Mesenteric Lymph Node Samples Research Articles

Detection of Extended- Spectrum Beta-Lactamase producing Escherichia coli from mesenteric lymph nodes of wild boars (Sus scrofa)

Wild boars (Sus scrofa) are increasing in several European countries, including Italy. In areas with intensive animal farming, like the Italian Emilia-Romagna region, they are likely to be exposed to antimicrobialresistant (AMR) bacteria of livestock origin. In 2017-2018, 108 mesenteric lymph nodes samples were collected from 108 wild boars hunted in Parma province, Emilia-Romagna region, to be tested for ESBL- and carbapenemase-producing Escherichia coli. One isolate (WB-21L) out of 108 (0.9%) was phenotypically confirmed as ESBLproducing E. coli. The strain WB-21L was tested by PCR for the genes blaSHV, blaCTX-M, blaTEM, blaAmpC, blaKPC, blaNDM, blaVIM, blaIMP, blaOXA-48, blaSPM, blaBIC, blaSIM, blaDIM, blaGIM, blaAIM, resulting positive for TEM β- lactamase. Resistance to ampicillin, amoxicillin/clavulanic acid, streptomycin, sulfasomidine, tetracycline and trimethoprim confirmed the multi-resistance nature of the strain WB-21L. Nine E.coli isolates showed resistance to meropenem by the Kirby Bauer test but none of them showed Meropenem MIC values indicative of resistance. In conclusion, the present study shows the presence of ESBL E. coli in wild boars and the possible risk of transfer to game meat handlers and consumers. Future studies are needed to better evaluate the sources of AMR bacteria in wildlife.

Short communication: Use of a new form of protected sodium butyrate to control Salmonella infection in fattening pigs

A field trial on a commercial pig farm was carried out to assess the efficacy of the addition in the diet of fattening pigs of a new form of sodium butyrate protected with a sodium salt of coconut fatty acid distillate (3 kg/ton of feed) to control Salmonella spp. infection. Around 50 pigs were assigned to treatment group and 50 kept as controls. During the fattening period pigs were monthly sampled (serum and feces), and after slaughter fecal and mesenteric lymph nodes samples were collected. No differences in the proportion of shedders were observed between the sodium butyrate and the control groups, but a significant reduction in the number of infected pigs (61% vs. 4%; p<0.01) and in the median ELISA Optical Density percentage values (55.9% vs. 19.4%; p<0.01) at slaughter was observed in pigs under treatment compared to the controls. In addition, an overall significant association between seropositivity and Salmonella shedding and infection was detected. Results from this study add more evidences on the positive effect of butyrate on the control of pig salmonellosis.

Experimental poisoning by Brachiaria decumbens in rabbits

ABSTRACT: Brachiaria spp. are important sources of forage for ruminants in Brazil, due to the easy cultivation, good resistance to drought, good adaptation to different soils and low maintenance cost. However, the ingestion of this grass has been related to photosensitization outbreaks in cattle and sheep with significant economic losses. The hepatotoxic effects related to the ingestion of grass are the formation of crystals and foamy macrophages due to the accumulation of toxic metabolites. The use of cattle and sheep in experiments involving the plant presents several obstacles in the ethical, economic and animal management. The objective of this study was to evaluate the sensitivity of rabbits as an experimental model for B. decumbens poisoning. Two experiments were carried out. In Experiment 1 four rabbits received the fresh plant in daily doses of 10, 20, 40 and 80g/kg body weight for 120 days. In Experiment 2 three rabbits received the fresh plant in amounts of 500g daily with duration of 210 days. The animals of Experiment 1 showed no clinical signs and no macroscopic and microscopic changes characteristic of B. decumbens poisoning. In Experiment 2 the animals also showed no clinical signs or significant macroscopic alterations. Histological analysis showed isolated foamy macrophages or present in random groups of cells in the liver and mesenteric lymph nodes. Samples of liver and mesenteric lymph nodes of the rabbits of Experiment 2 were submitted to the lectin-histochemistry technique. The WGA, sWGA and RCA lectins showed reactivity in foamy macrophages in both organs. This is the first study of our knowledge that demonstrates histopathological lesions caused expetimentally by Brachiaria spp. in rabbits, demonstrating its potential as an animal model.

