scholarly journals Enhancement of immunohistochemical detection of Salmonella in tissues of experimentally infected pigs

2015 ◽  
Vol 59 (3) ◽  
Author(s):  
J. Rieger ◽  
P. Janczyk ◽  
H. Hünigen ◽  
J. Plendl
Keyword(s):  
Lymph Nodes ◽  
Detection System ◽  
Intestinal Damage ◽  
Immunohistochemical Detection ◽  
Mesenteric Lymph Nodes ◽  
Tissue Samples ◽  
Serovar Typhimurium ◽  
Triton X ◽  
Foodborne Infections ◽  
Higher Sensitivity

<p><em>Salmonella</em> Typhimurium is one of the main pathogens compromising porcine and human health as well as food safety, because it is a prevailing source of foodborne infections due to contaminated pork. A prominent problem in the management of this bacteriosis is the number of subclinically infected carrier pigs. As very little is known concerning the mechanisms allowing <em>Salmonella</em> to persist in pigs, the objective of this study was to develop an immunohistochemical approach for the detection of salmonellae in tissue of pigs experimentally infected with <em>Salmonella</em> Typhimurium. Samples were obtained from a challenge trial in which piglets of the German Landrace were intragastrically infected with <em>Salmonella</em> enterica serovar Typhimurium DT104 (1.4-2.1x10<sup>10</sup> CFU). Piglets were sacrificed on days 2 and 28 post infection. Tissue samples of jejunum, ileum, colon, ileocecal mesenteric lymph nodes (<em>Lnn. ileocolici</em>), and tonsils (<em>Tonsilla veli palatini</em>) were fixed in Zamboni’s fixative and paraffin-embedded. Different immunohistochemical staining protocols were evaluated. <em>Salmonella</em> was detected in varying amounts in the tissues. Brown iron-containing pigments in the lymph nodes interfered with the identification of <em>Salmonella</em> if DAB was used as a staining reagent. Detergents like Triton X-100 or Saponin enhanced the sensitivity. It seems advisable not to use a detection system with brown staining for bacteria in an experimental setup involving intestinal damage including haemorrhage. The use of detergents appears to result in a higher sensitivity in the immunohistochemical detection of salmonellae.</p>

Mycobacterium avium subsp. avium in Lymph Nodes and Diaphragms of Pigs from One Infected Herd in the Czech Republic

2014 ◽  
Vol 77 (1) ◽  
pp. 141-144 ◽  
Author(s):  
PETR KRIZ ◽  
MARIJA KAEVSKA ◽  
IVA SLANA ◽  
IVA BARTEJSOVA ◽  
IVO PAVLIK
Keyword(s):  
Lymph Nodes ◽  
Mycobacterium Avium ◽  
Mycobacterium Avium Subsp ◽  
Mesenteric Lymph Nodes ◽  
Tissue Samples ◽  
The Czech Republic ◽  
Mesenteric Lymph ◽  
Qpcr Method ◽  
Pig Carcasses ◽  
Slaughtered Pigs

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 (102 to 103 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.

scholarly journals A mixed infection of Mycobacterium avium subsp. paratuberculosis and M. a. hominissuis in one red deer (Cervus elaphus) studied by IS900 BstEII and IS1245 PvuII RFLP analyses: a case report

2008 ◽  
Vol 53 (No. 8) ◽  
pp. 445-451 ◽  
Author(s):  
M. Moravkova ◽  
I. Trcka ◽  
J. Lamka ◽  
I. Pavlik
Keyword(s):  
Lymph Nodes ◽  
Mycobacterium Avium ◽  
Cervus Elaphus ◽  
Mixed Infection ◽  
Red Deer ◽  
Chronic Diarrhoea ◽  
Mesenteric Lymph Nodes ◽  
Tissue Samples ◽  
Mesenteric Lymph ◽  
Rflp Type

