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.     

Ectonucleotidase and adenosine deaminase as inflammatory marker in dairy cows naturally infected by Dictyocaulus viviparus

The aim of this study was to evaluate the influence of dictyocaulosis (mild or severe) on enzymes of NTPDase, 5′-nucleotidase, and adenosine deaminase (ADA) of dairy cows naturally infected by Dictyocaulus viviparus. Blood and faeces were collected from 22 dairy cows of the same farm to evaluate NTPDase (ATP and ADP substrate), 5′-nucleotidase, and ADA activities on days 0 (pre-treatment) and 10 (post-treatment). Seric activities of NTPDase (ATP substrate), 5′-nucleotidase, and ADA were lower (P < 0.05) in D. viviparus infected animals compared to uninfected cows. The number of D. viviparus larvae per gram of faeces varied among the animals, and they showed different degrees of severity according to respiratory clinical signs of the disease (cough and nasal discharge). Later, these cows were divided into two groups: those with mild (n = 10) and severe (n = 12) disease. Cows with severe disease showed higher NTPDase activity (ATP substrate) than those with mild disease (P ≤ 0.05). The opposite occurred with NTPDase (ADP substrate), 5′-nucleotidase, and ADA in cows with severe disease, that is, the enzymatic activity of these seric enzymes significantly decreased (P ≤ 0.05) compared to animals with mild disease. Infected animals showed reduced NTPDase activity (ATP and ADP substrate) after treatment. No enzymatic changes were observed for 5′-nucleotidase, and ADA pre- and post-treatment (P > 0.05). Based on these results, we conclude that dictyocaulosis alters NTPDase, 5′-nucleotidase, and ADA activities of cow naturally infected by the parasite, in consequence the enzymes act as inflammatory markers.     

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.     

Evaluation of Three Estrous Synchronization Protocols in Beef Heifers

Objectives of this study were to evaluate synchronization, conception, and pregnancy rates of heifers synchronized with melengestrol acetate (MGA)-prostaglandin F_2α (PGF_2α,), Select Synch, or Select Synch preceded by MGA (MGA-Select Synch). Heifers in the MGA-PGF_2α group (n = 209; BW = 378 kg) received MGA (0.5 mg/ d per heifer) for 14 d and PGF_2α (25 mg) 19 d later. Select Synch heifers (n = 213; BW = 374 kg) received gonadotropin-releasing hormone (GnRH; 100 μg) followed by PGF_2α (25 mg) 7 d later. The MGA-Select Synch heifers (n = 210; BW = 373 kg) were fed MGA (0.5 mg/d per heifer) for 7 d, GnRH (100 μg) the day following the last MGA feeding, and PGF_2α (25 mg) 7 d after GnRH. More (P<0.01) heifers were in estrus 1 to 4 d before PGF2a administration in both the Select Synch (20%) and MGA-Select Synch (24%) groups than in the MGA-PGF_2α (4%) group. Pregnancy rates for heifers in estrus early (d 1 to 4 before PGF_2α) were greater (P<0.05) for both Select Synch (55%) and MGA-Select Synch (63%) compared with MGA-PGF_2α heifers (18%). Synchronization rate (detected after PGF_2α) was greater (P<0.01) for MGA-PGF_2α heifers (86%) compared with Select Synch (66%) and MGA-Select Synch (68%) heifers; however, conception rate did not differ (P=0.13) and averaged 72, 63, and 62% for MGA-PGF_2α, Select Synch, and MGA-Select Synch heifers, respectively. Select Synch (52%), MGA-Select Synch (58%), and MGA-PGF_2α protocols (61%) provided similar (P=0.18) overall AI pregnancy rates; however, more heifers were in estrus before PGF_2α administration in protocols using GnRH.     

