Genetic polymorphism detection of two α-Casein genes in three Egyptian sheep breeds

Sheep milk is an excellent raw material for the milk processing industry especially in cheese production. The protein content and composition of sheep milk are important in the cheese manufacturing. The casein fraction of ruminant milk proteins consists of four caseins, namely αs1, αs2, β and κ-Casein. Casein genetic polymorphisms are important due to their effects on quantitative traits and technological properties of milk.This study aimed to detect the genetic polymorphism of αs1- and αs2-Casein genes in three native Egyptian sheep breeds; Rahmani, Barki and Ossimi. PCR-SSCP and PCR-RFLP were used to detect the genetic polymorphism of αs1-CN and αs2-CN genes, respectively.A 223-bp fragment of αs1-CN gene was amplified by PCR and SSCP results recorded the presence of three different patterns; TT, TC and CC; in 87 tested sheep animals. The sequence analysis of two homologous patterns showed a single nucleotide polymorphism (SNP) (T → C) at position 170. The frequencies of three patterns in the tested sheep breeds were 43.33%, 50.00%, and 6.67% in Rahmani; 83.33%, 13.33%, and 3.33% in Ossimi and 74.07%, 22.22%, and 3.70% in Barki, respectively. Our nucleotide sequences of αs1-CN T and C alleles were submitted to GenBank with the accession numbers KF018339 and KF018340, respectively.The restriction digestion of αs2-CN PCR product (1300-bp) by Tru1I endonuclease revealed three different genotypes; AA, AG and GG with frequencies of 66.67%, 30.00%, and 3.33% in Rahmani; 96.67%, 3.33%, and 0.00% in Ossimi and 96.15%, 3.85%, and 0.00% in Barki, respectively. The sequence analysis revealed the presence of a single nucleotide polymorphism (A → G) in intron 6 of αs2-CN gene. Our nucleotide sequence of αs2-CN gene was submitted to GenBank with the accession number JX080380.     

Patients Are Paying Too Much for Tuberculosis: A Direct Cost-Burden Evaluation in Burkina Faso

Background

Paying for health care may exclude poor people. Burkina Faso adopted the DOTS strategy implementing “free care” for Tuberculosis (TB) diagnosis and treatment. This should increase universal health coverage and help to overcome social and economic barriers to health access.

Methods

Straddling 2007 and 2008, in-depth interviews were conducted over a year among smear-positive pulmonary tuberculosis patients in six rural districts of Burkina Faso. Out-of-pocket expenses (direct costs) associated with TB were collected according to the different stages of their healthcare pathway.

Results

Median direct cost associated with TB was US$101 (n = 229) (i.e. 2.8 months of household income). Respectively 72% of patients incurred direct costs during the pre-diagnosis stage (i.e. self-medication, travel, traditional healers'' services), 95% during the diagnosis process (i.e. user fees, travel costs to various providers, extra sputum smears microscopy and chest radiology), 68% during the intensive treatment (i.e. medical and travel costs) and 50% during the continuation treatment (i.e. medical and travel costs). For the diagnosis stage, median direct costs already amounted to 35% of overall direct costs.

Conclusions

The patient care pathway analysis in rural Burkina Faso showed substantial direct costs and healthcare system delay within a “free care” policy for TB diagnosis and treatment. Whether in terms of redefining the free TB package or rationalizing the care pathway, serious efforts must be undertaken to make “free” health care more affordable for the patients. Locally relevant for TB, this case-study in Burkina Faso has a real potential to document how health programs'' weaknesses can be identified and solved.     

Dorsal and Ventral Hippocampus Modulate Autonomic Responses but Not Behavioral Consequences Associated to Acute Restraint Stress in Rats

Recent evidence has suggested that the dorsal (DH) and the ventral (VH) poles of the hippocampus are structurally, molecularly and functionally different regions. While the DH is preferentially involved in the modulation of spatial learning and memory, the VH modulates defensive behaviors related to anxiety. Acute restraint is an unavoidable stress situation that evokes marked and sustained autonomic changes, which are characterized by elevated blood pressure (BP), intense heart rate (HR) increases, skeletal muscle vasodilatation and cutaneous vasoconstriction, which are accompanied by a rapid skin temperature drop followed by body temperature increases. In addition to those autonomic responses, animals submitted to restraint also present behavioral changes, such as reduced exploration of the open arms of an elevated plus-maze (EPM), an anxiogenic-like effect. In the present work, we report a comparison between the effects of pharmacological inhibition of DH and VH neurotransmission on autonomic and behavioral responses evoked by acute restraint stress in rats. Bilateral microinjection of the unspecific synaptic blocker cobalt chloride (CoCl_2, 1mM) into the DH or VH attenuated BP and HR responses, as well as the decrease in the skin temperature, elicited by restraint stress exposure. Moreover, DH or VH inhibition before restraint did not change the delayed increased anxiety behavior observed 24 h later in the EPM. The present results demonstrate for the first time that both DH and VH mediate stress-induced autonomic responses to restraint but they are not involved in the modulation of the delayed emotional consequences elicited by such stress.     

