Relationship of Genetic Polymorphism For CYP19 Gen of Exon 3 With Quantity and Milk Composition in Domestic Female Goats

This study was carried out to investigate the effect of the genetic polymorphism of the third exon for CYP19 gene on the quantity and composition of milk in local female goats. Blood samples were collected from female goats in the fields of some breeder in Babil Governorate/Al-Qasim City. Milk composition were examined in the Public Health Laboratory at Al-Qasim Green University/College of Veterinary Medicine, the genetic bundle of the third axon of the CYP19 gene was isolated, which was 140 bp in size, and its genotyping was determined according to the different nucleotide sequences and their association with the quantity and milk composition in the samples which tested in the Bayolat Laboratory in Al-Diwaniyah Governorate. The proportions of the distribution of the genotypes of the CYP19 gene in a sample of local female goats TT, GG and TG were 33.34%, 26.66 and 40%, respectively. The results showed prevalence of the TG genotype for CYP19 gene in the Iraqi local goats on the pure genotypes TT and GG, which was a highly significant superior (P<0.01) in the percentages of fat, protein, lactose and CFPP, as well as the significant superiority of the mixed genotype significantly on the pure genotype in the percentages of solid non-fat and density. It could be concluded from the current study that the selection of females with TG genotype as source for future generations to obtain a generation selected for the desired traits.

Decreased susceptibility to dihydrofolate reductase inhibitors associated with genetic polymorphisms in Ugandan Plasmodium falciparum isolates

Background The Plasmodium falciparum dihydrofolate reductase (PfDHFR) inhibitors pyrimethamine and cycloguanil (the active metabolite of proguanil) have important roles in malaria chemoprevention, but drug resistance challenges their efficacies. A new compound, P218, was designed to overcome resistance, but drug susceptibility data for P. falciparum field isolates are limited. Methods We studied ex vivo PfDHFR inhibitor susceptibilities of 559 isolates from Tororo and Busia districts, Uganda from 2016-2020, sequenced 383 isolates, and assessed associations between genotypes and drug susceptibility phenotypes. Results Median IC50’s were 42,100 nM for pyrimethamine, 1,200 nM for cycloguanil, 13,000 nM for proguanil, and 0.6 nM for P218. Among sequenced isolates, three PfDHFR mutations, 51I (100%), 59R (93.7%), and 108N (100%), were very common, as previously seen in Uganda, and another mutation, 164L (12.8%), had moderate prevalence. Increasing numbers of mutations were associated with decreasing susceptibility to pyrimethamine, cycloguanil, and P218, but not proguanil, which does not act directly against PfDHFR. Differences in P218 susceptibilities were modest, with median IC50 1.4 nM for parasites with mixed genotype at position 164 and 5.7 nM for pure quadruple mutant (51I/59R/108N/164L) parasites. Conclusion Resistance-mediating PfDHFR mutations were common in Ugandan isolates, but P218 retained excellent activity against mutant parasites.

Genotyping and sero-virological characterisation of hepatitis B virus-infected blood donors in Central Eritrea

Aim To determine serological markers and genotypes profiles of HBV isolates in Central Eritrea. Materials & methods A total of 191 HBsAg sero-positive samples were randomly selected for the study from 23,232 screened blood donors from 2015 to 2017. Enzyme-linked immunosorbent assay was used to perform HBV serological markers screening, while genotypes were determined using type-specific primer-based multiplex-nested PCR. Results The median age of infected blood donors was 28.9 ± 10.26 years. Of 191 HBsAg reactive serum samples, 77.5% (148/191) were sero-positive for HBcAb-total, among which 99.3% (147/148) and 0.7% (1/148) were sero-positive for HBsAg and HBsAb, respectively. Interestingly, among 147 HBcAb-total/HBsAg reactive samples, 16 (10.9%) and 131 (77.9%) were sero-positive for HBeAg and HBeAb, respectively. For genotyping, 73 HBV isolates were successfully amplified and genotyped, with 59 (80.8%) had a mono-genotype and 14 (19.2%) had a mixed-genotype infection. Among HBV isolates with mono-genotypes: 39 (53.4%) D; 10 (13.7%) E; 6 (8.2%) A and 4 (5.5%) C. While five mixed-genotypes comprised: 6 (8.2%) C/D; 3 (4.1%) C/D/E; 2 (2.7%) each with genotype A/D and D/E; and 1 (1.4%) genotype B/D. Conclusion HBV genotype D is the predominant genotype, either as a HBV mono- or mixed-genotype infection, among blood donors in Eritrea.

