Calicophoron daubneyi (Paramphistomidae) in slaughtered cattle in Castilla y León (Spain)

The prevalence and aetiology of natural paramphistomosis was investigated in cattle slaughtered in the Castilla y León region (Spain) over a 3 year-period. The overall prevalence of positive animals was 6.20%. The parasite burden per animal ranged from 8 to 8005 (median = 144) and the ruminal atrium had the highest parasite burden whereas the ruminal dorsal sac the lowest. The prevalence and parasite burden increased with age while these parameters were lower in cattle under intensive management. Calicophoron daubneyi was the only Paramphistomidae species identified using morphoanatomical, histological and molecular methods in the studied animals.     

Risk factors associated with Neospora caninum seropositivity in randomly sampled Canadian dairy cows and herds

Our objective was to determine cow- and herd-level risk factors associated with seropositivity for Neospora caninum in a large number of randomly selected Canadian dairy herds, controlling for important confounding variables and co-infections with bovine leukemia virus (BLV), bovine viral diarrhea virus (BVDV) and Mycobacterium avium subspecies paratuberculosis (MAP). Serum samples were obtained from 30 randomly selected cows, where available, in 240 herds using monthly milk testing, within 6 of 10 provinces, and these samples were tested for antibodies against BLV, MAP and N. caninum using commercially available ELISA test kits. Five unvaccinated cattle >6 months old from each herd were tested for antibodies to BVDV using virus neutralization. Most herd-level predictors were obtained through personal interviews with questionnaires administrated to each farm manager. A mixed logistic-regression model was built using N. caninum serostatus at the cow-level as the outcome variable, with herd as a random effect and province as a fixed effect. A BLV seropositive cow was 1.50 times more likely to be seropositive for N. caninum than a BLV-seronegative cow, and this was the only cow-level variable to remain in the final model. Regarding herd-level variables, with “no on-farm dogs” as the baseline, “presence of dogs but not known to eat placentas and/or fetuses” increased the odds of seropositivity for N. caninum by a factor of 1.66. For “presence of dogs known to eat placentas and/or fetuses”, the odds ratio (OR) was 2.75, demonstrating a dose–response relationship. “Using embryo transfer” (OR = 0.69), “asking for a BVDV-negative test before introducing an animal” (OR = 0.30), “using monensin in dry cows” (OR = 0.71), and “heifers having nose-to-nose contact with calves” (OR = 0.73) were all dichotomous variables negatively associated with seropositivity for N. caninum. “Number of milk cows on the farm” (OR = 0.99), and “area (acres) used for forage production” (OR = 0.99) were continuous variables negatively associated with N. caninum seropositivity.     

Apparent seroprevalence,isolation and identification of risk factors for brucellosis among dairy cattle in Goa,India

Brucellosis is a highly contagious zoonotic infection affecting livestock and human beings. The disease has been reported worldwide except in few countries where it has been eradicated. The prevalence of brucellosis among cattle from 11 farms having a history of abortions was studied. A total of 481 samples comprising of blood, milk, vaginal swabs, vaginal discharges, placental tissues and fetal tissues were collected from 296 animals. Clinical samples were processed for the isolation of Brucella. Serum samples (n = 296) were tested by Rose Bengal Plate Test (RBPT) and indirect ELISA. A total of 90 (30.40%) and 123 (41.55%) samples were positive by RBPT and indirect ELISA, respectively. Also 27.02% samples were positive by both the tests. Brucella isolates (n = 8) were recovered from clinical samples using Brucella selective media. All the isolates demonstrated PCR amplification for the bcsp31 and IS711 genes. Amplification of Brucella abortus specific primer was demonstrated by all the isolates in AMOS PCR indicating isolates to be of either B. abortus biotype 1, 2 or 4. Risk factors for transmission of brucellosis among cattle population were studied by field surveys. It was observed that lack of awareness about brucellosis (OR = 8.739, P = 0.138) and inadequate floor space (OR = 0.278, P = 0.128) were crucial risk factors for transmission of bovine brucellosis.     