Vaccination with a genotype 1 modified live vaccine against porcine reproductive and respiratory syndrome virus significantly reduces viremia,viral shedding and transmission of the virus in a quasi-natural experimental model

The present study assessed the efficacy of vaccination against genotype 1 porcine reproductive and respiratory syndrome virus (PRRSV) in terms of reduction of the transmission. Ninety-eight 3-week-old piglets were divided in two groups: V (n = 40) and NV (n = 58) that were housed separately. V animals were vaccinated with a commercial genotype 1 PRRSV vaccine while NV were kept as controls. On day 35 post-vaccination, 14 NV pigs were separated and inoculated intranasally with 2 ml of a heterologous genotype 1 PRRSV isolate (“seeder” pigs, SP). The other V and NV animals were distributed in groups of 5 pigs each. Two days later, one SP was introduced into each pen to expose V and NV to PRRSV. Sentinel pigs were allocated in adjacent pens. Follow-up was of 21 days. All NV (30/30) became viremic after contact with SP while only 53% of V pigs were detected so (21/40, p < 0.05). Vaccination shortened viremia (12.2 ± 4 versus 3.7 ± 3.4 days in NV and V pigs, respectively, p < 0.01). The 50% survival time for becoming infected (Kaplan–Meier) for V was 21 days (CI_95% = 14.1–27.9) compared to 7 days (CI_95% = 5.2–8.7) for NV animals (p < 0.01). These differences were reflected in the R value as well: 2.78 (CI_95% = 2.13–3.43) for NV and 0.53 (CI_95% = 0.19–0.76) for V pigs (p < 0.05). All sentinel pigs (10/10) in pens adjacent to NV + SP pens got infected compared to 1/4 sentinel pigs allocated contiguous to a V + SP pen. These data show that vaccination of piglets significantly decrease parameters related to PRRSV transmission.     

Effect of supplemental sericea lespedeza leaf meal pellets on gastrointestinal nematode infection in grazing goats

Feeding sun-dried sericea lespedeza [SL; Lespedeza cuneata (Dum-Cours.) G. Don.] reduces gastrointestinal nematode (GIN) infection in goats fed in confinement, but effects of this forage when fed as a supplement to goats on pasture are unclear. A study was completed in which supplemental feeds (75 and 95% SL leaf meal pellets and a commercial pellet, all fed at 0.91 kg/head/day) were offered to thirty growing male Spanish goats (9 months old, 20.6 ± 2.8 kg, 10/treatment) grazing perennial warm-season grass pastures in Fort Valley, GA, from September to November, 2010. Fecal and blood samples were taken from individual animals weekly to determine fecal egg count (FEC) and packed cell volume (PCV), respectively, and animal weights were recorded at the start and end of the trial. After 11 weeks grazing, animals were slaughtered for recovery, counting, and speciation of adult GIN from the abomasum and small intestines. There was no difference in FEC between goats fed the 75 and 95% SL leaf meal pellets, but both groups had lower (P < 0.05) FEC than the goats fed the commercial pellets from days 35 to 77. The PCV values were not affected by the dietary treatments. Animal gain per day averaged 102.0, 77.2, and 53.3 g for goats fed 95% SL, commercial, and 75% SL pellets, respectively (P < 0.05). The 95% SL leaf meal pellet goats had 93.0 and 47.3% fewer (P < 0.05) total (male + female) adult Haemonchus contortus and Teladorsagia circumcincta, respectively, than control animals, while only male H. contortus were lower (47.6%; P < 0.05) in 75% SL-fed goats compared with commercial pellet-fed animals. Feeding supplemental SL leaf meal pellets improved animal performance (95% SL pellets) and reduced worm burdens (75 and 95% SL pellets) in young grazing goats and is a useful tool for natural GIN control in small ruminants.     