Influence of inoculation with plant growth promoting rhizobacteria (PGPR) on tomato plant growth and nematode reproduction under greenhouse conditions

Numerous species of soil bacteria which flourish in the rhizosphere of plants or around plant tissues stimulate plant growth and reduce nematode population by antagonistic behavior. These bacteria are collectively known as PGPR (plant growth promoting rhizobacteria). The effects of six isolates of PGPR Pseudomonas putida, Pseudomonas fluorescens, Serratia marcescens, Bacillus amyloliquefaciens, Bacillus subtilis and Bacillus cereus, were studied on tomato plant growth and root knot nematode reproduction after 45 days from nematode infection. The highest number of shoot dry weight/g (43.00 g) was detected in the plant treated with S. marcescens; then P. putida (34.33 g), B. amyloliquefaciens (31.66 g), P. fluorescens (30.0 g), B. subtilis (29.0 g), B. cereus (27.0 g) and nematode alone (untreated) 20 g/plant. While the highest number of plant height was observed when plant was treated with S. marcescens, P. fluorescens, P. putida, B. amyloliquefaciens and P. putida 52.66, 50.66, 48 and 48 cm respectively. No significant differences were seen between previous treatments but only had significant differences compared with untreated plant. The highest number of fruit/plant was observed when plants were treated with S. marcescens (10.66), then B. amyloliquefaciens (8.66), P. putida (8), P. fluorescens (8) and B. cereus (7.66). No significant differences between the last 4 treatments, but all had significant differences compared with untreated plants. The highest weight of plant yield (g) was observed with S. marcescens (319.6 g/plant) and the lowest weight of plant yield was observed in plants treated with nematode alone (untreated). On the other hand, the lowest numbers of J₂/10 g of soil (78), galls/root, (24.33) galls/root, egg masses/root (12.66) and egg/egg masses were observed in the plants treated with S. marcescens.     

Molecular Genetic Diversity in Populations of Fusarium pseudograminearum from Tunisia

Fusarium pseudograminearum is one of the major pathogens causing crown rot of wheat in the semi‐arid and arid areas in Tunisia. In this study, the molecular diversity of 74 isolates of F. pseudograminearum representing three populations from Tunisia and a set of isolates from the world collection was investigated. The potential mycotoxin‐producing ability was tested by PCR using primer pairs specific for the Tri3, Tri7 and Tri13 genes. Results indicated that all the isolates are potentially DON and/or 3‐AcDON producers. The mating‐type idiomorphs were identified using diagnostic PCR primer for MAT1‐1 and MAT1‐2. Both mating types were recovered from the same region and in some cases from the same field. Restriction analysis of the nuclear ribosomal DNA (nrDNA) intergenic spacer region (IGS) revealed 11 haplotypes, five of which were identified in the world collection. The analysis of population structure using the combined IGS and MAT data revealed that the total gene diversity (H_T = 0.108) was mostly attributable to diversity within populations (H_S = 0.102) and that the genetic differentiation among the four populations was low (G_ST = 0.09). The analysis of molecular variance (amova ) showed that 15% of the variability was between the Tunisian populations and the world collection. These findings indicate that quarantine measures should be in place to limit the introduction of new populations of F. pseudograminearum into Tunisia.     

Non-contiguous finished genome sequence and description of Bacteroides neonati sp. nov., a new species of anaerobic bacterium

Bacteroides neonati strain MS4^T, is the type strain of Bacteroides neonati sp. nov., a new species within the genus Bacteroides. This strain, whose genome is described here, was isolated from a premature neonate stool sample. B. neonati strain MS4^T is an obligate anaerobic Gram-negative bacillus. Here we describe the features of this organism, together with the complete genome sequence and annotation. The 5.03 Mbp long genome exhibits a G+C content of 43.53% and contains 4,415 protein-coding and 91 RNA genes, including 9 rRNA genes.     

Hepatitis C virus genotype 3a with phylogenetically distinct origin is circulating in Pakistan

Background

Hepatitis C virus (HCV) is one of the leading causes of viral hepatitis worldwide and its genotype 3a is predominant in vast areas of Pakistan.

Findings

The present study reports the first full sequence of HCV 3a isolate PK-1 from Pakistan. This nucleotide sequence was compared with six other HCV genotype 3a full length sequences from different regions of the world by using statistical methods of phylogenetic analysis.

Conclusion

The nucleotide difference of these seven sequences shows that HCV genotype 3a of phylogenetically distinct origin is circulating in Pakistan.