Olwen Golden

Fasciola hepatica is responsible for human disease and economic livestock loss on a global scale. Unlike the well characterized schistosomes, only the adult and juvenile stages of F. hepatica are implicated in disease, whereas the freely voided egg is not thought to contribute to host-parasite interactions. We investigated specific immune responses to soluble F. hepatica egg proteins (SFHEP), during a 14-week experimental infection, demonstrating significant increases in anti-SFHEP IgG1 (P = 0.001), transforming growth factor beta-1 (P = 0.008) and IL-10 (P < 0.001) titres at the onset of egg production. Western blot analysis of soluble SFHEP demonstrates that protein bands migrating at 61.6, 54.8 and 44 kDa become sero-reactive before the appearance of eggs within host faeces. Therefore, expression of some egg-associated proteins indicates progression to chronic disease. Antigenic bands were investigated through mass spectrometry, identifying a protein disulphide isomerase (PDI) (61.6 kDa), an enolase and ferritin-related proteins (54.8 kDa), and a cocktail of dehydrogenases (44 kDa). Biochemical analysis of egg secretions reveals proteolytic activity, which increases over time, indicating that proteases may be continually secreted during the course of egg maturation. The implications of egg-specific immune responses and proteolytic secretions are further discussed.

The increasing movement of people to wilderness areas, shrinking of wildlife habitats and the resulting urbanisation of wildlife has led to growing concerns about the transfer of parasitic diseases, particularly from contaminated faeces. Faecal samples from wild carnivores in Ireland were examined for the presence of protozoan and nematode parasites. Red fox (Vulpes vulpes) samples (n = 91) were positive for Uncinaria stenocephala (38%), Eucoleus aerophilus (26%), Toxocara canis (20%), Trichuris vulpis (4%) and Isospora-like oocysts (9%). Badger (Meles meles) samples (n = 50) were positive for Uncinaria criniformis (40%), E. aerophilus (6%) and Isospora-like oocysts (16%). No parasites were observed in pine marten (n = 48; Martes martes) faeces. Approximately 5% of American mink (Mustela vison) samples were positive for Cryptosporidium by polymerase chain reaction (identified as Cryptosporidium andersoni (n = 3) and 'mink' genotype (n = 1)). The results suggest that wild carnivores in Ireland have a range of parasites, although it is unclear from the present study to what extent these infections are associated with morbidity. While it can be expected that, via their faeces, wild carnivores contribute to the spread of these parasites, they are unlikely the primary source of environmental contamination. Therefore, they should not always be the principal target of control measures.

ABSTRACT Fasciola hepatica is a prevalent helminth parasite of livestock. Infection results in polarization of the host's immune response and generation of type 2 helper (Th2) immune responses, which are known to be inhibitory to Th1 responses. Bovine tuberculosis (BTB) is a bacterial disease of economic and zoonotic importance. Control polices for this disease rely on extensive annual testing and a test-and-slaughter policy. The correct diagnosis of BTB relies on cell-mediated immune responses. We established a model of coinfection of F. hepatica and Mycobacterium bovis BCG to examine the impact of helminth infection on correct diagnosis. We found the predictive capacity of tests to be compromised in coinfected animals and that F. hepatica infection altered macrophage function. Interleukin-4 and gamma interferon expression in whole-blood lymphocytes restimulated in vitro with M. bovis antigen was also altered in coinfected animals. These results raise the question of whether F. hepatica infection can affect the predictive capacity of tests for the diagnosis of BTB and possibly also influence susceptibility to BTB and other bacterial diseases. Further studies on the interplay between helminth infection and BTB are warranted.

Rectal swabs were collected from 147 household dogs and 35 household cats, including healthy animals, animals with gastrointestinal signs and animals with a variety of medical and surgical conditions. A combination of selective culture methods was used to optimise the recovery of Campylobacter species, and a PCR was used to confirm their isolation and to identify the species. The overall prevalence of Campylobacter species was 42·9 per cent in the cats and 41·5 per cent in the dogs. Campylobacter upsaliensis was the species most commonly isolated from the dogs and cats, and Campylobacter jejuni was the second most commonly isolated. Particularly high prevalences were detected in the few cats and dogs with diarrhoea, and in the cats and dogs that were six months old or younger.

