Development of specific PCR primers for identification and detection of <Emphasis Type="Italic">Phytophthora capsici</Emphasis> Leon

A PCR-based method was developed for the identification and detection of Phytophthora capsici in pepper plants. Three PCR primers (CAPFW, CAPRV1 and CAPRV2) specific for P. capsiciwere designed based on the sequence of its internal transcribed spacer regions. CAPFW/CAPRV1 amplify a 452 bp product from P. capsici DNA whereas CAPFW/CAPRV2 a 595 bp fragment; neither set amplifies DNA from pepper or several fungi pathogenic to pepper. In conventional (single-round) PCR, the limit of detection was 5 pg DNA for both primer sets, whereas in nested PCR the detection limit for both was of 0.5 fg. However, when the dilution series of target DNA were spiked with plant DNA, amplification declined two-fold in both conventional and nested PCR. The CAPFW/CAPRV2 set in conventional PCR was used to detect P. capsici DNA in inoculated plants. Detection occurred as soon as 8h post-inoculation in stem samples from infected but still symptomless plants. The method was also tested to detect fungal DNA in infected soils.     

一例猪附红细胞体与弓形体混合感染的诊治

附红细胞体病是由立克次氏体引起的一种散在的热性、溶血性入畜共患传染病，病猪以急性黄疸性贫血和发热为特征。弓形体病是由刚地弓形虫在多种动物和入的有核细胞内寄生引起的一种入畜共患的原虫病。本文通过对某猪场发生猪附红细胞体与弓形体混合感染的诊治，从发病情况、临床症状、剖检变化、实验室诊断、治疗等方面对该病进行了阐述。     

Arachidonic acid immunoregulation in lambs persistently infected with border disease virus

To evaluate arachidonic acid-related immunoregulatory mechanisms during long-term persistent pestivirus infection, we measured plasma contents of leukotriene C4 (LTC4), prostaglandin D2 (PGD2) and their plasma fatty acid (FA) precursor, arachidonic acid (AA), in six lambs with congenital border disease (BD). Significantly elevated average plasma LTC4 during the first half year of life was associated with increased PDG2 when compared to uninfected control lambs. Significantly elevated total plasma esterified AA stores suggest an effective BDV-mediated prostenoid immunostimulation. However, at 1 yr old, prostenoid secretion had fallen to normal (LTC4) or below normal (PGD2) levels. In contrast, there remained significantly elevated plasma esterified AA, present as available substrate for formation of these anti-viral immunoregulatory agents. These results suggested that preventing mobilization of AA from lipid stores for effective immune responses may be a viral strategy of BD virus that is associated with long term border disease effects.     

Effects of iron injection timing on suckling and subsequent nursery and growing-finishing performance and hematological criteria

Two experiments were conducted to evaluate the effects of Fe injection timing after birth on suckling and subsequent nursery and growing-finishing pig performance. The injectable Fe source used in both experiments was GleptoForte (Ceva Animal Health, LLC., Lenexa, KS). GleptoForte contains gleptoferron which is a Fe macromolecule complex. In Exp. 1, a total of 324 newborn pigs (DNA 241 × 600, initially 1.6 ± 0.04 kg body weight [BW]) within 27 litters were used. Two days after birth, all piglets were weighed, and six barrows and six gilts per litter were allotted to 1 of 6 treatments consisting of no Fe injection or 200 mg of injectable Fe provided in a single injection on d 2, 4, 6, 8, or 10 of age. Pigs were weaned (~21 d of age) and allotted to nursery pens with all pigs in each pen having received the same Fe treatment. In Exp. 2, a total of 1,892 newborn pigs (PIC 359 × C40; initially 1.5 ± 0.02 kg BW) within 172 litters were used. One day after birth, piglets were weighed, and 11 pigs within each litter were allotted to 1 of 6 treatments consisting of no Fe injection or 200 mg of injectable Fe provided on d 1, 3, 5, or 7 of age, or 200 mg on d 1 plus 200 mg on d 12 of age. Pigs were weaned (19 d of age) and placed in a commercial wean-to-finish facility in a total of 15 pens with equal representation of treatments in each pen. In both experiments, not providing an Fe injection after birth decreased (P < 0.05) preweaning average daily gain (ADG), weaning weight, and hemoglobin and hematocrit values compared with all other treatments. In Exp. 1, increasing the age that piglets received an Fe injection until 4 or 6 d after birth provided marginal evidence for an improvement (quadratic; P = 0.070) in preweaning ADG. For the nursery period, increasing the age that piglets received an Fe injection improved (quadratic; P = 0.013) d 80 BW, but there was no evidence of a difference (P > 0.10) in d 173 BW at the end of the grow-finish period. In Exp. 2, increasing the age that piglets received a 200 mg Fe injection showed no evidence of difference (P > 0.10) for subsequent nursery and growing-finishing ADG. In both experiments, hemoglobin and hematocrit values were decreased (linear; P < 0.05) at weaning with increasing age when pigs received an Fe injection. These experiments suggest that providing a 200 mg Fe injection within 7 d after farrowing is sufficient for optimizing preweaning and subsequent growth performance.     

Evaluation of tizanidine as a marker of canine CYP1A2 activity

The dog CYP1A2 enzyme is likely an important contributor to the metabolism of veterinary drugs. Dog CYP1A2 is expressed in liver, plus it is inducible and polymorphic, creating the potential for intersubject differences in pharmacokinetics. Hence, the ability to probe dog CYP1A2 activity and inhibition is relevant toward veterinary drug development and drug–drug interaction assessment. Previous studies have relied on human probes with questionable specificity for CYP1A2, so it was hypothesized that recombinant CYP1A2 could be used to find a specific CYP1A2 substrate. Intrinsic clearance experiments demonstrated that tizanidine was a substrate of CYP1A2. Profiling of tizanidine metabolites generated by CYP1A2 identified the imidazole metabolite that was detectable in dog plasma. The imidazole metabolite was subsequently used to evaluate tizanidine as a CYP1A2 probe. Co‐administration of the CYP1A inhibitor enrofloxacin with tizanidine significantly decreased (30%; n = 3) the formation of the imidazole metabolite vs. control experiments. As enrofloxacin is a weak inhibitor, further studies are required to confirm the sensitivity of tizanidine as an in vivo probe. However, tizanidine may be a more selective CYP1A2 probe than phenacetin when conducting in vitro studies due to the presence of other phenacetin‐metabolizing enzymes in dog liver microsomes.