Performance of spring-calving beef suckler cows and their progeny to slaughter on intensive and extensive grassland management systems

The performance of rotationally grazed beef suckler cows and their progeny to slaughter on two lowland grassland management systems differing in stocking rate (SR) and fertiliser nitrogen (N) level was compared over eight years. The two Systems were 1) Intensive (INT): SR of 0.56 (bull production) or 0.71 (steer production) ha cow^? 1 unit, 211 kg fertiliser N ha^? 1, two silage harvests, and 2) Extensive (EXT): SR of 0.69 (bull production) or 0.88 (steer production) ha cow^? 1 unit, 97 kg fertiliser N ha^? 1 and one staggered silage harvest. A cow unit was defined as a cow plus progeny to slaughter. On the silage harvesting area, the mean application rate for fertiliser N was 110 and 80 kg ha^? 1 for first and second harvests, respectively. Herbage dry matter digestibility both pre- and post-grazing was similar (P > 0.05) for the two systems, whereas herbage crude protein concentrations were generally significantly lower for the EXT than the INT system. There was no difference (P > 0.05) between the Systems in cow live weight, body condition score or their changes or in calf live weight gain from birth to weaning. Post-weaning, live weight gain, slaughter weight, carcass weight, kill-out proportion, estimated carcass gain, carcass conformation score or carcass fat score did not differ (P > 0.05) between the systems for heifer, steer or bull progeny. It can be concluded that similar animal performance levels can be expected in an extensive grassland-based suckler calf-to-beef system compatible with the EU, Rural Environmental Protection Scheme as that attained in a more intensive System comprising of both a moderately high SR (~ 1.25 higher) and fertiliser N application (~ 2.1 higher).     

Animal and Vegetation Response to Modified Intensive–Early Stocking on Shortgrass Rangeland

A comparison of animal gains and vegetation trends was made from 2002–2008 between a continuous season-long stocking (SLS) system and a modified intensive–early stocking system (IES) with late-season grazing (IES 1.6× + 1; 1.6 times the number of animals of the SLS system from May 1 to July 15, and 1 times the number of animals of SLS from July 15 to October 1) on shortgrass native rangeland of western Kansas. The continuous season-long stocked system placed animals at a density of 1.37 ha · steer^?1 from May through October, or 2.63 animal unit months (AUM) · ha^?1, whereas the intensive–early stocked system with late-season grazing (3.33 AUM · ha^?1) stocked pastures at 0.85 ha · steer^?1 from May through the middle of July, and then stocked pastures at 1.37 ha · steer^?1 for the remainder of the grazing season by removing the heaviest animals mid-July each yr. Average daily gains (0.78 vs. 0.70 kg · d^?1, P = 0.039) and total animal gain (58 vs. 52 kg, P = 0.042) were different between the continuous season-long stocked and the intensive–early stocked animals during the first half of the grazing season. No difference was found between average daily gain (0.61 vs. 0.62 kg · d^?1, P = 0.726) and total animal gain (48 vs. 49 kg, P = 0.711) for the continuous season-long stocked and intensive–early stocked with late-season grazing animals during the last half of the season. Total individual animal gain (106 vs. 101 kg, P = 0.154) and average daily gain (0.70 vs. 0.66 kg · d^?1, P = 0.152) was not different between the continuous season-long stocked and the intensive–early stocked system animals that were on pasture the entire grazing season. Total beef gain on a land-area basis (96 vs. 77 kg · ha^?1, P = 0.008) was greater for the modified intensive–early stocked system with late-season grazing with greater animal densities. Changes in residual biomass and most key vegetation components at the end of the grazing season were not different between the two systems.     

A field trial of infrared thermography as a non-invasive diagnostic tool for early detection of digital dermatitis in dairy cows

