Warthin-Starry Silver Stain Research Articles

Expression and clinical significance of IL-8 and Wnt2 in Helicobacter pylori-infected gastric cancer patients.

We assessed the expression levels of interleukin-8 (IL-8) and Wnt2 in Helicobacter pylori-infected gastric cancer (GC) patients, and investigated the association of these proteins with prognosis. Seventy-two GC patients, including 40 cases with H. pylori infection and 32 cases without H. pylori infection, were enrolled. Tumor and para-tumor normal tissues were collected. H. pylori infection was determined using the rapid urease test and Warthin-Starry silver staining. The expressions of IL-8 and Wnt2 were assessed using immunohistochemistry. The association between H. pylori infection, IL-8, Wnt2, and pathological and clinical data was analyzed. The relationship between H. pylori, IL-8, Wnt2, and prognosis was assessed using survival curves and regression analyses. The expression levels of IL-8 and Wnt2 in GC tissue significantly exceeded that in normal mucosa. The expression levels of IL-8 and Wnt2 were significantly higher in patients with H. pylori infection, than in those without H. pylori infection; and a positive correlation was observed between IL-8 and Wnt2 expressions. The positivity rate of IL-8 and Wnt2 in early-stage GC without lymph node metastasis markedly differed from that in advanced-stage GC with lymph node metastasis. Additionally, the survival rate of GC patients with positive IL-8 and Wnt2 expressions was significantly lower than in patients without such expression patterns. Clinical stage and Wnt2 were identified as independent prognostic factors of GC. IL-8 and Wnt2 were prognostic indicators for GC. H. pylori and IL-8 were not independent prognostic factors, whereas Wnt2 could independently serve as a prognostic factor for GC.

Leptospirosis in Wild Mice: An Ongoing Concern for the Azorean Islands

Background: Leptospirosis is a worldwide zoonotic infectious disease that affects Human and animals worldwide with increased risk in regions with temperate climates. Such is the case of the Azorean islands (a Portuguese archipelago with 9 islands) where this disease is considered endemic. Rodents of these islands have been considered the reservoir species for Leptospira and regulating measurements have been previously developed to control and prevent this disease in the archipelago. This study aims to determine a more recent infection rate of leptospirosis in the house-mouse (Mus musculus). Methodology & Findings: This retrospective study covering a period of 8 years (2011 to 2018), was performed by live capturing Mus musculus in 6 locations of S. Miguel Island (the most populated island of the archipelago). After euthanasia and necropsy of the mice, kidneys were surgically removed and processed in standard histological routine, and slides were stained with modified Warthin-Starry silver stain for spirochetes and scanned for the presence of Leptospires. It was possible to determine that from the 133 wild mice captured, 89 were positive for leptospirosis, representing a 66.91% infection rate. Conclusion: This study reveals that leptospirosis still represents an increased risk to Human lives and an uncalculated economical loss to the dairy farms of the Azorean islands.

IL-8 Links NF-κB and Wnt/β-Catenin Pathways in Persistent Inflammatory Response Induced by Chronic <i>Helicobacter pylori</i> Infection

