Protective potential of an attenuated Pasteurella multocida,which expresses only the N-terminal truncated fragment of P. multocida toxin

Pasteurella multocida serogroup D causes progressive atrophic rhinitis in pigs and produces a potent, intracellular, mitogenic toxin known as P. multocida toxin (PMT), which is encoded by the toxA gene. Highly toxic to cells, PMT is a poor antigen and becomes more immunogenic after its native structure has been destroyed. Previously, we found that the N-terminal fragment of PMT (N-PMT) can induce a strong immune response that is protective against wild-type challenge. Here, an attenuated P. multocida mutant expressing only N-PMT was developed and its protective effect was evaluated. The mutant provides protective immune responses against bacterial and toxin challenges, and so is a good live vaccine candidate.     

Contact and fumigant toxicity of oriental medicinal plant extracts against Dermanyssus gallinae (Acari: Dermanyssidae)

The acaricidal activity of methanolic extracts from 40 oriental medicinal plant species and a steam distillate of Cinnamomum camphora towards poultry house-collected adult Dermanyssus gallinae De Geer was examined using direct contact and vapour phase toxicity bioassays. Results were compared with those of 15 acaricides currently used. In filter paper contact toxicity bioassays using adult D. gallinae, C. camphora steam distillate (0.0051 mgcm(-2)) was the most toxic material, followed by extracts from Asarum sieboldii var. seoulens whole plant, Eugenia caryophyllata flower bud and Mentha arvensis var. piperascens whole plant (0.0063-0.0072 mgcm(-2)), based upon 24h LD(50) values. The acaricidal activity of these four plant preparations was almost comparable to that of profenofos (LD(50), 0.003 mgcm(-2)) but less effective than dichlorvos (LD(50), 0.0004 mgcm(-2)). The toxicity of Illicium verum fruit and Lysimachia davurica leaf extracts (0.09 mgcm(-2)) was almost comparable to that of benfuracarb, prothiofos, propoxur and fenthion (0.053-0.070mgcm(-2)). In vapour phase toxicity tests, these plant preparations were more effective in closed containers than in open ones, indicating that the mode of delivery of these plant extracts was largely a result of action in the vapour phase. Plants described herein merit further study as potential D. gallinae control agents.     

Glutathione content of in vivo and in vitro matured canine oocytes collected from different reproductive stages

Glutathione (GSH) concentrations of oocytes are considered as an important marker of the cytoplasmic maturation. The present study was designed to compare GSH concentrations of in vivo and in vitro matured canine oocytes. In vivo matured oocytes were collected 72 hr after ovulation by flushing fallopian tubes after laparotomy. Ovaries were collected from bitches with different reproductive stages, and collected oocytes were divided into 2 groups according to the size viz. < 120 microm and > 120 microm in diameter and cultured for 72 hr in Tissue Culture Medium-199 supplemented with 10% FBS, 2.2 mg/ml sodium bicarbonate, 2.0 microg/ml estrogen, 0.5 microg/ml FSH, 0.03 IU/ml hCG, and 1% penicillin-streptomycin solution in the presence or absence of 50 microM beta-mercaptoethanol. GSH concentrations were determined by the dithionitrobenzoic acid-glutathione disulfide (DTNB-GSSG) reductase recycling assay. GSH concentrations of immature canine oocytes were 2.9 and 3.8, 3.5 and 6.8, and 3.1 and 6.5 pM/oocyte for < 120 microm and > 120 microm in diameter oocyte groups at anestrous, follicular and luteal stage, respectively (P<0.05). In vivo matured oocytes had significantly higher GSH concentrations compared with in vitro matured oocytes. The GSH content was 19.2 pM/oocyte for in vivo matured oocytes, while 4.1 to 8.1 and 5.7 to 13.2 pM/oocyte for in vitro matured oocytes cultured in the absence or presence of beta-mercaptoethanol, respectively (P<0.05). Presence of beta-mercaptoethanol increased GSH synthesis in canine oocytes cultured in vitro, and oocytes collected from follicular and luteal stage was superior to anestrus oocytes.     

Comparison of polymerase chain reaction methods for the detection of <Emphasis Type="Italic">Theileria equi</Emphasis> infection using whole blood compared with pre-extracted DNA samples as PCR templates

Rapid, efficient, and reproducible procedures for isolating DNA before PCR gene amplification are essential for the diagnosis of piroplasms. In this study, we evaluated the ease and reliability of detecting Theileria equi by PCR using pre-extracted DNA samples (by QIAamp DNA Mini Kit and phenol-chloroform methods) compared with blood spotted on FTA cards as PCR templates. Although minimal variations in limit of detection were observed among the methods compared, overall, the use of pre-extracted DNA samples and blood spotted on FTA cards had comparable detection limits. These results indicate that T. equi infection can be efficiently detected directly from FTA cards by PCR without the need for pre-extraction of DNA from blood samples.     

