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 Sumice 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 Sumice 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 1.5kb with the dhfr1- aadA1 gene cassette were found in four isolates, followed by a 1kb integron with the aadA1 gene found in three isolates, and a 1.7kb 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.     

Genetic analysis of multi-drug resistance and the clonal dissemination of β-lactam resistance in Salmonella Infantis isolated from broilers

An epidemiologic study was conducted to investigate the incidence and characterize the antimicrobial resistance determinants, analyzing plasmid profiles, and establishing the genetic relationship among β-lactam-resistant isolates of Salmonella Infantis from broilers in Southern Japan. A total of 120 isolates were recovered from 56 flocks belonging to 44 holdings during 2004–2006. The percentages of resistance were as follows: ampicillin (24%), cephalothin (23%), cefoxitin (0%), ceftazidime (11%), cefotaxime (11%), chloramphenicol (0%), kanamycin (7.5%), ofloxacin (20%), oxytetracycline, streptomycin and sulfamethoxazole (100%) and trimethoprim (75%). The incidence of bla_TEM-encoded β-lactam resistance in 2004–2006 was significantly higher than in 1998–2003 (P < 0.001). BlnI-digested PFGE patterns generated two related clusters implicated in the dissemination of β-lactam resistance. Two types of plasmid profiles were observed and two plasmids of ca. 50 and 180-kb size were carried by β-lactam-resistant isolates. Streptomycin resistance was conferred by aadA1 (n = 116), aadA1-aadA2 (n = 1), and aadA1-strA-strB (n = 3). Resistances to kanamycin, oxytetracycline, sulfamethoxazole and trimethoprim were conferred by aphA1 (n = 9, 100%), tetA (n = 120, 100%) sul1 (n = 120, 100%) and dfrA5 (n = 90, 100%), respectively. Two types of class 1 integrons were detected: 1.0 kb (n = 120) and, 1.0/1.5 kb (n = 3). Integrons of 1.0/1.5 kb were found in isolates with the aadA1-strA-strB gene combination. For the first time, all S. Infantis isolates showed resistance to at least three classes of antimicrobial agents; and the intestinal tract of healthy poultry was a reservoir of the extended-spectrum cephalosporin-resistant isolates of serovar Infantis.     

Outbreak of anaplasmosis associated with novel genetic variants of Anaplasma marginale in a dairy cattle

During early lactation, dairy cows may present a transient immunosuppressive state and develop anaplasmosis caused by Anaplasma marginale. In this study, clinical anaplasmosis in dairy cattle in the Thrace region of Turkey was investigated with respect to within-herd prevalence, vertical transmission, and genetic diversity. In March and September 2015, thirty lactating cows showed primary clinical signs of anaplasmosis, including fever, anaemia, decreased milk yield, anorexia, and laboured breathing. Symptoms disappeared in most cows after administration of long-acting oxytetracycline, but nine of them (30%) died. Following diagnosis based on clinical signs, microscopy and molecular findings, blood samples were collected from apparently healthy lactating cows (n = 184), pregnant heifers (n = 39) and newborn calves (n = 24). DNA was extracted from each sample and analyzed for the presence of major surface proteins (MSPs) of A. marginale, followed by sequencing to assess diversity of isolates. Microscopic examination of erythrocytes revealed A. marginale inclusion bodies in symptomatic cows. Examination of thin blood smears showed 3.8% of the lactating, clinically asymptomatic, cows to be infected with A. marginale, while nPCR detected 31.0% positive. A. marginale infection was not detected in pregnant heifers by either method. Congenital infection was found in one calf by nPCR. This is the first report of transplacental transmission of A. marginale in Turkey. The MSP4 sequence analyses showed high genetic diversity among the isolates, presenting 97.6-99.6% homology at the amino acid level. The sequences of MSP1a amplicons revealed genetic diversity providing three new tandem repeats.     