Presence of Human G Rotavirus Genotypes Amongst Bovine Calves and its Combination with P Genotypes

Background: Bovine Rotavirus is one of the most important viral etiological agent responsible for causing neonatal diarrhea incurring severe economic loss to farmers. The presence of large genome size, segmented nature and absence of proof reading activity of RNA polymerase leads to frequent reassortment and thus emergence of new G and P types with ability of interspecies transmission. Methods: During an epidemiological study (July 2016 to July 2019) 200 diarrheic fecal samples were screened for Bovine Rotavirus A using ELISA and RNA-PAGE. Further, twenty two positive samples for RVA were subjected to molecular detection for VP6, VP4 and VP7 genes. Result: Ten (20/200) and 11(22/200) percent diarrheic fecal samples were found positive using ELISA and RNA-PAGE respectively. Twenty samples found positive in ELISA were also found positive in RNA-PAGE. Amongst which 22 (100%) samples were found positive for VP6, while 15 (68.18%) samples showed amplification for VP4 and VP7 gene. All Rotavirus A positive samples were genotyped by multiplex RT-PCR assay. G1G3 was found to be most predominant (53.33%) followed by G3 (26.66%), while one sample each showed the presence of G1G5 and G3G8 (6.66%). Ten samples showed mixed genotype (66.66%). One sample was non typeable (6.66%). Among the P types, P[11] was the most predominant (73.33%), while one sample each showed the presence of P[5] and P[5]P[11] (6.66%) and 02 samples were non typeable (13.33%). The G and P genotype combination determined in 12 samples were as follows; G3P [11] found in two samples (16.66%), G3P[5] in 01(08.33%), G1G5P[11] in 01(08.33%), G1G3P[11] in 07 (58.33%), while 01 (08.33%) sample had mixed genotype G1G3P[5]P[11] combination.

Biochemical and Virulence Characterization of Vibrio vulnificus Isolates From Clinical and Environmental Sources

Vibrio vulnificus is a deadly human pathogen for which infections occur via seafood consumption (foodborne) or direct contact with wounds. Virulence is not fully characterized for this organism; however, there is evidence of biochemical and genotypic correlations with virulence potential. In this study, biochemical profiles and virulence genotype, based on 16S rRNA gene (rrn) and virulence correlated gene (vcg) types, were determined for 30 clinical and 39 oyster isolates. Oyster isolates were more biochemically diverse than the clinical isolates, with four of the 20 tests producing variable (defined as 20–80% of isolates) results. Whereas, for clinical isolates only mannitol fermentation, which has previously been associated with virulence potential, varied among the isolates. Nearly half (43%) of clinical isolates were the more virulent genotype (rrnB/vcgC); this trend was consistent when only looking at clinical isolates from blood. The majority (64%) of oyster isolates were the less virulent genotype (rrnA or AB/vcgE). These data were used to select a sub-set of 27 isolates for virulence testing with a subcutaneously inoculated, iron-dextran treated mouse model. Based on the mouse model data, 11 isolates were non-lethal, whereas 16 isolates were lethal, indicating a potential for human infection. Within the non-lethal group there were eight oyster and three clinical isolates. Six of the non-lethal isolates were the less virulent genotype (rrnA/vcgE or rrnAB/vcgE) and two were rrnB/vcgC with the remaining two of mixed genotype (rrnAB/vcgC and rrnB/vcgE). Of the lethal isolates, five were oysters and 11 were clinical. Eight of the lethal isolates were the less virulent genotype and seven the more virulent genotype, with the remaining isolate a mixed genotype (rrnA/vcgC). A discordance between virulence genotype and individual mouse virulence parameters (liver infection, skin infection, skin lesion score, and body temperature) was observed; the variable most strongly associated with mouse virulence parameters was season (warm or cold conditions at time of strain isolation), with more virulent strains isolated from cold conditions. These results indicate that biochemical profiles and genotype are not significantly associated with virulence potential, as determined by a mouse model. However, a relationship with virulence potential and seasonality was observed.