Parasitic nematodes of the genus Anisakis are among the most important biological hazards associated with seafood. A survey of Anisakis spp. in European hake (Merluccius merluccius) was undertaken as this species is a staple of the Portuguese diet. Moreover, a literature review of cases of anisakiosis reported from Portugal, a country with one of the highest levels of fish consumption in the world, was also carried out. Seventy-five European hake caught in the Atlantic Ocean off the northern coast of Portugal were analyzed to determine the infection levels and site distribution of Anisakis spp. Isolated nematode larvae were identified to species level by molecular analysis. Two sets of samples were collected. Firstly, a total of 46 Anisakis spp. L3 larvae were collected with a prevalence of 76.7% (95% CI 61.5–91.8%) and intensity (mean ± SD, range) of 2.0 ± 1.2 (1–5). Most larvae were found on the liver (45.7%) and on the gonads (32.6%), but none in the muscle. The molecular analysi...

Five culture methods for the recovery of Campylobacter species (spp.) were evaluated on 361 rectal swabs collected from cats and dogs in Ireland. Speciation using PCR methods was performed on all isolates to assess the sensitivity of each culture method for isolation of Campylobacter spp., and to establish the prevalence of Campylobacter jejuni, C. coli, C. upsaliensis, C. lari and C. helveticus. Overall 163 of 361 (45.2%) samples were confirmed Campylobacter spp. positive. Direct plating onto modified charcoal cefoperazone deoxycholate agar (mCCDA) with cefoperazone, amphotericin and teicoplanin (CAT) selective supplement yielded a significantly higher prevalence of Campylobacter spp. (33.0%) than each of the other four methods (P < or = 0.05). This method was also the most sensitive method for isolation of C. upsaliensis compared with any of the other four methods used in the current study (P < or = 0.05). A direct plating method onto mCCDA agar with CCDA selective supplement and a filtration method onto blood agar after pre-enrichment in CAT supplemented broth yielded lower Campylobacter spp. prevalences of 19.7% and 17.5% respectively. A filtration method onto CAT agar and pre-enrichment in Preston broth before plating onto mCCDA agar were less sensitive for the isolation of Campylobacter spp. Speciation results of Campylobacter isolates revealed the majority of Campylobacter isolates were C. upsaliensis (50.0%) and C. jejuni (41.9%). A small number of isolates were C. coli (2.6%), C. lari (1.5%) and C. helveticus (1.1%). The overall detection of Campylobacter spp. in the 361 pets sampled was significantly increased by using a combination of isolation methods (P < or = 0.05), producing a more accurate determination of the prevalence of Campylobacter spp. in pets in Ireland and of the actual Campylobacter species. As the majority of Campylobacter spp. were recovered by direct plating onto mCCDA agar with CAT supplement, this method is the method of choice if only a single method is selected for isolation of the most common Campylobacter spp. detected in pets and humans.

Fasciola hepatica is responsible for human disease and economic livestock loss on a global scale. Unlike the well characterized schistosomes, only the adult and juvenile stages of F. hepatica are implicated in disease, whereas the freely voided egg is not thought to contribute to host-parasite interactions. We investigated specific immune responses to soluble F. hepatica egg proteins (SFHEP), during a 14-week experimental infection, demonstrating significant increases in anti-SFHEP IgG1 (P = 0.001), transforming growth factor beta-1 (P = 0.008) and IL-10 (P < 0.001) titres at the onset of egg production. Western blot analysis of soluble SFHEP demonstrates that protein bands migrating at 61.6, 54.8 and 44 kDa become sero-reactive before the appearance of eggs within host faeces. Therefore, expression of some egg-associated proteins indicates progression to chronic disease. Antigenic bands were investigated through mass spectrometry, identifying a protein disulphide isomerase (PDI) (61.6 kDa), an enolase and ferritin-related proteins (54.8 kDa), and a cocktail of dehydrogenases (44 kDa). Biochemical analysis of egg secretions reveals proteolytic activity, which increases over time, indicating that proteases may be continually secreted during the course of egg maturation. The implications of egg-specific immune responses and proteolytic secretions are further discussed.

Fasciola hepatica is a prevalent helminth parasite of livestock. Infection results in polarization of the host's immune response and generation of type 2 helper (Th2) immune responses, which are known to be inhibitory to Th1 responses. Bovine tuberculosis (BTB) is a bacterial disease of economic and zoonotic importance. Control polices for this disease rely on extensive annual testing and a test-and-slaughter policy. The correct diagnosis of BTB relies on cell-mediated immune responses. We established a model of coinfection of F. hepatica and Mycobacterium bovis BCG to examine the impact of helminth infection on correct diagnosis. We found the predictive capacity of tests to be compromised in coinfected animals and that F. hepatica infection altered macrophage function. Interleukin-4 and gamma interferon expression in whole-blood lymphocytes restimulated in vitro with M. bovis antigen was also altered in coinfected animals. These results raise the question of whether F. hepatica...