Bovine paramphistomosis in Galicia (Spain): Prevalence,intensity, aetiology and geospatial distribution of the infection

The present study explored various basic aspects of the epidemiology of paramphistomosis in Galicia, the main cattle producing region in Spain. In total, 589 cows from different farms located across the region were selected at random in the slaughterhouse for examination of the rumens and reticula for the presence of Paramphistomidae flukes. Paramphistomes were found in 111 of 589 necropsied cows (18.8%; 95% CI: 15.7–21.9%), with higher prevalences of infection in beef cows than in dairy cows (29.2% vs 13.9%). Although the number of flukes per animal was generally low (median = 266 flukes), some cows harboured large parasite burdens (up to 11,895 flukes), which may have harmful effects on their health or productivity. Cows with higher parasite burdens also excreted greater numbers of fluke eggs in their faeces, which suggests that heavily parasitized mature cows play an important role in the transmission of paramphistomosis. This role may be particularly important in Galicia, where the roe deer, which is the only wild ruminant in the study area, was found not to be a reservoir for the infection. The use of morpho-anatomical and molecular techniques applied to a large number of fluke specimens provided reliable confirmation that Calicophoron daubneyi is the only species of the family Paramphistomidae that parasitizes cattle in Galicia. The environmental data from the farms of origin of the necropsied cows were used in Bayesian geostatistical models to predict the probability of infection by C. daubneyi throughout the region. The results revealed the role of environmental risk factors in explaining the geographical heterogeneity in the probability of infection in beef and dairy cattle. These explanatory factors were used to construct predictive maps showing the areas with the highest predicted risk of infection as well as the uncertainty associated with the predictions.     

Risk factors for Salmonella spp in Portuguese breeding pigs using a multilevel analysis

Salmonella is the second most frequent cause of foodborne illness in the European Union (EU), so EU enforced legislation to achieve a reduction in Salmonella prevalence in the swine sector. To set the reduction target each country carried out a baseline survey to estimate Salmonella prevalence. The aim of our study was to identify risk factors for the presence of Salmonella in breeding pigs based on the data of the Baseline Study for Salmonella in Breeding Pigs in Portugal. In total, 1670 pen fecal samples from 167 herds were tested by culture and 170 samples tested positive. Along with the collection of the samples a survey was applied to collect information about the herd management and potential risk factors. Multilevel analysis was applied to the data using generalized linear mixed models and a logit link function. The outcome variable was the presence/absence of Salmonella in the pen fecal samples. The first level was assigned to the pen fecal samples and the second level to the herds. The results showed significant associations between Salmonella occurrence and the factors (p < 0.05): maternity pens versus mating pens (OR = 0.39, 95%CI: 0.24–0.63), feed from external or mixed source versus home source (OR = 2.81, 95%CI: 1.19–6.61), more than 10 animals per pen versus 10 animals per pen (OR = 2.02, 95%CI: 1.19–3.43), North Region versus Alentejo Region (OR = 3.86, 95%CI: 1.08–13.75), rodents control (OR = 0.23, 95%CI: 0.090–0.59), more than 90% of boars homebred or no boars versus more than 90% of boars from an external source (OR = 0.54, 95%CI: 0.3–0.97), semen from another herd versus semen from insemination centers (OR = 4.47, 95%CI: 1.38–14.43) and herds with a size of 170 or more sows (OR = 1.82, 95%CI: 1.04–3.19). This study offers very relevant information for both the Portuguese veterinary authorities and the pig farmers currently developing control programmes for Salmonella. This is the first study providing evidence for semen and boars source as risk factors for Salmonella in breeding pigs.     

NTPDase and 5′-nucleotidase as inflammatory markers in cattle naturally infected by Eurytrema coelomaticum

The aim of this study was to evaluate seric NTPDase and 5′nucleotidase activities of cattle naturally infected by Eurytrema coelomanticum, as well as to correlate them to histopathological lesions in the pancreas and the degree of parasitism. Blood samples and pancreas of 51 bovines were collected on a slaughterhouse in Southern Brazil: 33 from cattle naturally infected by E. coelomanticum (the Group A), and 18 from uninfected animals (the Group B). Infected animals showed an average of 532 parasites per pancreas. In the pancreatic histology, ducts displayed hyperplasia, stenosis, proliferation of fibrous tissue, and interstitial inflammatory infiltration of lymphocytes. The serum from infected animals showed an increase in NTPDase activity when ATP was used as substrate (P < 0.001). For the ADP substrate, there was no difference between groups regarding NTPDase activity (P = 0.37), as well as 5′-nucleotidase activity (P = 0.27). Correlating NTPDase activity (ATP substrate) with the degree of histopathological lesions (rho = 0.66, P < 0.001) and the parasitic load on the pancreas (rho = 0.65, P < 0.001), a positive correlation was observed. Similar results were found between the degree of histopathological lesions and NTPDase activity (ADP substrate; rho = 0.29, P = 0.03), and 5′nucleotidase activity (rho = 0.35, P = 0.01). Based on the results of NTPDase and 5′nucleotidase enzymes in cattle naturally infected by E. coleomanticum, it is possible to suggest that these enzymes are involved in the modulation of inflammation, and they can act as markers of inflammatory response.     

