scholarly journals Transmission and Serial Propagation ofEnterocytozoon bieneusi from Humans and Rhesus Macaques in Gnotobiotic Piglets

1998 ◽  
Vol 66 (11) ◽  
pp. 5515-5519 ◽  
Author(s):  
Ivanela Kondova ◽  
Keith Mansfield ◽  
Michael A. Buckholt ◽  
Barry Stein ◽  
Giovanni Widmer ◽  
...  
Keyword(s):  
Animal Model ◽  
In Situ Hybridization ◽  
Chronic Diarrhea ◽  
Rhesus Macaques ◽  
Scientific Progress ◽  
Enterocytozoon Bieneusi ◽  
Oral Challenge ◽  
Infected Piglet ◽  
Gnotobiotic Piglets

ABSTRACT For over a decade Enterocytozoon bieneusi infections in people with AIDS have been linked with chronic diarrhea and wasting. The slow scientific progress in treating these infections is attributed to the inability of investigators to cultivate the parasite, which has also precluded evaluation of effective therapies. We report here successful serial transmissions of E. bieneusi from patients with AIDS and from macaques with AIDS to immunosuppressed gnotobiotic piglets. One infected piglet was still excreting spores at necropsy 50 days after an oral challenge. Spores in feces were detected microscopically by trichrome stain and by PCR and within enterocytes by in situ hybridization and immunohistochemistry. E. bieneusiinfection induced no symptoms. The development of an animal model forE. bieneusi will open up new opportunities for investigating this parasite.

scholarly journals Prevalence of Enterocytozoon bieneusi in Swine: an 18-Month Survey at a Slaughterhouse in Massachusetts

2002 ◽  
Vol 68 (5) ◽  
pp. 2595-2599 ◽  
Author(s):  
Michael A. Buckholt ◽  
John H. Lee ◽  
Saul Tzipori
Keyword(s):  
In Situ Hybridization ◽  
Internal Transcribed Spacer ◽  
Enterocytozoon Bieneusi ◽  
Type D ◽  
Gnotobiotic Piglets

ABSTRACT Slaughterhouse pig samples were analyzed by PCR for Enterocytozoon bieneusi infection. Thirty-two percent were found to be positive, with rates being higher over the summer months. Three isolates from pigs were identical in their ribosomal internal transcribed spacer sequence to human E. bieneusi type D, two were identical to type F (from a pig), and nine were previously unreported. The viability of these spores was demonstrated by their ability to infect gnotobiotic piglets. The presence of the infection in liver was shown by in situ hybridization.

scholarly journals Localization of Persistent Enterocytozoon bieneusiInfection in Normal Rhesus Macaques (Macaca mulatta) to the Hepatobiliary Tree

1998 ◽  
Vol 36 (8) ◽  
pp. 2336-2338 ◽  
Author(s):  
Keith G. Mansfield ◽  
Angela Carville ◽  
Daniel Hebert ◽  
Laura Chalifoux ◽  
Daniel Shvetz ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Macaca Mulatta ◽  
Rhesus Macaques ◽  
Clinical Signs ◽  
Enterocytozoon Bieneusi ◽  
Proximal Jejunum ◽  
Microsporidian Parasite ◽  
Immunodeficiency Virus ◽  
History Of

Enterocytozoon bieneusi is the most common microsporidian parasite recognized in human patients with AIDS. Recently, we identified a virtually identical organism causing a spontaneous infection associated with hepatobiliary and intestinal disease in simian immunodeficiency virus (SIV)-infected macaques. To examine the natural history of the infection, we examined captive rhesus macaques for E. bieneusi by PCR, in situ hybridization, and cytochemical techniques. PCR performed on fecal DNA detected enterocytozoon infection in 22 (16.7%) of 131 normal rhesus macaques (Macaca mulatta), compared to 18 (33.8%) of 53 rhesus macaques experimentally inoculated with SIV. In normal rhesus macaques, persistence of infection was demonstrated for up to 262 days and was usually not associated with clinical signs. In six of seven normal rhesus animals, E. bieneusi was detected by PCR in bile obtained through percutaneous cholecystocentesis but not by in situ hybridization performed on endoscopic biopsies of duodenum and proximal jejunum.

scholarly journals Enteric viruses nucleic acids distribution along the digestive tract of rhesus macaques with idiopathic chronic diarrhea

2021 ◽  
Author(s):  
Eric Delwart ◽  
David Merriam ◽  
Amir Ardeshir ◽  
Eda Altan ◽  
Yanpeng Li ◽  
...  
Keyword(s):  
Digestive Tract ◽  
Chronic Diarrhea ◽  
Rhesus Macaques ◽  
Distal Colon ◽  
Viral Metagenomics ◽  
Dna Viruses ◽  
Mucosal Scraping ◽  
Rna And Dna ◽  
Lower Digestive Tract