Helicobacter pylori (H. pylori) infection can cause persistent inflammatory response in human gastric mucosal epithelial cells, which may result in the occurrence of cancer. However, the underlying mechanism of carcinogenesis has not been elucidated yet. Herein, we established the models of chronic H. pylori infection in GES-1 cells and C57BL/6J mice. Interleukin 8 (IL-8) level was detected by ELISA. The expression of NF-κB p65, IL-8, Wnt2 and β-catenin mRNA and proteins was evaluated by real-time PCR, Western blotting, immunofluorescence staining, and immunohistochemistry. The infection of H. pylori in mice was evaluated by rapid urease test, HE staining and Warthin‒Starry silver staining. The morphological changes of gastric mucosa were observed by electron microscopy. Our results showed that in H. pylori infected gastric mucosal cells along with activation of NF-κB signaling pathway and increase of IL-8 level, the expression of Wnt2 was also increased significantly, which preliminarily indicates that IL-8 can positively regulate the expression of Wnt2. Studies in chronic H. pylori infected C57BL/6J mice models showed that there was an increased incidence of premalignant lesions in the gastric mucosa tissue. Through comparing changes of gastric mucosal cell ultrastructure and analyzing the relationship between NF-κB signaling pathway and Wnt2 expression, we found that H. pylori infection activated NF-κB signal pathways, and the massive release of IL-8 was positively correlated with the high expression of Wnt2 protein. Subsequently, the activated Wnt/β-catenin signal pathways may be involved in the malignant transformation of gastric mucosal cells. Collectively, H. pylori chronic infection may continuously lead to persistent inflammatory response: activate NF-κB pathway, promote IL-8 release and thereby activate Wnt/β-catenin pathway. IL-8 probably plays an important role of a linker in coupling these two signal pathways.

An observational study to assess the Helicobacter pylori infection rates in patients with cholelithiasis

Background: Cholelithiasis and chronic cholecystitis cholelithiasis is fairly common all around the world. Since chronic inflammation causes repeated trauma to the gallbladder mucosa and DNA damage led by high or abnormal bile acid exposure, gallstones are closely related to the development of hepatobiliary cancers. After cholecystectomy, some patients experience ongoing upper abdomen pain, which is distressing for the surgeons and raises the possibility of a concomitant upper gastrointestinal issue, which in most cases turned out to be peptic ulcer disease. Numerous studies have found various Helicobacter species in the gallbladder tissue, gallstones, and bile taken from the gallbladder. We conducted this study to assess the relationship between cholelithiasis and H. pylori infection in the stomach. Ascending infection of H. pylori from the stomach and duodenum to biliary tree may have function in development of gall stone production and hepatobiliary carcinomas. Methods: Hospital based cross sectional observational study on patients admitted in surgical wards of SMS Hospital Jaipur for laparoscopic cholecystectomy. All patients underwent UGI and gastric antral biopsy followed by laparoscopic cholecystectomy, gastric mucosa and GB mucosa were subjected to Warthin starry silver stain for detection of H. pylori. Results: Out of 116 patients who underwent upper GI endoscopy followed by laparoscopic cholecystectomy, 29 (25%) patients were positive for H. pylori infection. Conclusions: This study shows significant positive co-relation between H. pylori infection in stomach, gall bladder to chronic cholecystitis and cholelithiasis, which signifies gastric colonisation of H. pylori has it’s role in biliary pathologies through ascending infection, the effectiveness of H. pylori eradication therapy in preventing these pathologies need to be studied further.

Immunohistochemical detection of Treponema pallidum in skin samples with clinical and histopathological correlations and Warthin-Starry staining critical analysis

BackgroundSyphilis in its different phases may be a difficult diagnosis in clinical and histopathological grounds. ObjectivesThe present study objectives were to evaluate the detection and tissue distribution of Treponema pallidum in skin lesions of syphilis. MethodsA blinded diagnostic accuracy study was performed with immunohistochemistry and Warthin-Starry silver staining in skin samples from patients with syphilis and other diseases. Patients attended two tertiary hospitals between 2000 and 2019. Prevalence ratios (PR) and 95% confidence intervals (95% CI) were calculated for the association between immunohistochemistry positivity and clinical-histopathological variables. ResultsThirty-eight patients with syphilis and their 40 biopsy specimens were included in the study. Thirty-six skin samples were used as non-syphilis controls. The Warthin-Starry technique was unable to accurately demonstrate bacteria in all samples. Immunohistochemistry showed spirochetes only in skin samples from patients with syphilis (24/40) with 60% sensitivity (95% CI 44.8‒75.2). Specificity was 100% and accuracy, 78.9% (95% CI 69.8‒88.1). Most cases had spirochetes in both dermis and epidermis and there was a high bacterial load. Study limitationsCorrelation between immunohistochemistry and clinical or histopathological characteristics was observed but was limited statistically due to the small sample size. ConclusionsSpirochetes were promptly seen in an immunohistochemistry protocol, which can contribute to the diagnosis of syphilis in skin biopsy samples. On the other hand, the Warthin-Starry technique showed to be of no practical value.