Natural killer T cell ligand alpha-galactosylceramide protects against gut ischemia reperfusion-induced organ injury in mice

Background & AimsGut ischemia reperfusion (I/R) injury is a life-threatening condition. The immune response plays an important role in I/R-induced organ injury. Alpha-galactosylceramide (α-GalCer) is a potent natural killer T (NKT) cell stimulator. Activation of NKT cells by α-GalCer has been shown to reduce I/R-induced injury in the liver and heart. However, whether α-GalCer has any protective effects on gut I/R-induced organ injury remained unknown. The aim of this study was to test the hypothesis that α-GalCer attenuates gut I/R-induced local and remote organ injury through modulating immune responses. MethodsGut ischemia was induced by placing a microvascular clip across the superior mesenteric artery for 30 min in male adult mice. After removing the clip, α-GalCer (2 µg/mouse) or normal saline containing 0.5% Tween 20 (Vehicle) was administered intraperitoneally. Blood, gut, lung and mesenteric lymph node (MLN) samples were collected 4 h after reperfusion to detect bacterial translocation, tight junction protein, tissue damage, edema, apoptosis, IL-4, IL-10, IFN-γ and TNF-α levels. Resultsα-GalCer significantly reduced bacterial translocation to the MLN, restored tight junction protein and attenuated gut and lung injury after gut I/R. α-GalCer markedly stimulated the production of IL-4, IL-10 and IFN-γ, but had no obvious effects on TNF-α production in gut I/R mice. Pretreatment with anti-CD1d, IL-4 or IL-10, but not IFN-γ blocking antibodies abolished the protective effects of α-GalCer in gut I/R. Conclusionsα-GalCer treatment improved gut barrier function and attenuated gut and lung injury after gut I/R. The beneficial effects of α-GalCer in gut I/R were NKT cell dependent and mediated through upregulation of IL-4 and IL-10. Thus, activation of NKT cells by α-GalCer may serve as a novel option in the treatment of gut I/R injury.

Pathomorphology of peripheral organs of immunogenesis in cats with spontaneous feline infectious peritonitis

This article presents the results of pathomorphological research on the spleen and mesenteric lymph nodes of 23 dead cats aged from 3 months to 7 years, which in their lifetimes (according to anamnesis, clinical signs, laboratory-instrumental methods of examination and VetExpert FCoV Ab express test) had been diagnosed with infectious peritonitis. All the animals were domestic. Blood was drawn from the subcutaneous vein of the forelimb of the diseased cats. We determined ESR, morphological parameters of blood and content of hemoglobin. For histological and histochemical examinations, we selected samples of spleen and mesenteric lymph nodes, which were fixated in 10% aqueous solution of neutral formalin, Carnoy’s and Bouin’s solutions. The prepared histological sections were stained using haematoxylin and eosin, Van Gieson’s stain, methyl green-pironin stain (Brashe), PAS-reaction (McManus), alcian blue and Congo red. Hematological parameters during 3 weeks of clinical progression of the disease among the cats demonstrated a decrease in the hemoglobin content and in the number of erythrocytes and leukocytes. Possible decrease in the number of lymphocytes indicated the development of an immune-deficiency state. Also, during the development of disease, the animals had possible increase in ESR, which indicated the development of an inflammatory process in the organism and decrease in the number of thrombocytes, which conditioned development of disseminated intravascular coagulation. The anatomical pathology autopsy showed that in most animals the spleen was diminished in size, the surface of the organ was tuberous, the capsule was wrinkled and mat, the edges were sharpened. The histostructural change was accompanied by a sharp depletion of the lymph nodes and reduction in the number of micro- and macrophages, which indicated the reduction of white pulp, rapid inhibition of the activity of T- and B-lymphocytes, plasmacytic and macrophage reaction and manifested in development of immune-deficient condition of the organism. In this process, the reticular carcass of the lymph nodes saturated with PAS-positive and eosinophilic masses was clearly manifested, which indicated formation of fibrinoid. In the spleen of 5 individuals, during staining using Congo red, we found deposition of amyloid masses both in the intima of the blood vessels and along the reticulary fibers of the lymph nodes. In the cytoplasm of macrophages, we found pyroninophilic formations. In two cases, we observed blood accumulation of red pulp and bleeding following the reduction of white pulp, and in one case fibrinogenous perisplenitis. In the mesenteric lymph nodes of most of the cats which had suffered from infectious peritonitis, we determined that edema, exposure of the reticular soft skeleton (stroma) of adrenal and paracortical zones, dilation of the border and central sinuses and thrombosis of vessels were followed by steep decrease in the number of T- and B-lymphocytes, plasma cells, micro- and macrophages, which indicated the development of atrophic processes of lympoid tissue and immune-suppression. In three cases, in mesenteric lymph nodes of cats, we determined development of sinus histiocytosis. The changes determined in the spleen and lymph nodes of the cats which had suffered from FIP indicate immune-suppressed condition and steep decrease in the functional ability of the organs and organism in general.

