The effect of human recombinant interleukin-2 on the porcine immune response to a pseudorabies virus subunit vaccine

The effect of human recombinant interleukin-2 (rIL-2) as an immune enhancing agent was evaluated in pigs vaccinated with a pseudorabies virus subunit vaccine (SV). Two groups of three pigs received two 25 micrograms doses of SV given 3 weeks apart. One group received 10(5) kg-1 day-1 of rIL-2 subcutaneously over two 5-day periods beginning on the day of the first and second vaccine inoculation. Six other pigs were immunized with two 5 micrograms doses of SV. Three of these pigs were treated as above with rIL-2. The effect of treatment was evaluated by comparing: the humoral response; the cell-mediated immune (CMI) response as measured by lymphocyte blastogenesis before and after virus challenge; and the weight response and virus excretion pattern after challenge with virulent pseudorabies virus (PRV). The humoral antibody response as detected by the serum virus neutralization (SN) assay and the enzyme linked immunosorbent assay (ELISA) was consistently higher in rIL-2 treated pigs than in non-treated pigs. These differences were significant (P less than 0.05) among high vaccine dose pigs prior to virus challenge when measured by the SN assay and during the anamnestic response period between days 3 and 10 after challenge when measured by both the SN assay and the ELISA. No differences were detected between treatment groups in the weight response, virus excretion pattern or the CMI response. These results suggest that human rIL-2 may have enhanced the immune response of pigs to the subunit vaccine.     

Epidemiological investigation of the prevalence and features of postweaning multisystemic wasting syndrome in Japan.

To investigate the prevalence and features of postweaning multisystemic wasting syndrome (PMWS) in Japan, an epidemiological study was conducted in 692 weaned pigs with various clinical signs, commonly including wasting or weight loss, collected from 129 swine farms between 2000 and 2003. The presence of PMWS was diagnosed by the detection of characteristic histological lesions and moderate to large amounts of porcine circovirus type 2 (PCV2) antigen within the lesions in multiple lymphoid tissues. Postweaning multisystemic wasting syndrome was positive in 23.4% of pigs (162/692) over the course of the study, and occurred in 50.4% of the farms (65/129). Mortality in 30-120-day-old pigs in the farms positive for PMWS varied from 0.1 to 32.0%. No significant difference in mortality was seen between PMWS-positive and -negative farms (P = 0.1). However, mortality was significantly higher in the PMWS-positive farms where PMWS was diagnosed in more than 50% of the pigs examined compared to farms negative for PMWS (P = 0.02). These findings indicate that PMWS has spread widely in Japan. Moreover it may exist in variable forms in swine farms, including an epidemic form or a subtle endemic or sporadic form. A case-control study suggested that risk factors for the occurrence of PMWS include porcine reproductive and respiratory syndrome (PRRS) pneumonias and Mycoplasma hyorhinis infection.     

Relationship between the first ovulation within three weeks postpartum and subsequent ovarian cycles and fertility in high producing dairy cows

The aim of this study was to investigate the relationship between the first ovulation within 3 weeks postpartum and subsequent ovarian cycles and fertility in high producing dairy cattle in Hokkaido, Japan. In Experiment 1, 110 cows (44 primiparous and 66 multiparous) were used to determine the effects of the first ovulation within 3 weeks postpartum on subsequent ovarian cycles. Milk samples were collected twice weekly from 7 to 100 days postpartum. The first ovulation was identified by an increase in milk progesterone (P4) to more than 1 ng/ml within 3 weeks postpartum. The numbers of cows showing ovulation and anovulation within 3 weeks postpartum were 31 (70.5%) and 13 (29.5%) in the primiparous cows and 35 (53.0%) and 31 (47.0%) in the multiparous cows, respectively. The patterns of ovarian resumption after calving were classified into two types (normal ovarian cycles and abnormal ovarian cycles) on the basis of milk P4 concentrations. Initiation of normal ovarian function in cows ovulated within 3 weeks postpartum occurred earlier than in anovulated cows regardless of the number of calvings (primiparous, 27.8 days vs. 44.4 days; multiparous, 30.6 days vs. 55.7 days; P<0.01). Out of the multiparous cows that ovulated within 3 weeks postpartum, initiation of normal ovarian function followed by a normal luteal phase was earlier than when it was followed by an abnormal luteal phase (25.5 days vs. 40.4 days; P<0.05). Milk P4 concentrations after the first ovulation were lower than those after the second ovulation in both the primiparous and multiparous cows (P<0.05). In Experiment 2, 22 multiparous cows were used to determine the effects of the first ovulation within 3 weeks postpartum on subsequent fertility. Blood samples were collected once a week from 0 to 3 weeks postpartum. The interval from parturition to first service in ovulated cows was shorter than in anovulated cows (68.4 days vs. 94.8 days; P<0.05). The conception rate by 100 days after calving tended to be higher in ovulated cows than in anovulated cows (50.0% vs. 16.7%, P=0.09). In conclusion, our data strongly suggests that ovulation within 3 weeks postpartum is a crucial phenomenon for subsequent resumption of ovarian function and conception, and thus it can be used as an index of subsequent reproductive performance.     