IL-8 Links NF-κB and Wnt/β-Catenin Pathways in Persistent Inflammatory Response Induced by Chronic Helicobacter pylori Infection

Helicobacter pylori (H. pylori) infection can cause persistent inflammatory response in human gastric mucosal epithelial cells, which may result in the occurrence of cancer. However, the underlying mechanism of carcinogenesis has not been elucidated yet. Herein, we established the models of chronic H. pylori infection in GES-1 cells and C57BL/6J mice. Interleukin 8 (IL-8) level was detected by ELISA. The expression of NF-κB p65, IL-8, Wnt2 and β-catenin mRNA and proteins was evaluated by real-time PCR, Western blotting, immunofluorescence staining, and immunohistochemistry. The infection of H. pylori in mice was evaluated by rapid urease test, H&E staining and Warthin-Starry silver staining. The morphological changes of gastric mucosa were observed by electron microscopy. Our results showed that in H. pylori infected gastric mucosal cells along with activation of NF-κB signaling pathway and increase of IL-8 level, the expression of Wnt2 was also increased significantly, which preliminarily indicates that IL-8 can positively regulate the expression of Wnt2. Studies in chronic H. pylori infected C57BL/6J mice models showed that there was an increased incidence of premalignant lesions in the gastric mucosa tissue. Through comparing changes of gastric mucosal cell ultrastructure and analyzing the relationship between NF-κB signaling pathway and Wnt2 expression, we found that H. pylori infection activated NF-κB signal pathways, and the massive release of IL-8 was positively correlated with the high expression of Wnt2 protein. Subsequently, the activated Wnt/β-catenin signal pathways may be involved in the malignant transformation of gastric mucosal cells. Collectively, H. pylori chronic infection may continuously lead to persistent inflammatory response: activate NF-κB pathway, promote IL-8 release and thereby activate Wnt/β-catenin pathway. IL-8 probably plays an important role of a linker in coupling these two signal pathways.

Coptis, Pinellia, and Scutellaria as a promising new drug combination for treatment of Helicobacter pylori infection.

Helicobacter pylori (H. pylori) is the most important infectious agent and plays an important role in the progression of chronic gastritis and the development of gastric cancer. To identify efficient therapeutic agents or strategies that can treat H. pylori infection. We performed literature analysis, experimental validation, and network pharmacology. First, traditional Chinese medicine (TCM) prescriptions for the treatment of H. pylori infection were obtained from the China National Knowledge Infrastructure, China Biology Medicine, China Science and Technology Journal Database, and WanFang databases. In addition, we conducted a relevant search by Reference Citation Analysis (RCA) (https://www.referencecitationanalysis.com). Next, we used TCM Inheritance Support System V2.5 to identify core drug combinations in the TCM prescriptions. Then, an H. pylori-associated chronic mouse model of gastritis was established. The antibacterial properties and anti-inflammatory potential of the core drug combination were evaluated by the rapid urease test, modified Warthin-Starry silver staining, histopathological analysis, and enzyme linked immunosorbent assay. Finally, the active compounds, hub targets, and potential signaling pathways associated with the core drug combination were analyzed by network pharmacology. The TCM treatment of H. pylori was mainly based on reinforcing the healthy Qi and eliminating pathogenic factors by simultaneously applying pungent dispersing, bitter descending, cold and warm drugs. The combination of Coptis, Pinellia, and Scutellaria (CPS) was identified as the core drug combination from 207 prescriptions and 168 herbs. This drug combination eradicated H. pylori, alleviated the gastric pathology induced by H. pylori infection, and reduced the expression levels of tumor necrosis factor-α (P = 0.024) and interleukin-1β (P = 0.001). Moreover, a total of 35 compounds and 2807 targets of CPS were identified using online databases. Nine key compounds (tenaxin I, neobaicalein, norwogonin, skullcapflavone II, baicalein, 5,8,2'-trihydroxy-7-methoxyflavone, acacetin, panicolin, and wogonin) and nine hub target proteins (EGFR, PTGS2, STAT3, MAPK3, MAPK8, HSP90AA1, MAPK1, MMP9, and MTOR) were further explored. Seventy-seven signaling pathways were correlated with H. pylori-induced inflammation and carcinogenesis. In summary, we showed that CPS is the core drug combination for treating H. pylori infection. Animal experiments demonstrated that CPS has bacteriostatic properties and can reduce the release of inflammatory cytokines in the gastric mucosa. Network pharmacology predictions further revealed that CPS showed complex chemical compositions with multi-target and multi-pathway regulatory mechanisms. Although the results derived from network pharmacology are not necessarily comprehensive, they still expand our understanding of CPS for treating H. pylori infection.