AbstractIdiopathic chronic diarrhea (ICD) is a common clinical condition in captive rhesus macaques, claiming 33% of medical culls (i.e. deaths unrelated to research). Using viral metagenomics we characterized the eukaryotic virome in digestive tract tissues collected at necropsy from nine animals with ICD. We show the presence of multiple viruses in the Parvoviridae and Picornaviridae family. We then compared the distribution of viral reads in the stomach, duodenum, jejunum, ileum, and the proximal, transverse, and distal colons. Tissues and mucosal scraping from the same locations showed closely related results while different gut tissues from the same animal varied widely. Picornavirus reads were generally more abundant in the lower digestive tract, particularly in the descending (distal) colon. Parvoviruses were more abundant in the upper reach particularly in the stomach. In situ hydridization (ISH) of fixed tissues showed punctuated staining for both these RNA and DNA viruses in the distal colon. Parvovirus ISH staining was also detected in the stomach/duodenum/jejunum in distinct oval-shaped structures. Therefore, the location of enteric viral nucleic acid differed widely between different viral families and along the length of the digestive tract.

In situ hybridization: A molecular approach for the diagnosis of the microsporidian parasite Enterocytozoon bieneusi

1999 ◽  
Vol 30 (1) ◽  
pp. 54-58 ◽  
Author(s):  
Jorge N Velasquez ◽  
Silvana Carnevale ◽  
Jorge H Labbe ◽  
Agustin Chertcoff ◽  
Marta G Cabrera ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Enterocytozoon Bieneusi ◽  
Molecular Approach ◽  
Microsporidian Parasite

scholarly journals Generation of Thyrotropin-Releasing Hormone Receptor 1-Deficient Mice as an Animal Model of Central Hypothyroidism

2004 ◽  
Vol 18 (6) ◽  
pp. 1450-1460 ◽  
Author(s):  
Roland Rabeler ◽  
Jens Mittag ◽  
Lars Geffers ◽  
Ulrich Rüther ◽  
Michael Leitges ◽  
...  
Keyword(s):  
Animal Model ◽  
In Situ Hybridization ◽  
Mrna Expression ◽  
Pcr Analysis ◽  
Target Cells ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Central Hypothyroidism ◽  
R1 Gene

Abstract To provide an animal model of central hypothyroidism, mice deficient in the TRH-receptor 1 (TRH-R1) gene were generated by homologous recombination. The pituitaries of TRH-R1−/− mice are devoid of any TRH-binding capacity, demonstrating that TRH-R1 is the only receptor localized on TRH target cells of the pituitary. With the exception of some retardation in growth rate, TRH-R1−/− mice appear normal, but compared with control animals they exhibit a considerable decrease in serum T3, T4, and prolactin (PRL) levels but not in serum TSH levels. In situ hybridization histochemistry and real-time RT-PCR analysis revealed that in adult TRH-R1−/− animals TSHβ-mRNA expression is not impaired whereas PRL mRNA and GH mRNA levels are considerably reduced compared with control mice. The numbers of thyrotropes, somatotropes, and lactotropes, however, are not affected by the deletion of the TRH-R1 gene. The mutant mice are fertile, and the dams nourish their pups well, indicating that TRH is not a decisive factor for suckling-induced PRL release. In situ hybridization and quantitative RT-PCR analysis, furthermore, revealed that, as in control animals, pituitary PRL-mRNA expression in TRH-R1−/− is considerably increased during lactation, albeit strongly reduced as compared with lactating control animals.

scholarly journals Rabbit Genital Tissue Is Susceptible to Infection by Rabbit Oral Papillomavirus: an Animal Model for a Genital Tissue-Targeting Papillomavirus

1998 ◽  
Vol 72 (6) ◽  
pp. 5239-5244 ◽  
Author(s):  
Stephen B. Harvey ◽  
Nancy M. Cladel ◽  
Lynn R. Budgeon ◽  
Patricia A. Welsh ◽  
James W. Griffith ◽  
...  
Keyword(s):  
Animal Model ◽  
In Situ Hybridization ◽  
Adult Rabbit ◽  
Viral Capsid Antigen ◽  
Tissue Targeting ◽  
Direct Inoculation ◽  
Domestic Rabbits ◽  
Immunohistological Staining ◽  
Genital Tissue