First molecular epidemiological study of Mycobacterium avium subsp. paratuberculosis in cattle and buffalo from different regions of Brazil.

Paratuberculosis is an incurable disease in ruminants with great worldwide economic impact, caused by Mycobacterium avium subsp. paratuberculosis (MAP). The objective of this study was to carry out a study of the molecular epidemiology of the MAP using the restriction enzyme analysis (REA) technique of IS1311 MAP region in biological samples of feces, intestinal tissue, and mesenteric lymph nodes of cattle and buffaloes from six Brazilian states. In total, 109 samples of feces and tissues of cattle and buffaloes were collected from animal paratuberculosis suspected. Twenty-five samples were positive in the detection of the DNA of the IS900 region of MAP and it was possible to type 18 strains in the analysis of the region IS1311, being 100% of them identified as belonging to subtype Bison MAP strain. This is the first epidemiological molecular study of MAP in Brazil. The results indicate that paratuberculosis is widespread in cattle and in buffaloes in several regions of Brazil, and the subtype Bison MAP strain was the only one identified in the samples analyzed in this study, demonstrating the similarity between the strains from different states tested. These results provide the necessary support for the implementation of paratuberculosis control strategies in cattle and buffaloes in Brazil.

Birth weight affects immune response of piglets during the peri-weaning period

Abstract Relationships between birth weight and a range of immune markers were studied in piglets during the peri-weaning period. A total of 47 sows and their respective litters adjusted to 12 piglets were used. Piglets were weighed after birth and within each litter 2 low weight (LW) and 2 high weight (HW) animals were identified: the average weight of the 2 groups was 1.12 and 1.81 ± 0.17 kg, respectively. Within litter, one couple of LW and HW piglets was sacrificed at 16 and 23 d of age, two days after weaning at day 21, to characterise different leukocyte subsets in mesenteric lymph nodes (MLN) and blood by flow cytometry. Jejunal mucosal and MLN samples were also collected to measure the expression of genes involved in immune response by qPCR. The percentage of blood γδT cells was higher in HW piglets before and after weaning (P=0.01 and P<0.001, respectively). At 16 d, LW animals had higher amounts of CD4+CD8+ T cells and dendritic cells in the blood (P=0.01 and P=0.04, respectively) and in MLN (P=0.02 and P=0.009 respectively), and lower count of blood cytotoxic T cells (P=0.01) than HW. Moreover, jejunal expression of TNFα was reduced in LW piglets (P=0.02), while MLN expression of CCL23, BMP2 and SPP1 was increased (P=0.003, P=0.002 and P=0.02, respectively). Two days after weaning (d 23), HW piglets had higher percentages of T cells (P=0.001) and lower amount of non-T cells (P=0.03) and natural killer cells (P=0.04) in blood compared to LW animals. These data revealed that piglet birth weight affects the proportion of leukocyte subsets in different tissues and the expression of genes involved in the immunity. Globally, our results suggest that the development of the immune system after birth differed between LW and HW piglets.