Use of digital pathology and artificial intelligence for the diagnosis of Helicobacter pylori in gastric biopsies.

A common source of concern about digital pathology (DP) is that limited resolution could be a reason for an increased risk of malpractice. A frequent question being raised about this technology is whether it can be used to reliably detect Helicobacter pylori (HP) in gastric biopsies, which can be a significant burden in routine work. The main goal of this work is to show that a reliable diagnosis of HP infection can be made by DP even at low magnification. The secondary goal is to demonstrate that artificial intelligence (AI) algorithms can diagnose HP infections on virtual slides with sufficient accuracy. The method we propose is based on the Warthin-Starry (W-S) silver stain which allows faster detection of HP in virtual slides. A software tool, based on regular expressions, performed a specific search to select 679 biopsies on which a W-S stain was done. From this dataset 185 virtual slides were selected to be assessed by WSI and compared with microscopy slide readings. To determine whether HP infections could be accurately diagnosed with machine learning. AI was used as a service (AIaaS) on a neural network-based web platform trained with 468 images. A test dataset of 210 images was used to assess the classifier performance. In 185 gastric biopsies read with DP we recorded only 4 false positives and 4 false negatives with an overall agreement of 95.6%. Compared with microscopy, defined as the "gold standard" for the diagnosis of HP infections, WSI had a sensitivity and specificity of 0.95 and 0.96, respectively. The ROC curve of our AI classifier generated on a testing dataset of 210 images had an AUC of 0.938. This study demonstrates that DP and AI can be used to reliably identify HP at 20X resolution.

Diagnostic Performance of glmM Gene and Histological Stains for Detection of <i>Helicobacter pylori</i> in Gastric Biopsy from Patients Admitted to Wad Madani Teaching Hospital, Sudan

Helicobacter pylori is the microbial agent most responsible for gastro-duodenal ulcer and chronic gastritis, which can develop into carcinoma of the stomach. This study was performed in Wad Medani Teaching Hospital, Sudan to detect Helicobacter pylori in stomach samples, and evaluate the performance of the tests used, which were histological stains and PCR. Gastric biopsies were obtained from 105 referred patients during endoscopy, and fixed specimens examined by haematoxylin-eosin and Warthin-Starry silver stains, while DNA was extracted for glmM gene amplification. Epigastric pain was the most common symptom at 78% (82/105) and chronic gastritis recorded with 71% (68/105) of endoscopy results. Warthin-Starry silver stain gave 31% (33/105) as positive for Helicobacter pylori followed by glmM gene 27% (28/105) and haematoxylin-eosin 24% (25/105). The study indicated good performance of histological staining and high specificity of glmM gene in detection of Helicobacter pylori from gastric biopsies.