ABSTRACT Rabbit oral papillomavirus (ROPV) is a mucosatropic papillomavirus which naturally infects oral mucosal sites of domestic rabbits. In this study, we tested the hypothesis that rabbit genital mucosa is also susceptible to ROPV infection by using the athymic mouse xenograft system and adult immunocompetent rabbits. Subrenal xenografts of ROPV-infected rabbit vulvar and penile sheath tissues were strongly positive for ROPV infection by histologic, in situ hybridization, and Southern analyses. Direct inoculation of adult rabbit penises with infectious ROPV produced small raised lesions of approximately 1 by 1 by 1 mm that were ROPV positive by both in situ hybridization and Southern analyses and were also viral capsid antigen positive by immunohistological staining. Infection of rabbit genital tissues with ROPV may be a useful animal model for the study of genital tissue-targeting papillomaviruses.

scholarly journals Infectious Agent and Immune Response Characteristics of Chronic Enterocolitis in Captive Rhesus Macaques

2003 ◽  
Vol 71 (7) ◽  
pp. 4079-4086 ◽  
Author(s):  
Karol Sestak ◽  
Christopher K. Merritt ◽  
Juan Borda ◽  
Elizabeth Saylor ◽  
Shelle R. Schwamberger ◽  
...  
Keyword(s):  
Clinical Symptoms ◽  
Chronic Diarrhea ◽  
Rhesus Macaques ◽  
Clinical Syndrome ◽  
Enterocytozoon Bieneusi ◽  
Enteric Pathogens ◽  
Lymphoid Tissues ◽  
Campylobacter Coli ◽  
Host Immune System ◽  
Necrosis Factor Alpha

ABSTRACT Chronic enterocolitis is the leading cause of morbidity in colonies of captive rhesus macaques (Macaca mulatta). This study's aim was to identify the common enteric pathogens frequently associated with chronic enterocolitis in normal, immunocompetent rhesus monkeys and to elucidate the influence of this clinical syndrome on the host immune system. We analyzed the fecal specimens from 100 rhesus macaques with or without clinical symptoms of chronic diarrhea. Retrospective analysis revealed an increased incidence of Campylobacter spp. (Campylobacter coli and Campylobacter jejuni), Shigella flexneri, Yersinia enterocolitica, adenovirus, and Strongyloides fulleborni in samples collected from animals with chronic diarrhea (P < 0.05). The presence of additional enteric pathogens, such as Escherichia coli, carrying the eaeA intimin or Stx2c Shiga toxin virulence genes, Balantidium coli, Giardia lamblia, Enterocytozoon bieneusi, and Trichuris trichiura was found in all animals regardless of whether diarrhea was present. In addition, the upregulation of interleukin-1α (IL-1α), IL-3, and tumor necrosis factor alpha cytokine genes, accompanied by an increased presence of activated (CD4+ CD69+) T lymphocytes was found in gut-associated lymphoid tissues collected from animals with chronic enterocolitis and diarrhea in comparison with clinically healthy controls (P < 0.05). These data indicate that chronic enterocolitis and diarrhea are associated, in part, with a variety of enteric pathogens and highlight the importance of defining the microbiological status of nonhuman primates used for infectious disease studies. The data also suggest that chronic colitis in rhesus macaques may have potential as a model of inflammatory bowel disease in humans.

scholarly journals Effect of 17beta-estradiol on hypothalamic GnRH-II gene expression in the female rhesus macaque

2004 ◽  
Vol 33 (1) ◽  
pp. 145-153 ◽  
Author(s):  
VS Densmore ◽  
HF Urbanski
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Mrna Expression ◽  
Rhesus Macaques ◽  
Molecular Forms ◽  
Stimulatory Action ◽  
Medial Basal Hypothalamus ◽  
Hybridization Histochemistry ◽  
In Situ Hybridization Histochemistry

The hypothalamus of rhesus macaques expresses two molecular forms of gonadotropin-releasing hormone (GnRH-I and GnRH-II). However, it is unclear whether these two neuropeptides play similar roles in the control of reproductive neuroendocrine function, especially in the context of positive and negative estrogen feedback. To address this issue, in situ hybridization histochemistry was used to compare the effect of 17beta-estradiol (E) on the expression of GnRH-I and GnRH-II mRNA in the medial basal hypothalamus (MBH) of adult female macaques. GnRH-I mRNA expression was found to be significantly (P<0.01) more abundant in ovariectomized (ovx) animals compared with ovariectomized E-treated (ovx+E) animals. In marked contrast, GnRH-II mRNA expression was found to be significantly (P<0.05) more abundant in ovx+E animals than in the ovx animals. To help elucidate how E exerts this stimulatory action on GnRH-II gene expression, hypothalamic sections were subsequently double labeled using a combination of immunohistochemisty for estrogen receptor (ER) -alpha or -beta and in situ hybridization histochemistry for GnRH-II. Approximately 50% of the GnRH-II positive cells in the MBH were found to express ERbeta, but none expressed ERalpha. Taken together, these data give credence to a novel pathway by which E may control the primate neuroendocrine reproductive axis, one that involves stimulation of GnRH-II release via an ERbeta-mediated mechanism.

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