A mixed infection with <i>Mycobacterium avium</i> subsp. <i>paratuberculosis</i> (<i>MAP</i>) and <i>Mycobacterium avium</i> subsp. <i>hominissuis</i> (<i>MAH</i>) in one naturally infected red deer stag from a game park is described. The animal was euthanized because of symptoms of poor condition, weight loss and chronic diarrhoea. In spite of that, pathological lesions were observed only in the mesenteric lymph nodes, which were five to ten times enlarged with confluent caseous granulomas of 1 to 10 mm in size. Mycobacteria were isolated from all studied samples: a mixed infection of <i>MAP</i> and <i>MAH</i> was confirmed by multiplex PCR for the detection of IS <i>900</i>, IS<i>901</i>1, IS<i>1245</i> and <i>dnaJ</i>. MAP</i> of the identical IS<i>900</i> <i>BstE</i>II RFLP type C1 was isolated from all tissue samples and faeces. <i>MAH</i> isolates were detected in six examined tissue samples, including three mesenteric lymph nodes with caseous granulomas. Only minor differences in the band numbers and position of four different IS<i>1245</i> <i>Pvu</i>II RFLP patterns of <i>MAH</i> isolates were found. It follows from these results that red deer may potentially be infected with <i>MAH</i>, when a <i>MAP</i> infection is under way.

scholarly journals Global transcriptional response of porcine mesenteric lymph nodes to Salmonella enterica serovar Typhimurium

Genomics ◽  
2007 ◽  
Vol 90 (1) ◽  
pp. 72-84 ◽  
Author(s):  
Yanfang Wang ◽  
Long Qu ◽  
Jolita J. Uthe ◽  
Shawn M.D. Bearson ◽  
Daniel Kuhar ◽  
...  
Keyword(s):  
Lymph Nodes ◽  
Salmonella Enterica ◽  
Salmonella Enterica Serovar Typhimurium ◽  
Transcriptional Response ◽  
Mesenteric Lymph Nodes ◽  
Mesenteric Lymph ◽  
Serovar Typhimurium

scholarly journals Oral Immunization with ATP-Dependent Protease-Deficient Mutants Protects Mice against Subsequent Oral Challenge with Virulent Salmonella enterica Serovar Typhimurium

2003 ◽  
Vol 71 (1) ◽  
pp. 30-39 ◽  
Author(s):  
Hidenori Matsui ◽  
Masato Suzuki ◽  
Yasunori Isshiki ◽  
Chie Kodama ◽  
Masahiro Eguchi ◽  
...  
Keyword(s):  
Lymph Nodes ◽  
Salmonella Enterica ◽  
Peyer's Patches ◽  
Oral Challenge ◽  
Secretory Iga ◽  
Peyer’S Patches ◽  
Lon Protease ◽  
Mesenteric Lymph Nodes ◽  
Mesenteric Lymph ◽  
Serovar Typhimurium

ABSTRACT We evaluated the efficacy of mutants with a deletion of the stress response protease gene as candidates for live oral vaccine strains against Salmonella infection through infection studies with mice by using a Salmonella enterica serovar Typhimurium mutant with a disruption of the ClpXP or Lon protease. In vitro, the ClpXP protease regulates flagellum synthesis and the ClpXP-deficient mutant strain exhibits hyperflagellated bacterial cells (T. Tomoyasu et al., J. Bacteriol. 184:645-653, 2002). On the other hand, the Lon protease negatively regulates the efficacy of invading epithelial cells and the expression of invasion genes (A. Takaya et al., J. Bacteriol. 184:224-232, 2002). When 5-week-old BALB/c mice were orally administered 5 × 108 CFU of the ClpXP- or Lon-deficient strain, bacteria were detected with 103 to 104 CFU in the spleen, mesenteric lymph nodes, Peyer's patches, and cecum 1 week after inoculation and the bacteria then decreased gradually in each tissue. Significant increases of lipopolysaccharide-specific immunoglobulin G (IgG) and secretory IgA were detected at week 4 and maintained until at least week 12 after inoculation in serum and bile, respectively. Immunization with the ClpXP- or Lon-deficient strain protected mice against oral challenge with the serovar Typhimurium virulent strain. Both the challenged virulent and immunized avirulent salmonellae were completely cleared from the spleen, mesenteric lymph nodes, Peyer's patches, and even cecum 5 days after the challenge. These data indicate that Salmonella with a disruption of the ATP-dependent protease ClpXP or Lon can be useful in developing a live vaccine strain.

scholarly journals Mesenteric Lymph Nodes Confine Dendritic Cell-Mediated Dissemination of Salmonella enterica Serovar Typhimurium and Limit Systemic Disease in Mice

2009 ◽  
Vol 77 (8) ◽  
pp. 3170-3180 ◽  
Author(s):  
Sabrina Voedisch ◽  
Christian Koenecke ◽  
Sascha David ◽  
Heike Herbrand ◽  
Reinhold Förster ◽  
...  
Keyword(s):  
Dendritic Cell ◽  
Lymph Nodes ◽  
Salmonella Enterica ◽  
Systemic Disease ◽  
Mouse Strains ◽  
Surgical Removal ◽  
Mesenteric Lymph Nodes ◽  
Mesenteric Lymph ◽  
Serovar Typhimurium

