Typical Pathological Lesions Research Articles

Utility of navigation system-guided submandibular gland core needle biopsy in the diagnosis of immunoglobulin G4-related sialadenitis

Pathological diagnosis is important for the definite diagnosis of immunoglobulin G4-related sialadenitis (IgG4-RS). Core needle biopsy (CNB) is a scarless technique; however the pathological heterogeneity of IgG4-RS (a particular feature of this disease) could be the potential cause of the inferior diagnostic capability of submandibular gland CNB (SMG-CNB) for IgG4-RS. The aim of this study was to explore technical improvements in SMG-CNB and improve its diagnostic power in IgG4-RS diagnosis. Eighteen patients clinically suspected for IgG4-RS were enrolled and underwent both SMG-CNB and SMG surgical biopsy. A navigation system (Brainlab) was employed during SMG-CNB to obtain representative samples and avoid blood vessel injury. Histopathological and immunopathological findings for the SMG-CNB samples were in good concordance with SMG surgical biopsy. There was no statistically significant difference between SMG-CNB and SMG surgical biopsy in IgG-positive cell count (132.4 ± 59.3 vs 132.2 ± 47.5, P = 0.99), IgG4-positive cell count (102.2 ± 39.7 vs 97.2 ± 27.6, P = 0.67), or IgG4-positive/IgG-positive cell count ratio (78.6% ± 0.1% vs 75.2% ± 0.1%, P = 0.29). A moderate or strong significant correlation was found between SMG-CNB and SMG surgical biopsy for these cell counts and ratio (all P < 0.01). The diagnostic consistency of SMG-CNB and SMG surgical biopsy was 100%. The Brainlab navigation system may assist in collecting representative SMG-CNB samples from typical pathological lesions. Tissues obtained from SMG-CNB are sufficient for the pathological diagnosis of IgG4-RS. Standardized SMG-CNB is expected to replace SMG surgical biopsy for IgG4-RS diagnosis.

Tau seeds from patients induce progressive supranuclear palsy pathology and symptoms in primates.

Progressive supranuclear palsy is a primary tauopathy affecting both neurons and glia and is responsible for both motor and cognitive symptoms. Recently, it has been suggested that progressive supranuclear palsy tauopathy may spread in the brain from cell to cell in a 'prion-like' manner. However, direct experimental evidence of this phenomenon, and its consequences on brain functions, is still lacking in primates. In this study, we first derived sarkosyl-insoluble tau fractions from post-mortem brains of patients with progressive supranuclear palsy. We also isolated the same fraction from age-matched control brains. Compared to control extracts, the in vitro characterization of progressive supranuclear palsy-tau fractions demonstrated a high seeding activity in P301S-tau expressing cells, displaying after incubation abnormally phosphorylated (AT8- and AT100-positivity), misfolded, filamentous (pentameric formyl thiophene acetic acid positive) and sarkosyl-insoluble tau. We bilaterally injected two male rhesus macaques in the supranigral area with this fraction of progressive supranuclear palsy-tau proteopathic seeds, and two other macaques with the control fraction. The quantitative analysis of kinematic features revealed that progressive supranuclear palsy-tau injected macaques exhibited symptoms suggestive of parkinsonism as early as 6 months after injection, remaining present until euthanasia at 18 months. An object retrieval task showed the progressive appearance of a significant dysexecutive syndrome in progressive supranuclear palsy-tau injected macaques compared to controls. We found AT8-positive staining and 4R-tau inclusions only in progressive supranuclear palsy-tau injected macaques. Characteristic pathological hallmarks of progressive supranuclear palsy, including globose and neurofibrillary tangles, tufted astrocytes and coiled bodies, were found close to the injection sites but also in connected brain regions that are known to be affected in progressive supranuclear palsy (striatum, pallidum, thalamus). Interestingly, while glial AT8-positive lesions were the most frequent near the injection site, we found mainly neuronal inclusions in the remote brain area, consistent with a neuronal transsynaptic spreading of the disease. Our results demonstrate that progressive supranuclear palsy patient-derived tau aggregates can induce motor and behavioural impairments in non-human primates related to the prion-like seeding and spreading of typical pathological progressive supranuclear palsy lesions. This pilot study paves the way for supporting progressive supranuclear palsy-tau injected macaque as a relevant animal model to accelerate drug development targeting this rare and fatal neurodegenerative disease.

