Selection of the most appropriate medium for assessing motility and DNA uptake of bovine spermatozoa

The aim of this study was to select the best medium to maintain sperm motility during sperm-DNA incubation and assess the DNA uptake by spermatozoa of Iranian Holstein bulls and its effects on sperm motility. Frozen sperms from an Iranian Holstein bull were thawed and centrifuged. Motile sperms were separated through Puresperm gradient [40/80%] followed by two times washing in SP-TALP medium. Then, sperms were washed once [PBS, Opti-MEM and SP-TALP] and incubated with DNA in each media followed by sperm motility estimation. The plasmid pEGFP-C1 was linearized and incubated with sperms at 37°C for 1 hour. Sperm-DNA mixture was treated with DNase I and the sperm pellet was washed with PBS. DNA extraction from sperms and supernatants from the last washing were used as template for PCR. Data was analyzed using SAS package and mean comparisons between sperm motility in different media were performed. Sperm motility after incubation in PBS, Opti-MEM and SP-TALP were 40[ +/- 2.89], 2[ +/- 1.53] and 54[ +/- 4.41] percent, respectively. PCR results from transfected sperms indicated that EGFP transgene internalized into the bovine sperms and DNaseI treatment could not eliminate it. In conclusion the best medium for sperm and DNA incubation was SPTALP. The DNA not only could attach to the post acrosomal region of spermatozoa but also could integrate into it. So bovine spermatozoa can be used as transgene carrier into oocyte

Typhoid fever: an epidemiologic survey of suspected cases referred to diagnostic laboratories in Rasht, 2002-2003

Typhoid fever has been endemic in Iran throughout its history. This Study was conducted in response to the rising number of cases reported from Rasht [capital of Gilan province in northern Iran]. we performed a descriptive study on 2031 suspected cases referred to city's diagnostic laboratories in 2002 and 2003. A [definite] diagnosis was defined as Vidal test titers rising 80-fold or more over a period of one week, or a positive bacteriologic test. We completed questionnaires for the definite cases, and used the EPI-INFO computer package to analyze the data. The serologic prevalence was 3.94%, and 25% of the seropositive individuals had positive bacterial cultures. The geometric mean of the reciprocal titers [GMRT] was 1: 1 80. The prevalence was the same in both sexes and among all age groups. Factors showing a statistically significant relationship with prevalence include occupation [the greatest frequency was seen in housewives], education level, place of residence, and source of drinking water. We conclude that high prevalence rates coupled with the ecological conditions in this area could favor the occurrence of salmonellosis outbreaks: the issue requires more extensive research for further clarification