Paired analysis of the microbiota of surface mucus and whole-tissue specimens in patients with chronic rhinosinusitis.

BACKGROUND: The role of bacteria in the pathogenesis of chronic rhinosinusitis (CRS) remains uncertain. Recent evidence suggests that bacteria are able to establish microcolonies within the underlying mucosa. However, to date there has been no systematic comparison of bacterial community composition and diversity in the surface mucosa with that of the underlying tissue. METHODS: Paired swabs and whole-tissue samples were collected from the middle meatus of 9 patients with CRS undergoing endoscopic sinus surgery. The bacterial composition and diversity of the samples were determined using 16S rRNA gene amplicon pyrosequencing. RESULTS: The bacterial communities of both swabs and tissues were dominated by known residents of the sinonasal cavity such as Staphylococcus, Corynebacterium, Prevotella, and Peptoniphilus. Although bacterial diversity (richness) did not differ between the 2 groups of samples, there were significant differences in the composition of bacterial communities. Molecular analyses revealed a large amount of interpersonal variation between patients. CONCLUSION: Swab and tissue samples revealed similar bacterial diversity to each other and to that of other microbiota studies reported in the CRS literature. However, bacterial composition was significantly different between the 2 sample types, even though the tissue biopsies also comprise bacteria from the surface. We speculate that the bacteria on the surface seed the underlying tissue via the damaged epithelium in CRS patients, which over time develops into a distinct bacterial community.

Chronic rhinosinusitis and cystic fibrosis: the interaction between sinus bacteria and mucosal immunity.

BACKGROUND: Chronic rhinosinusitis (CRS) is highly prevalent in cystic fibrosis (CF) patients, in whom a close correlation exists between the microbiology of the upper and lower respiratory tracts. We have reported intramucosal bacterial microcolonies in the sinus mucosa from idiopathic CRS patients and have made observations suggesting that these may result from mucosal immunotolerance secondary to altered macrophage function. In this study, we sought to determine whether intramucosal microcolonies exist in the mucosa of CF patients with CRS, and to investigate the associated mucosal immunology. METHODS: Mucus swabs and tissue biopsies were taken from 9 patients with CF undergoing functional endoscopic sinus surgery (FESS) for CRS, 11 with idiopathic CRS undergoing FESS, and 9 with normal sinuses having transnasal pituitary surgery. Microbiology samples were taken for culture and intramucosal microcolonies were sought using Gram staining. Mucosal immune cells were identified using fluorescent immunohistochemistry. RESULTS: Positive culture rates were similar between CRS patients and controls, but there were significantly more intramucosal microcolonies in the CRS groups (8/9 CF-CRS, 7/11 CRS), compared to controls (1/9). Furthermore, the biodensity of intramucosal microcolonies was significantly higher in CF-CRS than idiopathic CRS. Mirroring the microbiological observations, the number of CD163+ macrophages was significantly increased in CF-CRS compared to idiopathic CRS (p = 0.03). CONCLUSION: Intramucosal bacteria exist within the sinus mucosa of patients with CF, and in significantly greater numbers than in idiopathic CRS patients. We speculate that intramucosal microcolonies may also exist in the lower respiratory tract mucosa in CF and play a role in disease recalcitrance.

The interaction between bacteria and mucosal immunity in chronic rhinosinusitis: a prospective cross-sectional analysis.

BACKGROUND: We have detected intramucosal bacteria within the sinus mucosa of patients with chronic rhinosinusitis (CRS), but our attempts at characterizing these did not yield any discernible genotypic or phenotypic differences from surface bacteria. We hypothesized that the presence of intramucosal microcolonies reflected host mucosal immune dysfunction. This study characterizes the activation status of T cells, B cells, and macrophages in the sinus mucosa of patients with CRS and controls and determines the impact of bacteria on mucosal immunology. METHODS: Swabs and mucosal biopsy specimens were taken from 27 patients with CRS undergoing sinus surgery and 9 patients with normal sinuses having transnasal pituitary surgery. Microcolonies were detected using Gram staining, and the immune cells were characterized by immunohistochemical techniques. RESULTS: Swab culture rates for Staphylococcus aureus were similar between CRS and controls. However, there were significantly more intramucosal microcolonies in CRS (59% versus 11%) than in controls (p = 0.02). There were significantly more immune cells in CRS. Percentage of activated T and B cells were similar between CRS and controls, but there were significantly more CD163(+) M2 macrophages in patients with CRS (p = 0.0004). Furthermore, percentage of CD163(+) macrophages showed a positive correlation with disease severity. The presence of bacteria had no impact on immunology or disease severity. CONCLUSION: Tolerance of intramucosal microcolonies in CRS may reflect altered macrophage function in the host mucosa. The clinical severity of CRS is also dependent on the host mucosa immune dysfunction, rather than the presence of intramucosal microcolonies.

