Molecular Detection and Identification of Zoonotic Microsporidia Spore in Fecal Samples of Some Animals with Close-Contact to Human

Background: Microsporidia species are obligatory intracellular agents that can infect all major animal groups including mammals, birds, fishes and insects. Whereas worldwide human infection reports are increasing, the cognition of sources of infection particularly zoonotic transmission could be helpful. We aimed to detect zoonotic microsporidia spore in fecal samples from some animals with close – contact to human. Methods: Overall, 142 fecal samples were collected from animals with closed-contact to human, during 2012-2013. Trichrome – blue staining were performed and DNA was then extracted from samples, identified positive, microscopically. Nested PCR was also carried out with primers targeting SSU rRNA gene and PCR products were sequenced. Results: From 142 stool samples, microsporidia spores have been observed microscopically in 15 (10.56%) samples. En. cuniculi was found in the faces of 3 (15%) small white mice and 1 (10%) laboratory rabbits(totally 2.81%). Moreover, E. bieneusi was detected in 3 (10%) samples of sheep, 2 (5.12%) cattle, 1 (10%) rabbit, 3 (11.53%) cats and 2 (11.76%) ownership dogs (totally 7.74%). Phylogenetic analysis showed interesting data. This is the first study in Iran, which identified E. bieneusi and En. Cuniculi in fecal samples of laboratory animals with close – contact to human as well as domesticated animal and analyzed them in phylogenetic tree. Conclusion: E. bieneusi is the most prevalent microsporidia species in animals. Our results can also alert us about potentially zoonotic transmission of microsporidiosis.Key Words: Laboratory animals, Enterocytozoon bieneusi, Encephalitozoon cuniculi, Zoonotic transmission     

Evidence for a Leukocyte Adhesion Factor Produced by the Early Embryo

PROBLEM : To determine if the embryo may induce adhesive molecules needed for implantation. METHOD : Determination of whether platelet rosetting around lymphocytes might occur when exposed to sera from pregnant, but not nonpregnant patients and from culture fluid from embryos but not oocytes. RESULTS : 90.2% of women with positive sera beta human chorionic gonadotropin (P-hCG) levels taken at least 12 days postovulation demonstrated platelet rosette factor (PRF) vs only 18.7% when β-hCG was negative. Using mid-luteal phase sera in women receiving hCG injection 1 wk before, 64.7% had positive PRF when serial β-hCG levels were positive as did 100% of samples taken from in vitro fertilization (IVF) patients; however, only 15.3% were positive with negative serial hCG levels. Culture media from fertilized oocytes and embryos tested positive for PRF, but follicular fluid and media from unfertilized oocytes were negative. CONCLUSION : The early embryo secretes a factor(s) that gains access to maternal serum and promotes increased lymphocyte/platelet adhesiveness.     

EVALUATION OF RISK FACTORS AFFECTING ANASTOMOTIC LEAKAGE AFTER REPAIR OF ESOPHAGEAL ATRESIA

Background:

Anastomotic leak are reported among neonates who underwent esophageal atresia.

Aim:

To find risk factors of anastomotic leakage in patients underwent esophageal repair.

Methods:

All cases with esophageal atresia were included. In this case control study, patients were classified in two groups according to presence or absence of anastomotic leaks. Duration of study was 10 years.

Results:

Sixty-one cases were included. Mean±SD age at time of surgery in patients with leakage and without leakage was 9.50±7.25 and 8.83±6.93 respectively (p=.670). Blood transfusion and two layer anastomosis had significant correlation with anastomotic leakage.

Conclusion:

Blood transfusion and double layer anastomosis are associated with higher rate of anastomotic leakage.     

Effect of freeze–thawing procedure on chromatin stability, morphological alteration and membrane integrity of human spermatozoa in fertile and subfertile men

Cryopreservation is known to impair sperm motility and decrease the fertilization rate by detrimental effects on acrosomal structure and acrosin activity. However, the consequences of cryopreservation on the integrity of the sperm nucleus, chromatin stability and centrosome are less clear. The present study was designed to determine the effect of the freeze-thawing procedure on chromatin condensation (aniline blue staining) and the morphology (strict criteria) and membrane integrity of human spermatozoa. The structural and functional characteristics of the sperm plasma membrane were measured by the eosin-test and hypo-osmotic swelling test which were done separately. Sperm cryopreservation was performed on semen samples from two groups of men classified as fertile (n = 20) and subfertile (n = 72), based on their reproductive history and semen analysis according to WHO guidelines. The mean percentage of condensed chromatin, morphologically normal spermatozoa and membrane integrity in all semen samples investigated (n = 92) decreased significantly (p = 0.0001) after freeze-thawing, in comparison to the value observed prior to freezing. By comparing the semen samples between fertile and subfertile patients, significantly (p = 0.0009) greater damage was demonstrated in the subfertile than in the fertile group. Furthermore, no significant difference was observed between the two groups with regard to the morphological alteration and structural as well as functional damage of the sperm membrane. In conclusion, the freeze-thawing procedure significantly affects chromatin structure and sperm morphology, especially in the head and the tail regions, and this may explain the lower fertilization rate and IVF/ICSI outcome when frozen-thawed spermatozoa are used. In addition, this study demonstrates that chromatin condensation is a sensitive parameter for the evaluation of cryodamage of semen samples from fertile and subfertile patients, though subfertile patients with very poor semen characteristics have yet to be studied. It is therefore recommended that chromatin condensation be used as an additional parameter for the assessment of sperm quality after freeze-thawing.