In Vivo Biofilm Formation of Pathogenic Leptospira spp. in the Vitreous Humor of Horses with Recurrent Uveitis.

Equine recurrent uveitis (ERU) causes painful inflammatory attacks and oftentimes blindness in the affected eyes. The disease is considered a late sequela of systemic leptospirosis. The most effective therapy is the surgical removal of the vitreous (vitrectomy), which is not only therapeutic, but provides vitreous material that can be assessed diagnostically. For example, the lipL32 gene, culturable Leptospira spp., and anti-Leptospira antibodies have all been detected in vitreous samples obtained from eyes with chronic ERU. Despite this clear evidence of leptospiral involvement, the systemic administration of antibiotics in infected horses is ineffective at resolving ERU. This syndrome of chronic recurrent inflammation, which is unresponsive to antibiotic therapy, combined with apparent bacteria evading the immune response, is consistent with a biofilm-associated infection. The purpose of this study, therefore, was to detect the in vivo biofilm formation of Leptospira spp. in vitreous samples collected during vitrectomy and examined using a Warthin-Starry silver stain and immunohistochemistry. All known steps of biofilm formation were visualized in these samples, including individual Leptospira spp., leptospiral microcolonies and dense roundish accumulations of Leptospira spp. In many instances spirochetes were surrounded by an extracellular substance. Taken together, data from the present study show that ERU is a biofilm-associated intraocular leptospiral infection, which best explains the typical clinical course.

Hepatosplenic Cat Scratch Disease: Description of Two Cases Undergoing Contrast-Enhanced Ultrasound for Diagnosis and Follow-Up and Systematic Literature Review.

Cat scratch disease (CSD) is a disease usually characterized by self-limited lymphadenopathy of the young man. Rarely CSD, however, can manifest itself as an unusual hepatosplenic form (HS-CSD) in immunocompetent patients. HS-CSD diagnosis is generally based on clinical features, imaging, and serologies, but sensitivity of serologies is very variable, like that of other diagnostic methods, as Warthin-Starry silver stain and isthology. Also there are no specific markers for the follow-up. The use of the CEUS (abdominal contrast-enhanced ultrasound) in HS-CSD is not previously described in literature examined, but we think that CEUS can be of help to diagnosis and follow-up of these patients, even after an initial CT scan, because it is a sensitive method, as seen in other diseases associated with granulomas, such as sarcoidosis. We describe 2 new cases of HS-CSD, and we performed a systematic review of the clinical cases reported in the past 10 years in the literature associated to an analysis of clinical, diagnostic, and therapeutic aspects of the disease.

Dissemination kinetics and pathology of canine Leptospira icterohaemorrhagiae isolate in a guinea pig infection model

ABSTRACT Despite significant progress made in elucidating the mechanism of acute human leptospirosis in different organs, there is a paucity of information in organs such as the heart, pancreas, brain, and adrenal gland. This study was designed to establish leptospire dissemination kinetics and patho-morphological changes associated with these orangs in the guinea pig infection model using cultural isolation (CI), polymerase chain reaction (PCR), Warthin Starry silver stain (WSss), immunohistochemistry (IH), and transmission electron microscopy (TEM). Twenty guinea pigs were inoculated intra-peritoneally with a low dosage of 1 × 107 Leptospira interrogans serovar Icterohaemorrhagiae and 10 as control using distilled water. The guinea pigs were sacrificed at post-infection day (p.i.d.) ½, 1, 3, 5, and 7 followed by the harvest of the brain, pancreas, adrenal gland, heart, lungs, liver, kidneys, and spleen for CI, PCR, HE, WSss, IH, and TEM evaluations. The study revealed early dissemination of Leptospira organism in the brain, heart, pancreas, and adrenal gland and exerted various histopathological changes that were not explicitly elucidated in previous studies. This study revealed that the virulent pathogenic isolate of Leptospira organism obtained from clinically infected dog mimicked the same clinical manifestations, gross and histopathological changes especially in organs that were not previously evaluated.