Comparative efficacy evaluation of different CSF vaccines in pigs with CSF maternally derived antibodies

Classical swine fever (CSF) is a highly contagious and important swine disease in China. Sporadic outbreaks with mild clinical signs are still being reported despite massive vaccination with the CSF C-strain vaccine. One possible reason for vaccine failure could be interference from maternally derived antibodies (MDAs) during vaccination in the field. The aim of this study was to evaluate the efficacy of different CSF vaccines in the presence of MDAs and to assess the different vaccination schemes in the field. The results demonstrated that vaccination with a single dose of C-strain-PK vaccine protected pigs against severe clinical signs and significantly reduced viremia. The impact of MDAs was negligible. The interference was also mild during a prime and boost vaccination scheme using the C-strain-ST vaccine. In contrast, a significant influence of MDAs on the efficacy of the subunit E2 vaccine in a one-dose vaccination scheme was observed, with pigs showing severe clinical signs, CSF-associated death, typical pathological lesions and a high level of viremia after challenge, despite robust E2 antibody induction. A field vaccination and challenge study further confirmed the superior effectiveness of a single dose of C-strain-PK vaccine in the presence of MDAs in comparison to a routine prime and boost vaccination scheme applied in the field, with pigs having fever, chronic signs, significant viremia and shedding after challenge. Delaying the vaccination time from the age of 28 days to 45 days, when MDA was low, was beneficial for improving the clinical protection and immunity induced by vaccines. Altogether, the results presented here emphasize that a high-quality vaccine and a scientific design of the vaccination scheme based on serological surveillance are essential pillars to control and eliminate CSF in China.

Co-infection of Candidatus Piscichlamydia Trichopodus (Order Chlamydiales) and Henneguya sp. (Myxosporea, Myxobolidae) in Snakeskin Gourami Trichopodus pectoralis (Regan 1910).

The present study describes a simultaneous infection of a novel Chlamydia-like organism (CLO) with a Myxozoa parasite, Henneguya sp. in snakeskin gourami Trichopodus pectoralis in Thailand. A new CLO is proposed “Candidatus Piscichlamydia trichopodus” (CPT) based on 16S rRNA phylogenetic analysis. Systemic intracellular CPT infection was confirmed by histological examination, in situ hybridization, PCR assay, and sequencing of 16S rRNA. This novel pathogen belongs to the order Chlamydiales but differs in certain aspects from other species. The histopathological changes associated with CPT infection were different from the typical pathological lesions of epitheliocystis caused by previously known CLO. Unlike other CLO, CPT localized in the connective tissue rather than in the epithelial cells and formed smaller clumps of intracellular bacteria that stained dark blue with hematoxylin. On the other hand, typical myxospores of the genus Henneguya with tails were observed in the gill sections. Infection with Henneguya sp. resulted in extensive destruction of the gill filaments, most likely leading to respiratory distress. Due to the frequency of co-infections and the unavailability of culture methods for CLO and Henneguya sp., it was difficult to determine which pathogens were directly responsible for the associated mortality. However, co-infections may increase the negative impact on the host and the severity of the disease. Given the commercial importance of the snakeskin gourami and its significant aquaculture potential, the findings of this study are important for further studies on disease prevention.

Water-soluble coenzyme Q10 provides better protection than lipid-soluble coenzyme Q10 in a rat model of chronic tacrolimus nephropathy.

