Bivariate flow cytometry of farm animal chromosomes: a potential tool for gene mapping

Bivariate flow karyotypes of chromosomes from sheep, cattle and pig lymphocytes and from a cattle-mouse somatic cell hybrid line were obtained using a dual laser fluorescence-activated cell sorter (FACS). Pig chromosomes were resolved into 19-20 peaks, indicating that most, if not all, pig chromosomes could be separated by this technique. Sheep chromosomes showed incomplete separation but three clear peaks, presumably representing the three large metacentric chromosomes, plus five other clusters were obtained. Cattle chromosomes showed poor separation but about four peaks could be distinguished, indicating that certain chromosomes could be sorted in this species. The use of cattle-mouse hybrids may enable other individual cattle chromosomes to be obtained. It is concluded that FACS separation will be a useful additional tool for gene mapping.     

Application of a reverse dot blot DNA–DNA hydridization method to quantify host‐feeding tendencies of two sibling species in the Anopheles gambiae complex

A DNA–DNA hybridization method, reverse dot blot analysis (RDBA), was used to identify Anopheles gambiae s.s. and Anopheles arabiensis (Diptera: Culicidae) hosts. Of 299 blood‐fed and semi‐gravid An. gambiae s.l. collected from Kisian, Kenya, 244 individuals were identifiable to species; of these, 69.5% were An. arabiensis and 29.5% were An. gambiae s.s. Host identifications with RDBA were comparable with those of conventional polymerase chain reaction (PCR) followed by direct sequencing of amplicons of the vertebrate mitochondrial cytochrome b gene. Of the 174 amplicon‐producing samples used to compare these two methods, 147 were identifiable by direct sequencing and 139 of these were identifiable by RDBA. Anopheles arabiensis bloodmeals were mostly (94.6%) bovine in origin, whereas An. gambiae s.s. fed upon humans more than 91.8% of the time. Tests by RDBA detected that two of 112 An. arabiensis contained blood from more than one host species, whereas PCR and direct sequencing did not. Recent use of insecticide‐treated bednets in Kisian is likely to have caused the shift in the dominant vector species from An. gambiae s.s. to An. arabiensis. Reverse dot blot analysis provides an opportunity to study changes in host‐feeding by members of the An. gambiae complex in response to the broadening distribution of vector control measures targeting host‐selection behaviours.     

Long-term response and recovery to nutrient addition of a partitioned arctic lake

1. To study the bottom‐up linkages in arctic lakes, we treated one side of a partitioned lake with inorganic nitrogen and phosphorus for a 6‐week period each summer for 6 years starting in the summer of 1985. We took a variety of weekly measurements to determine the impact of the nutrient loading on the lake and continued weekly measurements for 2–6 years after the cessation of nutrient loading to observe the recovery of the treated side. The loading rates (2.91 mmol N m^?2 day^?1 and 0.23 mmol P m^?2 day^?1) were five times the calculated loading rates for Toolik Lake, located nearby. 2. In all 6 years of nutrient addition, phytoplankton biomass and productivity were greater in the treated sector than the reference sector. In the first 4 years of nutrient addition there was no flux of phosphorus from the mineral‐rich sediments. This changed in the last 2 years of nutrient addition as phosphorus was released to the lake. 3. The response of the animal community to increased plant production was mixed. One of the four macro‐zooplankton species (Daphnia longiremis) increased in number by about twofold in the first 5 years. However, the copepod Cyclops scutifer showed no response during the treatment phase of the study. The benthic invertebrate response was also mixed. After a 2‐year lag time the snail Lymnaea elodes increased in the treated lake sector but chironomids did not. 4. Ecosystem response to fertilisation was not controlled solely by nutrient addition because phosphorus was not recycled from the sediments until the last 2 years of nutrient addition. Phytoplankton still showed the effects of nutrient addition in the recovery period and the hypolimnion of the treated sector was still anaerobic starting at 6 m in 1996.