The interaction between bacteria and mucosal immunity in chronic rhinosinusitis: A prospective cross-sectional analysis.

BACKGROUND: We have detected intramucosal bacteria within the sinus mucosa of patients with chronic rhinosinusitis (CRS), but our attempts at characterizing these did not yield any discernible genotypic or phenotypic differences from surface bacteria. We hypothesized that the presence of intramucosal microcolonies reflected host mucosal immune dysfunction. This study characterizes the activation status of T cells, B cells, and macrophages in the sinus mucosa of patients with CRS and controls and determines the impact of bacteria on mucosal immunology. METHODS: Swabs and mucosal biopsy specimens were taken from 27 patients with CRS undergoing sinus surgery and 9 patients with normal sinuses having transnasal pituitary surgery. Microcolonies were detected using Gram staining, and the immune cells were characterized by immunohistochemical techniques. RESULTS: Swab culture rates for Staphylococcus aureus were similar between CRS and controls. However, there were significantly more intramucosal microcolonies in CRS (59% versus 11%) than in controls (p = 0.02). There were significantly more immune cells in CRS. Percentage of activated T and B cells were similar between CRS and controls, but there were significantly more CD163+ M2 macrophages in patients with CRS (p = 0.0004). Furthermore, percentage of CD163+ macrophages showed a positive correlation with disease severity. The presence of bacteria had no impact on immunology or disease severity. CONCLUSION: Tolerance of intramucosal microcolonies in CRS may reflect altered macrophage function in the host mucosa. The clinical severity of CRS is also dependent on the host mucosa immune dysfunction, rather than the presence of intramucosal microcolonies.

Supraclavicular artery island flap for reconstructing defects in the head and neck region.

PURPOSE OF REVIEW: The supraclavicular artery island flap is a rotation flap that offers a versatile reconstructive option for head and neck defects. Recent anatomical studies have improved our understanding of the vascular supply of the supraclavicular artery island flap. Furthermore, several published large series describe the utility and reliability of this flap. In this article, we review the scientific literature describing the vascular anatomy of the supraclavicular artery island flap, its clinical application, and limitations in reconstructing defects in the head and neck region. RECENT FINDINGS: The vascular anatomy and surface markings, optimal flap design, surgical techniques employed to improve reliability, and aesthetic and functional outcomes of the supraclavicular artery island flap in head and neck reconstruction are well documented in the literature. SUMMARY: The supraclavicular artery island flap offers a versatile and well tolerated option in reconstruction of head and neck defects with several advantages over more traditional regional flaps and distant-free flaps.

Malignant lymphoma in african lions (panthera leo).

Malignant lymphoma has become an increasingly recognized problem in African lions (Panthera leo). Eleven African lions (9 male and 2 female) with clinical signs and gross and microscopic lesions of malignant lymphoma were evaluated in this study. All animals were older adults, ranging in age from 14 to 19 years. Immunohistochemically, 10 of the 11 lions had T-cell lymphomas (CD3(+), CD79a(-)), and 1 lion was diagnosed with a B-cell lymphoma (CD3(-), CD79a(+)). The spleen appeared to be the primary site of neoplastic growth in all T-cell lymphomas, with involvement of the liver (6/11) and regional lymph nodes (5/11) also commonly observed. The B-cell lymphoma affected the peripheral lymph nodes, liver, and spleen. According to the current veterinary and human World Health Organization classification of hematopoietic neoplasms, T-cell lymphoma subtypes included peripheral T-cell lymphoma (4/11), precursor (acute) T-cell lymphoblastic lymphoma/leukemia (2/11), chronic T-cell lymphocytic lymphoma/leukemia (3/11), and T-zone lymphoma (1/11). The single B-cell lymphoma subtype was consistent with diffuse large B-cell lymphoma. Feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) testing by immunohistochemistry on sections of malignant lymphoma was negative for all 11 lions. One lion was seropositive for FeLV. In contrast to domestic and exotic cats, in which B-cell lymphomas are more common than T-cell lymphomas, African lions in this study had malignant lymphomas that were primarily of T-cell origin. Neither FeLV nor FIV, important causes of malignant lymphoma in domestic cats, seems to be significant in the pathogenesis of malignant lymphoma in African lions.