Effect of dietary nutmeg oil on heat‐stress tolerance‐related parameters in Korean native chicken reared under hot temperature

This study investigated the effect of dietary nutmeg oil (NO) on growth performance, blood parameters, lipid peroxidation and heat shock protein (HSP) 70 expression in Korean native chicken (KNC) reared under hot temperature. We allocated 273 meat‐type KNCs (Hanhyup‐3, 4‐week‐old, body weight [BW] = 539.93 ± 1.75 g) to the following three treatments with seven replicate pens (13 birds/pen) per treatment. Three treatment diets were as follows: (a) Control, basal diet without NO supplementation; (b) NO 250; and (c) NO 500, basal diet supplemented with 250 and 500 ppm NO respectively. Diets and water were provided ad libitum throughout the 6‐week feeding trial. During overall period (0–6 weeks), no differences (p > 0.05) were observed in BW gain (BWG), feed intake (FI) and feed conversion rate (FCR) among treatments. However, the FI at 0–3 weeks decreased (p < 0.05) quadratically with increasing NO levels. Most blood parameters did not differ (p > 0.05) among treatments, although the monocyte level of the NO 500 group was considerably lower (p > 0.05) than that of the other groups. Furthermore, dietary NO did not affect serum triglyceride, cholesterol, total protein, albumin, calcium, phosphorus and alanine aminotransferase (ALT) levels (p > 0.05); however, it linearly decreased serum aspartate aminotransferase (AST) level (p < 0.05). Additionally, serum malondialdehyde (MDA) concentration decreased (p < 0.05) and heart MDA concentration was lower (p = 0.08) with increasing dietary NO supplementation. After a 3‐hr heat (35°C) challenge, the rectal temperature (RT) reduced (p < 0.05) linearly with increasing NO levels. Dietary NO did not affect liver HSP70 (p > 0.05) gene expression. In conclusion, NO potentially enhanced the ability of chickens to alleviate heat stress. Furthermore, our findings suggest that lipid oxidation inhibition by dietary NO likely mediated the enhanced heat‐stress tolerance of the chickens.     

The nutritional value of degermed,dehulled corn for pigs and its impact on the gastrointestinal tract and nutrient excretion

Three experiments were designed to assess the feeding value and potential environmental benefits of feeding degermed, dehulled corn, a low fiber by-product originating from the corn dry milling process, to pigs. Twelve 27-kg (SE = 0.8) barrows were used in Exp. 1 to measure the apparent fecal digestibility of DM, GE and N of degermed, dehulled corn compared with corn grain. Two diets were formulated to contain either 96.4% of degermed, dehulled corn or corn grain plus supplemental vitamins and minerals. Digestibilities of DM, GE, and N were greater in degermed, dehulled corn (96.2, 96.0, and 93.6%, respectively) compared with corn grain (89.0, 89.0, and 78.4%, respectively) (P < 0.01). Overall, a 67 and 29% reduction in DM and N excretion, respectively, was observed. In Exp. 2, eight 70-kg (SE =1.8) barrows were surgically fitted with ileal cannulae and fed the same diets as in Exp. 1, to measure the ileal digestibility of nutrients in degermed, dehulled corn. Ileal digestibility of DM, energy, and N was 13, 15, and 7% greater in degermed, dehulled corn (P < 0.05). Apparent ileal digestibility coefficients of leucine, methionine, and phenylalanine were greater in degermed, dehulled corn compared with corn grain (P < 0.05) while a trend for a lower tryptophan digestibility in degermed, dehulled corn was observed (P = 0.067). In Experiment 3, 96 nursery pigs with an initial average BW of 8.8 kg (SE = 0.08), fed a starter diet formulated with degermed, dehulled corn or corn grain as the major grain source, were used in a 28-d growth performance study. At the end of the study, 24 pigs (1 pig per pen) were sacrificed and gastrointestinal tract measurements were taken. Daily growth rates of pigs were the same between diets (0.64 kg/d). A trend for reduced feed intake (P = 0.073) in pigs fed degermed, dehulled corn led to a 4% improvement in gain to feed (P < 0.05). Feeding degermed, dehulled corn had no effect on gut fill, gastrointestinal tract weight, or liver weight (P > 0.05). Ileal villus lengths and crypt depths were not affected by feeding degermed, dehulled corn although ileal villus widths were greater in pigs fed corn grain. Results from these trials suggest that corn processed to remove poorly digestible fiber fractions provides more digestible nutrients than corn grain. As a result, degermed, dehulled corn reduces fecal and N excretion, thus providing a means to reduce nutrient excretion.     

Comparative study on calretinin immunoreactivity in gerbil and rat retina

Expression of calretinin in retina has been ascribed to multiple biological and functional aspects in the visual system. In this study, we examined the distribution patterns of calretinin immunoreactivity in gerbil and rat retina. In the gerbil, calretinin immunoreactivity was present in bipolar and amacrine cells of the inner nuclear layer and in neurones of the ganglion cell layer. In the rat, amacrine and ganglion cells showed calretinin immunoreactivity, but bipolar cells did not contain calretinin immunoreactivity. In both species, calretinin immunoreactivity was absent in cones, cone bipolars, and horizontal cells. In conclusion, gerbil as well as rat has a rod-dominant retina. The differences in calretinin expression between rat and gerbil require further investigations under various functional and developmental conditions.