Investigations of the proteolytic Gram Negative Psychrotrophs(GNP) bacteria was the basal objective of this study. A conventional diagnostic PCR technique based on three pairs of primers including SerAgeneto amplify an 950 bp fragment of Acinetobacter sppDNA,serAgene for/A. hydrophila ( 650bp): and aprgene for/S. marcescens (500bps) was done.In the present study the 29 bacterial isolates obtained from 100 cows raw milk samples were collected randomly from healthy cows with different age and breed present in different farms of Thi-Qar province, previously refrigerated for 72 hr. These isolates subjected to cultural-based enrichment and PCR-based identification .The present results revealed that the raw milk GNP contamination overall ratio was 29% . Acinetobacter spp were the most predominant bacteria (16%) among the studied GNP contaminants, while A.hydrophila appeared in a ratio of 7% and S. marcescens showed the lower ratio ( 6%).,the results of the studied genes product of GNP bacteria was considered to be highly statistically significant (P>0.001).The distribution of studied GNP according to age ,parturition number and breed of studied animals was investigated. The effect of these factors on the PCR-based identification results was considered to be not statistically significant(P>0.05) however, the higher overall ratio(29.1%) for cow raw milk contamination was observed in raw milk of cows between <3 -< 9 year of age. In general cows at first age group (<3 -< 9 year) showed high ratio of raw milk contamination(7.6 and6.3%) with GNP bacteria (Serratia marscense and Aremonase hydrophila respectively)..Concerning the number of parturition, cows with high numbers of parturition( ˃ 6-<12) showed high overall ratio(38.5%) of contamination also high ratio of Acinetobacter spp(23.1%) ,Aremonase hydrophila(7.7%) and Serratia marscense 162 (7.7%) were observed in the same group of cows. According to breed, a high overall ratio of GNP bacterial contamination was observed in 40% of crossbred cow raw milk followed by Friesian cows(32.1%).Beside that raw milk of crossbred cows showed a high ratio of contamination with Acinetobacter spp (20%) and Aremonase hydrophila(13.3%) while Serratia marscense appeared as a higher contaminant in Friesian cows raw milk with the ratio of its contamination was10.7%.
Ninety six for each healthy(n=96)and atopic,(n=96)individuals duals from the same geographical region, paired by sex and age, their sera specific IgE antibodies were estimated by enzyme linked immune sorbent assay test (ELISA) and genotyped by polymerase chain reaction based onHLA-DQB1*0602, HLA-DQB1*0604 andHLA-DRB1*12.The specific IgE based on ELISA results revealed that Out of 96 only59 (61.5%) of atopic patients were sensitive to CR allergen. The association between sensitivity to CR allergen and age was considered to be not statistically significant (P>0.05).However the higher rate of CR allergens sensitivity(62.9%) was observed in first age group(<45 year) of atopic patients. In contrast the effect of sex on sensitivity to CR allergens was considered to be statistically significant (p<0.05) and the higher rate of sensitivity(75.6%) was observed in atopic patients males.The overall differences in the HLA-DQB1*0602, HLA-DQB1*0604 and HLA-DRB1*12 alleles frequency between patients and controlswerestatistically (p<0.05).According to the results of risk factors statistical analysis values(p:value = 0.0001; 210 highly significant (p<0.01) concerning the effect of the age and sex. In general the allele HLA-DRB1*12 was not observed in both atopic patients and controls in contrast HLA-DQB1*0602 was present in atopic patients only while HLA-DQB1*0604 appeared in both patients and controls with different frequencies The older atopic patients showed higher frequency (61.8% ) for the HLA-DQB1*0602 allele . In contrast higher frequency of HLA-DQB1*0604 allele occurred in younger patients(40.3%) .According to sex ,the higher frequency of HLA-QB1*0602allele was observed in males patients(31.7%)while the allele HLA-DQB1*0604 higher frequency(17.9 %)was observed in the females of the control group.The overall frequency of HLA-DQB1*0602(60.4%)orHLA-DQB1*0604(39.6%) as a single allele was observed in the seropositive or seronegative atopic patients.The seropositive showed higher frequency(35.6 and 15.3%) for HLA-DQB1*0602and HLA-DQB1*0604 respectively.
The local Streptomyces sp. strain showed an ability to produce antimicrobial metabolite active against standard strains, in primary and secondary screening. The produced antibiotic was extracted, purified and identified as a peptide antibiotic produced about 1.4g/L in 7 days incubation period, and its LD 50 was 5500. There was an inverse effect for orange acridine dye on the grown colonies number of S. sp., the 28 g/ml dye concentration was chosen as the best concentration because it led to colonies killing by 95%. Plasmid DNA extracted from S. sp. and then transformed to E. coli pBR 322, the E. coli pBR 322 showed negative results against the standard strains in primary screening before plasmid DNA transformation, while transformed E. coli pBR322 showed positive results. The antibiotic produced by trans. E. coli pBR322 was extracted, purified and identified by the same ways, which gave the same antibiotic produced by S. sp. with an increase of 2.2 g/L in the quantity and shorter period of time (2 days).
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