Ophidiomycosis in Red Cornsnakes (Pantherophis guttatus): Potential Roles of Brumation and Temperature on Pathogenesis and Transmission

2020 ◽  
Vol 57 (6) ◽  
pp. 825-837
Author(s):  
Christina M. McKenzie ◽  
Paul T. Oesterle ◽  
Brian Stevens ◽  
Leonard Shirose ◽  
Gabriela F. Mastromonaco ◽  
...  
Keyword(s):  
Experimental Model ◽  
Quantitative Polymerase Chain Reaction ◽  
Horizontal Transmission ◽  
Skin Lesions ◽  
Future Studies ◽  
Contact Transmission ◽  
Granulomatous Dermatitis ◽  
Polymerase Chain ◽  
Snake Fungal Disease ◽  
Free Ranging

Ophidiomycosis (snake fungal disease) is caused by the fungus Ophidiomyces ophiodiicola. As ophidiomycosis is difficult to study in free-ranging snakes, a reliable experimental model is needed to investigate transmission, pathogenesis, morbidity, and mortality, and the effects of brumation and temperature on disease development. Our objective was to develop such a model via subcutaneous injection of O. ophiodiicola conidia in red cornsnakes ( Pantherophis guttatus). The model was used to evaluate transmission and the effects of brumation and temperature in co-housed inoculated and noninoculated snakes. All 23 inoculated snakes developed lesions consistent with ophidiomycosis, including heterophilic and granulomatous dermatitis, cellulitis, and myositis, and embolic fungal granulomas throughout the liver and the coelomic connective tissue in 21/23 (91%). In the inoculated snakes, 21% of skin swabs, 37% of exuvia, and all liver samples tested positive by qPCR (quantitative polymerase chain reaction) for O. ophiodiicola. A post brumation skin swab from 1/12 noninoculated snakes that brumated in contact with inoculated snakes tested positive by qPCR, suggesting possible contact transmission. That snake had microscopic skin lesions consistent with ophidiomycosis, but no visible fungal elements. Of the 23 inoculated snakes, 20 (87%) died over the 70-day experiment, with ophidiomycosis considered the primary cause of death; 12 (52%) of the inoculated snakes died during brumation. Overall, this experimental model of ophidiomycosis reproduced skin lesions analogous to those of many natural cases, and internal lesions similar to the most severe natural cases. The study provides tentative experimental evidence for horizontal transmission in brumation, and offers a tool for future studies of this widespread snake disease.

scholarly journals Ophidiomycosis surveillance of snakes in Georgia, USA reveals new host species and taxonomic associations with disease

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Ellen Haynes ◽  
Houston C. Chandler ◽  
Benjamin S. Stegenga ◽  
Laura Adamovicz ◽  
Emilie Ospina ◽  
...  
Keyword(s):  
Multinomial Logistic Regression ◽  
Skin Lesions ◽  
Bipartite Network ◽  
Eastern United States ◽  
New Host ◽  
Temporal And Spatial ◽  
Snake Fungal Disease ◽  
Nerodia Erythrogaster ◽  
Free Ranging ◽  
New Host Species

Abstract Ophidiomycosis (snake fungal disease) is caused by the fungus Ophidiomyces ophiodiicola and threatens snake health worldwide. It has been documented throughout the eastern United States and severe cases have recently been reported in Georgia, USA. To evaluate disease distribution and prevalence in this state, 786 free-ranging snakes were examined for skin lesions consistent with ophidiomycosis and swabbed to detect O. ophiodiicola DNA using qPCR. Sampled snakes represented 34 species and 4 families; 27.5% had skin lesions, 13.3% were positive for O. ophiodiicola DNA, and 77.8% of the qPCR positive individuals had skin lesions. This is the first report of O. ophiodiicola in five of the 22 species that were qPCR positive. Multinomial logistic regression modeling indicated that Drymarchon couperi had a higher relative risk of apparent ophidiomycosis (lesions present and qPCR positive), and the best models predicting qPCR result and ophidiomycosis category included individual factors and excluded temporal and spatial factors. Phylogeny-based bipartite network analysis showed that Nerodia erythrogaster, Nerodia taxispilota, and D. couperi had the highest prevalence of apparent ophidiomycosis; this category was more prevalent in the subfamily Colubrinae and less prevalent in Natricinae. These results provide important information about ophidiomycosis epidemiology, which has implications for snake conservation.