Background/AimsCoenzyme Q10 (CoQ10), is a promising antioxidant; however, low bioavailability owing to lipid-solubility is a limiting factor. We developed water-soluble CoQ10 (CoQ10-W) and compared its effects with conventional lipid-soluble CoQ10 (CoQ10-L) in an experimental model of chronic tacrolimus (Tac) nephropathy.MethodsCoQ10-W was developed from a glycyrrhizic-carnitine mixed layer CoQ10 micelle based on acyltransferases. Chronic nephropathy was induced in rats with 28-day Tac treatment; they were concomitantly treated with CoQ10-L or CoQ10-W. CoQ10 level in plasma and kidney were measured using liquid chromatography–mass spectrometry. CoQ10-W and CoQ10-L effects on Tac-induced nephropathy were assessed in terms of renal function, histopathology, oxidative stress, and apoptotic cell death. Their effects on cell viability and reactive oxygen species (ROS) production were assessed in cultured proximal tubular cells, human kidney 2 (HK-2) cells.ResultsThe plasma CoQ10 level was significantly higher in the CoQ10-W group than in the CoQ10-L group. Tac treatment caused renal dysfunction, typical pathologic lesions, and oxidative stress markers. Serum creatinine was restored in the Tac + CoQ10-L or CoQ10-W groups compared with that in the Tac group. CoQ10-W administration reduced oxidative stress and apoptosis markers. Mitochondrial ultrastructure assessment revealed that the addition of CoQ10-L or CoQ10-W with Tac increased mitochondrial size and number than Tac treatment alone. In vitro investigations revealed that both CoQ10-L and CoQ10-W improved cell viability and reduced ROS production in the Tac-induced HK-2 cell injury.ConclusionsCoQ10-W has a better therapeutic effect in Tac-induced renal injury than conventional CoQ10-L, possibly associated with improved CoQ10 bioavailability

P1572EFFECTIVENESS OF COENZYME Q10-MICELLE COMPARED WITH COENZYME Q10 ON TACROLIMUS-INDUCED RENAL INJURY

Abstract Background and Aims We and others have recently demonstrated that Coenzyme Q10 (CoQ10) has protective effects against diabetes mellitus and various types of renal injury. This study investigated whether CoQ10-micelle treatment would affords superior renoprotection compared with CoQ10 in the governing tacrolimus (Tacrolimus)-induced renal injury in the rats. Method Male adult Sprague-dawley Rats were treated daily with Tacrolimus (1.5mg/kg/day, subcutaneous), CoQ10 (20mg/kg/day, oral), and CoQ10-micelle (20 mg/kg/day, oral) for 4 weeks. The effects of CoQ10 orCoQ10-micelle on Tac-induced renal injury were assessed in terms of renal function, histopathology, oxidative stress and apoptotic cell death. Results After 4 weeks of Tacrolimus treatment to rats caused renal dysfunction, typical pathologic lesions, and oxidative stress marker. The serum creatinine was reduced by Tac co-treatment with CoQ10 or CoQ10-micelle groups compared with the Tac and VH group (0.31 ± 0.03 in the VH group vs. 0.43 ± 0.041 in the Tac group vs.0.37 ± 0.031 in the Tac+CoQ10 group 0.30 ± 0.02123 in the Tac+CoQ10-micellegroup; 1P&amp;lt;0.05 vs. VH. 2P&amp;lt;0.05 vs. TAC. . 3P&amp;lt;0.05 vs. TAC+C.) The administration of CoQ10-micelle improved renal immunoreactivity, which was accompanied by reductions in oxidative stress and apoptosis. Assessment of the mitochondrial ultrastructure by electron microscopy revealed that tacrolimus co-treatment with CoQ10-micelle increased the size and number of mitochondria more than co-treatment with CoQ10, compared with that induced by TAC treatment alone. Conclusion These findings suggest that both CoQ10 and CoQ10-micelle effectively attenuates Tac-induced renal injury, and CoQ10-micelle provides more benefits than that of CoQ10.