An unmatched case controlled study of clinicopathologic abnormalities in dogs with Bartonella infection

We compared clinicopathologic findings in dogs with Bartonella infection to Bartonella spp. negative dogs suspected of a vector-borne disease. Cases (n = 47) and controls (n = 93) were selected on the basis of positive or negative enrichment culture PCR results, respectively. Signalment, clinicopathologic findings and treatments were extracted from medical records. DNA sequencing identified Bartonella henselae (n = 28, 59.6%), Bartonella vinsonii subsp. berkhoffii (n = 20, 42.6%), Bartonella koehlerae (n = 3, 6.4%), Bartonella volans-like (n = 3, 6.4%) and Bartonella bovis (n = 1, 2.1%). There were no significant differences in age, breed, size, sex or neuter status between cases and controls. Dogs infected with Bartonella sp. often had a history of weight loss [OR = 2.82; 95% CI: 1.08–7.56] and were hypoglobulinemic [OR = 4.26; 95% CI: 1.31–14.41]. With the exception of weight loss and hypoglobulinemia, clinicopathologic abnormalities in Bartonella-infected dogs in this study were similar to dogs suspected of other vector-borne infections.     

Western European epidemiological survey for parvovirus and coronavirus infections in dogs

An epidemiological survey for canine parvovirus (CPV) and canine coronavirus (CCoV) infections was conducted in Western Europe. A total of 156 faecal samples were collected from dogs with diarrhoea in Spain (n = 47), Italy (n = 39), France (n = 26), Germany (n = 21), the United Kingdom (n = 8), Belgium (n = 10), and the Netherlands (n = 5). Using molecular assays for virus detection and characterisation, CPV and CCoV were found to be widespread in European dog populations, either alone or in mixed infections. In agreement with previous reports, the original type CPV-2 was shown not to circulate in European dogs. The recently identified virus variant CPV-2c was predominant in Italy and Germany and present at high rates in Spain and France but was not detected in the UK or Belgium. Except for the UK, CCoV genotype I was identified in all European countries involved in the survey, albeit at a lower prevalence rates than CCoV genotype II.     

Antimicrobial resistant Escherichia coli isolates in cattle and house sparrows on two Czech dairy farms

Rectal smears of calves, cows and young bulls, as well as cloacal smears of house sparrows (Passer domesticus), from farms at the villages of Šumice and Troskotovice, Czech Republic, were examined for E. coli resistant to 12 antimicrobials. The resistant isolates were tested for antimicrobial-resistance genes and integrons. Totals of 40% (n = 183), 3% (n = 95), 0% (n = 33), and 9% (n = 54) of Escherichia coli isolates from calves, cows, young bulls and house sparrows, respectively, were antimicrobial resistant. The following genes were identified in cattle E. coli isolates: tetA, tetB (isolates resistant to tetracycline), bla_TEM (beta-lactams), strA, aadA (streptomycin), sul1, sul2 (sulphonamides), and cat, floR (chloramphenicol). Seven of 16 antimicrobial-resistant calf isolates from the Šumice farm possessed class 1 integrons with the aadA1 gene cassette integrated, 1 kb in size. On the Troskotovice farm, eight of 57 antimicrobial-resistant calf isolates possessed class 1 integrons. Integrons of 1.5 kb with the dhfr1- aadA1 gene cassette were found in four isolates, followed by a 1 kb integron with the aadA1 gene found in three isolates, and a 1.7 kb integron with the dhfr17-aadA5 gene cassette and the phenotype ASSuTSxtNaCipCCfG. The prevalence of resistant E. coli in calves compared to adult cattle was much higher and probably was influenced by oral antimicrobial usage in calves, feeding with milk and colostrum from treated cows, as well as mechanisms unrelated to antimicrobial drug selection. Although house sparrows lived together with the cattle and came into contact with cattle waste on the farm, they were not infected by resistant E. coli isolates with the same characteristics as those found in cattle.     