ABSTRACT In humans with typhoid fever or in mouse strains susceptible to Salmonella enterica serovar Typhimurium (S. Typhimurium) infection, bacteria gain access to extraintestinal tissues, causing severe systemic disease. Here we show that in the gut-draining mesenteric lymph nodes (MLN), the majority of S. Typhimurium-carrying cells show dendritic-cell (DC) morphology and express the DC marker CD11c, indicating that S. Typhimurium bacteria are transported to the MLN by migratory DCs. In vivo FLT-3L-induced expansion of DCs, as well as stimulation of DC migration by Toll-like receptor agonists, results in increased numbers of S. Typhimurium bacteria reaching the MLN. Conversely, genetically impaired DC migration in chemokine receptor CCR7-deficient mice reduces the number of S. Typhimurium bacteria reaching the MLN. This indicates that transport of S. Typhimurium from the intestine into the MLN is limited by the number of migratory DCs carrying S. Typhimurium bacteria. In contrast, modulation of DC migration does not affect the number of S. Typhimurium bacteria reaching systemic tissues, indicating that DC-bound transport of S. Typhimurium does not substantially contribute to systemic S. Typhimurium infection. Surgical removal of the MLN results in increased numbers of S. Typhimurium bacteria reaching systemic sites early after infection, thereby rendering otherwise resistant mice susceptible to fatal systemic disease development. This suggests that the MLN provide a vital barrier shielding systemic compartments from DC-mediated dissemination of S. Typhimurium. Thus, confinement of S. Typhimurium in gut-associated lymphoid tissue and MLN delays massive extraintestinal dissemination and at the same time allows for the establishment of protective adaptive immune responses.

scholarly journals IgA production, coliforms analysis and intestinal mucosa morphology of piglets that received probiotics with viable or inactivated cells

2007 ◽  
Vol 27 (6) ◽  
pp. 241-245 ◽  
Author(s):  
Maria A. Martins Rodrigues ◽  
Deise A. de Oliveira Silva ◽  
Ernesto A. Taketomi ◽  
Francisco J. Hernandez-Blazquez
Keyword(s):  
Lymph Nodes ◽  
Lactobacillus Acidophilus ◽  
Serum Levels ◽  
Bifidobacterium Bifidum ◽  
Control Group ◽  
Mesenteric Lymph Nodes ◽  
Tissue Samples ◽  
Mesenteric Lymph ◽  
Health And Development ◽  
Iga Production

Two types of probiotics were used in piglets. One product is a mixed culture of viable Lactobacillus acidophilus, Enterococcus faecium e Bifidobacterium bifidum. The second product is composed of inactivated Lactobacillus acidophilus cells. The piglets received two weekly oral doses for 30 days while a control group did not receive probiotics. All piglets were euthanized at the 30th day of life and the mesenteric lymph nodes, the small intestine, and blood samples were collected. The tissue samples were studied by light microscopy and the blood serum was analyzed by ELISA method. The treatment with the probiotic with viable cells produced higher serum levels of IgA (P<0.05) and more IgA expressing cells were found in the mesenteric lymph nodes than observed in the inactivated cells treatment or control groups (P<0.05). Also, intestinal villi were longer, crypts were deeper (P<0.05) and fecal coliform count was lower than found in the inactivated product (P<0.05). These results suggest that viable probiotics are more efficient than inactivated probiotics to induce immunostimulation and intestinal modifications in piglets, thus improving their health and development.

scholarly journals Salmonella enterica Serovar Typhimurium Travels to Mesenteric Lymph Nodes Both with Host Cells and Autonomously

2018 ◽  
Vol 202 (1) ◽  
pp. 260-267 ◽  
Author(s):  
Alberto Bravo-Blas ◽  
Lotta Utriainen ◽  
Slater L. Clay ◽  
Verena Kästele ◽  
Vuk Cerovic ◽  
...  
Keyword(s):  
Lymph Nodes ◽  
Salmonella Enterica ◽  
Salmonella Enterica Serovar Typhimurium ◽  
Host Cells ◽  
Mesenteric Lymph Nodes ◽  
Mesenteric Lymph ◽  
Serovar Typhimurium

scholarly journals Isolation of Mycobacterium avium subsp. paratuberculosis from mouflon in Bulgaria

