Western blot analysis of Leishmania infantum antigens using sera from pentamidine-treated dogs

Pentamidine treatment has been used successfully to establish immune cellular responses in recovered dogs. In this paper, we examined the appearance and disappearance of antibodies over time against crude Leishmania antigens and purified gp63 or gp70 proteins in sera from cured dogs using a Western blotting technique.Following the treatment, a pattern of antibody specificities to parasite antigens was observed in the sera of cured dogs. Antibodies to gp63 and gp70 were maintained after cure. In addition, the reaction with a 26 kD band observed during the clinical phase was no longer recognized by sera taken from recovery dogs. Interestingly, two proteins, 85 and 110 kD, not observed during the patent phase were detected by sera taken from treated dogs. Such patterns of antibody specificities to various parasite antigens might represent a useful parameter to determine the actual phase of the disease process.     

Biological activity of immunostimulatory CpG DNA motifs in domestic animals

Bacterial DNA contains a much higher frequency of CpG dinucleotides than are present in mammalian DNA. Furthermore, bacterial CpG dinucleotides are often not methylated. It is thought that these two features in combination with specific flanking bases constitute a CpG motif that is recognized as a "danger" signal by the innate immune system of mammals and therefore an immune response is induced when these motifs are encountered. These immunostimulatory activities of bacterial CpG DNA can also be achieved with synthetic CpG oligodeoxynucleotides (ODN). Recognition of CpG motifs by the innate immune system requires engagement of Toll-like receptor 9 (TLR-9), which induces cell signaling and subsequently triggers a pro-inflammatory cytokine response and a predominantly Th1-type immune response. CpG ODN-induced innate and adaptive immune responses can result in protection in various mouse models of disease. Based on these observations, clinical trials are currently underway in humans to evaluate CpG ODN therapies for cancer, allergy and infectious disease. However, potential applications for immunostimulatory CpG ODN in species of veterinary importance are just being explored. In this review, we will highlight what is presently known about the immunostimulatory effects of CpG ODN in domestic animals.     

Comparison of antibody values in sera of pigs vaccinated with a subunit or an attenuated vaccine against classical swine fever

Ten pigs, aged 85 days, were vaccinated with a subunit vaccine containing 32 g of classical swine fever virus glycoprotein E2 (gp E2) (group 1), and a further 10 pigs were vaccinated with a C strain vaccine (10^4±0.15 TCID₅₀/ml), produced by amplification in minipig kidney (MPK) cell culture (group 2). Nine non-vaccinated pigs served as a control group (group 3). Serum samples were collected before (day 0) and at 4, 10, 21 and 28 days after vaccination and were analysed by two commercially available enzyme immunoassays and by a neutralizing peroxidase-linked assay (NPLA). At the same times, peripheral blood was taken for determining the total leukocyte count and the body temperature was taken daily. Antibodies were not detected in serum samples collected before vaccination (day 0), and no side-effects that could be connected with vaccination were observed during the trial. Ten days after vaccination 6/10 pigs vaccinated with the subunit vaccine were seropositive. On days 21 and 28, the ratios of serologically positive to vaccinated pigs were 9/10 and 10/10, respectively. Four of the ten pigs that were vaccinated with the C strain vaccine were positive on day 21 and 9/10 on day 28. However, the results of the NPLA showed that only 4/10 pigs had an antibody titre >1:32 at the end of the trial in both the vaccinated groups, even though the subunit vaccine initiated an earlier and higher level of neutralizing antibodies than the vaccine produced from the C strain. Challenge was performed 28 days after vaccination on four randomly selected pigs from both vaccinated groups. The pigs survived the challenge without showing any clinical signs of classical swine fever (CSF), while two nonvaccinated control pigs died on the 10th and 12th days after infection.     

Pregnancy Rate in Zebu Cows with Two Different Postpartum Intervals Exposed to a Two-Bull Rotational System

In order to evaluate the reproductive effect of a rotational breeding system, the pregnancy rate of 69 Zebu cows was assessed, 32 late postpartum (211±93 days postpartum, LP) and 37 early postpartum (averaging 63±8 days after calving, EP). In the LP group, 60% (19/32) were cycling before exposure to the bulls but only 38% (14/37) in the EP group. The two groups were dissimilar from the start (p<0.05). The cows were exposed in a rotational breeding system, with six Brahman bulls with previous sexual experience paired off in three groups (A–B, C–D and E–F). The cows were exposed to each pair of bulls for a period of 3 weeks, with one week in between each period, when the females were without the presence of a male. In the LP group, the pair of bulls A–B obtained a pregnancy rate of 72% (23/32), whereas for bulls C–D the pregnancy rate was 33% (3/9) and for bulls E–F the pregnancy rate was 67% (4/6). In contrast, for the EP cows, bulls A–B obtained a pregnancy rate of 32% (12/37), bulls C–D a pregnancy rate of 67% (12/18) and for bulls E–F a pregnancy rate of 55% (6/11). The percentage pregnancy obtained using the pair of bulls A–B differed (p<0.05) between LP and EP, whereas for bulls C–D and E–F it was similar. The pregnancy rate was different (p<0.05) in the first 3 weeks of mating for the LP group, the highest number of cows becoming pregnant during the first 3 weeks of the study. In contrast, the highest number of pregnancies in EP occurred evenly during the last 6 weeks. This study suggests that the reproductive performance of pairs of bulls in a 9-week rotational programme with the overall pregnancy rate is similar (94% in the LP and 81% in the EP). However, the time taken for the females to become pregnant, and hence the performance of the bulls, is related to the average number of days postpartum for the cows.     

Comparison of milk production of dairy cows kept in cold and warm loose-housing systems

A retrospective cohort study was conducted to test whether the lactation curves of cows kept in cold loose-housing systems (CLHs) were the same as for cows in warm loose-housing systems (WLHs) in the Nordic countries. Approximately 40 000 test-day records from 5366 Ayrshire or Black and White cows kept on 38 CLHs and 166 WLHs in Finland during 1996 and 1997 were used. Analysis used a random-coefficient model (correcting for parity, breed and calving-year-season and the correlation-structure between test-days of the same cow and cows of the same herd). Cows in a CLH produced up to 1 l less milk per test-day, but this difference was not statistically significant. Surprisingly, the difference in milk yield was not affected by calving-year-season, parity or breed.