Disseminated microsporidiosis due to Encephalitozoon hellem in an Egyptian fruit bat (Rousettus aegyptiacus).

Disseminated microsporidiosis was diagnosed in an adult female Egyptian fruit bat that died unexpectedly in a zoo. Gross findings, which were minimal, included poor body condition, bilateral renomegaly, and mottling of the liver. Histopathological lesions, which were particularly pronounced in the urogenital tract and liver, consisted primarily of inflammation associated with intracytoplasmic microsporidian spores. Polymerase chain reaction -based methods were used to establish the identity of the microsporidian as Encephalitozoon hellem. E. hellem is an emerging cause of human and avian disease, manifested mainly as opportunistic infection in immunosuppressed patients. This report describes the first documented case of E. hellem in a non-human mammalian species.

Hepatocellular carcinoma in black-tailed prairie dogs (Cynomys ludivicianus): tumor morphology and immunohistochemistry for hepadnavirus core and surface antigens.

From 1994 to 2002, tissues from 61 prairie dogs were submitted to Northwest ZooPath for histopathology. Of these, 12 (20%) had hepatocellular carcinoma (HCC). Three were pets submitted from private veterinary practices. The others were submitted from zoos in the United States. All were adults, ranging from young adult to 7 years of age, with average age of 5.1 years. The most common clinical signs were weight loss, lethargy, palpable abdominal mass, and respiratory difficulty. All tumors were well-differentiated HCCs in which four histologic patterns were recognized. The trabecular pattern was predominant in nine tumors, and the pseudoglandular pattern was predominant in two tumors. The pelioid pattern was also represented in eight tumors. A papillary pattern was present in one case. In seven cases vacuolar change resembling lipidosis was present in the neoplastic hepatocytes of both primary and metastatic tumors. Anaplasia was mild to moderate in most tumors, but a marked degree of anaplasia was noted in the metastatic foci of the case with papillary differentiation. Metastasis to lung was noted in five cases. One of these also had metastasis to the spleen, and another had metastasis to heart and mediastinum. In two cases there was concurrent hepatitis and in two cases, cirrhosis. All tumors and nonneoplastic liver stained negatively for woodchuck hepatitis virus surface and core antigens, and orcein and Victoria blue positive staining of hepatocytes typical of hepadnavirus infection in humans and woodchucks was not present. HCC is apparently common in captive prairie dogs. The hepatic neoplasia observed in prairie dogs was similar to that associated with hepadnaviral infection in humans, woodchucks, and ground squirrels, but no direct evidence of hepadnaviral infection was detected. The rate of metastasis in captive prairie dogs was higher than that reported in woodchucks.

Tyzzer's disease in a neonatal rainbow lorikeet (Trichoglossus haematodus).

A captive-born 8-day-old male rainbow lorikeet (Trichoglossus haematodus) was found dead. Histologically, there were necrotizing hepatitis, myocarditis, and ventriculitis. Silver stain revealed argyrophilic filamentous bacilli within hepatocytes, smooth myofibers of the gizzard, and cardiac myofibers surrounding foci of necrosis. Immunohistochemistry using anti-Clostridium piliforme RT and MSK strain antisera reacted positively against bacilli within hepatocytes, cardiac myofibers, smooth myofibers of the gizzard, and splenic and intestinal macrophages. Polymerase chain reaction (PCR) assay of paraffin-embedded liver, heart, gizzard, spleen, and small intestine amplified the 196-bp DNA fragment specific to 16S ribosomal RNA of C. piliforme. The results of histopathology, immunohistochemistry, and PCR are consistent with C. piliforme infection in this lorikeet.

A disease resembling inclusion body disease of boid snakes in captive palm vipers (Bothriechis marchi).