Frequency of virulence factors in Escherichia coli isolated from suckling pigs with diarrhoea in China

Escherichia coli-associated diarrhoea is an important disease adversely affecting the pig industry. This study was conducted to investigate the frequency of virulence factors expressed by E. coli strains isolated from suckling pigs with diarrhoea in China. A total of 381 E. coli strains, obtained from 290 faecal samples from pigs on 38 farms, were tested for fimbriae (K88, K99, 987P, F41, F18, F17), non-fimbrial adhesins (AIDA-I, paa, CS31A, eae, saa), enterotoxin (LT-I, LT-II, STa, STb, EAST1), Shiga toxin (Stx1, Stx2, Stx2e), pathogenicity islands (HPI, LEE), α-haemolysin (hlyA), afa8 gene cluster (afaD, afaE) and sepA genes by PCR. Out of the 381 isolates, 206 carried at least one virulence gene. Of the 206 virulence positive isolates, the virulence factor genes detected were EAST1 (n = 120), irp2 (n = 59), paa (n = 50), STb (n = 41), AIDA-I (n = 34), LT-I (n = 23), ler (n = 11), hlyA (n = 9), K88 (n = 8), eae (n = 8), STa (n = 7), sepA (n = 6), F18 (n = 5), afaD (n = 3), afaE (n = 3), K99 (n = 2) and Stx2e (n = 1), with most isolates carrying multiple virulence genes. These results demonstrate that relatively few isolates from the study population express K88, K99, LT-I or STa, but that EAST1 (58%), irp2 (29%), AIDA-I (16.5%), paa (24%) and STb (20%) are frequent virulence factors expressed by E. coli strains isolated from suckling pigs with diarrhoea in China.     

Characterization of integrons in multiple antimicrobial resistant Escherichia coli isolates from bovine endometritis

To assess the prevalence of antimicrobial resistance and three classes of integrons in Escherichia coli (E. coli) strains (n = 57) isolated from bovine endometritis in Inner Mongolia of China, antimicrobial susceptibility and the presence of three types of integrons were characterized. Most isolates were susceptible to ceftiofur, furazolidone, ciprofloxacin and enrofloxacin, while 57 isolates were all resistant to sulfamethoxydiazine and trimethoprim. High resistant incidence rates were exhibited to sulfadiazine, tetracycline, oxytetracycline, cefazolin, chloramphenicol. Forty-six of 57 E. coli strains were resistant to above 10 antibiotics (80.70%). The integrase gene and gene cassettes of integrons were amplified by PCR. DNA sequencing and analysis were used to identify the genetic content of the integron-variable regions. Neither class II nor class III integron was detected, while 36.8% (n = 21) of the isolates were positive for the presence of intI1 gene. Analysis of gene cassettes revealed that six gene cassettes were found, which encoded resistance to trimethoprim (dhfr, dhfrI, dfrA17) and aminoglycosides (aadA1, aadA2, aadA5). Among them, the gene cassette array dfrA17–aadA5 was found most prevalent (66.7%). The resistance profile of positive-integron isolates was relatively broad and they were resistant to more than eight antimicrobials (n ? 8). The correlation analysis revealed the incidence of integrons among the isolates were related to the multiple antibiotic resistance profile, indicating integrons play an important role in the dissemination and spread of the antimicrobial resistant strains.     

Evaluation of Low-Stress Herding and Supplement Placement for Managing Cattle Grazing in Riparian and Upland Areas

