Alternate pathway of infection with Hepatozoon americanum and the epidemiologic importance of predation.

BACKGROUND: The range of American canine hepatozoonosis (ACH) is expanding from the southern USA northward. Transmission of Hepatozoon americanum occurs by ingestion of infected Gulf Coast ticks, Amblyomma maculatum. The source of the protozoan for the tick remains undetermined; infected dogs are unusual hosts for the tick. OBJECTIVE: Compare possible sources of infection by field investigations of 2 multiple-dog outbreaks of ACH. ANIMALS: Twenty-eight privately owned dogs (Canis familiaris), 1 coyote (Canis latrans), 31 wild-trapped cotton rats (Sigmodon hispidus), 24 wild-trapped field mice (Peromyscus leucopus), and 9 wild-caught rabbits (Sylvilagus spp.) from sites in eastern Oklahoma were monitored for hepatozoonosis. Six laboratory-raised cotton rats (S. hispidus), 6 Sprague-Dawley rats (Rattus norvegicus), 6 C57BL/6J-Lystbg-J/J mice (Mus musculus), 6 outbred white mice (M. musculus), 6 New Zealand white rabbits (Oryctolagus cuniculus), and 2 dogs were acquired through commercial vendors for experimental transmission trials of H. americanum. METHODS: Four of 15 dogs in a rural neighborhood and 5/12 hunting Beagles were confirmed to be infected by blood smear examination, muscle biopsy, and polymerase chain reaction assay of the 18S rRNA gene of Hepatozoon species. Histories and tick host preferences led to field collections of common prey of canids and experimental transmission trials of H. americanum to selected prey (M. musculus, S. hispidus, R. norvegicus, and O. cuniculus). RESULTS: Dogs with ready access to prey (4/15 dogs) or that were fed prey retrieved from hunts (5/12 hunting Beagles) became infected, providing evidence that predation is an important epidemiologic component of ACH infection. Experimental transmission studies identified a quiescent, infectious stage (cystozoite) of the parasite that provides an alternate mode of transmission to canids through predation, demonstrating that cotton rats, mice, and rabbits but not brown rats may act as paratenic hosts of H. americanum. CONCLUSIONS AND CLINICAL IMPORTANCE: Predation of prey harboring infected A. maculatum or containing cystozoites of H. americanum in their tissues provide 2 modes of transmission of ACH to dogs, putting unconfined dogs at increased risk of infection in endemic areas.

Bacterial fermentation in the gastrointestinal tract of non-ruminants: influence of fermented feeds and fermentable carbohydrates.

The search for alternatives to in-feed antibiotics in animal nutrition has highlighted the role dietary modulation can play in improving gut health. Current antibiotic replacement strategies have involved the use of microbes beneficial to health (probiotics) or fermentable carbohydrates (prebiotics) or both (synbiotics). The present review recognises the contribution of fermented feeds and fermentable carbohydrates in improving the gut environment in non-ruminants. It proposes the screening of probiotic bacteria for the production of fermented feeds and supplementation of these feeds with fermentable carbohydrates prior to feeding animals. It is suggested that the term 'fermbiotics' should be used to describe this intervention strategy.

The effect of copper on the death rate of Salmonella typhimurium DT104:30 in food substrates acidified with organic acids.

AIMS: The objective of this study was to investigate the effect of copper ions on the survival of Salmonella typhimurium DT104:30 in acidified liquid food substrates. METHODS AND RESULTS: The decimal reduction time (Dvalue) of Salm. typhimurium DT104:30 was determined in acidified liquid pig feed (LPF) and skimmed milk (SM) containing a range of copper concentrations (0-50 ppm). As copper concentration increased, the death rate of Salm. typhimurium DT104:30 increased. In LPF acidified with 150 mmol l-1 lactic acid the presence of 50 ppm copper resulted in a 10-fold increase in the death rate. CONCLUSIONS: The presence of copper salts in acidic liquid food substrates significantly increases the death rate of Salm. typhimurium DT104:30. SIGNIFICANCE AND IMPACT OF THE STUDY: This finding could influence policy on levels of copper in pig feed.