Infrared thermography (IRT) was used to detect digital dermatitis (DD) prior to routine claw trimming. A total of 1192 IRT observations were collected from 149 cows on eight farms. All cows were housed in tie-stalls. The maximal surface temperatures of the coronary band (CB) region and skin (S) of the fore and rear feet (mean value of the maximal surface temperatures of both digits for each foot separately, CB_max and S_max) were assessed. Grouping was performed at the foot level (presence of DD, n = 99; absence, n = 304), or at the cow level (all four feet healthy, n = 24) or where there was at least one DD lesion on the rear feet, n = 37). For individual cows (n = 61), IRT temperature difference was determined by subtracting the mean sum of CB_max and S_max of the rear feet from that of the fore feet.Feet with DD had higher CB_max and S_max (P < 0.001) than healthy feet. S_max was significantly higher in feet with infectious DD lesions (M-stage: M2 + M4; n = 15) than in those with non-infectious M-lesions (M1 + M3; n = 84) (P = 0.03), but this was not the case for CB_max (P = 0.12). At the cow level, an optimal cut-off value for detecting DD of 0.99 °C (IRT temperature difference between rear and front feet) yielded a sensitivity of 89.1% and a specificity of 66.6%. The results indicate that IRT may be a useful non-invasive diagnostic tool to screen for the presence of DD in dairy cows by measuring CB_max and S_max.     

The influence of spasmolytic agents on heart rate variability and gastrointestinal motility in normal horses

The effects of hyoscine-N-butylbromide (hyoscine) and propantheline-bromide (propantheline) on heart rate (HR), HR variability (HRV) and gastrointestinal tract (GIT) contractions in the normal horse were determined. Five adult horses had ECG recordings for 180 min after treatment with propantheline (100 mg), hyoscine (120 mg) or saline. Both propantheline and hyoscine reduced GIT sounds, with propantheline having a longer duration of effect (⩾120 min). Both drugs elevated HR relative to the control baseline period (P < 0.05), with the effects of propantheline again being of longer duration. HRV analysis indicated that propantheline suppressed Total Power (P < 0.05), and both the high frequency (HF) and low frequency (LF) components of the power spectral analysis for up to 60–90 min post treatment. Hyoscine had no effect on HRV Total Power but reduced the HF component for 30 min after drug injection. Time domain variables correlated with Total Power and HF data (P < 0.01). The marked effect of these compounds on parasympathetic control of cardiac and GIT function in normal horses should be taken into consideration when evaluating a clinical response to these agents.     

The Effects of Dietary Zinc Concentration and Source on Yearling Bull Growth and Fertility

Yearling Angus bulls (n = 325) were allotted by BW into six pens of equal size. One of three dietary treatments consisting of 1) dietary Zn at 40 ppm, all supplied by zinc sulfate (ZnS); 2) dietary Zn at 40 ppm with 33.3% supplied by zinc proteinate and 66.6% supplied by ZnS (ZnPS); or 3) dietary Zn at 60 ppm, all supplied by ZnS (ZnHi) was delivered to two pen replicates. Initial and final liver biopsies were collected and analyzed for Zn concentration. Scrotal circumference and BW measures were collected at the start and conclusion of the trial. Semen from bulls intended for public sale (n = 167) was collected and evaluated for motility and morphological abnormalities. Bulls with percent normal sperm cell counts < 70% or with motility scores less than fair (motility scores = poor, fair, good, and very good) were considered classification deferred (CD). Following 126 d of treatment, bulls on the ZnHi treatment had a greater (P=0.06) increase in liver Zn concentration than bulls on the ZnS treatment, but not bulls on the ZnPS treatment (P=0.18) (‒9.8, 1.2, and 20.6 ppm for ZnS, ZnPS, and Zn Hi, respectively). No differences in scrotal circumference or ADG were detected. Bulls receiving ZnPS had higher (P<0.05) percent normal sperm cells than did bulls receiving ZnS, but not bulls receiving ZnHi (55.5, 68.9, and 62.5% for ZnS, ZnPS, and ZnHi, respectively). The frequency of CD was less (P<0.05) for bulls on ZnPS and ZnHi treatments than for bulls on the ZnS treatment. In all fertility measures observed in this trial, bulls receiving the ZnPS rated greatest, followed by bulls on ZnHi, and lastly by bulls on ZnS.     

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.     

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.     