Management practices are often needed to ensure that riparian areas are not heavily grazed by livestock. A study was conducted in Montana during midsummer to evaluate the efficacy of low-stress herding and supplement placement to manage cattle grazing in riparian areas. Three treatments were evaluated in three pastures over a 3-yr period in a Latin-square design (n = 9). Each year, naïve 2-yr-old cows with calves were randomly assigned to the three treatments: 1) free-roaming control, 2) herding from perennial streams to upland target areas, and 3) herding to upland sites with low-moisture block supplements. Stubble heights along the focal stream were higher (P = 0.07) in pastures when cattle were herded (mean ± SE, 23 ± 2 cm) than in controls (15 ± 3 cm). Global positioning system telemetry data showed that herding reduced the time cows spent near (< 100 m) perennial streams (P = 0.01) and increased the use of higher elevations (P = 0.07) compared with controls. Evening visual observations provided some evidence that free-roaming cows (44% ± 19%) were in riparian areas more frequently (P = 0.11) than herded cows (23% ± 6%). Fecal abundance along the focal stream was less (P = 0.07) with herding (61.9 ±  kg · ha⁻¹) than in controls (113.2 ±  kg · ha⁻¹). Forage utilization within 600 m of supplement sites was greater (P = 0.06) when cows were herded to low-moisture blocks (18% ± 6%) compared with controls and herding alone (8% ± 2%). Moving cattle to uplands at midday using low-stress herding is an effective tool to reduce use of riparian areas. Herding cattle to low-moisture blocks can increase grazing of nearby upland forage but may not provide additional reduction in cattle use of riparian areas compared with herding alone.     

Molecular characterization of quinolone resistance mechanisms and extended-spectrum β-lactamase production in Escherichia coli isolated from dogs

The increasing prevalence of antimicrobial resistances is now a worldwide problem. Investigating the mechanisms by which pets harboring resistant strains may receive and/or transfer resistance determinants is essential to better understanding how owners and pets can interact safely. Here, we characterized the genetic determinants conferring resistance to β-lactams and quinolones in 38 multidrug-resistant Escherichia coli isolated from fecal samples of dogs, through PCR and sequencing. The most frequent genotype included the β-lactamase groups TEM (n = 5), and both TEM + CTX-M-1 (n = 5). Within the CTX-M group, we identified the genes CTX-M-32, CTX-M-1, CTX-M-15, CTX-M-55/79, CTX-M-14 and CTX-M-2/44. Thirty isolates resistant to ciprofloxacin presented two mutations in the gyrA gene and one or two mutations in the parC gene. A mutation in gyrA (reported here for the first time), due to a transversion and transition (TCG → GTG) originating a substitution of a serine by a valine in position 83 was also detected. The plasmid-encoded quinolone resistance gene, qnrs1, was detected in three isolates. Dogs can be a reservoir of genetic determinants conferring antimicrobial resistance and thus may play an important role in the spread of antimicrobial resistance to humans and other co-habitant animals.     

Characterisation of ceftiofur resistance in swine bacterial pathogens

A PCR based method was developed for the identification of ceftiofur resistance genes (bla_CMY-2, bla_TEM-1, and ampC) in swine bacterial pathogens. Using this method, the ceftiofur resistant (n = 76) and susceptible (n = 45) strains of Bordetella bronchiseptica, Salmonella spp., Escherichia coli, and Pasteurella multocida were screened for the presence of these three genes. The resistant genes were detected in 70% (bla_TEM-1), 68% (bla_CMY-2) and 45% (ampC) of the resistant isolates and in 18% (bla_TEM-1), 27% (bla_CMY-2), and 36% (ampC) of the susceptible isolates. Results obtained in the present study showed widespread distribution of these three resistance genes in ceftiofur-resistant swine pathogens. It was also observed that more pathogens are acquiring these resistance genes.     

Effect of restricted suckling on physiological and behavioural stress parameters in dual-purpose cattle in the tropics

The present study evaluated the effect of restricted suckling (RS) on some physiological and behavioural stress measures compared to artificial rearing (AR). Thirty Holstein × Zebu cows and their calves were randomly assigned to one of two treatments. Cows were milked once daily. All calves were tethered in front of their dams during milking permitting physical contact between them in order to stimulate milk ejection, starting at 5 days of age. Calves in the RS treatment were allowed to suckle during 60 min daily, 30 min after milking and 30 min in the afternoon, while calves in the AR treatment were fed milk from teat bottles at the same times. Data were collected from day 4 to day 9, while treatments started on day 5. Heart and respiratory rates, body weight and temperature were measured (in that order) 15 min before entry into the milking parlour. Blood samples and pedometer readings were collected about 50 min later, immediately on return to their home pens. Higher heart rate in calves was found in the RS group than in the AR group. However, cortisol concentration was higher (P < 0.05) in the AR than in the RS group. No difference was found in average weight gain, milk consumption, number of steps counted, body temperature and respiratory rate in RS and AR calves. AR induced significantly (P < 0.05) higher level of serum cortisol concentration in the cows than did RS. No difference (P > 0.05) was found in body temperature, heart and respiratory rates in RS and AR treatments. It was concluded that suckling and nursing reduces some stress parameters in calves and cows in the AR treatment.     