Pathomorphological characteristic of immune organs for circovirus infection of piglets

The article elucidates the results of pathoanatomical and histological searches of peripheral immune organs for circovirus infection of piglets. In one of the farms of Transcarpathian region among the pigs of 3–4 months of age were found clinically ill animals, blood was taken, serum test was performed and circiiros was diagnosed. The pathoanatomical autopsy of 12 corpses of piglets was done. During the autopsy of corpses macroscopic changes were studied and samples of mesenteric lymph nodes and spleen were taken, which were fixed in 10% neutral formalin, dehydrated in alcohols of increasing concentration, followed by sealing and filling with paraffin. From paraffin blocks, histological cuts were made, stained with hematoxylin and eosin, using the Van Gizon method. Light microscopy and photographing were performed using a Leica DM-2500 (Switzerland) microscope and Leica DFC450C cameras and the Leica Application Suite Version 4.4 software. For macroscopic examination, in all cases, hyperplasia of the spleen and serous-hemorrhagic lymphadenitis have been established. For histological examination of mesenteric lymph nodes in sick piglets with cirrhosis against a background of severe congestive hyperemia, diapedeous hemorrhages, exudation, there was a sharp decrease in the number of lymphocytes in the lymph nodes, activation of reticuloendothelial cells, transformation of them into epithelioid and formation of giant cells. The detection of polycarocytes in the lymph nodes of sick piglets on the circus is a morphological confirmation of the cytopathic effect of the virus. The histological search of the spleen of pigs for circovirus infection on the background of blood filling, edema of trabeculae, plasma permeation of vascular walls was marked by an intensive decrease in the number of white pulp cells, as evidenced by a sharp decrease in the lymph nodes themselves. The revealed histostructural changes indicated a depletion of the spleen, suppression of its generative function, and an immunosuppressive state of one of the organs of immunogenesis.

Goat paratuberculosis in Shiraz: Histopathological and molecular approaches.

In the present study, Mycobacterium avium subsp. paratuberculosis (MAP) was investigated in goats slaughtered in Shiraz abattoir using histopathological examinations and polymerase chain reaction (PCR). Ilium and mesenteric lymph node samples from 66 suspected goat carcasses to Johne’s disease were collected. Among 66 examined slaughtered goats, nine (13.63%) goats were positive for MAP in both histopathological and PCR examinations. Eight goats were positive in PCR method while no lesion related to Johne’s disease was observed in their histopathological sections. All positive goats in histopathological examination were also positive in PCR. Based on the results of PCR, the detection rate of MAP in Shiraz abattoir was 25.80% (17 goats). According to the present findings, although both histopathological and PCR methods are appropriate for detecting Johne’s disease, PCR is more sensitive than histopathological examination.

Characterization of Salmonella isolated from apparently healthy slaughtered cattle and retail beef in Hawassa, southern Ethiopia

