An evaluation of interventions for reducing the risk of PRRSV introduction to filtered farms via retrograde air movement through idle fans

Porcine reproductive and respiratory syndrome virus (PRRSV) is an economically significant pathogen of pigs that can be transported via the airborne route out to 9.1 km. To reduce this risk, large swine facilities have started to implement systems to filter contaminated incoming air. A proposed means of air filtration failure is the retrograde movement of air (back-drafting) from the external environment into the animal air space through non-filtered points such as idle wall fans; however, this risk has not been validated. Therefore, the purpose of this study was threefold: (1) to prove that PRRSV introduction via retrograde air movement through idle fans is a true risk; (2) to determine the minimum retrograde air velocity necessary to introduce PRRSV to an animal airspace from an external source; and (3) to evaluate the efficacy of different interventions designed to reduce this risk. A retrograde air movement model was used to test a range of velocities and interventions, including a standard plastic shutter, a plastic shutter plus a canvas cover, a nylon air chute, an aluminum shutter plus an air chute and a double shutter system. Results indicated that retrograde air movement is a real risk for PRRSV introduction to a filtered air space; however, it required a velocity of 0.76 m/s. In addition, while all the interventions designed to reduce this risk were superior when compared to a standard plastic shutter, significant differences were detected between treatments.     

Emerging avian pathogenic Escherichia coli strains belonging to clonal groups O111:H4-D-ST2085 and O111:H4-D-ST117 with high virulence-gene content and zoonotic potential

The present study characterizes, for the first time, two emerging avian pathogenic Escherichia coli (APEC) clonal groups of serogroup O111: O111:H4-D-ST117 and O111:H4-D-ST2085. The clonal group O111:H4-D-ST117 was already present in APEC strains isolated between 1991 and 2000, and was still present in strains isolated between 2004 and 2009, showing long time evolution according to the virulence-gene differences and macrorestriction profiles. Among ST117 strains, two virulence profiles could be distinguished: papG II-positive tsh-negative strains which satisfied criteria for extraintestinal pathogenic E. coli (ExPEC), and papG II-negative tsh-positive strains without ExPEC status. Interestingly, we have detected a human septicemic O111:H4-D-ST117 ExPEC strain isolated from a hemocultive in 2000 whose macrorestriction profile showed >85% similarity with four APEC strains of the study. The clonal group O111:H4-D-ST2085 was exclusively detected in 17 APEC strains isolated in 2008 and 2009, and showed short time evolution based on its homogeneity since all were nalidixic acid-resistant, all had ExPEC status, and most carried papG II and tsh genes. From the clinical point of view, O111:H4-D-ST2085 seems a successful clonal group that could be the result of the epidemiological evolution of O111:H4-D-ST117. Due to the increasing prevalence of both clonal groups among clinical APEC isolates, their high virulence-gene content, and zoonotic potential, we suggest them as possible candidates for the development of a future vaccine against avian colibacillosis.     

Effect of including distillers dried grains with solubles in the diet, with or without antimicrobial regimen, on the ability of growing pigs to resist a Lawsonia intracellularis challenge

A disease challenge experiment was conducted to determine if including 10% dried distillers grains with solubles (DDGS) in the diet, with or without antimicrobial supplementation, reduces the incidence or severity, or both, of intestinal lesions in growing pigs after an Lawsonia intracellularis challenge. One hundred 17-d-old weaned pigs were blocked by sex, ancestry, and BW and randomly allotted to 1 of 5 treatment groups: negative control, unchallenged, corn-soy diet; positive control, challenged, corn-soy diet; 10% DDGS diet, challenged; positive control with antimicrobial regimen, challenged; and 10% DDGS diet with antimicrobial regimen, challenged. For antimicrobial-supplemented treatments, diets contained 33 ppm bacitracin methylene disalicylate throughout the experiment, with chlortetracycline (Aureomycin) pulsed at 550 ppm from d 3 prechallenge to d 11 postchallenge. Challenged pigs were orally inoculated with 8.0 x 10(8) L. intracellularis organisms after a 4-wk prechallenge period. On d 21 postchallenge, pigs were euthanized, lesions of intestinal mucosa were evaluated, and ileal tissue samples were analyzed by immunohistochemistry to determine the presence and proliferation rate of L. intracellularis. Compared with other dietary treatments, feeding a diet containing 10% DDGS reduced ileum and colon lesion length and prevalence (P < 0.05) and reduced severity of lesions in the ileum (P < 0.05) and colon (P < 0.10) in challenged pigs. Compared with other challenged pigs, those fed the diet containing the antimicrobial regimen had a lower prevalence and severity of lesions in the jejunum (P < 0.05) and tended to have reduced total tract lesion length (P = 0.11). Compared with other challenged pigs, pigs on the 10% DDGS diet with antimicrobial regimen exhibited no differences in length, severity, or prevalence of lesions (P > 0.15), but fecal shedding of L. intracellularis was reduced on d 14 postchallenge (P < 0.05). No dietary effects on fecal shedding were observed by d 20 postchallenge (P > 0.10). The proportion of cells infected with L. intracellularis was reduced when DDGS (P = 0.05) or antimicrobial (P = 0.10) diets were fed. Under the conditions of this experiment, dietary inclusion of 10% DDGS appears to provide some benefit to growing pigs subjected to a moderate L. intracellularis challenge, similar to those of a currently approved antimicrobial regimen.     