Comparative pharmacokinetics of an injectable cephalexin suspension in beef cattle

This study describes and compares the pharmacokinetics of a single 7.5 mg/kg dose of cephalexin monohydrate oil-based 20% suspension after its administrations to six cows by the intramuscular (i.m.) and subcutaneous (s.c.) routes, and to five calves by the i.m. route. Significantly (P < 0.05) higher peak plasma concentrations (5.6 ± 0.79 μg/ml versus 3.93 ± 1.24 μg/ml) and lower half-life (1.81 ± 0.56 h versus 4.21 ± 0.82 h) and mean residence time (4.12 ± 1.07 h versus 6.63 ± 0.85 h) were obtained after i.m. administration when compared to the s.c. administration to cows. No differences were found between pharmacokinetic parameters calculated for cows and calves. Cephalexin plasma concentrations remained above 0.5–0.75 μg/ml for 11–14 h and 8–9 h after the s.c. and i.m. administrations, respectively. Thus, route of administration may be an important issue to be considered when calculating dosage schedules for successful treatments and safe withdrawal times for veterinary medicines.     

Effects of early weaning and breed on calf performance and carcass and meat quality in autumn-born bull calves

This study assessed the effects of age at weaning (early weaning at 90 d or traditional weaning at 150 d) and breed (Parda de Montaña or Pirenaica) on calf performance and carcass and meat quality in autumn-calving beef cattle. At calving, 14 Parda de Montaña and 14 Pirenaica cow–calf pairs were randomly assigned to one of two weaning treatments, and kept indoors during lactation. After weaning, calves were fed an intensive diet until slaughter at 450 kg. The interaction between age at weaning and breed was not significant for any of the parameters studied. From 90 d to 150 d, early weaned calves had greater ADG (P = 0.001) and IGF-I concentrations (P = 0.001) than traditionally weaned calves, but their leptin concentrations were similar (P = 0.15). During the finishing phase, performance, daily feed intake, and efficiency did not differ between treatments. Early weaning did not affect age at slaughter, carcass weight, fatness score, fat colour, and meat quality, but improved carcass conformation (P = 0.04). Early weaned calves had greater total DMI (P = 0.002) with greater concomitant feed costs (P = 0.001) and yielded a slightly greater income than traditionally weaned calves; therefore, economic margins did not differ. Parda de Montaña calves tended to have greater ADG from birth to 90 d and were heavier at 90 d (P = 0.01) than were Pirenaica calves. From 90 d to 150 d, performance and IGF-I and leptin concentrations did not differ between breeds; thus, Parda de Montaña calves remained heavier at 150 d. During the finishing phase, at times, weight gains of Parda de Montaña and Pirenaica calves differed, but the overall performance, feed intake, and efficiency of the two breeds were similar. Pirenaica calves had heavier carcasses (P = 0.04) with greater conformation scores (P = 0.04) than Parda de Montaña calves; thus, income per carcass was greater for the former than the latter (P = 0.007). As feed costs were similar for both breeds, the economic margin of Pirenaica calves was greater than that of Parda de Montaña calves (P = 0.01). In conclusion, in both breeds weaning strategies had similar effects on performance and carcass and meat quality; however, from an economic point of view, and considering only the costs associated with the calf, raising Pirenaica calves would be more profitable, at either age at weaning.     