Oestradiol enhances plasma growth hormone and insulin-like growth factor-I concentrations and increased the expression of their receptors mRNAs in the liver of ovariectomized cows

Many metabolic hormones, growth hormone (GH), insulin-like growth factor-I (IGF-I) and insulin affect ovarian functions. However, whether ovarian steroid hormones affect metabolic hormones in cattle remains unknown. This study aimed to determine the effect of sex steroids on the plasma profiles of GH, IGF-I and insulin and their receptors in the liver and adipose tissues of dairy cows. Ovariectomized cows (n = 14) were randomly divided into four groups: control group (n = 3) was treated with saline on Day 0; oestradiol (E2) group (n = 3), with saline and 1 mg oestradiol benzoate (EB) on Day 0 and 5, respectively; progesterone (P4) group (n = 4) with two CIDRs (Pfizer Inc., Tokyo, Japan) from Day 0; and E2 + P4 group (n = 4) with two CIDRs on Day 0 that were removed on Day 6 and were immediately injected with 1 mg EB. The animals were euthanized after the experiment, and liver and adipose tissues samples were quantitatively analysed using real-time PCR for the expression of mRNA for the GH (GHR), IGF-I (IGFR-I) and insulin (IR) receptor mRNAs. Oestradiol benzoate significantly increased the number of peaks (p < 0.05), pulse amplitude (p < 0.05) and area under the curve (AUC; p < 0.01) for plasma GH; moreover, it increased plasma IGF-I concentration (p < 0.05), but it had no effect on the plasma insulin profile. P4 significantly decreased the AUC (p < 0.01), compared with the control group, whereas it did not affect the number of peaks and the amplitude of GH pulses. P4 + E2 did not affect the GH pulse profile. E2 increased the mRNA expression of GHR, IGFR-I and IR in the liver (p < 0.05), whereas both P4 and E2 + P4 did not change their expressions. Our results provide evidence that the metabolic and reproductive endocrine axes may regulate each other to ensure optimal reproductive and metabolic function.     

Electrical velocimetry for noninvasive cardiac output and stroke volume variation measurements in dogs undergoing cardiovascular surgery

Objective

To compare electrical velocimetry (EV) noninvasive measures of cardiac output (CO) and stroke volume variation (SVV) in dogs undergoing cardiovascular surgery with those obtained with the conventional thermodilution technique using a pulmonary artery catheter.

Development of a new selective medium for isolation of methicillin-resistant Staphylococcus aureus

A new selective medium containing cephem antibiotics was developed for isolation of methicillin-resistant Staphylococcus aureus (MRSA). MRSA colonies on a medium containing ceftazidime (CAZ) were most easily identifiable and a medium containing cefoperazone (CPZ) was superior in suppressing the growth of other bacteria. With the medium containing a couple of CAZ and CPZ, MRSA and methicillin-resistant coagulase-negative staphylococci (MRCNS) were detected from 2 and 1 of 15 chicken meat samples respectively. The MRSA and MRCNS recovery test showed that the medium was effective for MRSA isolation, suppressing the growth of other bacteria efficiently. These results suggested that the medium containing a couple of CAZ and CPZ was useful for MRSA detection from foods and animals.     