The present study was conducted to determine the occurrence, serotype distribution and antimicrobial resistance of Salmonella serotypes in slaughtered cattle, slaughterhouse environment and retail beef. Cecal content (CC), mesenteric lymph nodes (MLN), spleen and carcass swab (CS) samples (each, n=150) were collected from 150 cattle slaughtered at Hawassa municipality slaughterhouse. Floor swab specimens (SHFS) were collected on 11 occasions from the slaughterhouse, and 100 beef samples were collected from 100 butcher shops. The samples were cultured for Salmonella, following standard procedures. A total of 14 Salmonella isolates belonging to 3 serotypes namely Salmonella enterica serotype Muenchen (4 isolates), S. enterica serotype 1,4,5,12:i:- (5) and S. enterica serotype Korovi (5) were recovered. All of the 5 S. enterica serotype 1,4,5,12:i:- isolates belonged to phage type 120. Four (2.7%) of the slaughtered cattle carried Salmonella in their CC and/or MLN, while none of the spleen samples were positive for Salmonella. Salmonella was isolated from four (2.7%) CC and two (1.3%) MLN samples. Out of the total of 150 CS samples, two (1.3%) were found contaminated with Salmonella, while 4 (4%) of the 100 beef samples obtained from butcher shops yielded Salmonella. Two of the 11 (18%) SHFS were positive for Salmonella. All the four isolates from beef were S. enterica serotype Muenchen, while both of the isolates from MLN were S. enterica serotype1,4,5,12:i:-. Both S. enterica serotype1,4,5,12:i:- and S. enterica serotype Korovi were isolated from CC, CS and slaughterhouse environment. All the 14 isolates recovered during the study were tested and found pan-susceptible to a panel of 14 antimicrobials. The present study helped to update the information on the occurrence, serotype distribution and antibiogram of Salmonella in slaughter cattle and beef in Ethiopia.

Effect of the addition of protected sodium butyrate to the feed on Salmonella spp. infection dynamics in fattening pigs

Organic acids (OA) are seen as an alternative to antibiotics to reduce the burden of enteropathogens. Two replicates of a field trial were carried out to assess the effect of the addition of protected sodium butyrate (PSB) to the feed (dose of 3kg/T) along the fattening period on the dynamics of Salmonella spp. infection in pigs. In each trial, around 50 pigs were assigned to a treatment group (TG) and 50 kept as controls (CG). Pigs were serologically monitored monthly and on-farm fecal samples and fecal and mesenteric lymph nodes (MLN) samples at slaughter were collected. In the first replicate, pigs became probably infected with Salmonella before the addition of PSB to the feed, but despite of that an overall lower proportion of shedders along the end of fattening period and lower seroprevalence before slaughter (50% vs. 89.6%; P <0.001) was observed in the TG. In the second replicate, few pigs shed Salmonella during the trial, which precluded the finding of significant differences between groups for both Salmonella infection and shedding, but the seroprevalence at slaughter in the CG was again significantly higher than in the TG (31.1% vs. 13.7%, respectively; P=0.02) and it was related to a higher proportion of shedders and MLN-positive pigs. When results from both trials were analyzed together, a significant increasing risk of shedding in the CG was observed at 90days of fattening and at slaughter, and an overall significant decreasing trend in OD% values and thus in seroprevalence was also observed when pigs approached to slaughter. In conclusion, the dietary administration of this PSB during the whole fattening period was able to reduce significantly the seroprevalence in the TG, which may reflect a positive effect on the control of Salmonella at the end of the fattening period.

Imbalance of intestinal immune function in piglets infected by porcine circovirus type 2 during the fetal period.

Porcine circovirus type 2- (PCV2-) associated reproductive disorders and enteritis have commonly been observed on PCV2-contaminated pig farms in recent years. In order to investigate disorders of intestinal immunity in piglets infected by PCV2 during the fetal period, 9 PCV2b-infected piglets and 6 non-infected piglets at one day of age were selected and euthanised prior to suckling. Samples of mesenteric lymph nodes (MLNs) and duodena were collected to investigate factors related to intestinal immunity and to detect lymphocytic apoptosis. The results indicated that there were no significant changes in the levels of IL-2, IL-10 and transforming growth factor-β (TGF-β) in the PCV2b-infected piglets but IFN-γ levels were significantly lower (P < 0.01) and IL-4 levels were significantly higher (P < 0.05) in infected piglets than in the controls. Furthermore, lymphocytic apoptosis increased in PCV2b-infected piglets and CD4+ to CD8+ ratios were lower in these piglets than in the controls. These findings suggest vertical transmission of PCV2b to fetuses, leading to an imbalance of intestinal immune function in piglets.

