Diagnostic value of rK39 dipstick in zoonotic visceral leishmaniasis in Turkey

K39 is a repetitive immunodominant epitope in a kinesin-related protein expressed predominantly in the amastigotes of visceral Leishmania spp. Enzyme immunoassays of patient's sera with recombinant K39 (rK39) proved to be highly specific and sensitive for diagnosis of active visceral leishmaniasis (VL, kala-azar). The same assays in dipstick format were also found effective for diagnosis of both human VL (HVL) and canine VL (CanVL) in most endemic areas of these diseases. Fifty-eight human patients and 22 dogs, clinically suspected of kala-azar, were screened with rK39 dipstick in comparison with the conventional methods of diagnosis, i.e., microscopic examinations of bone marrow and lymph node aspirates and immunofluorescent antibody tests (IFAT), respectively. Sixteen patients and 12 dogs were found to be rK39 dipstick positive. The results were corroborated with those of parasitological examinations, except 1, rK39-positive but smear-negative, case in each group. IFAT of the 2 discordant cases gave positive results. The rK39 dipstick is thus reliable for diagnosis of both HVL and canVL cases in Turkey.     

A novel method for transuterine identification of piglets

Implantable microchips provide a secure, permanent and unique identification of individual animals. When performing fetal intervention studies in experimental animal models easy and secure identification of fetuses is desirable, as having test and control groups within the same uterus reduces the total number of animals used in a study. The aims of this study were: (1) to establish a protocol to identify porcine fetuses in utero by microchip implantation and (2) to assess postnatally whether clinical or pathological reactions to the implant occurred. Two Danish Landrace/Danish Large White crossbred sows at day 100 of gestation were used. The sows were sedated with azaperone and induced with propofol intravenously. Anaesthesia was maintained with isoflurane and oxygen. Antibiotics were administered intramuscularly (i.m.) at induction and analgesia was given pre-, intra- and postoperatively. A laparotomy was performed and the uterus exteriorized. The rump of the first fetus was recognized through the uterine wall and the thigh muscle of the fetus was fixed between the thumb and the forefinger. The microchip was then implanted into the fetus at an angle of 45 degrees i.m. in the lateral hindleg using an insertion device with a 12G needle. The same procedure was done in every fetus. The uterus was returned to the abdomen and the abdominal wall closed. The sows gave birth to 24 liveborn piglets and one stillborn. None of the liveborn piglets were limping at the time of birth and no visible cutaneous or palpable reactions on the hindlegs were observed. Following euthanasia, the microchip was easily localized and no macroscopic reactions at the implantation site were seen. None of the piglets had more than one microchip implanted. Histology showed a chronic mild foreign body granulomatous inflammatory response with peripheral eosinophils surrounding the microchip. No inflammation was evident in the adjacent muscles. It is concluded that transuterine identification of piglets two weeks before delivery is feasible using a microchip implant as an effective, easy and reliable method for identification of individuals after birth.     

Fetal gene transfer by transuterine injection of cationic liposome-DNA complexes

In utero injection of cationic liposome-DNA complexes (CLDCs) containing chloramphenicol acetyltransferase, beta-galactosidase (beta-gal), or human granulocyte colony-stimulating factor (hG-CSF) expression plasmids produced high-level gene expression in fetal rats. Tissues adjacent to the injection site exhibited the highest levels of gene expression. Chloramphenicol acetyltransferase expression persisted for at least 14 days and was reexpressed following postnatal reinjection of CLDCs. Intraperitoneal administration of the hG-CSF gene produced high serum hG-CSF levels. X-gal staining demonstrated widespread beta-gal expression in multiple fetal tissues and cell types. No toxic or inflammatory responses were observed, nor was there evidence of fetal-maternal or maternal-fetal gene transfer, suggesting that CLDCs may provide a useful alternative to viral vectors for in utero gene transfer.