Development of PCR Primers for the Detection of Porcine DNA in Feed Using mtATP6 as the Target Sequence

Yoshida, Tomotaro; Nomura, Tetsuya; Shinoda, Naoki; Kusama, Toyoko; Kadowaki, Koh-ichi; Sugiura, Katsuaki

doi:10.3358/shokueishi.50.89

Cited by 12 publications

(9 citation statements)

References 16 publications

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“…Total RNA from spleen was extracted using RNeasy Mini Kit (Qiagen, Courtaboeuf, France) as previously described . The pig primer sequences were previously described . qPCR was performed using an ABI Prism 7300 sequence detector system (Applied Biosystems, Villebon‐sur‐Yvette, France) using SYBR‐green fluorescence (Master Mix Quantifast Syber Green; Qiagen) for quantification.…”

Section: Methodsmentioning

confidence: 99%

“…10 The pig primer sequences were previously described. 12 qPCR was performed using an ABI Prism 7300 sequence detector system (Applied Biosystems, Villebon-sur-Yvette, France) using SYBR-green fluorescence (Master Mix Quantifast Syber Green; Qiagen) for quantification. Results were presented as mRNA expression in the control, placebo and EPIT groups.…”

Section: Immune Parametersmentioning

confidence: 99%

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Treatment of gastric eosinophilia by epicutaneous immunotherapy in piglets sensitized to peanuts

Mondoulet

Kalach

Dhelft

et al. 2017

Clin Experimental Allergy

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Summary Background Eosinophilic gastrointestinal disorders (EGIDs) are hypersensitivity disorders frequently triggered by food allergy and manifested by mucosal eosinophilic infiltration at any level of the gastrointestinal tract. This study established a model of gastric eosinophilia in peanut‐sensitized piglets to evaluate the efficacy of epicutaneous immunotherapy (EPIT) for its treatment. Methods Experiments were carried out in piglets first sensitized by three intra‐peritoneal injections of peanut protein extract (PPE) with adjuvant, and then given PPE orally for 10 days, a sequence leading to gastric eosinophilia assessed by endoscopy. For 3 months, eight piglets received active EPIT, using Viaskin® loaded with PPE, applied daily on the ear, while eight received placebo EPIT (Placebo). Piglets were exposed to a second 10‐day period of PPE orally. Lesions were scored by endoscopy on the last day of PPE exposure. After killing, all parts of the digestive tract were analysed by a pathologist unaware of the piglets’ status. IgE response was measured, and mechanistic parameters were analysed in the spleen. Results After sensitization, a significant increase of total IgE was observed in sensitized compared to naive animals (61.1 ± 13.3 vs 27.8 ± 6 ng/mL, P < .01). Following oral intake of PPE, sensitized piglets developed moderate gastritis compared to naive piglets (1.5 vs 1.0, median score). After 3 months of immunotherapy, median IgE was significantly reduced in EPIT vs placebo piglets (61.4 ± 16.3 vs 105.9 ± 25.6 ng/mL, P < .01). Active EPIT significantly reduced gastric mucosal lesions induced by PPE oral intake (macroscopic score 0 [0‐2] vs 2 [1‐3], P < .01, respectively, active vs placebo) and gastric mucosa eosinophils counts (239 eosinophils/mm2 [59‐645] vs 2554 eosinophils/mm2 [462‐8057], P < .01, respectively active vs placebo). GATA‐3, IL‐5 and eotaxin mRNA expression decreased significantly after EPIT (P < .05). Conclusions This study describes a large animal model of gastric eosinophil in peanut‐sensitized piglets. Utilizing this model, we demonstrated the efficacy of EPIT in treating peanut‐induced EGIDs.