Gene expression profiling of follicular lymphoma and normal germinal center B cells using cDNA arrays

Blood ◽  
2002 ◽  
Vol 99 (1) ◽  
pp. 282-289 ◽  
Author(s):  
Hervé Husson ◽  
Elizabeth G. Carideo ◽  
Donna Neuberg ◽  
Joachim Schultze ◽  
Olivier Munoz ◽  
...  
Keyword(s):  
Gene Expression ◽  
B Cells ◽  
Germinal Center ◽  
Quantitative Polymerase Chain Reaction ◽  
Differentially Expressed Gene ◽  
P Value ◽  
Future Studies ◽  
Polymerase Chain ◽  
Follicular Lymphomas ◽  
Necrosis Factor

Follicular lymphomas (FLs) are neoplastic counterparts of normal germinal center (GC) B cells. FLs are characterized by t(14;18) with deregulation of the Bcl-2 (BCL2) gene. The presence of t(14;18) and overexpression of Bcl-2 is necessary, but not sufficient, to cause this disease. An array containing 588 complementary DNAs (cDNAs) was used to compare the gene expression between GC B cells and FL cells. To specifically monitor genes expressed in normal GC B and FL cells and not the entire tissue compartment, normal and malignant B cells were purified from tissues. Using the array, 37 genes were up-regulated and 28 were down-regulated in FL cells as compared to normal GC B cells. The expression level of each differentially expressed gene was verified by quantitative polymerase chain reaction. Following these studies 24 genes were up-regulated and 8 genes down-regulated with a P value less than .1. Included among the genes that were up-regulated in FLs were cell cycle regulator proteins CDK10, p120, p21CIP1, and p16INK4A; transcription factors/regulators Pax-5 and Id-2, which are involved in normal B-cell development; and genes involved in cell-cell interactions, tumor necrosis factor, interleukin-2Rγ (IL-2Rγ), and IL-4Rα. Among the genes that were down-regulated in FLs wereMRP8 and MRP14, which are involved in adhesion. Interestingly, several of these genes are localized within chromosomal regions already described to be altered in FLs. These findings provide a basis for future studies into the pathogenesis and pathophysiology of FL and may lead to the identification of potential therapeutic targets as well as antigens for immunotherapeutic strategies.

scholarly journals Molecular screening of Bartonella in free-ranging capybaras (Hydrochoerus hydrochaeris) from Paraná State, Southern Brazil

2022 ◽  
Vol 43 (2) ◽  
pp. 889-894
Author(s):  
Nelson Jessé Rodrigues dos Santos ◽  
◽  
Renan Bressianini do Amaral ◽  
Luiz Ricardo Gonçalves ◽  
Rogério Ribas Lange ◽  
...  
Keyword(s):  
Quantitative Polymerase Chain Reaction ◽  
Animal Health ◽  
Surveillance Program ◽  
Southern Brazil ◽  
Molecular Screening ◽  
Gapdh Gene ◽  
Hydrochoerus Hydrochaeris ◽  
Polymerase Chain ◽  
Free Ranging ◽  
Active Surveillance Program

Bartonella is an emerging group of facultative intracellular bacteria causing circulatory and systemic disorders. Hosts for Bartonella are mostly mammals, specifically rodents, having a growing number of Bartonella species related to their infection. Capybaras (Hydrochoerus hydrochaeris) are abundant native rodents of Brazil, commonly found in urban parks. In the present study, we aimed to perform molecular screening of capybaras for Bartonella spp. Blood samples were collected from 17 free-ranging animals captured in Paraná State, Southern Brazil. None of the collected samples tested positive for the Bartonella-nuoG gene by quantitative polymerase chain reaction (qPCR), although all of them successfully amplified the mammal endogenous glyceraldehyde-3-phosphate dehydrogenase (gapdh) gene. Additionally, all animals were infested exclusively by Amblyomma dubitatum ticks at the time of sampling. This study was part of an active surveillance program, which is critical for monitoring animal health status, particularly in capybaras.