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Organic acids (OA) are seen as an alternative to antibiotics to reduce the burden of enteropathogens. Two replicates of a field trial were carried out to assess the effect of the addition of protected sodium butyrate (PSB) to the feed (dose of 3 kg/T) along the fattening period on the dynamics of Salmonella spp. infection in pigs. In each trial, around 50 pigs were assigned to a treatment group (TG) and 50 kept as controls (CG). Pigs were serologically monitored monthly and on-farm fecal samples and fecal and mesenteric lymph nodes (MLN) samples at slaughter were collected. In the first replicate, pigs became probably infected with Salmonella before the addition of PSB to the feed, but despite of that an overall lower proportion of shedders along the end of fattening period and lower seroprevalence before slaughter (50% vs. 89.6%; P < 0.001) was observed in the TG. In the second replicate, few pigs shed Salmonella during the trial, which precluded the finding of significant differences between groups for both Salmonella infection and shedding, but the seroprevalence at slaughter in the CG was again significantly higher than in the TG (31.1% vs. 13.7%, respectively; P = 0.02) and it was related to a higher proportion of shedders and MLN-positive pigs. When results from both trials were analyzed together, a significant increasing risk of shedding in the CG was observed at 90 days of fattening and at slaughter, and an overall significant decreasing trend in OD% values and thus in seroprevalence was also observed when pigs approached to slaughter. In conclusion, the dietary administration of this PSB during the whole fattening period was able to reduce significantly the seroprevalence in the TG, which may reflect a positive effect on the control of Salmonella at the end of the fattening period.

Vertical Transmission of PCV2b to Fetuses in Sows Intramuscularly Infected with PCV2b.

In order to investigate whether PCV2b can be transmitted across the placenta in sows thereby causing vertical infection of fetuses, six sows in 75 day of pregnancy were either intramuscularly inoculated with a PCV2b culture supernatant (n = 4) or mock infected with cell culture supernatant (n = 2). At parturition, 3 newborn piglets from each litter were randomly selected and euthanatized prior to suckling. Samples of the mesenteric lymph nodes, spleens, kidneys, hearts and lungs were collected for PCR, histopathological and immunohistochemical assays. The results showed that the newborn piglets from PCV2b-infected sows had histopathological lesions in the tested tissues. Moreover, PCV2b antigen and DNA were detected as well. These findings therefore suggested that porcine circovirus type 2b can be transmitted across the placenta of sows, thereby leading to PCV2b vertical infection of the fetuses.

Characterization of New Zealand White Rabbit Gut-Associated Lymphoid Tissues and Use as Viral Oncology Animal Model.

Rabbits have served as a valuable animal model for the pathogenesis of various human diseases, including those related to agents that gain entry through the gastrointestinal tract such as human T cell leukemia virus type 1. However, limited information is available regarding the spatial distribution and phenotypic characterization of major rabbit leukocyte populations in mucosa-associated lymphoid tissues. Herein, we describe the spatial distribution and phenotypic characterization of leukocytes from gut-associated lymphoid tissues (GALT) from 12-week-old New Zealand White rabbits. Our data indicate that rabbits have similar distribution of leukocyte subsets as humans, both in the GALT inductive and effector sites and in mesenteric lymph nodes, spleen, and peripheral blood. GALT inductive sites, including appendix, cecal tonsil, Peyer's patches, and ileocecal plaque, had variable B cell/T cell ratios (ranging from 4.0 to 0.8) with a predominance of CD4 T cells within the T cell population in all four tissues. Intraepithelial and lamina propria compartments contained mostly T cells, with CD4 T cells predominating in the lamina propria compartment and CD8 T cells predominating in the intraepithelial compartment. Mesenteric lymph node, peripheral blood, and splenic samples contained approximately equal percentages of B cells and T cells, with a high proportion of CD4 T cells compared with CD8 T cells. Collectively, our data indicate that New Zealand White rabbits are comparable with humans throughout their GALT and support future studies that use the rabbit model to study human gut-associated disease or infectious agents that gain entry by the oral route.