Purification of N-acetyllactosamine-binding activity from the porcine sperm membrane: possible involvement of an ADAM complex in the carbohydrate-binding activity of sperm

Although the importance of carbohydrate recognition by sperm during egg zona pellucida binding has been widely reported, the sperm molecular species that recognize the carbohydrates are poorly characterized. Our previous cytochemical study indicated that two kinds of carbohydrate-binding proteins are expressed on porcine sperm heads-one recognizes N-acetyllactosamine (Galβ1-4GlcNAc-), and the other recognizes the Lewis X structure (Galβ1-4(Fucα1-3)GlcNAc-). For this report, we used proteomic techniques to characterize the sperm proteins that bind N-acetyllactosamine. Porcine sperm plasma membrane was solubilized with a detergent solution and subjected to sequential chromatography with dextran sulfate agarose, affinity, and hydroxyapatite, and the binding activities in the eluates were monitored by a solid-phase binding assay. The tryptic peptides of two proteins most likely associated with the binding activities were subjected to tandem mass spectrometry sequencing. A subsequent database search identified one of the two proteins as predicted disintegrin and metalloprotease domain-containing protein 20-like (XP_003128672). The other protein was identified as disintegrin and metalloprotease domain-containing protein 5 (AB613817) by database searches for homologous amino acid sequences, cDNA cloning, nucleotide sequencing and nucleotide database searches. Furthermore, two-dimensional blue native/SDS-PAGE demonstrated that they formed a variety of non-covalent complexes. Therefore, these ADAM complexes probably are responsible for the N-acetyllactosamine-binding activity. An affinity-purified fraction containing these ADAM complexes showed zona pellucida-binding activity, though the activity was relatively weak, and the presence of another zona pellucida-binding protein that probably works in concert with these ADAM complexes was suggested. Immunofluorescence testing suggested that ADAM20-like was localized on the anterior part of the sperm plasma membrane.     

Methodology for the study of the hypothalamic-pituitary hormone secretion in cattle

Studies on the neuroregulatory mechanisms on the secretion of anterior pituitary (AP) hormones in domestic animals are important because nearly all complex physiological and metabolic processes are regulated by the AP hormones. To examine them, this article considers in vivo approaches such as the techniques of intrahypothalamic injection, intracerebroventricular injection, push-pull perfusion, and microdialysis, which have been employed in our own research group for the study in cattle. Also, in vitro approaches such as bovine AP cell culture and the AP explants superfusion system are described. This article clarifies the potential of neuroendocrine study techniques in cattle.     

The effects of l‐DOPA and sulpiride on growth hormone secretion at different injection times in Holstein steers

The effects of l ‐DOPA, a precursor of dopamine (DA), and sulpiride, a D₂‐type DA receptor blocker, on growth hormone (GH) and prolactin (PRL) secretion were investigated in steers. Eight Holstein steers (212.8 ± 7.8 kg body weight) were used. Lighting conditions were 12:12 L:D (lights on: 06.00–18.00 hours). Blood samplings were performed during the daytime (11.00–15.00 hours) and nighttime (23.00–03.00 hours). Intravenous injections of drugs or saline were performed at 12.00 hour for the daytime and 00.00 hour for the nighttime, respectively. Plasma GH and PRL concentrations were determined by radioimmunoassay. l ‐DOPA did not alter the GH secretion when it was injected at 12.00 hour (spontaneous GH level at its peak). On the other hand, l ‐DOPA increased GH secretion at 00.00 hour (GH level at its trough). Injection of sulpiride suppressed GH secretion at 12.00 hour but did not affect GH levels at 00.00 hour. l ‐DOPA inhibited and sulpiride stimulated PRL release during both periods. These results suggest that dopaminergic neurons have stimulatory action on GH secretion and inhibitory action on PRL secretion in cattle. In addition, injection time should be considered to evaluate the exact effects on GH secretion due to its ultradian rhythm of GH secretion in cattle.