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.     

A Multistate Investigation of Antibiotic‐Resistant Salmonella enterica Serotype I 4,[5],12:i:‐ Infections as Part of an International Outbreak Associated with Frozen Feeder Rodents

While most human Salmonella infections result from exposure to contaminated foods, an estimated 11% of all Salmonella infections are attributed to animal exposures, including both direct animal handling and indirect exposures such as cleaning cages and handling contaminated pet food. This report describes the epidemiologic, environmental and laboratory investigations conducted in the United States as part of the response to an international outbreak of tetracycline‐resistant Salmonella enterica serotype I 4,[5],12:i:‐ infections with over 500 illnesses occurring from 2008 to 2010. This investigation found that illness due to the outbreak strain was significantly associated with exposure to pet reptiles and frozen feeder rodents used as food for pet reptiles. Salmonella isolates indistinguishable from the outbreak strain were isolated from a frozen feeder mice‐fed reptile owned by a case patient, as well as from frozen feeder mice and environmental samples collected from a rodent producing facility (Company A). An international voluntary recall of all Company A produced frozen feeder animals sold between May 2009 and July 2010 occurred. Only 13% of cases in our investigation were aware of the association between Salmonella infection and mice or rats. Consumers, the pet industry, healthcare providers and veterinarians need to be aware of the potential health risk posed by feeder rodents, whether live or frozen. Frozen feeder rodent producers, suppliers and distributors should follow the animal food labelling requirements as described in 21 CFR §501.5, and all packages of frozen feeder rodents should include safe handling instructions. Persons should wash their hands thoroughly with soap and water after handling live or frozen feeder rodents, as well as reptiles or anything in the area where the animals live. Continued opportunities exist for public health officials, the pet industry, veterinarians and consumers to work together to prevent salmonellosis associated with pet food, pets and other animals.     

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.