Angiostrongylus vasorum in Great Britain: a nationwide postal questionnaire survey of veterinary practices.

The lungworm, Angiostrongylus vasorum, was first reported in indigenous dogs in southwestern England in 1980 and has since been recorded in Wales, southeastern England and, more recently, in the West Midlands, northern England and Scotland. The nationwide distribution of the parasite was evaluated using a postal questionnaire sent to 3950 small animal practices during 2009. Information was sought on the location of each practice, awareness of the parasite locally, number of cases diagnosed over the past year and whether diagnosis was based on clinical signs alone or supported by additional tests. 1419 practices returned a usable response, the majority being located in a city/town. Nearly one-third of responding practices were aware of the parasite locally, 20.7 per cent had seen at least one confirmed case and 0.3 per cent >20 confirmed cases over the past year. The most widely used tests were faecal examination and any type of imaging. Existing clusters of infection were detected in southeastern England and south Wales; infection was also found to be widespread in central England, though patchy in northern England and Scotland. Using distribution of clinical cases as an indicator of parasite distribution, this study confirmed that A. vasorum has spread beyond traditional UK endemic foci.

Associations between blood gastrin, ghrelin, leptin, pepsinogen and Ostertagia ostertagi antibody concentrations and voluntary feed intake in calves exposed to a trickle infection with O. ostertagi.

Twenty-five, castrated male Holstein-cross calves, between 4 and 5 months of age, weighing 156.5+/-12.2 kg and reared under conditions designed to minimise the risk of parasitic infection, were allocated to one of the five treatment groups on the basis of initial bodyweight. The groups were (1) ad libitum (ad lib) fed controls (ALC); (2) ad lib fed infected (INF) and treated with topical eprinomectin on Day 56; (3) controls pair-fed with the INF group (PFC); (4) ad lib fed controls treated with eprinomectin on Days 0 and 56 (E-ALC) and (5) ad lib fed, infected and treated with eprinomectin on Days 0 and 56 (E-INF). Infection comprised a trickle infection with the equivalent of 10,000 larvae of Ostertagia ostertagi per day from Day 0 to Day 56 and the study concluded on Day 77. Parameters measured throughout the study included: liveweight, feed intake, faecal egg counts; plasma pepsinogen, gastrin, ghrelin and leptin; plasma antibodies to adult O. ostertagi. No significant differences in feed intake or liveweight gain were observed between any of the different groups, a finding thought to result from the high quality of feed offered. Significant differences between the INF and control groups however were observed in faecal egg counts, plasma pepsinogen, gastrin and O. ostertagi antibodies, which were all elevated, and leptin, which was reduced. Values of these parameters for the E-INF group were intermediate between the INF and ALC groups. Plasma ghrelin showed no association with either feed intake or parasitism. Further studies are needed to fully elucidate the roles of various biochemical and neuroendocrine mediators for inappetence in ruminants with parasitic gastroenteritis.

Angiostrongylus vasorum and Eucoleus aerophilus in foxes (Vulpes vulpes) in Great Britain.

The nematode parasite Angiostrongylus vasorum is a source of increasing concern in several parts of the world, where it causes significant disease in dogs. Wild canids, especially foxes, are likely to have a role in the epidemiology of canine infection, and the parasite could also affect fox health and population dynamics. The heart and pulmonary vasculature of 546 foxes culled mostly by gamekeepers in Great Britain in 2005-2006 were examined by dissection and a modified flushing technique. Forty foxes were found to be infected, giving an overall prevalence in the UK fox population of 7.3% (5.3-9.9). Prevalence varied widely between regions, from 0% (0-3) in Scotland and northern England to 23% (16-32) in south-east England. This closely matches the perceived incidence of disease in dogs, which is commonly diagnosed in the south-east but rarely in the north. In the Midlands, where disease has recently appeared in dogs, prevalence in foxes was 4.8% (2-11). Close geographical overlap of parasite distribution in foxes and dogs does not necessarily indicate an important wildlife reservoir of infection, but does suggest that A. vasorum might be spreading northwards. The hearts of infected foxes had thicker right ventricles than those of uninfected foxes, suggesting that the parasite could affect fox health and fitness. Burdens ranged from 1 to 59 adult nematodes. Sex, age and body condition were not significantly associated with infection. Eucoleus aerophilus and Crenosoma vulpis, nematode parasites of the respiratory system, were found in 213 and 11 foxes respectively, with slightly higher prevalence of E. aerophilus in the south and east. No specimens of the heartworm Dirofilaria immitis were found, giving an upper 95% confidence interval for prevalence of 0.84%.

