Introduction. The centrifugal-flotation technique allows all nematode species and life stages to be separated from root debris and residual soil particles to facilitate their observation. The principle of the method applied, key advantages, starting plant material and time required are presented. Materials and methods. Necessary laboratory materials, and details of the 19 steps required for extracting nematodes from plant tissue for counting are described. Possible troubleshooting is explained.

The number of cells in a preimplantation embryo is directly correlated to the health and viability of the embryo. There are currently no methods to count the number of cells in late-stage preimplantation embryos noninvasively. We assessed the ability of optical quadrature microscopy (OQM) to count the number of cells in mouse preimplantation embryos noninvasively. First, to test for possible light toxicity, we exposed two-cell mouse embryos to OQM and differential interference contrast (DIC) microscopy and assessed their ability to develop to the blastocyst stage. We found no inhibition of development from either mode of microscopy for up to 2 h of light exposure. We also imaged eight-cell to morula stage mouse preimplantation embryos by OQM nd developed two methods for counting the number of cells. The contour signature method (CSM) used OQM images alone and the phase subtraction method (PSM) used both OQM and DIC images. We compared both methods to standard cell counting techniques and found that the PSM was superior to all other noninvasive cell counting methods. Our work on mouse embryos should be applicable to human embryos. The ability to correctly count the number of cells in human preimplantation embryos could lead to the transfer of fewer embryos in in vitro fertilization (IVF) clinics and consequently a lower rate of high-risk multiple-infant births.

Somatic cell counting by the Coulter Counter method, California mastitis test and bacteriological examination were performed on milk samples taken at fortnightly intervals throughout lactation from healthy primiparous and iniiltiparous ewes of the Chios and Karagouniki breeds and from healthy ewes in five grazing flocks in northern Greece.

The proportion of the bacteriologically positive milk samples was 0·22. The prevalence of subclinical mastitis ranged, between the flocks, from 29 to 43% of the ewes. Subclinical mastitis occurred more frequently in Karagouniki than in Chios eives and in multiparous than in primiparous ewes of both breeds. Coagulase-negative staphylococci and S. aureus were isolated in 44 and 33% of the bacteriologically postive samples, respectively. No parity, breed or flock differences in normal somatic cell count were noticed, but stage of lactation influenced normal somatic cell count both in primiparous and multiparous ewes. The results of this investigation also suggest that (a) ovine milk samples with somatic cell count more than 1000 × 103 cells per ml, should be tested bacteriologically, (b) somatic cell count values between 1000 and 2000 × 103 cells per ml should cause suspicion about the presence of coagulase-negative staphylococci in the milk, and (c) the presence of S. aureus in ewe's milk results in a significant increase in somatic cell count, but the wide range of the values does not permit us to suggest any threshold value. The significant correlation between the results of the Coulter Counter somatic cell count method and the Califortiia mastitis test, indicates that the latter can be used for the determination of somatic cell count in ewe's milk.