Between April 1998 and June 1999, 8 palm vipers (Bothriechis marchi) were diagnosed with a disease similar to inclusion body disease (IBD) of boids. Six palm vipers were captive bred, and 2 were wild caught. All of the vipers were adults at the time of death. Three palm vipers were found dead with no premonitory clinical signs, and 5 had anorexia plus possibly 1 of the following clinical signs: regurgitation, paresis, and dehydration. Histologically, all snakes had intracytoplasmic, round to oval, single to multiple eosinophilic inclusion bodies in hepatocytes and renal tubular epithelial cells. Inclusion bodies were distributed among other organs with varying frequency. Common concurrent histologic lesions were urate nephrosis, septic thrombi, and hepatocellular degeneration. Ultrastructurally, inclusions had features similar to inclusions in boid snakes with IBD.

Spontaneous tumours in captive African hedgehogs (Atelerix albiventris): a retrospective study.

Forty tumours were diagnosed in 35 (53%) of 66 captive African hedgehogs documented at Northwest ZooPath (NZP) between 1994 and 1999. Three hedgehogs had more than one type of tumour and the remaining 32 had a single type. Of the 35 hedgehogs with tumours, 14 were female, 11 were male, and 10 were of unknown gender; 21 were from zoological parks and 14 were privately owned. Twenty of the hedgehogs with tumours were adult (>1 year old) with a median age of 3.5 years (range 2-5.5 years); 15, of unreported age, were classified as adult. Thirty-four (85%) of the 40 tumours were classified as malignant and six (15%) as benign. The integumentary, haemolymphatic, digestive and endocrine systems were common sites for tumours. The most common tumours were mammary gland adenocarcinoma, lympho-sarcoma and oral squamous cell carcinoma.

Cardiomyopathy in captive African hedgehogs (Atelerix albiventris).

From 1994 to 1999, 16 captive African hedgehogs (Atelerix albiventris), from among 42 necropsy cases, were diagnosed with cardiomyopathy. The incidence of cardiomyopathy in this study population was 38%. Fourteen of 16 hedgehogs with cardiomyopathy were males and all hedgehogs were adult (>1 year old). Nine hedgehogs exhibited 1 or more of the following clinical signs before death: heart murmur, lethargy, icterus, moist rales, anorexia, dyspnea, dehydration, and weight loss. The remaining 7 hedgehogs died without premonitory clinical signs. Gross findings were cardiomegaly (6 cases), hepatomegaly (5 cases), pulmonary edema (5 cases), pulmonary congestion (4 cases), hydrothorax (3 cases), pulmonary infarct (1 case), renal infarcts (1 case), ascites (1 case), and 5 cases showed no changes. Histologic lesions were found mainly within the left ventricular myocardium and consisted primarily of myodegeneration, myonecrosis, atrophy, hypertrophy, and disarray of myofibers. All hedgehogs with cardiomyopathy had myocardial fibrosis, myocardial edema, or both. Other common histopathologic findings were acute and chronic passive congestion of the lungs, acute passive congestion of the liver, renal tubular necrosis, vascular thrombosis, splenic extramedullary hematopoiesis, and hepatic lipidosis. This is the first report of cardiomyopathy in African hedgehogs.

Coronavirus-associated epizootic catarrhal enteritis in ferrets.

OBJECTIVE: To characterize clinical signs and lesions and identify the etiologic agent associated with epizootic catarrhal enteritis in domestic ferrets. DESIGN: Cross-sectional study. ANIMALS: 119 ferrets with epizootic diarrhea of presumed viral cause and 5 control ferrets. PROCEDURE: Clinical records and biopsy or necropsy specimens of ferrets with presumed epizootic catarrhal enteritis were reviewed. Immunohistochemical staining for coronavirus antigen was performed on paraffin-embedded tissues from approximately 10% of affected ferrets to identify viral antigen and determine its distribution. Transmission electron microscopy was performed on fecal samples and sections of jejunum. Virus isolation studies as well as immunofluorescent tests for other similar viruses were performed. RESULTS: Characteristic microscopic lesions consistent with intestinal coronavirus infection (vacuolar degeneration and necrosis of villus enterocytes; villus atrophy, fusion, and blunting; and lymphocytic enteritis) were consistently detected in affected ferrets. Coronavirus particles were identified in feces and jejunal enterocytes by use of transmission electron microscopy. Immunohistochemical staining of jejunal sections revealed coronavirus antigens. Antigen staining was not detected in healthy ferrets or ferrets with other gastrointestinal tract diseases. Virus isolation was unsuccessful, and other similar viruses were not detected. CONCLUSIONS AND CLINICAL RELEVANCE: Results strongly implicate a coronavirus as the causative agent of epizootic catarrhal enteritis in ferrets. Diagnosis may be made on the basis of a combination of historical, clinical, and microscopic findings.

