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Abstract

An HPLC-DAD method was developed for the determination of formaldehyde in animal feed and silage. The method is based on the determination of the product of chemical reaction between formaldehyde and 2,4-dinitrophenylhydrazine. A 3 g of feed or silage were extracted with Milli-Q water with phosphoric acid and next formaldehyde was derivative with the use 2,4-dinitrophenyl- hydrazine in acetronitrile solution. The extract was purified with 0.45 µm syringe filters and separeted on Zorbax Eclipse XDB C18 column and detection was carried out at 360 nm. Formal- dehyde was eluted with a mobile phase consisting of acetonitrile/water in isocratic elution. This method provided average recoveries of 90.6% to 102.2%, with CVs of 2.6% to 6.4% for feed and from 91.3% to 108.7% with CVs of 1.1% to 4.1% for silage in the ranged of 50 to 1000 mg/kg feeds and silage. The LOD and LOQ for formaldehyde in feed and silage ranged from 1.6 to 2.6 and 2.7 to 5.7 mg/kg, respectively. The methodology was applied for the analysis of feed and silage samples collected from poultry, pigs and cows farms.

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Authors and Affiliations

E. Patyra
K. Kwiatek
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Abstract

This paper addresses the issue of the development of FOR DREAD THAT – a negative purpose subordinator in the history of the English language. The theoretical foundation of this work are the mechanisms of grammaticalisation suggested by Heine and Kuteva in many works of theirs. The gathered material shows that the development of this relatively rarely used subordinator constitutes a case of a typical grammaticalisation whose rise might have been the result of analogy with FOR FEAR THAT.

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Authors and Affiliations

Andrzej M. Łęcki
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Abstract

A highly immunogenic nucleotide fragment (195bp) was selected from the Mhp183 gene of Mycoplasma hyopneumoniae using information software technology and was named Mhp183195bp. Three Mhp183195bp were linked to form a new nucleotide sequence called Mhp183615bp. Mhp183615bp was directly synthesized and cloned into a pET100 vector and expressed in Escherichia coli. After purification, the proteins were successfully validated using SDS-PAGE and Western blot. BALB/c mice were injected with purified proteins on the first, eighth, and fifteenth days of feeding, respectively; serum samples were collected from mice on the day of immunization and on the 22nd day after immunization. The antibody level in mouse serum was detected by Western blotting using purified expressed proteins as antigens. IL-2, TNF-α and IFN-γ were simultaneously detected in mouse serum by ELISA. The 30 kDa protein was successfully expressed and reacted specifically with the specific serum Mhp His-Tag mouse monoclonal antibody and pig antibody. The expressed recombinant protein was immunogenic. The expression levels of IFN-γ, IL-2 and TNF-α were found to be significantly higher on day 22 than in the control group. This study suggests that the expressed recombinant protein could be used as one of the novel vaccine candidates for Mhp.
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Bibliography


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Authors and Affiliations

M. Xu
1
J. Zheng
1
S. Hu
1
G. Wang
1

  1. College of Veterinary Medicine, Hunan Agricultural University, Road 1#, Changsha, 410000, China

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