Identification and assessment of pathogenicity of a naturally reassorted infectious bursal disease virus from Henan, China

Infectious bursal disease virus (IBDV) is still a vital etiological agent in poultry farms. IBDV outbreaks occasionally occur due to the presence of very virulent, reassortment or variant strains. Vaccine immunization has played crucial roles in IBD control for decades. However, survival pressure of IBDV from the vaccine immunization also increases the reassortments of circulating viruses. In this study, an IBDV strain was isolated from several broiler farms in Henan Province, central part of China, and named IBDV HN strain. Based on the results of RT-PCR, sequencing and phylogenic analyses of VP1 and VP2 genes, the IBDV HN strain is a novel reassortment strain in the Henan region. Segment A of this strain appears to originate from the very virulent IBDV strain, while segment B comes from the other field reassortment strains. This may be the result of natural reassortant of virus circulating in the field. About 60% (6/10) of experimentally infected specific pathogen-free chickens died after 3 to 5 d post-infection with typical symptom and pathological lesions. The IBDV HN strain was prone to horizontal transmission, which poses a serious threat to the chicken industry. Further investigation on the prevalence, virulence, and evolution of HN strain IBDV will provide a foundation for the prevention and control of the disease in this region.

First report of dermatophilosis in wild European red squirrels (Sciurus vulgaris)

The carcases of two wild red squirrels (Sciurus vulgaris) from Anglesey, North Wales, which had severe skin lesions, were presented to the Animal and Plant Health Agency. One was found...

Virulence evaluation of classical swine fever virus subgenotype 2.1 and 2.2 isolates circulating in China

Classical swine fever (CSF) remains an important pig disease in China, where it usually presents with mild or atypical clinical manifestations, with large scale outbreaks rarely seen. This has led to speculation about the possible circulation of viral strains of low virulence. To investigate this possibility, five field isolates within the predominant genotype 2 (2.1b, 2.1c, 2.1 h and 2.2) were evaluated and compared by experimental infection of naturally farrowed but colostrum-deprived piglets. All infected piglets displayed clinical signs, including persistent high fever, depression, anorexia, dyspnea, conjunctivitis, constipation, and hesitant gait. Typical pathological lesions, including pulmonary edema, hemorrhagic or cellulosic exudation, and swelling and hemorrhage of lymph nodes, were observed. Viremia and Erns protein expression in the blood of all infected animals were detectable from 3 to 5 days post infection (DPI), their presence correlating with the onset of fever, clinical signs and leukopenia. E2 antibody did not develop in any of the field CSFV-infected piglets during the disease course, while Erns antibody was detectable in 4–56% of infected animals at various time points. Mortalities ranged from 20 to 80% within 21 DPI, progressing to 100% by 43 DPI. Based on clinical scores and fatalities within 21 DPI, 2 of the 5 field isolates were classified as of moderate virulence and 3 of high virulence; i.e., no field isolates of low virulence were identified. The study has provided data supporting the use of these isolates as challenge viruses to evaluate the efficacy of current CSF vaccines.

Intervention mechanisms of mesenchymal stem cells in juvenile mice model of acute lung injury induced by lipopolysaccharide