Stocking Rate and Fuels Reduction Effects on Beef Cattle Diet Composition and Quality

An experiment was conducted to evaluate the influence of forest fuels reduction on diet quality, botanical composition, relative preference, and foraging efficiency of beef cattle grazing at different stocking rates. A split plot factorial design was used, with whole plots (3 ha) being fuel reduced or no treatment (control), and split plots (1 ha) within whole plots were grazed to three levels of forage utilization; (low) 3 heifers · ha^?1, (moderate) 6 heifers · ha^?1, (high) 9 heifers · ha^?1, with a 48-h grazing duration. Grazing treatments were applied in August of 2005 and 2006. Cattle diet composition and masticate samples were collected during 20-min grazing bouts using six ruminally cannulated cows in each experimental unit. Relative preference indices indicated a strong preference for grass regardless of treatment and stocking rate. Grass consumption was lower in control pastures (P < 0.05) and tended (P < 0.095) to decrease with increased stocking rates. Shrub use was higher in control pastures displaying a quadratic effect (P < 0.05) due to stocking, whereas shrub use increased with stocking rate across all treatments. Cattle grazing control pastures consumed diets higher in crude protein compared to cattle grazing treated pastures (P < 0.05). In vitro dry matter digestibility values were lower (P < 0.05) in control sites and tended (P = 0.10) to decrease with increased stocking rates. In both control and treated pastures, bites per minute and grams consumed per minute declined (P = 0.003) with increased stocking, indicating foraging efficiency of cattle decreases with increased stocking rates. Our data indicated cattle grazing late season grand fir habitat types have a strong preference for grasses regardless of treatment or stocking rate. However, as stocking rate increased in both control and treated pastures, grass consumption decreased, shrub consumption increased, and foraging efficiency decreased.     

Entamoeba infections in different populations of dogs in an endemic area of Lahore,Pakistan

Entamoeba histolytica, a protozoan parasite that affects humans and other primates all over the world. It is a common waterborne pathogen in endemic areas that have fecal oral transmission cycle. The aim of the present study was to examine the prevalence of E. histolytica and other Entamoeba species cysts in three different dog populations. Fecal samples from 600 dogs were collected and processed to detect Entamoeba cysts using the triple fecal test (light microscopy) and fecal antigens of E. histolytica were detected using a fecal antigen ELISA (TechLab E. histolytica II). Because it is impossible to differentiate E. histolytica from Entamoeba dispar and E. moshkovskii, using light microscopy we referred to all cysts morphologically consistent with E. histolytica as E. histolytica/dispar/moskovskii to reflect this uncertainty. Samples from 197 household dogs without clinical signs, 122 samples from household dogs exhibiting clinical signs of diarrhea, dysentery and vomiting and 281 stray dogs with no specific clinical signs were examined. Entamoeba histolytica-like cysts were observed in 94 (15.6%, 95% CI = ±3.88) by triple fecal test microscopy and E. histolytica antigens were demonstrated in 66 (11%, 95% CI = ±4.41) by fecal antigen ELISA in 600 fecal samples. Significant differences (P ≤ 0.05) in prevalence were found between the three populations. Twenty (10.1%, 95% CI = ±7.86) and 11 (5.6%, 95% CI = ±7.70) of 197 fecal samples from household dogs without clinical signs were positive by microscopy and by antigen ELISA, respectively. Twenty-nine (23.8%, 95% CI = ±6.58) and 23 (18.8%, 95% CI = ±7.81) of 122 the fecal samples from household dogs with clinical signs were positive by microscopy and by antigen ELISA, respectively. Forty-five (16.01%, 95% CI = ±5.62) and 32 (11.3%, 95% CI = ±6.38) of 281 fecal samples from stray dogs were positive by microscopy and by fecal antigen ELISA, respectively. Dogs from the youngest age group (6 months to 1 year) were more likely to be E. histolytica antigen positive than were dogs from the other two older age groups, with a significant difference (P ≤ 0.05) between all age groups. Statistically, no significant (P ≥ 0.05) difference of prevalence was seen in male and female dogs. The local dogs had the highest prevalence rate of E. histolytica antigens (36 of 246, 14.2%, 95% CI = ±6.32) followed by imported breeds (11 of 115, 9.5%, 95% CI = ±10.4) and crossbred (19 of 239, 8.3%, 95% CI = ±7.47), indicating a significant (P ≤ 0.05) trend of positivity between various breeds of dogs. These findings suggest that dogs may play an important role in the epidemiology of this pathogen.     

