle were examined for the presence of Salmonella in faeces, mesenteric lymph nodes, abdominal and diaphragmatic muscles. Salmonellae were cultured from 23 (7.1%) of the animals. Salmonellae were isolated from 2 (3.1%) and 3 (4.5%) of 65 pooled faecal and mesenteric lymph node samples, respectively. Nine (2.8%) abdominal muscle and 10 (3.1%) diaphragmatic muscle samples (n = 323 of each) were contaminated by Salmonella. About 60% of the serovars identified in the abdominal and diaphragmatic muscles were also detected from faeces and mesenteric lymph node samples. The five different serovars isolated were Salmonella mishmarhaemek (48%) S. typhimurium (20%). S. enteritidis (12%), S. guildford (12%) and S. dublin (48%). The antimicrobial resistance profiles of 25 of the Salmonella isolates with 17 antimicrobials showed that 52%, (13/25) of them were resistant to three or more antimicrobials. Both strains of Salmonella (S. mishmarhaemek and S. typhimurium) showed multiple resistance to ampicillin, sulfamethoxazole and ticarcillin.

Transdiaphragmatic Actinobacillus actinomycetemcomitans infection: case report.

To our knowledge, we report the first case of transdiaphragmatic Actinobacillus actinomycetemcomitans infection in which the patient presented with an abdominal tumor. The clinical manifestation of this infection was similar to that of Actinomyces israelii infection and was initially misinterpreted as a malignancy. The actual diagnosis was made following prolonged culture of the biopsy specimen of the involved tissue. The pathogen was susceptible to penicillin, ampicillin, tetracycline, cephalosporins, metronidazole, aminoglycosides, and quinolones. The patient was treated with a combination of amoxicillin and norfloxacin, and 3 months later his condition had clinically improved.

Effects of viral infection on contraction of the diaphragm in mice.

Isometric contractile properties of isolated phrenic nerve-diaphragm muscle preparations were used to study the effects of picornavirus infections on diaphragm muscle function. Properties of muscles from virus-inoculated and control mice were similar during brief contractions. However, when subjected to a series of fatiguing contractions by indirect or direct stimulation, muscles of mice inoculated with a paralytic variant of encephalomyocarditis (EMC) virus showed a greater rate of fatigue and a reduced capacity to recover from fatigue than did muscles from uninoculated control mice or muscles from mice inoculated with a nonparalytic coxsackievirus B3 (CVB3). Mice paralyzed by EMC virus infection had high titers of virus in the brain and similar titers of virus in diaphragm muscle as found in diaphragm muscles of CVB3-inoculated mice. The results indicate that EMC virus infection of mice leads to increased fatigability of the diaphragm muscle and that there are both neural and muscular components of this enhanced fatigue.

Increased survival from peritonitis after blockade of transdiaphragmatic absorption of bacteria.

Based upon the knowledge that bacteria in the peritoneal cavity gain access to circulating blood by way of transdiaphragmatic absorption into lymph, a study was carried out to determine whether this absorption protects or endangers the host. Blockade of absorption produced by intraperitoneal (IP) injection of platelet rich plasma (PRP) or by scarification of the peritoneal surface of the diaphragm increased survival time in rats with double colonic perforation from 20 per cent in control rats to 86 and 93 per cent, respectively. Each of these maneuvers significantly increased the number of negative blood culture findings.

[Detection of Trichinella spiralis infection in spontaneously infected pigs intended for slaughter using the enzyme-linked immunosorbent assay (ELISA) (author's transl)]. / De detectie van Trichinella spiralis infecties bij natuurlijk geïnfecteerde slachtvarkens met behulp van de enzyme-linked immunosorbent assay (ELISA)

In a study of seventy-seven pigs intended for saughter, from two farms on which Trichinella infections had previously been detected by trichinoscopy, the sensitivity of the enzyme-linked immunosorbent assay (ELISA) was compared with that of two direct (trichinoscopy and digestion) and three indirect (serological) methods of detection (fluorescent antibody technique, counter-electrophoresis and Ouchterlony's agar gel diffusion method). In five out of seventy-seven pigs, the digestion procedure revealed the presence of larvae of Trichinella spiralis in the diaphragm. Four of these pigs showed a very high degree of infection, whereas one showed a very low degree of infection. Infection was shown to be present in all five infected pigs using the ELISA method, whereas the findings were only positive in the four severely infected pigs when trichinoscopy, the fluorescent antibody technique, counter-electrophoresis and Ouchterlony's method were used. The ELISA was sufficiently sensitive to detect Trichinella infection even in those pigs which showed a low degree of infection.

[Isolation of Toxoplasma strains via the digestive method]. / Izolirane na shtamove toksoplazmi chrez digestivniia metod

Tested was the effectiveness of the digestive method for the isolation of Toxoplasma strains. This method was used to study the musculature of the diaphragm in 80 sheep, 40 cattle, and 80 pigs. Isolated were one strain that was pathogenic and 3 apathogenic strains for albino mice. They were found only in the diaphragm of sheep. It is believed that the digestive method is effective for the isolation of Toxoplasma strains.