Epidemiology of subclinical dairy cow nematode infections on five farms in England in 2002 and a comparison with results from 1978 to 1979.

A detailed study of the epidemiology of subclinical nematode infections in adult dairy cows was conducted on five well-managed English dairy farms during the 2002 grazing season. These same farms had also participated in a similar study in 1978-1979 and thus provided a unique opportunity to compare the epidemiological findings after an interval of over two decades. Common factors, including the prevalence of infection, faecal worm egg output, pasture larval count and nematode genera present, and estimated daily larval intake at pasture, were compared between the two studies. Subclinical roundworm burdens, as judged by faecal egg counts, were widespread in dairy cows in 2002, but the prevalence of animals with patent infections and the magnitude of the worm egg output were significantly lower than in 1978-1979. Both the prevalence of infection and mean faecal worm egg output exhibited a marked seasonal peak during the summer months in 2002. Pasture larval numbers were, however, nearly three times higher in 2002 with Ostertagia dominating the nematode genera to which cows were exposed at pasture in both surveys. Factors considered likely to account for differences in parasite epidemiology include the marked increase in herd productivity over the intervening period, the increased proportion of Holstein genetics with the consequent increase in milk yield and feed intake, changes in grassland management and increased stocking rate seen on the majority of farms.

Vagal and splanchnic afferent nerves are not essential for anorexia associated with abomasal parasitism in sheep.

Heavy burdens of the abomasal nematode, Ostertagia (Telodorsagia) circumcincta, in growing lambs result in a reduction in liveweight gain due largely to a drop in voluntary feed intake. The present study investigated: (1) the role of subdiaphragmatic vagal and non-vagal visceral afferent nerves in mediating a reduction in voluntary feed intake, using subdiaphragmatic vagal deafferentation (vagotomy) either alone or in combination with coeliac-superior mesenteric ganglionectomy (vagotomy and sympathectomy); and (2) the association between appetite, abomasal pH, selected blood values (amidated gastrin (G-17-amide), glycine-extended gastrin (G-17-Gly), pepsinogen and leptin) and worm burden, in sheep experimentally infected with 100,000 O. circumcincta infective larvae per os. Neither vagotomy alone nor vagotomy and sympathectomy in combination adversely affected the establishment or course of development of the parasite burden, when compared with a control group subject to sham surgery. Furthermore, neither surgical procedure prevented the drop in appetite seen 5-10 days post-infection, although combined vagotomy and sympathectomy did reduce voluntary feed intake prior to the start of the study. Ostertagia infection resulted in a significant increase in abomasal pH in all three groups, which was accompanied by an increase in blood G-17-amide and in G-17-Gly, the latter reported for the first time in parasitized ruminants. There were no significant differences in blood leptin, also reported for the first time in parasitized sheep, either between groups or in comparison with pre-infection levels, though weak negative correlations were established between blood leptin and appetite from day 5 to the end of the study in all three groups and a positive correlation with blood G-17-amide in the control group over the same period. These data suggest that neither intact subdiaphragmatic vagal afferent nerves or coeliac-superior mesenteric ganglion fibres, nor changes in circulating gastrin and leptin concentrations play a major role in mediating the hypophagic effects of O. circumcincta in parasitized sheep.

Field trial of a Caryospora species vaccine for controlling clinical coccidiosis in falcons.

A Caryospora species vaccine was prepared and used in an attempt to prevent infection and associated morbidity in falcons. A blind field trial was conducted, involving a vaccinated group of 20 birds and two control groups of seven and four birds, which were subsequently challenged with a live mixed-species vaccine. There was a statistically significant reduction in morbidity and shedding of oocysts in the vaccinated group compared with the control groups.

Effects of Ostertagia ostertagi and omeprazole treatment on feed intake and gastrin-related responses in the calf.