Molecular biological identification of Babesia,Theileria, and Anaplasma species in cattle in Egypt using PCR assays,gene sequence analysis and a novel DNA microarray

In this preliminary study, a novel DNA microarray system was tested for the diagnosis of bovine piroplasmosis and anaplasmosis in comparison with microscopy and PCR assay results. In the Dakahlia Governorate, Egypt, 164 cattle were investigated for the presence of piroplasms and Anaplasma species. All investigated cattle were clinically examined. Blood samples were screened for the presence of blood parasites using microscopy and PCR assays. Seventy-one animals were acutely ill, whereas 93 were apparently healthy. In acutely ill cattle, Babesia/Theileria species (n = 11) and Anaplasma marginale (n = 10) were detected. Mixed infections with Babesia/Theileria spp. and A. marginale were present in two further cases. A. marginale infections were also detected in apparently healthy subjects (n = 23). The results of PCR assays were confirmed by DNA sequencing. All samples that were positive by PCR for Babesia/Theileria spp. gave also positive results in the microarray analysis. The microarray chips identified Babesia bovis (n = 12) and Babesia bigemina (n = 2). Cattle with babesiosis were likely to have hemoglobinuria and nervous signs when compared to those with anaplasmosis that frequently had bloody feces. We conclude that clinical examination in combination with microscopy are still very useful in diagnosing acute cases of babesiosis and anaplasmosis, but a combination of molecular biological diagnostic assays will detect even asymptomatic carriers. In perspective, parallel detection of Babesia/Theileria spp. and A. marginale infections using a single microarray system will be a valuable improvement.     

Evaluation of long-term antibody responses to two inactivated bovine viral diarrhoea virus (BVDV) vaccines

The aim of the present study was to determine the serological response of heifers after vaccination with two inactivated bovine viral diarrhoea virus (BVDV) vaccines by means of various ELISA tests. Three dairy farms were selected from the Galicia region of Spain. In each herd, a batch of heifers to be vaccinated for the first time was selected and followed for 15 months. Heifers from farm 1 (n = 25) were vaccinated with Vaccine A, whereas heifers from farm 2 (n = 16) were vaccinated with Vaccine B. Heifers from farm 3 (n = 17), where no BVDV vaccines were used, acted as controls. Blood samples were analyzed periodically for BVDV antibodies, using five commercial ELISAs, based on BVDV p80 antigen or whole virus.At the end of the study, none of the animals vaccinated with Vaccine A seroconverted according to p80 antibody status, whereas up to 80% tested positive by ELISA against whole virus antigen. For the animals vaccinated with Vaccine B, 2/16 animals seroconverted according to p80 antibody ELISAs, whereas all had seroconverted according to the ELISA against whole virus antigen. In most cases, based on the use of ELISAs to detect specific antibodies against the p80 protein, at 15 months post-vaccination with inactivated BVDV vaccines the responses did not seem to interfere with detection of antibody to BVDV infection. However, the finding of a small proportion of vaccinated animals seropositive against BVDV p80 antigen suggests that antibodies that interfere with diagnosis of BVDV infection within the herd could exist, even when using p80 ELISAs.     

Neospora caninum DNA detection by TaqMan real-time PCR assay in experimentally infected pregnant heifers