Matrine Regulates Th1/Th2 Balance to Treat Eczema by Upregulating Interferon-γ

2020 ◽  
Vol 20 (6) ◽  
pp. 3378-3386 ◽  
Author(s):  
Ling Zhang ◽  
Sucai Lin ◽  
Yongping Zheng ◽  
Haitang Wang
Keyword(s):  
Quantitative Polymerase Chain Reaction ◽  
Skin Lesions ◽  
Interferon Γ ◽  
Enzyme Linked Immunosorbent Assay ◽  
High Dose ◽  
Inflammatory Factor ◽  
Chain Reaction ◽  
Treatment Groups ◽  
Polymerase Chain ◽  
Effective Medication

Although matrine (C15H22N2O) has been confirmed to be an effective medication in the treatment of eczema, the mechanisms by which it does so are still unclear. In this study, the mechanisms by which matrine treats eczema were investigated by using oxymatrine, a matrine derivative, to treat guinea pigs with eczema. The differences between the treatment groups in this study were statistically significant (P < 0.05). We prepare nanoparticles to extract RNA. The results showed that the treatment with a high dose of oxymatrine (high-dose OMT) reduced the damage done by eczema to guinea pig skin tissue (i.e., skin lesions). The high-dose OMT inhibited the expression of pro-inflammatory factor proteins, as quantified by enzyme-linked immunosorbent assay (ELISA). The high-dose OMT also increased Th1 and CD4+TGFβ+ levels, as measured by flow cytometry. Examination of skin lesions showed that the high-dose OMT alleviated the symptoms of eczema. We used magnetic nanobeads to extract nucleic acids for detection with quantitative polymerase chain reaction (qPCR), and found that the high-dose OMT improved the expression of pro-inflammatory factor genes. Using western blot analysis, we also found that the high-dose OMT was able to regulate the expression levels of IFN-γ and TGF-β proteins. Our experimental results indicated that matrine treats eczema by upregulating IFN-γ and downregulating TGF-β levels to regulate the Th1/Th2 balance.

Guinea pig seborrheic dermatitis model of Malassezia restricta and the utility of luliconazole

2019 ◽  
Vol 58 (6) ◽  
pp. 820-826 ◽  
Author(s):  
Hiroyasu Koga ◽  
Yukimi Munechika ◽  
Hiroko Matsumoto ◽  
Yasuko Nanjoh ◽  
Kazutoshi Harada ◽  
...  
Keyword(s):  
Guinea Pig ◽  
Quantitative Polymerase Chain Reaction ◽  
Seborrheic Dermatitis ◽  
Skin Lesions ◽  
Antifungal Drugs ◽  
Pathogenic Factor ◽  
Skin Manifestations ◽  
Polymerase Chain ◽  
Guinea Pig Model ◽  
Malassezia Restricta

Abstract Seborrheic dermatitis (SD) is a multifactorial disease in which Malassezia restricta has been proposed as the predominant pathogenic factor. However, experimental evidence supporting this hypothesis is limited. A guinea pig SD model using a clinical isolate of M. restricta was used to elucidate the pathogenicity of M. restricta. Also, the efficacy of 1% luliconazole (LLCZ) cream, a topical imidazole derivative, against M. restricta was compared with that of a 2% ketoconazole (KCZ) cream in the same guinea pig model. Dorsal skin hairs of guinea pig were clipped and treated with M. restricta by single or repeated inoculations without occlusion. Skin manifestations were examined macroscopically and histologically. A quantitative polymerase chain reaction (PCR) assay was also performed for mycological evaluation. An inflammatory response mimicking SD occurred after repeated as well as single inoculation but not in abraded skin. The inflammation score attained its maximum on day 11 and persisted until day 52. The yeast form of the fungal elements was distributed on the surface of stratum corneum and around the follicular orifices, and an epidermal and dermal histological reaction was observed. Application of 1% LLCZ or 2% KCZ cream significantly improved the skin manifestations and decreased the quantity of M. restricta rDNA in the skin lesions. The efficacy of topical antifungal drugs suggested that M. restricta is a pathogenic factor contributing to SD.