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Section: Methodsmentioning

confidence: 99%

Section: Immune Parametersmentioning

confidence: 99%

Treatment of gastric eosinophilia by epicutaneous immunotherapy in piglets sensitized to peanuts

Mondoulet

Kalach

Dhelft

et al. 2017

Clin Experimental Allergy

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“…In Japan, PCR identification of species-specific, animal group-specific, and plant DNA is employed as part of feed ban in place for the control of BSE. The PCR method is of fundamental importance for the detection of prohibited animal-derived material in feed [92] and could be employed as part of inspection of meat and other products of animal origin intended for human consumption.…”

Section: Infectivity Of Prion Diseases and Amyloid Proteinsmentioning

confidence: 99%

A Concise Review of Amyloidosis in Animals

Woldemeskel

2012

Veterinary Medicine International

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Amyloidosis refers to a group of protein misfolding diseases characterized by deposition of a particular amyloid protein in various organs and tissues of animals and humans. Various types and clinical forms of amyloidosis, in which the pathology and pathogenesis is diverse depending upon the underlying causes and species affected, are reported in domestic and wild animals. The clinical findings are also quite variable consequent to the variation of the tissues and organs involved and the extent of functional disruption of the affected organs in various animal species. The affected organs may be enlarged and exhibit variable pallor grossly, or the amyloid deposit may be discernible only after microscopic examination of the affected tissues. Amyloid appears as a pale eosinophilic homogenous extracellular deposit in tissues. However, microscopic examination and Congo red staining with green birefringence under polarized light are needed to confirm amyloid and differentiate it from other apparently similar extracellular deposits such as collagen and fibrin. Identifying the type of amyloid deposit needs immunohistochemical staining, ultrastructural characterization of the amyloid fibril, and if feasible also genetic studies of the involved species for clinical and prognostic purposes. This paper provides a concise review of the occurrence of amyloidosis in domestic and wild animals.

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“…Moreover, significant transcriptional down‐regulation of C‐KIT , differentiation marker of spermatogonia (An et al, ), was detected in porcine SSCs cultured in 0.2% (w/v) agarose‐based 3D hydrogels compared with those in 2D microenvironments (Supporting Information Figure S3). Differentiation into spermatozoa still occurred in recipient mouse testis transplanted with undifferentiated porcine SSCs maintained in 0.2% (w/v) agarose‐based 3D hydrogels, as determined by the presence of spermatozoa with a round‐shaped head (Honaramooz et al, ) (Supporting Information Figure S4A) and a mitochondria containing pig‐specific mtDNA (Yoshida et al, ) (Supporting Information Figure S4B). From these results, porcine SSCs are maintained in an undifferentiated state more effectively in 3D than in 2D culture microenvironments.…”

Section: Resultsmentioning

confidence: 99%

Porcine spermatogonial stem cells self‐renew effectively in a three dimensional culture microenvironment

Park

Kim

et al. 2017

Cell Biology International

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Generally, self-renewal of spermatogonial stem cells (SSCs) is maintained in vivo in a three-dimensional (3D) microenvironment consisting of the seminiferous tubule basement membrane, indicating the importance of the 3D microenvironment for in vitro culture of SSCs. Here, we report a 3D culture microenvironment that effectively maintains porcine SSC self-renewal during culture. Porcine SSCs were cultured in an agarose-based 3D hydrogel and in 2D culture plates either with or without feeder cells. Subsequently, the effects of 3D culture on the maintenance of undifferentiated SSCs were identified by analyzing cell colony formation and morphology, AP activity, and transcriptional and translational regulation of self-renewal-related genes and the effects on proliferation by analyzing cell viability and single cell-derived colony number. The 3D culture microenvironment constructed using a 0.2% (w/v) agarose-based 3D hydrogel showed the strongest maintenance of porcine SSC self-renewal and induced significant improvements in proliferation compared with 2D culture microenvironments. These results demonstrate that self-renewal of porcine SSCs can be maintained more effectively in a 3D than in a 2D culture microenvironment. Moreover, this will play a significant role in developing novel culture systems for SSCs derived from diverse species in the future, which will contribute to SSC-related research.

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Development of PCR Primers for the Detection of Porcine DNA in Feed Using mtATP6 as the Target Sequence

Cited by 12 publications

References 16 publications

Treatment of gastric eosinophilia by epicutaneous immunotherapy in piglets sensitized to peanuts

Treatment of gastric eosinophilia by epicutaneous immunotherapy in piglets sensitized to peanuts

A Concise Review of Amyloidosis in Animals

Porcine spermatogonial stem cells self‐renew effectively in a three dimensional culture microenvironment

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