Molecular and biological characterisation of Cryptosporidium in pigs

OBJECTIVE: Genetic and biological characterisation of 12 isolates of Cryptosporidium from pigs and comparing them with Cryptosporidium isolates from humans and cattle. DESIGN: Cryptosporidium isolates from pigs were compared with those obtained from human and cattle using rDNA sequence analysis. The infectivity of two of the porcine isolates was determined in neonatal mice and the clinical history of the infected pigs recorded. RESULTS: Pig-derived isolates of Cryptosporidium exhibited two distinct genotypes; a porcine genotype and a bovine genotype, which is common to cattle and other livestock. The porcine genotype did not produce any infection in neonatal mice whereas the bovine genotype did. CONCLUSION: Two distinct genetically and biologically differing strains of Cryptosporidium appeared to be associated with acute diarrhoea in pigs. Whether Cryptosporidium was a primary or secondary pathogen is unclear but warrants further investigation. As the bovine genotype is known to infect humans, the results suggest that pigs can act as reservoirs of cryptosporidial infections for humans and other live-stock. The zoonotic potential of the pig-adapted genotype is uncertain and requires further study.     

Movement of dieldrin through soils: II.—In sloping troughs and soil columns

A glass-lined trough was divided into six compartments by vertical transverse partitions and filled with soil to a height of 5 cm above the top of each partition. The base of the box and the surface of the soil sloped forwards one end with a gradient of 1 in 3·7. Dieldrin was applied (22 kg active ingredient/ha) to the soil surface in the uppermost compartment and the movement of dieldrin into leachate and down the slope was followed for 17 weeks. During this time 19·5 cm of rain fell. Less than 0·02% of the dieldrin appeared in the leachate and 99% of this was collected during the first 9 weeks, mainly from the uppermost compartment. Very little dieldrin moved down the slope and it was not detected in the leachate from compartments other than the treated one during 9–17 weeks after treatment. In other experiments, 3 cylinders were filled respectively with sandy loam, heavy clay loam and peat which were transferred from the field as intact cores. Three other cylinders were filled with soils of the same types but broken up. Approximately 2% of the dieldrin leached through one column but usually it was much less than 0·1%. The largest amounts of dieldrin were leached down columns of heavy clay loam, intermediate amounts down columns of peat, and least through sandy loam. Ten times as much dieldrin leached down the columns of intact soil as through those with broken soil. The results indicate that the movement of dieldrin from treated soil into water systems by leaching is limited and is unlikely to be a major pathway for the contamination of water.     

Effect of activation methods for bovine oocytes after intracytoplasmic injection

A principal nuclear transfer procedure is to inject a donor cell into the perivitelline space in an enucleated oocyte and then electric fusion is performed (cell fusion method). The effects of activation methods in reconstructed oocytes for the serum-starved somatic cell cloning procedure were investigated in this study by means of intracytoplasmic injection (i.c.i.). Bovine oocytes were enucleated at 18-22 h for in vitro maturation, and subsequently the nucleus of cumulus cell collected from Japanese Black Bulls (JBCC) after 5-7 days of starved culture was injected into the recipient cytoplast with a piezo-micromanipulator. At 1 h after i.c.i., reconstructed oocytes were stimulated with ethanol (ET) or calcium ionophore (CaI) as the first activation treatment, followed by cycloheximide (CHX) or 6-dimethylaminopurin (DMAP) treatment as the second activation. In the experiment on the first activation method, the proportion of reconstructed oocytes developing to the blastocyst stage was significantly (p<0.01) higher in the ET activation method than that with CaI (10.5% and 4.7%, respectively). And the experiment on the second activation method after ET treatment showed similar proportions of blastocyst development in both CHX and DMAP treatments (5.9% and 2.8%, respectively). The present results indicated that combined activation treatment with ET and CHX was efficient for reconstructed bovine oocytes by i.c.i.