Granulomatous Dermatitis due to Malassezia sympodialis

2011 ◽  
Vol 135 (9) ◽  
pp. 1085-1087 ◽  
Author(s):  
Harsha B Desai ◽  
Philip L Perkins ◽  
Gary W Procop
Keyword(s):  
Culture Media ◽  
Skin Lesions ◽  
Etiologic Agent ◽  
Chest Roentgenogram ◽  
Test Results ◽  
Malassezia Pachydermatis ◽  
Malassezia Species ◽  
Granulomatous Dermatitis ◽  
Lipid Supplementation ◽  
Polymerase Chain

A 67-year-old man, with multiple skin lesions that appeared over 2 years, had biopsies that disclosed granulomatous dermatitis with associated small yeasts. The urinary antigen test results were negative for Histoplasma infection; cultures from the biopsies did not grow any fungi or other potential pathogens. The chest roentgenogram results were normal. Morphologic examination revealed features of a Malassezia species. Broad-range fungal polymerase chain reaction and DNA sequencing disclosed that the infecting fungus was Malassezia sympodialis, a lipid-dependent yeast. This report supports one other case report that Malassezia species may cause granulomatous dermatitis; in the previous case, the etiologic agent was Malassezia pachydermatis, a nonlipid-dependent species. We recommend the use of lipid-supplemented culture media for specimens from patients with granulomatous dermatitis because several Malassezia species are dependent on lipid; the absence of lipid supplementation in routine cultures likely explains the negative culture results for this patient. This, to our knowledge, is the first report of granulomatous dermatitis caused by M sympodialis.

Investigation of duck plague virus in hoar areas of Bangladesh

2020 ◽  
Vol 26 (1-2) ◽  
pp. 73-78
Author(s):  
A Hossen ◽  
MH Rahman ◽  
MZ Ali ◽  
MA Yousuf ◽  
MZ Hassan ◽  
...  
Keyword(s):  
Cytopathic Effect ◽  
Clinical Sample ◽  
Chorioallantoic Membrane ◽  
Clinical Samples ◽  
Isolation And Identification ◽  
Duck Plague Virus ◽  
Pcr Method ◽  
Polymerase Chain ◽  
The Individual ◽  
Free Ranging

Duck plague (DP) is the most important infectious disease of geese, ducks and free-ranging water birds. The present study was conducted to determine the prevalence of duck plague virus followed by isolation and identification. For these purposes, a total of 155 cloacal swabs samples were collected randomly from duck of different haor areas of Bangladesh including 45 (41 surveillance and 4 clinical) samples from Netrokona; 42 (40 surveillance and 2 clinical) samples from Kishoregonj; 30 samples from Brahmanbaria and 38 samples from Sunamganj. The samples were processed and pooled (1:5 ratio) for initial screening of target polymerase gene of duck plague virus by polymerase chain reaction (PCR) method. All the samples of a positive pool were then tested individually for identifying the individual positive samples. The result showed that out of 155 samples, 41 (26.45%) were found positive in which 17 were from Netrokona, where 15 (36.58%) were from surveillance samples and 2 (50%) were from clinical sample; 16 were from Kishoregonj, where 14 (35%) were from surveillance samples and 2 (100%) were from clinical sample; 2 (6.6%) were from Brahmanbaria and 5 (13.15%) were from Sunamganj. These positive samples were inoculated into 9-10 days embryonated duck eggs (EDE) through chorioallantoic membrane (CAM) route for the isolation of virus. The EDE died earlier was also chilled, and in a similar way, the CAMs were collected and again performed PCR for id entification of virus. Out of 41 PCR positive samples, 26 samples were isolated and reconfirmed by PCR. Subsequently, DPV was isolated in primary duck embryo fibroblasts cell culture and confirmed by observing cytopathic effect (CPE). Bang. J. Livs. Res. Vol. 26 (1&2), 2019: P. 73-78