Infection with the bovine abomasal nematode, Ostertagia ostertagi, results in a loss of acid-secreting parietal cells and an increase in gastric pH. The effects of an experimental infection with Ostertagia and/or daily treatment with omeprazole (OMP) at 2mgkg(-1) bodyweight for four consecutive days (experiment days 24-27, inclusive) on voluntary feed intake, blood and tissue gastrin concentrations, abomasal G-cell numbers, gastric pH, and blood cholecystokinin (CCK) and pepsinogen concentrations were investigated in the calf. Ostertagia-infected calves demonstrated a significant drop in feed intake between days 24 and 27 post-infection (38%; P<0.001) and in G-cell numbers (42%; P<0.05) and significant increases in abomasal pH (P<0.001), fundic mucosal weight (99%; P<0.01), and blood gastrin (P<0.05) and pepsinogen (P<0.0001). OMP treatment of worm-free animals resulted in a significant drop in intake between days 24 and 27 (30%; P<0.001) and in G-cell numbers (17%; P<0.05) and significant increases in abomasal pH (P<0.01) and blood gastrin (P<0.001). OMP treatment of Ostertagia-infected animals with an existing hypergastrinaemia had no effect on feed intake, abomasal pH, blood gastrin or pepsinogen or abomasal G-cell numbers. Blood CCK concentrations were also unaffected by either Ostertagia infection or OMP treatment. These data suggest that: (a) the depression in feed intake associated with OMP in worm-free calves was not due to a side effect of drug treatment; (b) inappetance in Ostertagia-infected animals is closely associated with the parasite-induced hypergastrinaemia; and (c) the elevation in abomasal pH was a major factor responsible for the elevated blood gastrin concentrations seen in parasitised and OMP-treated animals.

Experiments showing that electromagnetic fields can be used to treat inflammatory diseases.

While it is well known that electromagnetic fields (EMFs) can induce repair of non-healing bone fractures, EMF therapy remains confined to orthopedic clinics mainly because the biological and physical mechanisms underlying the therapy are unknown. However, it is generally believed that non-invasive, EMF therapy might have a broad, albeit currently unrecognized clinical potential. In support of this view, we report that 0.1 mT, 60 Hz EMFs induce a 20% mean-increase in anti-CD3 binding to T cell receptors (TcRs) of Jurkat cells, a T lymphocyte cell line. Additionally, we show that 60 Hz sinusoidal EMFs and a commercial bone healing EMF modulate signal transduction pathways that regulate lymphocyte proliferation and that are normally triggered by activating the Jurkat TcR. Similar EMF effects are shown in human peripheral blood lymphocytes (hPBLs), exposed to EMFs in culture and in rat PBLs, when donor animals are exposed to a bone healing field (21 days, 4 hr/day). Although we do not yet satisfactorily understand the differences we obtain in cell and animal based experiments, our findings clearly demonstrate that EMFs can regulate lymphocyte proliferation in vitro and in vivo. Since T cells are key modulators of inflammation, the development of EMF based therapeutic devices to regulate their activity can be expected to provide important tools to treat numerous human inflammatory diseases such as psoriasis and arthritis.

Models of the uniformity of electro-magnetic fields generated for biological experiments by Merritt coils.

Electromagnetic field (EMF) producing wire coils were described by Merritt et al, Rev. Sci. Instrum. 54 (7), 1983. Merritt coils produce large volume EMFs in which statistical numbers of biological experiments are performed. We build and use Merritt coils for cell/animal studies and are developing therapeutic EMF systems. Here we present models illustrating the EMFs produced by our coils and discuss the criteria that should be applied to the use of Merritt and other coils to achieve valid experimental results. In a companion paper at this meeting Nindl et al, describe biological experiments, using these Merritt coils, showing that EMFs may be useful in treating many inflammatory disease states. Although the large-volume EMFs produced by Merritt coils are convenient for biological experiments the EMFs are not perfectly uniform and the deviations can be a significant source of experimental error. The orientation and size of experimental objects are key contributors to these deviations. To evaluate our Merritt coils we solved the Biot-Savart law explicitly for ideal 3-coil and 4-coil Merritt systems and compared these theoretical EMFs with those of our systems. We present a detailed examination of deviations in magnetic field amplitude, as well as magnetic field direction, as a function of location within the coils. We find that spherically shaped experimental sets minimize these deviations. We developed simple formulae for accurately predicting deviations associated with Merritt coils.

Cold stress-induced modulation of cell immunity during acute Toxoplasma gondii infection in mice.