S1749 Human Intestinal Spirochetosis: An Atypical Cause of Symptomatic Colonic Ulceration

INTRODUCTION: This case demonstrates an atypical presentation of intestinal spirochetosis in a patient with HIV on anti-retroviral therapy with adequate viral suppression. CASE DESCRIPTION/METHODS: A 56-year-old male presented with one day of hematochezia and abdominal cramping. He had a past medical history of HIV on Highly Active Antiretroviral Therapy with recent undetectable viral load and CD4 count of 1225. On initial assessment, he was tachycardic with frank blood on rectal examination. Hemoglobin was 11.8 with a baseline of 14.7. Computed Tomography scan of the abdomen and pelvis was significant for contrast extravasation in the cecum consistent with active gastrointestinal bleed. His hemoglobin further downtrended to 9.4, but he remained hemodynamically stable. Colonoscopy revealed a solitary twenty mm ulcer in the cecum without active bleeding. Biopsy obtained from the ulcer edge demonstrated intestinal spirochetosis (Warthin-Starry stain positive) and focal acute colitis with lamina propria hemorrhage. Patient remained clinically stable without further episodes of hematochezia and was discharged before biopsies resulted. Unfortunately, he was lost to follow-up. DISCUSSION: Human intestinal spirochetosis is rare in developed countries but when present, they are often found in homosexual males and those with HIV. Stool colonization rates in homosexual men in the US have ranged between 20.6 and 62.5%. It is most commonly caused by Brachyspira aalborgi or Brachyspira pilosicoli. Patients are often asymptomatic but can present with abdominal pain and watery diarrhea. Endoscopic findings are usually non-specific since mucosa may be normal, polypoid, or mildly erythematous. There are only case reports of colonic ulceration. Diagnosis is made by histology with hematoxylin-eosin and Warthin-Starry silver stains which demonstrate spirochetal adhesion along the border of the intercryptal epithelial layer. Treatment is reserved for immunocompromised and symptomatic patients and consists of metronidazole for 10 days with resolution of symptoms. Complete mucosal healing has been observed for those who opt for repeat endoscopy. Our patient is unique since he had sudden large-volume hematochezia without diarrhea and a rather discrete endoscopic finding despite viral suppression and a robust CD4 count. Therefore, clinical suspicion for intestinal spirochetosis should be moderately high in patients with these specific risk factors.Figure 1.: Clean-based cecal ulcer.Figure 2.: Warthin-Starry silver stain of cecal ulcer edge biopsy highlighting spirochetes on the surface epithelium.

Analysis of microbial changes in the tonsillar formalin-fixed paraffin-embedded tissue of Chinese patients with IgA nephropathy

BackgroundImmunoglobulin A nephropathy (IgAN) is a prevalent chronic glomerular disease contribution to end-stage renal failure (ESRD). The tonsillar microbiota is closely associated with IgAN diseases based on the mucosal immune response. However, the composition and function of in tonsillar microbiota in participant patients with IgAN remains unknown. In this study, we detected the tonsillar microbiota changes of IgAN patients in Heilongjiang province located in northeast China. Material and methodsWe collected from 21 patients with IgAN and 16 patients with chronic tonsillitis (CT) who had undergone tonsillectomy previously. Histological review of all samples from formalin-fixed paraffin-embedded (FFPE) tissue were performed. Extracted DNA from FFPE tissue blocks, after that V4 regions of 16S ribosomal RNA (rRNA) sequencing and comparative analyses of tonsillar flora between two groups were performed. The statistical analysis used the SPSS version of 21. ResultsVisualization of microorganisms by Gram and Warthin-Starry (WS) silver stains, preliminarily observed the morphological characteristics of microbiome in FFPE tissue cases, such as bacteria or fungi. Tonsillar FFPE samples from the IgAN patients and CT controls showed significant differences in tonsillar microbial certain compositions and functions. We found that there were eight dominant genera that can be available to distinguish IgAN patients from CT controls. Compared with CT controls, at genus level, the relative abundances of Methylocaldum and unclassified_f_Prevotellaceae were significantly higher, while the abundances of Anaerosphaera, Halomonas, Trichococcus, Peptostreptococcus, norank_f_Synergistaceae and unclassified_k_norank_d_Bacteria were significantly lower in IgAN patients. Principal co-ordinates analysis (PCOA) distinguished IgAN patients from CT controls, and receiver operating characteristic (ROC) curves analysis confirmed that the diagnosis of disease has certain diagnostic significance. In addition, Functional analysis revealed that partly Enzymes and KOs were increased in the IgAN patients. ConclusionsHistological screening results were very helpful for further gene sequencing, not only to supplement the observation of bacterial morphology and structure, but also to prepare for subsequent gene sequencing and bioinformatics analysis. We elucidated subtle relevance between changes in tonsillar microbiota and IgAN patients, which can be utilized to predict the incidence of IgAN disease. In addition, we predicted that some enzymes, and KOs were closely related to IgAN.