2020 ◽  
Vol 9 (5-6) ◽  
pp. 665-670
Author(s):  
T. Savova ◽  
R. Petrova ◽  
V. Valcheva ◽  
M. Bonovska ◽  
H. Najdenski
Keyword(s):  
Lymph Nodes ◽  
Mycobacterium Avium ◽  
Intestinal Wall ◽  
Mesenteric Lymph Nodes ◽  
Tissue Samples ◽  
Diffuse Infiltration ◽  
Domestic Ruminants ◽  
Mesenteric Lymph ◽  
Faecal Samples ◽  
Depth Study

Mycobacterium avium subsp. paratuberculosis (MAP) is the etiological agent of paratuberculosis (John’s disease) mainly in large and small domestic and wild ruminants, and suspected causative agent in human Crohn’s disease. In Bulgaria, paratuberculosis is still poorly researched in both groups of ruminants. We present results of the first in-depth study of mouflon, grown free in one hunting reserve in the Western region of the country. The aim was to prove the presence of MAP in diagnostic materials from regularly hunted or dead mouflon suspected for paratuberculosis. Small intestine and mesenteric lymph nodes (MLN) from 12 hunted and 4 dead mouflon and 10 faecal samples (Fc) were studied in the period of 2009–2013. Typical for paratuberculosis pathomorphological lesions were observed in four mouflon (of 16 examined). The intestinal wall was thickened, strongly folded and soft, with severe hyperemia. The MLN were enlarged, soft, with marbled appearance. The affected section of the ileum showed hyperplasia of the mucous corion and submucosa with diffuse infiltration of epithelioid cells. Lymphadenopathy with atrophy of T and B lymphocytes areas was observed in the mesenteric lymph nodes. For bacteriological isolation of MAP, the tissue and faecal samples were decontaminated with NALC-NaOH, cultured in Middlebrook 7H9 Broth and on Herrold’s medium. The Ziehl–Neelsen stained smears and isolates were examined microscopically for acid-fast bacteria. Presence of MAP was observed in tissue samples of 4 (25%) mouflon and in 2 (20%) faecal samples. The same samples were confirmed by the IS900 PCR for the presence of specific for MAP fragments with a commercial amplification kit. The cases of paratuberculosis found at different times in the free-living mouflon in our study prove that the disease exists in Bulgaria and highlight the need for more serious control of the disease among wild and domestic ruminants.

scholarly journals Pretreatment of Mice with Streptomycin Provides a Salmonella enterica Serovar Typhimurium Colitis Model That Allows Analysis of Both Pathogen and Host

2003 ◽  
Vol 71 (5) ◽  
pp. 2839-2858 ◽  
Author(s):  
Manja Barthel ◽  
Siegfried Hapfelmeier ◽  
Leticia Quintanilla-Martínez ◽  
Marcus Kremer ◽  
Manfred Rohde ◽  
...  
Keyword(s):  
Lymph Nodes ◽  
Salmonella Enterica ◽  
Molecular Mechanisms ◽  
Intestinal Inflammation ◽  
Intercellular Adhesion Molecule ◽  
Infection Model ◽  
Intercellular Adhesion Molecule 1 ◽  
Mesenteric Lymph Nodes ◽  
Receptor Knockout Mouse ◽  
Serovar Typhimurium

ABSTRACT Salmonella enterica subspecies 1 serovar Typhimurium is a principal cause of human enterocolitis. For unknown reasons, in mice serovar Typhimurium does not provoke intestinal inflammation but rather targets the gut-associated lymphatic tissues and causes a systemic typhoid-like infection. The lack of a suitable murine model has limited the analysis of the pathogenetic mechanisms of intestinal salmonellosis. We describe here how streptomycin-pretreated mice provide a mouse model for serovar Typhimurium colitis. Serovar Typhimurium colitis in streptomycin-pretreated mice resembles many aspects of the human infection, including epithelial ulceration, edema, induction of intercellular adhesion molecule 1, and massive infiltration of PMN/CD18+ cells. This pathology is strongly dependent on protein translocation via the serovar Typhimurium SPI1 type III secretion system. Using a lymphotoxin β-receptor knockout mouse strain that lacks all lymph nodes and organized gut-associated lymphatic tissues, we demonstrate that Peyer's patches and mesenteric lymph nodes are dispensable for the initiation of murine serovar Typhimurium colitis. Our results demonstrate that streptomycin-pretreated mice offer a unique infection model that allows for the first time to use mutants of both the pathogen and the host to study the molecular mechanisms of enteric salmonellosis.

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