Neosporosis has been considered the main cause of abortion between the first and the second trimester of pregnancy in cattle. Therefore, the objective of this study was to identify the presence of Neospora caninum DNA obtained from experimental models based on the evaluation of different areas of the fetal nervous system and organs from heifers previously inoculated with NC-1 after or before insemination. This study was performed with Hereford × Nelore (n = 29) heifers and all animals were considered free of diseases at the beginning of the experiment. All animals were bred by fixed-time artificial insemination (TAI) and allocated as follows: (a) seronegative heifers subjected to TAI (TAI, n = 9), (b) heifers infected with N. caninun 60 days prior to TAI (NC-1 + TAI, n = 9), and (c) heifers submitted to TAI and infected with N. caninum 60 days later (TAI + NC-1, n = 11). The pregnancy was confirmed by transrectal ultrasonography 35 days after TAI and evaluated every 30 days until the end of gestation. Fetuses were collected surgically at 170 days of gestation, and immediately necropsied to remove tissues aseptically. Samples of the central nervous system (CNS), heart, kidney, lung, liver, skeletal muscle and caruncle were collected for DNA extraction. Days of gestation at abortion and interval from abortion to first insemination were examined by Student's t-test. At 35 days of gestation the pregnancy rates in the group NC-1 + TAI (4/9, 44.4%) was lower than in the control group (8/9, 88.8%, P < 0.05). At 60 days, the pregnancy rates in the NC-1 + TAI group (0/4, 0%) was lower compared to TAI + NC-1 (5/7, 71.4%) and control (6/8, 75.0%) groups (P < 0.05). Animals from the group NC-1 + TAI were re-inseminated 60 days after the first TAI. After pregnancy losses throughout the study, 5 animals (TAI), 3 animals (NC-1 + TAI) and 5 animals (TAI + NC-1) maintained pregnancy until 170 days of gestation. TaqMan RT-PCR demonstrated the presence of N. caninum DNA in the medulla and right posterior cortex in 3 out of 5 fetuses from the TAI + NC-1 group. We concluded that heifers infected after TAI had a higher incidence of the parasite at the fetus CNS. Identification of N. caninum by TaqMan RT-PCR would assist in the investigation of infection and in the evaluation of vaccines or therapeutic drugs to control neosporosis in cattle.     

The effect of a pressure wrap (ThunderShirt®) on heart rate and behavior in canines diagnosed with anxiety disorder

The effect of moderate-to-deep pressure on soft tissue has been studied in both humans and nonhuman animals with positive results shown in reducing anxiety and promoting relaxation. Canine anxiety disorders are often treated with medications and behavior modification programming. Pressure wraps are sometimes used to treat canine anxiety, but the effects of this investigation have not been rigorously investigated. The aims of this study were to investigate the use of a pressure wrap (ThunderShirt®; ThunderWorks, Durham, NC) on heart rate and behavior in dogs diagnosed with an anxiety disorder. Subjects were 90 dogs (39 males and 51 females) that were diagnosed with Separation Anxiety or Generalized Anxiety Disorder. Dogs were randomly assigned to 1 of the 3 groups, namely Experimental Group 1 (dogs wore the ThunderShirt® per manufacturer's recommendations), Experimental Group 2 (dogs wore the ThunderShirt® loosely without pressure), and Control Group (dogs had no specific treatment). Average and maximum heart rates were measured at baseline and after 15 minutes alone in the kennel. Dogs were video-recorded and analysis completed for 12 behaviors, namely pacing, panting, yawning, tongue-flicking, drooling, elimination, barking, stress whining, howling, licking, door orientation, and calmness. The results were as follows: Dogs in Experimental Group 1 showed significantly less increase from baseline in average heart rate than the Control Group dogs (P < 0.001) and dogs in Experimental Group 2 (P < 0.001). Dogs in Experimental Group 1did not differ significantly from Control Group dogs in maximum heart rate when all dogs were considered, but did differ significantly from the Control Group (P = 0.04) when only those dogs not currently on anxiety medication were considered. There were no statistically significant differences among groups on behavioral outcomes, except that the dogs in the Control Group were significantly more likely to orient toward the door than the dogs in both Experimental Groups 1 (OR = 7.46, P = 0.01) and 2 (OR = 4.42, P = 0.04). Dogs in the Experimental Group 1 presented a trend of less tongue-flicking (P = 0.06) and yawning (P = 0.07) stress reduction behaviors when compared with the other 2 groups. More research should be conducted with the ThunderShirt® on behavioral outcomes. The ThunderShirt® can be used with behavior modification programming and medication as an adjunct treatment option to assist dogs diagnosed with anxiety disorder to reduce heart rate.     