Objective To explore the changes in expression levels of nuclear factor (NF)-κB, tumor necrosis factor (TNF)-α in the lungs of juvenile mice with acute lung injury (ALI) induced by lipopolysaccharide (LPS). And observe the repair of lung damage after intervening with exogenous mesenchymal stem cells (MSCs). Methods Thirty male juvenile C57 mice were randomly divided into the control group, the ALI group, and the ALI+ MSCs group by the random number table method. Mice from each group were euthanized at 12 h and 48 h. The ALI model of juvenile mice was established by intraperitoneal injection of LPS 10 mg/kg. MSCs from mice bone marrow were isolated, cultured and amplified in vitro, and the MSCs (1×106/ml) 0.1 ml were given to mice via caudal vein. MSCs marker were identificated by flow cytometry. Pathomorphological changes of mice lung were observed under light microscope after Hematoxylin-Eosin staining. The protein expression changes of NF-κB, TNF-α were observed using immunohistochemistry. Results Compared with the control group, the protein expression levels of NF-κB, TNF-α were significantly higher at 12 h and 48 h in the lungs of the ALI group (P<0.05). While those in ALI + MSCs group were markedly lower at these time points than the ALI group[NF-κB: 12 h: (0.181±0.008) OD vs (0.203±0.008) OD, 48 h: (0.197±0.002) OD vs (0.210±0.005) OD; TNF-α: 12 h: (0.185±0.004) OD vs (0.201±0.011) OD, 48 h: (0.185±0.002) OD vs (0.215±0.009) OD] (P<0.05). Histopathological evalution showed that typical pathological inflammation lesions in the lung were observed in ALI group, including alveolar congestion, hemorrhage, edema, infiltration of neutrophils in the airspace or vessel wall, thickness of the alveolar wall; pathological changes were relieved obviously in ALI+ MSCs group. Conclusion The expression of NF-κB and TNF-α are increased in lung tissues in the juvenile mice model of ALI induced by LPS. MSCs can alleviate injury degree of ALI induced by LPS in mice, the mechanism of action may correlate with decreasing NF-κB and TNF-α content in lung tissue. Key words: Acute lung injury; Mesenchymal stem cells; Nuclear factor-κB; Tumor necrosis factor-α; Mice

Isolation and characterization of a subtype C avian metapneumovirus circulating in Muscovy ducks in China.

Subtype C avian metapneumovirus (aMPV-C), is an important pathogen that can cause egg-drop and acute respiratory diseases in poultry. To date, aMPV-C infection has not been documented in Muscovy ducks in China. Here, we isolated and characterized an aMPV-C, designated S-01, which has caused severe respiratory disease and noticeable egg drop in Muscovy duck flocks in south China since 2010. Electron microscopy showed that the isolate was an enveloped virus exhibiting multiple morphologies with a diameter of 20–500 nm. The S-01 strain was able to produce a typical cytopathic effect (CPE) on Vero cells and cause death in 10- to 11-day-old Muscovy duck embryos. In vivo infection of layer Muscovy ducks with the isolate resulted in typical clinical signs and pathological lesions similar to those seen in the original infected cases. We report the first complete genomic sequence of aMPV-C from Muscovy ducks. A phylogenetic analysis strongly suggested that the S-01 virus belongs to the aMPV-C family, sharing 92.3%-94.3% of nucleotide identity with that of aMPV-C, and was most closely related to the aMPV-C strains isolated from Muscovy ducks in France. The deduced eight main proteins (N, P, M, F, M2, SH, G and L) of the novel isolate shared higher identity with hMPV than with other aMPV (subtypes A, B and D). S-01 could bind a monoclonal antibody against the F protein of hMPV. Together, our results indicate that subtype-C aMPV has been circulating in Muscovy duck flocks in South China, and it is urgent for companies to develop new vaccines to control the spread of the virus in China.

S8-2 The role of hydrogen sulfide in diabetic nephropathy (DN)

Hydrogen sulfide (H 2 S), a third gasotransmitter, has been confirmed as playing a major role in diverse biological processes such as vascular relaxation, insulin secretion, inflammation and apoptosis. Cystathionine β -Synthase (CBS) and cystathionine γ -lyase (CSE) are major biosynthetic enzymes of H 2 S. Much interest has been focused on H 2 S as a potent vasorelaxative substance, since CSE knockout mice developed hypertension. In the normal kidney, both CBS and CSE are expressed in the proximal tubules, but not in the glomeruli or distal tubules. Administration of the H 2 S donor sodium hydrosulfide (NaHS) increased peritubular capillary (PTC) diameter and blood flow. In the models of DN, we reported that hyperglycemia reduces the expression of eNOS in the glomeruli, thereby reducing NO production and inducing typical pathological glomerular lesions of DN. However, the precise roles of H 2 S in DN have not been fully identified. In a spontaneous diabetic model CSE expression was markedly reduced, whereas CBS expression was unaffected. Progressive DN showed vasoconstriction and a loss of blood flow in PTC that was ameliorated by the NaHS treatment. The biological profile of CSE resembles that of eNOS. These findings suggest that both H 2 S and NO might share the role of regulation of tubulointerstitial microcirculation. H 2 S could represent a target of treatment for ischemic injury in DN.