Damar İçi Madde Bağımlılığı Olan ve Madde Bağımlısı Olmayan Hastalar Arasında Hepatit C Virus (HCV) Genotiplerinin Dağılımı

Genotype distribution of hepatitis C virus (HCV) can vary over the years between different patient groups and regions. The prevalence of intravenous drug users (IVDU) is known to increase in our country, yet there are a limited number of studies investigating the distribution of HCV genotypes in this group. These data are essential for monitorization of the changes in HCV epidemiology. The present study aimed to evaluate the five-year results of HCV genotyping among patients infected with HCV related to IVDU and unrelated to drug use. Plasma samples of 720 patients (HCV antibody, HCV RNA positive), which were sent to our laboratory for HCV genotyping between January 2014-March 2019 were analyzed. HCV RNA extraction from plasma samples was performed in the automated-extraction system of EZ1 advanced (Qiagen, Germany) using the EZ1 virus mini kit v2.0 (Qiagen, Germany). Amplicons were obtained by amplifying the 5’NCR and core gene region in the Rotorgene 6000 real-time PCR (Qiagen, Germany) device with the HCV RNA real-time quantitative 2.0 (NLM, Italy) kit. For the genotyping, a commercial line probe assay (LIPA) based on in vitro reverse hybridization GEN-C2.0 kit (NLM, Italy) which can distinguish 1, 2, 3, 4, 6 genotypes and 1a, 1b, 2a/c, 2b, 3a, 3b, 3c, 3k, 4a, 4b, 4c/d, 4e, 4f, 4h, 5a, 6a/b, 6g, 6f/q, 6m, 7a subtypes of HCV, based on variations in the 5’-NCR and core regions was used. HCV genotype distribution of 266 IVDU (93.2%: male; median age: 25 ± 6.82) and 454 non-drug users (51.3%: male; median age: 56.5 ± 16.06) were examined. In order of frequency in the group with IVDU; genotype 1a, 3a, 1b, 4c/d, 2b, 4, 3 were observed and genotype 1, 2a/c and mixed genotype (1+3a) were detected in one patient. In the group without IVDU, in order of frequency; genotype 1b, 1a, 3a, 1, 2a/c, 4 were observed and genotype 2b, 4c/d, 5a, 6a/b, 6 and mixed genotype (3+4) were detected in one patient. Genotypes 1a and 3a were significantly higher in the IVDU group (p< 0.00001, p< 0.00001), while 1b was significantly higher in patients without IVDU (p< 0.00001). Genotypes 1a and 3a were more common in young men (p< 0.00001, p= 0.000163), while 1b was higher in middleaged women (p< 0.00001). The incidence of genotypes 1b (p= 0.021) and 3a (p= 0.012) was higher in foreign nationals than the Turkish patients. When the HCV genotype distribution was examined by years, it was observed that the percentages of genotype 1b and 1a were decreasing, while the percentage of genotype 3a was increasing. As a result, in this study, HCV genotype distribution among IVDU was observed to be different from the general population without IVDU. It was found that genotypes 1a and 3a were more common in the IVDU group. As in the other regions of our country, genotype 1b was found most frequently in the general population. Genotype 3a increases significantly compared to years. In our study, the determination of genotypes existing in different parts of the world may be due to the foreign nationals living in our city and our region is a tourism center. It is also necessary to investigate whether there is an increase in IVDU over the years.