The impact of dystocia on dairy calf health,welfare, performance and survival

Up to one-third of dairy calves are born after dystocia and this is a major cause of calf mortality. This study investigated the neonatal physiology, survival, health and subsequent growth of dairy calves following dystocia and is the first longitudinal study to analyse multiple effects and to look beyond the perinatal period.A total of 455 live born Holstein calves (N: No assistance, n = 360; FN: Farmer assistance but normally presented calf, n = 82; FM: Farmer assistance of malpresented calf, n = 13) were followed from birth to first service (heifers) or until leaving the farm (bulls). Compared to N calves, FN and FM animals had higher salivary cortisol concentrations at day 1 (P < 0.001) and FN calves had lower passive immune transfer (P = 0.03). Dystocia had no biologically significant impact on rectal temperature throughout the first 4 days (P > 0.05). During the first 60 days, FM calves had a higher proportion of days with non-routine health treatments (P < 0.05) and, by the time of weaning, mortality in FN and FM heifers was higher than in N calves (2.8×; P < 0.01). However, in surviving calves, growth to first service was not affected by dystocia category (P > 0.05).Calves which survive dystocia experience lower passive immunity transfer, higher mortality and higher indicators of physiological stress. Such calves have poorer welfare in the neonatal period and possibly beyond. Strategies need to be implemented to improve the subsequent health and welfare of such calves and to lower the incidence of dystocia.     

Method of Supplementation May Affect Cattle Grazing Patterns

Supplement placement can be used to manipulate livestock grazing patterns. The objective of this case study was to compare the effect of low-moisture blocks (LMB) and range cake (barley-based cylindrical cubes, 2 cm in diameter, and 2 to 8 cm long) supplementation on cattle grazing patterns in Montana foothill rangeland. One group of nonlactating cows (n = 79) was fed cake 3 times per week (1.8 kg · cow⁻¹ · feeding⁻¹), and the other group (n = 81) had continuous access to LMB in separate pastures using a crossover design. Movement patterns of cows were recorded with global positioning system collars during four periods (2 wk · period⁻¹) during autumn. Range cake was fed on accessible areas, and LMB were placed in higher and steeper terrain. Intake of LMB averaged (mean ± SE) 318 ± 50 g · d⁻¹. Cows fed LMB (8.07° ± 0.20°) were observed on steeper slopes (P = 0.08) than cows fed range cake (6.96° ± 0.19°). Forage utilization decreased as slope increased to a greater degree when range cake was fed than when LMB was fed (P = 0.001). Cows spent more time (P = 0.05) within 100 m of LMB (274 ± 23 min · d⁻¹) than at range cake feeding sites (67 ± 24 min · d⁻¹). Strategic placement of LMB on high, steep terrain appears to be a more practical and effective approach than traditional hand-feeding range cake on intermediate terrain to improve uniformity of cattle grazing on rugged rangeland.     

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.     

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.     

Administration of anesthetic and analgesic prevent the suppression of many leukocyte responses following surgical castration and physical dehorning