Isolation and characterization of a subtype C avian metapneumovirus circulating in Muscovy ducks in China

Subtype C avian metapneumovirus (aMPV-C), is an important pathogen that can cause egg-drop and acute respiratory diseases in poultry. To date, aMPV-C infection has not been documented in Muscovy ducks in China. Here, we isolated and characterized an aMPV-C, designated S-01, which has caused severe respiratory disease and noticeable egg drop in Muscovy duck flocks in south China since 2010. Electron microscopy showed that the isolate was an enveloped virus exhibiting multiple morphologies with a diameter of 20–500 nm. The S-01 strain was able to produce a typical cytopathic effect (CPE) on Vero cells and cause death in 10- to 11-day-old Muscovy duck embryos. In vivo infection of layer Muscovy ducks with the isolate resulted in typical clinical signs and pathological lesions similar to those seen in the original infected cases. We report the first complete genomic sequence of aMPV-C from Muscovy ducks. A phylogenetic analysis strongly suggested that the S-01 virus belongs to the aMPV-C family, sharing 92.3%-94.3% of nucleotide identity with that of aMPV-C, and was most closely related to the aMPV-C strains isolated from Muscovy ducks in France. The deduced eight main proteins (N, P, M, F, M2, SH, G and L) of the novel isolate shared higher identity with hMPV than with other aMPV (subtypes A, B and D). S-01 could bind a monoclonal antibody against the F protein of hMPV. Together, our results indicate that subtype-C aMPV has been circulating in Muscovy duck flocks in South China, and it is urgent for companies to develop new vaccines to control the spread of the virus in China.

Ginseng Treatment Attenuates Chronic Cyclosporine Nephropathy via Reducing Oxidative Stress in an Experimental Mouse Model

Background: This study was performed to investigate whether ginseng extract has a protective effect in an experimental mouse model of chronic cyclosporine (CsA) nephropathy. Methods: Mice were treated with CsA (30 mg/kg/day, subcutaneously) with or without Korean red ginseng extract (KRG) (0.2, 0.4 g/kg/day, orally) on a 0.01% salt diet for 4 weeks. The effect of KRG on CsA-induced renal injury was evaluated by assessing renal function and pathology, mediators of inflammation, tubulointerstitial fibrosis and apoptotic cell death. Using an in vitro model, we also examined the effect of KRG on CsA-treated proximal tubular cells (HK-2). Oxidative stress was measured by assessing 8-hydroxy-2′-deoxyguanosine (8-OHdG) levels in 24-hour urine, tissue sections, and culture media. Results: Four weeks of CsA treatment caused renal dysfunction, typical pathologic lesions and apoptotic cell death. KRG treatment reduced serum creatinine and blood urea nitrogen and histopathology and increased creatinine clearance. Proinflammatory and profibrotic molecules such as induced nitric oxide synthase, cytokines, transforming growth factor (TGF)-β1 and TGF-β1-inducible gene h3 and apoptotic cell death, also decreased with KRG treatment. Consistent with these results, in vitro studies showed that addition of KRG protected against CsA-induced morphological changes, cytotoxicity, inflammation, and apoptotic cell death as demonstrated by annexin V binding. These changes were accompanied by decrease in the level of 8-OHdG in urine and culture supernatant after KRG treatment. Conclusion: The results of our in vivo and in vitro studies demonstrate that KRG has a protective effect in CsA-induced renal injury via reducing oxidative stress.