AN IMPROVED METHOD FOR INHIBITOR FREE NUCLEIC ACIDS FROM POLYPHENOL RICH LEAVES OF MUSA SPP

2017 ◽  
Vol 23 (1) ◽  
Author(s):  
N.NANDHA KUMAR ◽  
K. SOURIANATHA SUNDARAM ◽  
D. SUDHAKAR ◽  
K.K. KUMAR
Keyword(s):  
Polymerase Chain Reaction ◽  
Quantitative Polymerase Chain Reaction ◽  
Proteinase K ◽  
Chain Reaction ◽  
Banana Plant ◽  
High Molecular Weight Dna ◽  
Musa Spp ◽  
Leaf Tissues ◽  
Polymerase Chain

Excessive presence of polysaccharides, polyphenol and secondary metabolites in banana plant affects the quality of DNA and it leads to difficult in isolating good quality of DNA. An optimized modified CTAB protocol for the isolation of high quality and quantity of DNA obtained from banana leaf tissues has been developed. In this protocol a slight increased salt (NaCl) concentration (2.0M) was used in the extraction buffer. Polyvinylpyrrolidone (PVP) and Octanol were used for the removal of polyphenols and polymerase chain reaction (PCR) inhibitors. Proteins like various enzymes were degraded by Proteinase K and removed by centrifugation from plant extract during the isolation process resulting in pure genomic DNA, ready to use in downstream applications including PCR, quantitative polymerase chain reaction (qPCR), ligation, restriction and sequencing. This protocol yielded a high molecular weight DNA isolated from polyphenols rich leaves of Musa spp which was free from contamination and colour. The average yields of total DNA from leaf ranged from 917.4 to 1860.9 ng/ìL. This modified CTAB protocol reported here is less time consuming 4-5h, reproducible and can be used for a broad spectrum of plant species which have polyphenol and polysaccharide compounds.

Pim-1 is up-regulated by constitutively activated FLT3 and plays a role in FLT3-mediated cell survival

Blood ◽  
2005 ◽  
Vol 105 (4) ◽  
pp. 1759-1767 ◽  
Author(s):  
Kyu-Tae Kim ◽  
Kristin Baird ◽  
Joon-Young Ahn ◽  
Paul Meltzer ◽  
Michael Lilly ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Quantitative Polymerase Chain Reaction ◽  
Colony Growth ◽  
Dominant Negative ◽  
Internal Tandem Duplication ◽  
Downstream Target ◽  
Before And After ◽  
Flt3 Expression ◽  
Polymerase Chain ◽  
Expression Microarrays

AbstractConstitutively activating internal tandem duplication (ITD) mutations of the receptor tyrosine kinase FLT3 (Fms-like tyrosine kinase 3) play an important role in leukemogenesis, and their presence is associated with poor prognosis in acute myeloid leukemia (AML). To better understand FLT3 signaling in leukemogenesis, we have examined the changes in gene expression induced by FLT3/ITD or constitutively activated wild-type FLT3 expression. Microarrays were used with RNA harvested before and after inhibition of FLT3 signaling. Pim-1 was found to be one of the most significantly down-regulated genes upon FLT3 inhibition. Pim-1 is a proto-oncogene and is known to be up-regulated by signal transducer and activator of transcription 5 (STAT5), which itself is a downstream target of FLT3 signaling. Quantitative polymerase chain reaction (QPCR) confirmed the microarray results and demonstrated approximately 10-fold decreases in Pim-1 expression in response to FLT3 inhibition. Pim-1 protein also decreased rapidly in parallel with decreasing autophosphorylation activity of FLT3. Enforced expression of either the 44-kDa or 33-kDa Pim-1 isotypes resulted in increased resistance to FLT3 inhibition-mediated cytotoxicity and apoptosis. In contrast, expression of a dominant-negative Pim-1 construct accelerated cytotoxicity in response to FLT3 inhibition and inhibited colony growth of FLT3/ITD-transformed BaF3 cells. These findings demonstrate that constitutively activated FLT3 signaling up-regulates Pim-1 expression in leukemia cells. This up-regulation contributes to the proliferative and antiapoptotic pathways induced by FLT3 signaling. (Blood. 2005;105: 1759-1767)

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