The regulation mechanism of apoptosis by visfatin in the mesenteric lymph nodes of LPS-treated rats.

Visfatin is an adipocytokine displaying multiple functional properties, which plays a role in the regulation of cell apoptosis and inflammation by an as yet unidentified mechanism. The aim of the present study was to determine if visfatin is involved in apoptosis pathway induced by LPS in rat Mesenteric lymph nodes (MLNs). Experimental rats were divided into four groups and MLNs samples were collected from each group. The morphological changes of the MLNs were examined by histological imaging. CD68 and ENPP1 were detected with immunohistochemistry and Western Blot. Apoptosis was evaluated with TUNEL and Flow Cytometry, the mRNA levels of the apoptosis-related genes were detected by qRT-PCR, and the protein levels of the apoptotic-related factors were detected by western blot. The main results showed that visfatin could significantly increase the macrophages in MLNs and prevent cell apoptosis from LPS-induced mesenteric lymph nodes, activate apoptotic signaling pathways and regulate the mRNA levels of the apoptosis-related genes. Visfatin had a pro-apoptotic effect on normal MLNs, whereas it exerted an anti-apoptotic effect during LPS-induced cell apoptosis in rat MLNs. In short, visfatin plays a dual role in the apoptosis in rat MLNs, which is mediated by both the mitochondrial apoptotic pathway and the death-receptor apoptotic pathway.

Intrauterine Infection by Mycobacterium avium subsp. paratuberculosis in buffalo (Bubalus bubalis)

Intrauterine Infection by Mycobacterium avium subsp. paratuberculosis in buffalo (Bubalus bubalis) This study reports the detection of Mycobacterium avium subsp. paratuberculosis (Map) in the uterus and fetus of a buffalo (Bubalus bubalis). Samples of the buffalo cow’s ileum, mesenteric lymph node, uterus and placenta as well as various fetal organs were collected. Quantitative real-time polymerase chain reaction (qPCR), histopathology (hematoxylin-eosin (HE) staining) and Ziehl-Neelsen (ZN) staining were performed. Histopathological analysis revealed lesions consistent with paratuberculosis (PTB) in the mesenteric lymph node and small intestine, and ZN staining revealed the presence of acid-alcohol resistant bacilli (AARB). Histological lesions were not identified in fetal tissues. qPCR indicated that uterus, mesenteric lymph node and small intestine samples from the buffalo cow were positive for Map. In contrast, in the fetus, only samples from the digestive tract were positive for Map based on qPCR data. These results indicate that Map is present in various buffalo organs and tissues, including within the reproductive system. Therefore, the intrauterine transmission of Map may occur in buffaloes and could represent an important source of infection.

Scrapie diagnosis in a goat and four Santa Inês sheep from the same herd in Brazil