Measles Viral Infection in PD-1 Gene Knockout Mice

Purpose: Subacute sclerosing panencephalitis (SSPE) is a neurodegerative disease due to persistent measles virus infection. We investigated the role of programmed death-1 (PD-1) molecule which is related with chronic viral infection in developing SSPE in mouse. Methods: We adopt the , , and wild type BALB/c 3 week old mice to make an animal model of SSPE by injecting measles virus (SSPE strain) intraventricularly. Three months after infusion of virus, the mice were sacrificed and examined if the typical pathologic lesions had been progressed. The sera were collected from each group of mice and the serum level of IL-21 was measured with ELISA kit. Results: The necrotic lesions on white matter and gliosis were found in focal areas in wild type BALB/c. The extent of lesion was smaller in heterotype BALB/c. Scanty lesions were found in mice. The sera level of IL-21 was not elevated in all three groups. Conclusion: Our data suggest that the PD-1 molecule may play a role in persistent viral infection.

Detection of a putative hemolysin operon, hhdBA, of Haemophilus parasuis from pigs with Glässer disease

The aim of the current study was to investigate whether polymerase chain reaction amplification of 16S ribosomal (r)RNA and a putative hemolysin gene operon, hhdBA, can be used to monitor live pigs for the presence of Haemophilus parasuis and predict the virulence of the strains present. Nasal cavity swabs were taken from 30 live, healthy, 1- to 8-week-old pigs on a weekly cycle from a commercial Thai nursery pig herd. A total of 27 of these pigs (90%) tested positive for H. parasuis as early as week 1 of age. None of the H. parasuis-positive samples from healthy pigs was positive for the hhdBA genes. At the same pig nursery, swab samples from nasal cavity, tonsil, trachea, and lung, and exudate samples from pleural/peritoneal cavity were taken from 30 dead pigs displaying typical pathological lesions consistent with Glässer disease. Twenty-two of 140 samples (15.7%) taken from 30 diseased pigs yielded a positive result for H. parasuis. Samples from the exudate (27%) yielded the most positive results, followed by lung, tracheal swab, tonsil, and nasal swab, respectively. Out of 22 positive samples, 12 samples (54.5%) harbored hhdA and/or hhdB genes. Detection rates of hhdA were higher than hhdB. None of the H. parasuis-positive samples taken from nasal cavity of diseased pigs tested positive for hhdBA genes. More work is required to determine if the detection of hhdBA genes is useful for identifying the virulence potential of H. parasuis field isolates.

Immunopathology and cytokine responses in commercial broiler chickens with gangrenous dermatitis

Gangrenous dermatitis (GD) is an emerging disease of increasing economic importance in poultry resulting from infection by Clostridium septicum and Clostridium perfringens type A. Lack of a reproducible disease model has been a major obstacle in understanding the immunopathology of GD. To gain better understanding of host–pathogen interactions in GD infection, we evaluated various immune parameters in two groups of birds from a recent commercial outbreak of GD, the first showing typical disease signs and pathological lesions (GD-like birds) and the second lacking clinical signs (GD-free birds). Our results revealed that GD-like birds showed: reduced T-cell and B-cell mitogen-stimulated lymphoproliferation; higher levels of serum nitric oxide and α-1-acid glycoprotein; greater numbers of K55+, K1+, CD8+, and MHC class II+ intradermal lymphocytes, and increased K55+, K1+, CD8+, TCR1+, TCR2+, Bu1+, and MHC class II+ intestinal intraepithelial lymphocytes; and increased levels of mRNAs encoding proinflammatory cytokines and chemokines in skin compared with GD-free chickens. These results provide the first evidence of altered systemic and local (skin and intestine) immune responses in GD pathogenesis in chickens.