Mammary gland tumors in captive African hedgehogs.

From December 1995 to July 1999, eight mammary gland tumors were diagnosed in eight adult captive female African hedgehogs (Atelerix albiventris). The tumors presented as single or multiple subcutaneous masses along the cranial or caudal abdomen that varied in size for each hedgehog. Histologically, seven of eight (88%) mammary gland tumors were malignant. Tumors were classified as solid (4 cases), tubular (2 cases), and papillary (2 cases). Seven tumors had infiltrated into the surrounding stroma and three tumors had histologic evidence of neoplastic vascular invasion. Three hedgehogs had concurrent neoplasms. These are believed to be the first reported cases of mammary gland tumors in African hedgehogs.

Subcutaneous atypical mycobacteriosis in captive tiger quolls (Dasyurus maculatus).

From July 1989 to October 1998, 9/37 (24%) adult captive tiger quolls (Dasyurus maculatus) were diagnosed with atypical mycobacterial infection involving the subcutis and skin. Females were more often affected than males (seven females, two males). Grossly, lesions presented as focal thickenings, plaques, and abscesses within the subcutis, often with fistulous tracts. The subcutis and skin overlying cervical and thoracic regions were the primary sites of infection. Cytology of subcutaneous impression smears from all nine affected tiger quolls revealed pyogranulomatous inflammation admixed with several acid-fast bacilli. Histologically, all tiger quolls had nodular to diffuse pyogranulomatous panniculitis and cellulitis. Small numbers of acid-fast bacilli were noted histologically in 7/9 (78%) animals. Skin cultures from seven tiger quolls were positive for one or more different Runyon group IV mycobacteria. The disease described in these tiger quolls is similar to subcutaneous atypical mycobacteriosis of humans and domestic animals.

Necropsy and histopathologic findings in 14 African hedgehogs (Atelerix albiventris): a retrospective study.

From fiscal years 1992 through 1996, 14 African hedgehog (Atelerix albiventris) cases were submitted to the Animal Disease Diagnostic Laboratory at Purdue University. The most common diagnoses were splenic extramedullary hematopoiesis (91%), hepatic lipidosis (50%), renal disease (50%), and neoplastic disease (29%). Other less frequent necropsy findings were myocarditis (21%), colitis (14%), bacterial septicemia (14%), and pneumonia (14%). The data indicate that splenic extramedullary hematopoiesis, hepatic lipidosis, renal disease, and neoplasms are frequent postmortem findings in hedgehogs.

Intestinal lymphosarcoma in captive African hedgehogs.

Two captive adult female African hedgehogs (Atelerix albiventris) had inappetance and bloody diarrhea for several days prior to death. Both hedgehogs had ulceration of the small intestine and hepatic lipidosis. Histopathology revealed small intestinal lymphosarcoma with metastasis to the liver. Extracellular particles that had characteristics of retroviruses were observed associated with the surface of some neoplastic lymphoid cells by transmission electron microscopy. These are the first reported cases of intestinal lymphosarcoma in African hedgehogs.

Unilateral cerebral necrosis resembling feline ischemic encephalopathy in an African lion (Panthera leo).

In November 1996, a 14-yr-old captive male African lion (Panthera leo) had sudden onset of left-sided hemiparesis and mydriasis of the left eye. After 24 hr of supportive care, the lion showed no improvement and was euthanized. At necropsy, the right cerebral hemisphere was diffusely and irregularly swollen and malacic. Histopathology revealed extensive acute necrosis and edema of the portion of the right cerebral hemisphere that received blood from the right middle cerebral artery. Gross and histopathologic cerebral findings resembled those of feline ischemic encephalopathy.

Corynebacterial pneumonia in an African hedgehog.

A 3-mo-old, male African hedgehog (Atelerix albiventris) was anorectic and lethargic for a period of 3 days prior to death. Necropys revealed lungs that were diffusely firm, dark red, and dorsally adhered by fibrinous tags to the pericardial sac. Histopathology revealed necrosuppurative bronchopneumonia with pulmonary abscesses and suppurative pericarditis and myocarditis. A Corynebacterium sp. was isolated from the lungs. We believe this is the first reported case of corynebacterial pneumonia in an African hedgehog.