Infection with Toxoplasma gondii in the acute phase results in nonspecific suppression of immunologic function in mice and humans. The present study examined the effects of a physical stressor, i.e., cold stress (CS), on macrophage function (nitrite production, parasite survival) and splenic blastogenesis in the acute phase of murine T. gondii infection. In our stress paradigm, female BALB/c mice were placed in cold water (1 +/- 0.5 C), 5 min each day for 8 days. Nitrite production and parasite survival were measured in cultured peritoneal macrophages obtained from mice subjected to CS after in vivo activation with interferon-gamma/lipopolysaccharide (CS + ACT), and in vitro infection with T. gondii tachyzoites. Peritoneal macrophages from CS + ACT mice showed decreased nitrite production compared to control but activated cells (ACT). Spleen cell proliferation to in vitro stimulation with the mitogens concanavalin A (Con A) and anti-CD3, and Toxoplasma lysate antigen (TLA) was measured in splenocytes obtained from BALB/c mice during the acute phase of infection with T. gondii. Mice subjected to CS and infection (CS + INF) had maximum splenocyte proliferation on days 8 and 15 followed by a subsequent decline on day 28 postinoculation (PI). In contrast, infected mice not subjected to stress (INF) showed decreased splenocyte proliferation on days 8 and 15 followed by an increase on day 28 PI. The rate of mortality was decreased in the CS + INF compared to the INF group during acute infection. These results suggest that CS may alter the pathogenesis of T. gondii infection by modulating acute-phase responses, provoking a state of transient disequilibrium between the host and parasite.

Evaluation of flea control strategies using fipronil on cats in a controlled simulated home environment.

Three groups of six cats were kept in similar carpeted pens in which a self-replicating population of Ctenocephalides felis had been established. One group was left untreated, but the other groups were treated every 28th day with 0.5 ml of a 10 per cent fipronil spot-on formulation, and the cats in one of the treated groups also wore a methoprene collar. No fleas were found on any of the treated cats, either during the first 13 weeks of the study, when heavy flea burdens were developing in the control pen, or over the next 11 weeks when a declining number of fleas was present on the control group.

Identification of potential activators of proteinase-activated receptor-2.

In order to identify physiological activators of proteinase-activated receptor-2 (PAR-2), a peptide chloromethane inhibitor (biotinyl-Ser-Lys-Gly-Arg-CH2Cl) based on the cleavage site for activation of PAR-2 was synthesised and tested with 12 trypsin-like serine proteinases. The second-order rate constant (ki/Ki) for the formation of the covalent proteinase-inhibitor complex varied by 2 x 10(5)-fold between the proteinases. Biotinyl-Ser-Lys-Gly-Arg-CH2Cl reacted very rapidly with trypsin, acrosin from sperm and tryptase from mast cells: the ki/Ki values with these proteinases were greater than 10(5) M(-1) x s(-1). Thus, the specificity of these proteinases matched the sequence of the activation site of PAR-2 and it can be concluded that these proteinases are potential physiological activators of PAR-2.

Comparison of flea control strategies using imidacloprid or lufenuron on cats in a controlled simulated home environment.

OBJECTIVE: To compare the effect of monthly treatments with imidacloprid (an adulticide) or lufenuron (an insect development inhibitor) for protecting cats against Ctenocephalides felis felis in a simulated home environment. ANIMALS: 3 matched groups of 4 cats each. PROCEDURE: A self-propagating flea life cycle continuously exposing cats to 'natural' infestation was established in 3 pens. Small artificial infestations were later superimposed to mimic the effect of a cat roaming outdoors and acquiring extraneous fleas. One pen housed an untreated control group, and the other 2 pens housed cats treated every 28th day with an imidacloprid spot-on formulation or lufenuron suspension, respectively. Flea counts were performed at 14-day intervals for 112 days. RESULTS: Flea numbers increased on control cats around day 42 when mean counts on cats in the imidacloprid and lufenuron groups decreased by 100 and 86.8 percent, respectively. Fleas were not found on any imidacloprid-treated cat, but lufenuron-treated cats were consistently parasitized. CONCLUSIONS: Imidacloprid administered at monthly intervals maintained flea burdens below the limit of detection, whereas clinically important flea populations developed in the lufenuron treatment pen. CLINICAL RELEVANCE: Results from this experimental model suggest that flea populations within a home may be controlled by carefully timed on-host treatments with potent long-acting insecticides such as imidacloprid.