Immunohistochemistry and polymerase chain reaction for detection of Mycoplasma hyopneumoniae infection in piglets

Summary: Objective: Few studies exist that concentrate on the detection of M. hyopneumoniae by PCR in piglets. Most of these studies do not permit the differentiation of latent infections from infections with typical pathomorphological lesions. The aim of the study was to characterize the occurrence of M. hyopneumoniae infections in piglets by combining and evaluating the results of macroscopical, histological, molecularbiological and immunohistochemical examinations. The evaluation of the suitability of the immunohistochemistry (IHC) as an alternative or as a supplement to other tests was a further aim. The study-design does not allow any conclusions regarding the prevalence of early M. hyo - pneumoniae-infections in piglets. Material and methods: 96 clinically affected piglets from eight pre-selected herds with an increased risk for M. hyopneumoniae infection were chosen for necropsy. Macroscopical and histological examinations of the lungs were performed. BALF and lung tissue were taken for the detection of M. hyopneumoniae by nested PCR (nPCR) and IHC. Results: 16 of 33 nPCR-positive piglets were positive for antigen by IHC while 63 nPCR-negative piglets were also negative for antigen by IHC (kappa index = 0.55). Of the 17 piglets with a nPCR-positive but IHC-negative test result, 12 originated from one herd leading to the assumption that not all field strains were detectable with the monoclonal antibody utilized. Fisher’s exact test showed a significant association (p &lt; 0.0001) between detection of M. hyopneumoniae by nPCR in BALF or by IHC in lung tissue and the occurrence of typical pathological lesions. Conclusion: IHC with the monoclonal antibody used in this study is a suitable additional method but no alternative to the direct detection of M. hyopneumoniae by nPCR. Clinical relevance: In the case of the detection of M. hyopneumoniae in BALF by nPCR, typical pathological lesions can be expected with a higher probability than in nPCR-negative piglets.

Impact of natural sheep–goat transmission on detection and control of small ruminant lentivirus group C infections

Dissemination of small ruminant lentivirus (SRLV) infections in Norway is affected by the different control strategies used for maedi-visna virus (MVV) infections in sheep and caprine arthritis-encephalitis virus (CAEV) infections in goats. Here we investigated SRLV phylogenetic group variants in sheep. CAEV-like isolates, belonging to phylogenetic group C, were found among both seropositive sheep and goats in mixed flocks, in which sheep and goats are kept together. Intra-herd clustering confirmed that mixed flock animals were infected by the same virus variant, suggesting ongoing interspecies transmission. Few sheep flocks were found to be infected with the MVV-like phylogenetic group A. The apparent absence of SRLV group A type in goats is probably due to the MVV control programme and animal management practices. SRLV group C targets lungs and mammary glands in sheep, and induces typical SRLV pathological lesions. SRLV group C isolated from the sheep mammary glands suggested a productive infection and potential for transmission to offspring. SRLV group C was most prevalent among goats. A lower PCR sensitivity in seropositive sheep suggested a lower load of SRLV group C provirus in sheep than in goats. Higher genetic divergence of group C than in other SRLV groups and extensive heterogeneity among group C isolates in the matrix C-terminal region demonstrate the need for identifying conserved target regions when developing PCR protocols for SRLV detection. As sheep and goats may serve as reservoirs for all SRLV genogroup types, successful control programmes require inclusion of both species.

Iatrogenic Phospholipidosis Mimicking Fabry Disease

FABRY DISEASE is one of several well-characterized lipidoses with a renal phenotype. This X-linked disease is caused by inactivation of the lysosomal enzyme α-galactosidase A, which can result in intracellular accumulation of globotriaosylceramide within a variety of renal cells, including podocytes, glomerular endothelial cells, mesangial cells, tubular epithelium, interstitial foam cells, vascular myocytes, and vascular endothelium. 1 Sessa A. Meroni M. Battini G. et al. Renal pathological changes in Fabry disease. J Inherit Metab Dis. 2001; 24: 66-70 Crossref PubMed Scopus (64) Google Scholar In many cells, especially podocytes, prominent vacuolization of cell cytoplasm caused by the presence of intracellular lipid inclusions can be recognized at the light microscopic level. Ultrastructural analysis invariably shows intracellular osmiophilic structures, including myelin figures or zebra bodies that are highly characteristic of this disease. However, certain drugs, of which amiodarone and chloroquine are most notable, can cause cellular injury indistinguishable from classic Fabry disease. This needs to be kept in mind in the evaluation of typical pathological lesions.