Scrapie is a fatal and progressive transmissible spongiform encephalopathy (TSE) of natural occurrence in sheep and goats. The suspicion of scrapie may be based on clinical signs; however, the detection of pathological features of the prionic protein (PrP) in target tissues is necessary to diagnose the disease. The presence of an abnormal protein form (PrPSc) in lymphoreticular and nervous tissues is an important characteristic in diagnosis. This paper reports a case of scrapie in a flock of 55 Suffolk crossbred sheep, 19 Santa Inês sheep and 21 goats in the Mato Grosso state, midwestern Brazil. The animals were euthanized after the confirmation of a scrapie case with clinical signs in a Suffolk sheep in the same farm. Samples of brainstem at the level of the obex and lymphoid issues like palatine tonsils, mesenteric lymph nodes, third eyelid fixed in formalin 10% were processed for histological examination. Histological examination with hematoxylin and eosin did not show any microscopic changes in samples. Immunohistochemistry (IHC) examination to detect anti-prion PrPSc was performed in lymphoid tissues. Scrapie diagnosis was confirmed based on IHC positive results for PrPSc in lymphoid tissues of a crossbreed goat and four Santa Inês sheep, without any clinical scrapie signs. IHC showed positive staining in at least three lymphoid germinal centers in goat mesenteric lymph node, palatine tonsil, and third eyelid samples. The mesenteric lymph node, and tonsil samples of all sheep showed positive immunostaining, and only one sheep showed positive staining in lymphoid follicles in the third eyelid. Scrapie diagnosis using IHC in fixed samples of lymphoreticular tissue is technically feasible to detect the disease in both goats and sheep, as a form of pre-clinical diagnosis. The results indicate that the herd was infected by a sheep coming from another herd where scrapie had been diagnosed before.

The Effect of Curcumin on an Animal Intestinal Ischemia/Reperfusion Model for Bacterial Translocation and Inflammatory Response

Ischemia/reperfusion (IR) injury of the intestine is a major problem in abdominal pathological condition and is associated with a high morbidity and mortality. The purpose of the study is to investigate the effects of curcumin on the bacterial translocation incidence and inflammatory response in rats submitted to bowel ischemia reperfusion injury. Thirty-two Wistar albino rats with a weight of 200 to 250 g were used in the study. They were randomly divided into 3 groups (n = 10 for each group): sham only operated group(group I); IR group (group II); and IR + curcumin treatment group (group III). Curcumin (curcumin from Curcuma longa) 20 mg/kg/day was given orally to the curcumin group. All animals were given 109 E. Coli by orogastric intubation 12 hours before sampling. Seventy-two hours after the first operation, mesenteric lymph node and blood samples were obtained and cultured. Blood samples of 2 mL were obtained for a polymerase chain reaction study. A piece of terminal ileum was also sampled for histopathologic examination. Mesenteric lymph node and blood cultures of all control animals were positive for microbiological growth, and polymerase chain reaction results were positive in seven of the eight rats. Histopathologically, edema, vasodilatation and inflammatory cell infiltration were found to be less in the other groups in comparison to the control group. Curcumin reduced bacterial translocation in blood, hepatocellular damage, and plasma cytokine levels. Curcumin reduced the incidence of bacterial translocation in intestinal I/R. rats. These results suggest that Curcumin would be clinically useful in the treatment of intestinal I/R injury.

Acute infection with bovine viral diarrhea virus of low or high virulence leads to depletion and redistribution of WC1+ γδ T cells in lymphoid tissues of beef calves

The objective of this study was to determine the abundance and distribution of γδ T lymphocytes in lymphoid tissue during acute infection with high (HV) or low virulence (LV) non-cytopathic bovine viral diarrhea virus (BVDV) in beef calves. This study was performed using tissue samples from a previous experiment in which thirty beef calves were randomly assigned to 1 of 3 groups: LV [n=10; animals inoculated intranasally (IN) with LV BVDV-1a (strain SD-1)], HV [n=10; animals inoculated IN with HV BVDV-2 (strain 1373)], and control (n=10; animals inoculated with cell culture medium). On day 5 post inoculation, animals were euthanized, and samples from spleen and mesenteric lymph nodes (MLN) were collected to assess the abundance of WC1+ γδ T cells. A higher proportion of calves challenged with BVDV showed signs of apoptosis and cytophagy in MLN and spleen samples compared to the control group. A significantly lower number of γδ T cells was observed in spleen and MLN from calves in HV and LV groups than in the control calves (P<0.05). In conclusion, acute infection with HV or LV BVDV resulted in depletion of WC1+ γδ T cells in mucosal and systemic lymphoid tissues at five days after challenge in beef calves. This reduction in γδ T cells in the studied lymphoid tissues could be also due to lymphocyte trafficking to other tissues.