Combined Application of Immunohistochemistry and Warthin-Starry Silver Stain on the Pathologic Diagnosis of Cat Scratch Disease.

Cat scratch disease (CSD) is an infectious disorder caused primarily by the bacterium Bartonella henselae (B. henselae). Immunohistochemistry (IHC) and Warthin-Starry silver stain (WS) are considered to be indispensable to diagnose CSD in combination with morphologic characteristics. In this study, we retrieved and reviewed 46 cases of paraffin-embedded lymphadenitis with histologic and/or clinical suspicion of CSD between 2014 and 2018, and detected B. henselae by IHC and WS, respectively, and evaluated the application significance of IHC and WS for the detection of B. henselae and validated their values in the pathologic diagnosis of CSD. B. henselae was detected by IHC and WS; validation of 2 methods for detecting B. henselae was evaluated by sensitivity, specificity, false-positive rate, false-negative rate, precision, negative predictive value, and agreement rate. Microscopically, suppurative granulomas and/or multiple stellate microabscesses were observed in the accessory cortex of lymph nodes, especially near the subcapsule. Our results showed that 80.4% (37/46) of cases were positive for B. henselae by IHC, manifesting mainly punctuate, granular, or linear to outline the shape of bacteria. However, the positive rate of B. henselae by the WS method was 52.2% (24/46). There was a significant difference between IHC and WS (P=0.023). Moreover, a positive percentage of B. henselae was 97.8% (45/46), which was detected by the combined application of IHC and WS. The combination of IHC and WS exhibited high sensitivity (97.8%) and good agreement rate (86.5%). The combined application of the IHC and WS method may have important clinical advantages, which is with the highest sensitivity and agreement rate for pathologic diagnosis of CSD.

Histochemical and immunohistochemical detection of pathogenic leptospires serovars in tissues of some captive wildlife from a University zoo in Nigeria

ABSTRACTThe detection and documentation of pathogenic Leptospira serovars in wild captive and zoological garden animals are scarce in literature from Nigeria. The knowledge of the prevalence of prevalence of pathogenic Leptospira serovars in these animals as a zoonotic risk to workers, zoo visitors and the general public is essential. This investigation was carried out on archival kidney and liver samples of captive and Zoological Garden animals (66) of an institutional facility, submitted for necropsy to the Department of Veterinary Pathology between the periods of 2010–2015. The gross diagnosis reports were obtained from the necropsy records, detection of pathogenic Leptospira serovars was by Warthin Starry silver staining and immunohistochemistry techniques using standard methods. Six samples out of the sixty-six samples were positive for leptospira four samples were positive by silver stain method, while two samples were positive by immunohistochemistry. In this study, serovar Pomona and grippotyphosa were detected in the foxes while serovar Pomona was detected in the horse. This study has revealed the presence of pathogenic leptospires in some captive wild and zoological garden animals.