The objectives of the current research were to determine the physiological effects and responses of many leukocytes following surgical castration and/(or) physical dehorning and the influence of anesthetics and analgesics in 3-month-old calves. Eighty 3-month-old Holstein bull calves were completely randomized to treatments in a 2 × 2 × 2 factorial arrangement with castration, dehorning, and anesthetic/analgesic as the main effects. Peripheral blood samples were collected just before (0) and 0.5, 1.5, 2.5, 4, 6, 24, and 72 h after the respective procedure(s) and analyzed for total leukocyte and differential counts, as well as plasma cortisol and haptoglobin concentrations. Blood from the 0, 0.5 and 24 h collections were analyzed for many ex vivo leukocyte responses. Data were analyzed using a repeated measures analysis of variance with the fixed effects of treatment, time, and the interaction of treatment × time. Pre-planned contrasts were performed to determine the effect of (1) management procedure (castration and/(or) dehorning), (2) anesthetic/analgesic, and (3) were the management procedures additive. There were treatment × time interactions (P < 0.05) on plasma cortisol and haptoglobin concentrations as well as for total leukocyte and neutrophil concentrations in blood. Castration and dehorning increased cortisol concentrations and the effect of the procedures was additive (P < 0.02). Dehorning alone elicited a greater (P < 0.05) cortisol response than castration alone. In contrast, the leukocytosis and neutrophilia was greater (P < 0.01) among castrated calves. In addition, haptoglobin concentrations at 24 h after castration were elevated (P < 0.01) in calves that were castrated. Both castration and dehorning suppressed (P = 0.04) many leukocyte responses including the secretion of tumor necrosis factor-α when whole blood cultures were stimulated with lipopolysaccharide, surface expression of l-selectin on peripheral blood neutrophils, and the oxidative burst intensity of peripheral blood neutrophils when co-cultured with an Escherichia coli. The effects of castration and dehorning on blood leukocyte counts or any of the leukocyte responses were not additive (P > 0.23). Castration and dehorning effects of plasma haptoglobin concentrations tended (P = 0.10) to be additive at 72 h after the procedure(s). Prior administration of local anesthetic and a systemic analgesic attenuated (P < 0.001) the cortisol response and prevented (P = 0.03) the observed leukocytosis, neutrophilia, and leukocyte suppression. These data suggest that calves should be castrated and dehorned on the same day rather than spreading them out across two days and calves should be administered pain relief prior to performing either procedure.     

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.     

Antimicrobial resistance changes in enteric Escherichia coli of horses during hospitalisation: Resistance profiling of isolates

The aim of this study was to determine whether hospitalisation of horses leads to increased antimicrobial resistance in equine faecal Escherichia coli isolates. E. coli were cultured from faecal samples of horses on admission and after 7 days of hospitalisation; antimicrobial susceptibility was determined for eight antimicrobial agents. Resistance profiles of E. coli isolates were grouped into clusters, which were analysed to determine resistance patterns. Resistance to 7/8 antimicrobial agents and multi-drug resistance (MDR; resistance to ?3 antimicrobial classes) were significantly higher after 7 days of hospitalisation. Forty-eight resistance profiles were identified; 15/48 were present on day 0 only, 16/48 on day 7 only and 17/48 at both times of sampling. There was a significant association between day 7 profiles and resistance detected to an increased number of antimicrobial agents. Hospitalisation of horses for 7 days resulted in alterations in equine faecal E. coli antimicrobial resistance profiles.     

Differential expression of pro-inflammatory and anti-inflammatory cytokines during experimental infection with low or high virulence bovine viral diarrhea virus in beef calves

The objective was to compare the mRNA expression of pro-inflammatory (TNF-α, IL-1β, IFN-γ, IL-2, IL-12, IL-15) and anti-inflammatory (IL-4, IL-10, TGF-β) cytokines, after experimental infection with low or high virulence noncytopathic (ncp) bovine viral diarrhea virus (BVDV). Thirty BVDV-naïve, beef calves were intranasally inoculated with low (LV; n = 10, SD-1) or high (HV; n = 10, 1373) virulence ncp BVDV or with BVDV-free cell culture medium (Control, n = 10). Calves were euthanized on day 5 post-inoculation, and tracheo-bronchial lymph node and spleen samples were collected for mRNA expression through quantitative-RT-PCR. mRNA levels of pro-inflammatory (TNF-α, IL-1β, IL-2, IFN-γ) and anti-inflammatory (IL-4 and IL-10) cytokines were up-regulated in tracheo-bronchial lymph nodes of HV, but not in LV, compared to the control group (P < 0.05). IL-12 mRNA level was up-regulated in tracheo-bronchial lymph nodes of both LV and HV groups (P ≤ 0.05). A significant up-regulation of IL-15 mRNA was observed in tracheo-bronchial lymph nodes for LV calves (P < 0.002), but not for HV calves. Experimental inoculation with BVDV-2 1373 stimulated significant mRNA expression of pro-inflammatory and anti-inflammatory cytokines. In contrast, inoculation with BVDV-1a SD-1 only resulted in up-regulation of IL-12 and IL-15 mRNA, which is associated with activation of macrophages and NK cells during innate immune response.