Gasterophilus spp. infections in horses from northern and central Kazakhstan

A cross-sectional survey was performed to obtain current data on the gastrointestinal myiasis of horses in the provinces of Kostanay, Akmola and Karagandy, northern and central Kazakhstan. The stomach, small intestine and rectum of 148 slaughter horses were examined for Gasterophilus spp. larvae during a 26-month study period. All horses were infected with 2nd and 3rd stage larvae (mean intensity: 803 ± 350), and 22% of them harboured >1000 Gasterophilus spp. larvae each. Four species were identified: G. intestinalis (prevalence: 100%; mean intensity: 361 ± 240 larvae), G. haemorrhoidalis (100%; 353 ± 191), G. nasalis (100%; 73 ± 36) and G. pecorum (91.2%; 18 ± 10). Horses aged < 2 years were higher infected with Gasterophilus larvae than 2–4 years old animals. Both the prevalence and extremely high intensity of Gasterophilus infections of horses in these Kazakh regions suggest respective control measurements to improve the health and performance of the animals and to increase the economic income of horse owners.     

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.     

Oxidative stress in dairy cows seropositives for Neospora caninum

Bovine neosporosis is caused by the protozoan Neospora caninum and is one of the major causes of abortion in cows. Cattle are intermediate hosts of this parasite and may have asymptomatic or symptomatic infections. Therefore, the aim of this study was to evaluate oxidative stress marker reactive oxygen species (ROS), thiobarbituric reactive acid substances (TBARS) levels, glutathione S-transferase (GST), adenosine deaminase (ADA), and butyrylcholinesterase (BChE) activities in dairy cows seropositives for N. caninum (asymptomatic or symptomatic). Dairy cows (n = 90) were tested by immunofluorescent antibody assay (IFA) for N. caninum and divided accordingly into three groups: the group A (seronegatives, n = 30), the group B (seropositives and asymptomatic, n = 30), and the group C (seropositives and symptomatic, n = 30). It was observed increased levels of TBARS and reduced (P < 0.05) BChE activity in seropositives either asymptomatic or symptomatic animals. ROS levels and ADA activity increased, and GST activity decreased (P < 0.05) only in seropositives symptomatic dairy cows (the group C) compared to seronegatives dairy cows (the group A). Based on these results, it was observed that seropositive animals showed cell damage associated with oxidative stress and inflammation, mainly in those with symptomatic infections. Increased seric ROS levels and BChE activity may have influenced N. caninum pathogenesis in symptomatic animals due to increased cell damage and exacerbated inflammatory response, leading to the development of clinical signs.     

Evaluation of three immunoserological techniques in the detection of porcine trichinellosis

Three immunoserological tests (IST) used for the detection of porcine trichinellosis, immunofluorescence (IF), enzyme-inmunoanalysis (EIA), and Western blot (WB), were compared. Three groups of animals were analyzed: Group 1, animals naturally infected with parasite burdens (PB) of <1 muscle larvae (ML)/g (n = 18); Group 2, animals naturally infected with PB of ≥2 ML/g (n = 23); Group 3, animals raised and home-slaughtered on farms in Argentina (n = 59). Animals from Groups 1 and 2 were identified in outbreaks and were analyzed by individual artificial digestion (AD) of ≥30 g of muscle. Animals in Group 3 were subjected to AD of 5 g of muscle.The detection percentages in sera of swine with the lower PB were 100% for IF, 72% for EIA, and 50% for WB. Eighty-three percent of the animals were serologically positive by two or three techniques. In pigs with the higher PB, the detection percentage was similar for IF and EIA (100% vs. 91%, respectively), and was lower for the WB (61%). Ninety-six percent of the animals were serologically positive by two or three techniques. Group 3 animals had similar detection percentages for the three techniques (IF, 30%; EIA, 29%; WB, 42%). Twenty-five percent of the animals were serologically positive by two or three techniques. Two animals were positive by AD with PB of 0.33 and 2.4 ML/g, and were positive for IF and WB, or IF, EIA, and WB. Results indicate that the sensitivity of each technique depends on the PB, and always ranked in sensitivity as IF > EIA > WB. For the lower PB, the decrease in the sensitivity is more pronounced for the EIA. Although the WB has a low sensitivity, the detection of the specific bands for Trichinella spiralis makes it a useful confirmatory tool. Considering that more than 83% of the parasitologically positive animals had 2 or 3 positive serological results using the techniques tested here, for the diagnosis of porcine trichinellosis, pigs positive by two of these serological techniques must be regarded as truly infected pigs.     