Helicobacter pylori associated gastritis: A histopathological study of gastric mucosal biopsies

Background: Helicobacter pylori (H. pylori) is one of the major cause of gastrointestinal diseases. It is classified as class-I carcinogen & thus Its important to study the histological changes in H. pylori gastritis. Aim: To evaluate associated histological features and prevalence of H. pylori associated gastritis by histopathological method. Methods and Material: Biopsies from 75 patients with various upper gastrointestinal symptoms were studied. Study period was from August 2016 to August 2018. Each biopsy was stained with Haematoxylin & Eosin (H&E) & Warthin Starry silver stain. The slides were examined for H. pylori & various histological features. Results: H. pylori infection was high in the fourth decade with male predominance. 47/75 (62.6%) cases were H. pylori positive. Among H. pylori positive cases neutrophilia activity, mononuclear infiltrate, atrophy & intestinal metaplasia was seen in 93.6%, 100%, 14.9% & 2.2% cases respectively. Conclusion: Histopathological evaluation is the gold standard for diagnosing H. pylori infection. Furthermore, large scale studies are required to evaluate H. pylori associated gastritis to prevent morbidity & mortality

Case Report / Olgu Sunumu: Pyogranulomatous myocarditis and diaphragmatic myositis associated with Bartonella henselae in a cat from Turkey

A case of pyogranulomatous myocarditis and diaphragmatic myositis caused by Bartonella henselae in an 18 months old tricolor female cat in Turkey was reported in this study. The cat had flea infestation before clinical disease onset. The cat showed clinical findings including lethargy, anorexia, dyspnea, hypersalivation, and polydipsia. At necropsy, the myocardium contained numerous 1 to 2 mm, discrete to coalescing, white-grey nodular foci with approximately 250 ml of serosanguineous fluid in the thorax. By Warthin-Starry silver staining, argyrophilic pleomorphic coccobacilli were seen within the myocardial and diaphragmatic pyogranulomatous lesions. B. henselae DNA was detected in heart, kidney and liver tissues by targeting the riboflavin synthase C ( rib C) gene by PCR. Using immunofluorescence technique, B. henselae was also visualized in myocardial tissues. This report provides additional evidence in support of an etiological association between pyogranulomatous myocarditis and diaphragmatic myositis and B. henselae infection in cats.

Intestinal spirochetosis: An unusual cause of postantibiotic diarrhea.

(A) A blue fringe along the surface epithelium of the colonic mucosa on hematoxylin-eosin staining, a characteristic histology of intestinal spirochetosis. (B) Numerous spirochetes highlighted on Warthin-Starry silver staining.

Helicobacter spp. in domestic cats: identification and relationship with anatomical and histopathological gastric changes in animals of blood group A

ABSTRACT: The aim of this study was to evaluate the presence of gastric Helicobacter-like organisms and the endoscopic and histopathological changes in domestic cats with blood type A. Samples from the stomach antrum, body and fundus were collected from 32 mixed-breed stray domestic cats using gastroscopy. Urease testing and cytological analysis were performed in fresh samples. Tissue sections were processed and stained with hematoxylin and eosin (H&E) and the Warthin-Starry (WS) silver staining methods for histopathological examination. Helicobacter spp. were detected in 100% of samples subjected to silver staining and cytological analysis, and in 96.9% of samples subjected to urease testing. In 87.5% of the cats, mononuclear inflammatory-cell infiltrates were identified. The graduation and distribution of inflammatory infiltrates in these cats revealed mild (78.1%) to moderate (9.4%) inflammatory changes in at least one gastric region. These changes were independent of the colonization score. Hyperplasia of the lymphoid follicles was detected in three cats. Cats of blood group A are often colonized by Helicobacter spp. and the macroscopic and microscopic findings are consistent with studies in domestic cats reported to date, concluding that the most common blood group in cats is not associated with high susceptibility to symptomatic gastritis.