The effect of temperature on the growth and persistence of Salmonella in fermented liquid pig feed.

Two studies were conducted to investigate the effect of temperature on the fate of Salmonella enterica subsp. enterica serovar typhimurium DT104:30 in fermented liquid pig feed. These were (1) by co-inoculation of feed with S. typhimurium DT104:30 and Pediococcus pentosaceus, as the fermenting organism, and (2) by fermenting feed for 48, 72 or 96 h prior to inoculation with S. typhimurium DT104:30. In co-inoculated feed incubated at 20 degrees C, S. typhimurium DT104:30 persisted for at least 72 h. In contrast, in feed incubated at 30 degrees C, no S. typhimurium DT104:30 were detectable 48 h after inoculation. In prefermented feed, S. typhimurium DT104:30 died four to five times faster in feed maintained at 30 degrees C (D(value) 34-45 min) compared with feed maintained at 20 degrees C (D(value) 137-250 min). This was not entirely due to differences in lactic acid concentration as feed fermented for 72 or 96 h at 20 degrees C and feed fermented for 48 h at 30 degrees C contained similar concentrations of lactic acid (160-170 mM). Low numbers of S. typhimurium DT104:30 were still detectable in fermented feed 24 h after inoculation at 20 degrees C. In contrast, none were detectable 6-7 h after inoculation at 30 degrees C. The results of these studies indicate that it would be advisable for pig producers to control the temperature of liquid feed tanks to reduce the risk of Salmonella contamination.

Evaluation of a novel, natural oligosaccharide gum as a sustained-release and mucoadhesive component of calcitonin buccal tablets.

The objective of this study was to evaluate the gum from Hakea gibbosa (hakea) as a sustained-release and mucoadhesive component in buccal tablets for a model peptide, namely, salmon calcitonin. Flat-faced core tablets containing either 12 or 32 mg of hakea and 40 microg (200 IU) of salmon calcitonin (sCT) per tablet were formulated using a direct compression technique and were coated with Cutina on all but one face. The in vitro release profiles were sigmoidal in nature and according to a mathematical model indicated super Case II transport as the primary mechanism of release. The resulting plasma sCT and calcium concentrations were determined following both intravenous administration and buccal application of mucoadhesive tablets in rabbits. Following intravenous administration, the mean values determined for t(1/2) (alpha), t(1/2) (beta), V(d), and CL for sCT were 0.76 +/- 0.06 min, 67 +/- 18 min, 1484 +/- 454 mL/kg, and 19 +/- 2 mL/min.kg, respectively. Following the application of the mucoadhesive buccal tablets which contained 40 microg of sCT and either 12 or 32 mg of hakea, the calculated apparent bioavailability (F) and clearance (CL) were 37 +/- 6% and 19 +/- 3.3 mL/min.kg and 16 +/- 8% and 18 +/- 0.4 mL/min. kg, respectively. Serum calcium concentrations indicated that biologically active sCT was delivered across the rabbit buccal mucosa. The strength of mucoadhesion of the tablets was also quantitated in terms of the force of detachment as a function of time. The force of detachment for the mucoadhesive buccal tablets containing either 12 or 32 mg of hakea and 40 microg of sCT increased from 4.47 +/- 0.68 to 8.41 +/- 1.0 N and 8.23 +/- 1.62 to 14.98 +/- 1.63 N, respectively, from 5 to 90 min following application to excised rabbit intestinal mucosa. These results demonstrate that the novel, natural gum from Hakea gibbosa may be used to sustain the release of sCT from a unidirectional-release buccal tablet. The mechanism of in vitro release is likely to involve peptide diffusion/polymer dissolution. The mucoadhesive strength, as measured by the force of detachment, can be modulated by altering the amount of hakea in the tablet. The mucoadhesive buccal tablets described in this paper represent an improved transbuccal delivery system for therapeutic polypeptides.