Application of cattle slurry containing Mycobacterium avium subsp. paratuberculosis (MAP) to grassland soil and its effect on the relationship between MAP and free-living amoeba

Slurry from dairy farms is commonly used to fertilize crops and pastures. This mixture of manure, urine and water can harbor multiple microbial pathogens among which Mycobacterium avium subsp. paratuberculosis (MAP) is a major concern. Persistence of MAP in soil and infection of soil Acanthamoeba was evaluated by culture, real-time IS900 PCR, and by staining of amoeba with acid-fast and vital stains comparing soils irrigated with MAP-spiked or control dairy farm slurry. MAP DNA was detected in soil for the 8 month study duration. MAP was detected by PCR from more soil samples for plots receiving MAP-spiked slurry (n = 61/66) than from soils receiving control slurry (n = 10/66 samples). Vital stains verified that intracellular MAP in amoeba was viable. More MAP was found in amoeba at the end of the study than immediately after slurry application. There was no relationship between MAP presence in soil and in amoeba over time. Infection of amoeba by MAP provides a protected niche for the persistence and even possibly the replication of MAP in soils. As others have suggested, MAP-infected amoeba may act like a “Trojan horse” providing a means for persistence in soils and potentially a source of infection for grazing animals.     

Systemic and local immune response in pigs intradermally and intramuscularly injected with inactivated Mycoplasma hyopneumoniae vaccines

The systemic and respiratory local immune response induced by the intradermal administration of a commercial inactivated Mycoplasma hyopneumoniae whole-cell vaccine (Porcilis^® MHYO ID ONCE – MSD AH) in comparison with two commercial vaccines administered via the intramuscular route and a negative control (adjuvant only) was investigated. Forty conventional M. hyopneumoniae-free pigs were randomly assigned to four groups (ten animals each): Group A = intradermal administration of the test vaccine by using the needle-less IDAL^® vaccinator at a dose of 0.2 ml; Group B = intramuscular administration of a commercially available vaccine (vaccine B); Group C = intramuscular administration of the adjuvant only (2 ml of X-solve adjuvant); Group D = intramuscular administration of a commercially available vaccine (vaccine D). Pigs were vaccinated at 28 days of age. Blood and bronchoalveolar lavage (BAL) fluid samples were collected at vaccination (blood only), 4 and 8 weeks post-vaccination. Serum and BAL fluid were tested for the presence of antibodies by ELISA test. Peripheral blood monomorphonuclear cells (PBMC) were isolated to quantify the number of IFN-γ secreting cells by ELISpot. Moreover, cytokine gene expression from the BAL fluid was performed. Total antibodies against M. hyopneumoniae and specific IgG were detected in serum of intradermally and intramuscularly (vaccine B only) vaccinated pigs at 4 and 8 weeks post-vaccination. M. hyopneumoniae specific IgA were detected in BAL fluid from vaccinated animals (Groups A and B) but not from controls and animals vaccinated with the bacterin D (p < 0.05). Significantly higher gene expression of IL-10 was observed in the BAL fluid at week 8 post-vaccination in the intradermally vaccinated pigs (p < 0.05). The results support that the intradermal administration of an adjuvanted bacterin induces both systemic and mucosal immune responses. Moreover, the intramuscularly administered commercial vaccines each had a different ability to stimulate the immune response both systemically and locally.