The effects of farm management practices on liver fluke prevalence and the current internal parasite control measures employed on Irish dairy farms

Fasciolosis caused by Fasciola hepatica is responsible for major production losses in cattle farms. The objectives of this study were to assess the effect of farm management practices on liver fluke prevalence on Irish dairy farms and to document the current control measures against parasitic diseases. In total, 369 dairy farms throughout Ireland were sampled from October to December 2013, each providing a single bulk tank milk (BTM) sample for liver fluke antibody-detection ELISA testing and completing a questionnaire on their farm management. The analysis of samples showed that cows on 78% (n = 288) of dairy farms had been exposed to liver fluke. There was a difference (P < 0.05) between farms where cows were positive or negative for liver fluke antibodies in (a) the total number of adult dairy cows in herds, (b) the number of adult dairy cows contributing to BTM samples, and (c) the size of the total area of grassland, with positive farms having larger numbers in each case. There was no difference (P > 0.05) between positive and negative farms in (a) the grazing of dry cows together with replacement cows, (b) whether or not grazed grassland was mowed for conservation, (c) the type of drinking water provision system, (d) spreading of cattle manure on grassland or (e) for grazing season length (GSL; mean = 262.5 days). Also, there were differences (P < 0.001) between drainage statuses for GSL with farms on good drainage having longer GSL than moderately drained farms. The GSL for dairy cows on farms with good drainage was 11 days longer than for those with moderate drainage (P < 0.001). The percentage of farmers that used an active ingredient during the non-lactating period against liver fluke, gastrointestinal nematodes, lungworm, and rumen fluke was 96%, 85%, 77% and 90%, respectively. Albendazole was the most frequently used active ingredient for treatment against gastrointestinal nematodes (57%), liver fluke (40%) and lungworm (47%), respectively. There was a difference (P < 0.05) in the use of triclabendazole and albendazole between positive and negative farms, with triclabendazole use being more common in positive farms. This study highlighted differences in dairy management practices between Irish farms with dairy herds exposed or not exposed to liver fluke and stressed the need of fine-scale mapping of the disease patterns even at farm level to increase the accuracy of risk models. Also, comprehensive advice and professional support services to farmers on appropriate farm management practices are very important for an effective anthelmintic control strategy.     

Blood-feeding patterns of horse flies in the French Pyrenees

Horse flies can mechanically transmit Besnoitia besnoiti, the agent of bovine besnoitiosis. Although previously limited to enzootic areas, especially the French Pyrenees Mountains, bovine besnoitiosis is now considered a re-emerging disease in western Europe. To improve understanding of the role of horse flies as mechanical vectors, this study investigated their blood-feeding ecology in the eastern French Pyrenees, in two high-altitude summer pastures whose main domestic ungulates were cattle, and in a wildlife park with native fauna. Species-specific PCR assays were conducted to identify the sources of blood meals: wild boar, horse, cattle (or bison), sheep (or mouflon), goat, red deer, roe deer and izard (or Pyrenean chamois). In La Mouline pasture, tabanids (N = 20) fed on red deer (70%) and cattle (30%). In Mantet pasture, tabanids (N = 24) fed on cattle (52%), red deer (20%), wild boar (16%), horse (8%) and sheep (4%). In the wildlife park, Tabanus bromius (N = 32), the most abundant species collected, fed on red deer (85%), bison (9%) and wild boar (6%). Despite relatively high densities in both the pastures and in the wildlife park, small wild ungulates (izard, mouflon and roe deer) were not detected as a source of blood meals. Only two mixed blood meals were identified in two specimens of T. bromius: cattle/horse for the specimen collected in the pastures, and bison/wild boar for the specimen collected in the wildlife park. Our findings showed that tabanids display a level of opportunistic feeding behaviour, in addition to a preference for red deer, the latter being particularly true for Philipomyia aprica, the most abundant species collected in the pastures.     

Complex epidemiology and zoonotic potential for Cryptosporidium suis in rural Madagascar

Cryptosporidium spp. is the most important parasitic diarrheal agent in the world, is among the top four causes of moderate-to-severe diarrheal disease in young children in developing nations, and is problematic as an opportunistic co-infection with HIV. In addition, Cryptosporidium is a persistent challenge for livestock production. Despite its zoonotic potential, few studies have examined the ecology and epidemiology of this pathogen in rural systems characterized by high rates of overlap among humans, domesticated animals, and wildlife. To improve our understanding of the zoonotic potential of Cryptosporidium species in the rural tropics, we screened humans, livestock, peridomestic rodents, and wildlife using PCR-RFLP and sequencing-based approaches to distinguish species of Cryptosporidium in rural southeastern Madagascar. Cryptosporidium of multiple species/genotypes were apparent in this study system. Interestingly, C. suis was the dominant species of Cryptosporidium in the region, infecting humans (n = 1), cattle (n =  18), pigs (n= 3), and rodents (n = 1). The broad species range of C. suis and the lack of common cattle Cryptosporidium species (Cryptosporidium parvum and Cryptosporidium andersoni) in this system are unique. This report represents the fifth confirmed case of C. suis infection in humans, and the first case in Africa. Few rural human and livestock populations have been screened for Cryptosporidium using genus-specific genotyping methods. Consequently, C. suis may be more widespread in human and cattle populations than previously believed.     

Ivermectin and moxidectin resistance characterization by larval migration inhibition test in field isolates of Cooperia spp. in beef cattle,Mato Grosso do Sul,Brazil

Ivermectin (IVM) resistance of Cooperia spp. in cattle has become an increasing and global problem. The early detection of anthelmintic resistance (AR) is important to propose strategies to slow down the development of resistance and requires sensitive, reliable, economic high-throughput and practical tests. The purpose of the present study was to apply a larval migration inhibition test (LMIT) for evaluating IVM and MOX efficacy against well-characterized field isolates of Cooperia spp. infecting cattle in Brazil. Eight isolates were used for IVM and seven for MOX. The following EC50 values of IVM were observed for the isolates: susceptible, 1.16 ηmol; Nova Alvorada do Sul I, 4.09 ηmol (RF = 3.52); Campo Grande BNA, 3.57 ηmol (RF = 3.07); Campo Grande TBR, 4.09 ηmol (RF = 3,52); Nova Alvorada do Sul II, 2.50 ηmol (RF = 2.15); Bandeirantes, 11.35 ηmol (RF = 9.78); Campo Grande II, 6.03 ηmol (RF = 5.20); and Porto Mortinho, 8.63 ηmol (RF = 7.44). For MOX, the following EC50 values were observed: susceptible, 0.75 ηmol; Campo Grande BNA, 0.93 ηmol (RF = 1.24); Campo Grande TBR, 0.36 ηmol (RF = 0.48); Nova Alvorada do Sul II, 2.57 ηmol (RF = 3.42); Bandeirantes, 1.43 ηmol (RF = 1.90); Campo Grande II, 1.08 ηmol (RF = 1.44); and Porto Mortinho, 0.49 ηmol (RF = 0.65). The LMIT used in the present study can be a useful tool for in vitro evaluation of IVM, but not of MOX. However, such methodology cannot be used in large-scale studies yet. The isolates of Cooperia spp. showed various degrees of resistance to IVM, though remaining susceptible to MOX.     

Genetic diversity in Trypanosoma theileri from Sri Lankan cattle and water buffaloes

Trypanosoma theileri is a hemoprotozoan parasite that infects various ruminant species. We investigated the epidemiology of this parasite among cattle and water buffalo populations bred in Sri Lanka, using a diagnostic PCR assay based on the cathepsin L-like protein (CATL) gene. Blood DNA samples sourced from cattle (n = 316) and water buffaloes (n = 320) bred in different geographical areas of Sri Lanka were PCR screened for T. theileri. Parasite DNA was detected in cattle and water buffaloes alike in all the sampling locations. The overall T. theileri-positive rate was higher in water buffaloes (15.9%) than in cattle (7.6%). Subsequently, PCR amplicons were sequenced and the partial CATL sequences were phylogenetically analyzed. The identity values for the CATL gene were 89.6–99.7% among the cattle-derived sequences, compared with values of 90.7–100% for the buffalo-derived sequences. However, the cattle-derived sequences shared 88.2–100% identity values with those from buffaloes. In the phylogenetic tree, the Sri Lankan CATL gene sequences fell into two major clades (TthI and TthII), both of which contain CATL sequences from several other countries. Although most of the CATL sequences from Sri Lankan cattle and buffaloes clustered independently, two buffalo-derived sequences were observed to be closely related to those of the Sri Lankan cattle. Furthermore, a Sri Lankan buffalo sequence clustered with CATL gene sequences from Brazilian buffalo and Thai cattle. In addition to reporting the first PCR-based survey of T. theileri among Sri Lankan-bred cattle and water buffaloes, the present study found that some of the CATL gene fragments sourced from water buffaloes shared similarity with those determined from cattle in this country.