In vivo estimation of sheep carcass composition using real-time ultrasound with two probes of 5 and 7.5 MHz and image analysis

Ultrasonic measurements were taken on 46 sheep using a real-time ultrasound machine equipped with 2 probes (5 and 7.5 MHz). Measurements of subcutaneous fat thickness (SC) and muscle LM depth (MD) and area (MA) were taken at 2 locations: over the 13th thoracic vertebra (SC13, MD13, and MA13, respectively) and at the interval between the third and fourth lumbar vertebrae (SC34, MD34, and MA34, respectively). Fat thickness was also measured over the third sternebra of the sternum. The relationship between carcass and in vivo ultrasound measurements was high for all the measurements (r(2) between 0.54 and 0.96, P < 0.01). Concerning MD and SC, the 7.5 MHz probe estimates were consistently more precise than the 5-MHz estimates (r(2) increased between 0.09 and 0.13), but the reverse occurred with the MA estimates, although to a lesser extent. Estimates of carcass composition for muscle, subcutaneous fat, intermuscular fat, internal fat, and total fat based on BW explained a large amount of variation in muscle (87%), subcutaneous fat (85%), intermuscular fat (79%), internal fat (74%), and total fat (87%). In most cases (55 of 70) the introduction of one ultrasound measurement in addition to BW in the multiple regression equations further improved the explanation of variation for weight of carcass tissues, internal fat, and total fat. For carcass muscle estimation, the ultrasound measurements of muscle provided an increase of r(2) between 0.05 and 0.10 (P < 0.01). The SC13 and SC34 gave the best improvements in estimating subcutaneous fat, intermuscular fat, internal fat, and total fat (r(2) increased between 0.05 and 0.17; P < 0.01). Prediction of the proportions of the carcass components (internal and total fat from BW) was clearly lower than the prediction of the absolute amounts of these traits. Inclusion of one or more ultrasound measurements in addition to BW increased the predictive ability of the equations. Both probes were useful to estimate carcass muscle depth and area and fat depth, but the 7.5-MHz probe showed a greater ability to estimate depth. For all traits, the stepwise procedure demonstrated that the best fit was obtained with BW and one or more ultrasound measurement with the 7.5-MHz probe.     

An epidemiological evaluation of Mycobacterium bovis infections in wild game animals of the Spanish Mediterranean ecosystem

Recreational hunting of indigenous wild artiodactyls has been one of the most lucrative and rapidly growing industries in Western Spain over the last five years. In the absence of careful ecological management, one consequence of the commercial exploitation of this natural resource has been the appearance of outbreaks of infectious disease; most notably bovine tuberculosis. From the outset of the study in 1997, we have observed a steady increase in prevalence of Mycobacterium bovis (M. bovis) in both species reaching 1.74 (+/-0.17) in deer in 2002 and 2.32 (+/-0.24) in wild boar. The latter species seems to be most severely affected with pulmonary lesions appearing more chronic than those observed in deer. In this study, we describe the epidemiology of M. bovis in European wild boar (Sus scrofa) and Iberian red deer (Cervus elaphus hispanicus) in Extremadura (W. Spain); a region where there are large areas of natural habitat for these species.     

Chlamydophila psittaci in free-living Blue-fronted Amazon parrots (Amazona aestiva) and Hyacinth macaws (Anodorhynchus hyacinthinus) in the Pantanal of Mato Grosso do Sul, Brazil

Chlamydophila psittaci (C. psittaci) infection was evaluated in 77 free-living nestlings of Blue-fronted Amazon parrots (Amazona aestiva) and Hyacinth macaws (Anodorhynchus hyacinthinus) in the Pantanal of Mato Grosso do Sul, Brazil. Tracheal and cloacal swab samples from 32 wild parrot and 45 macaw nestlings were submitted to semi-nested PCR, while serum samples were submitted to complement fixation test (CFT). Although all 32 Amazon parrot serum samples were negative by CFT, cloacal swabs from two birds were positive for Chlamydophila DNA by semi-nested PCR (6.3%); these positive birds were 32 and 45 days old. In macaws, tracheal and cloacal swabs were positive in 8.9% and 26.7% of the samples, respectively. Complement-fixing antibodies were detected in 4.8% of the macaw nestlings; macaw nestlings with positive findings were between 33 and 88 days old. These results indicate widespread dissemination of this pathogen in the two evaluated psittacine populations. No birds had clinical signs suggestive of chlamydiosis. To the best of our knowledge, this is the first report on C. psittaci in free-living Blue-fronted Amazon parrots and Hyacinth macaws in Brazil.     

Comparison of different methods for diagnosis of porcine proliferative enteropathy

The objectives of this study were: (1) to compare 2 methods of serology; (2) to compare 3 histologic techniques; and (3) to compare 2 methods of detecting shedding in pigs experimentally challenged with Lawsonia intracellularis. The sensitivities of these tests were determined by the detection of infection. Forty 5-week-old pigs were inoculated on day 0 with intestinal homogenate from pigs with proliferative enteropathy (PE). Clinical evaluation was done on day 7 and daily from day 14 to 28 postinoculation. Fecal shedding of L. intracellularis was monitored by use of polymerase chain reaction (PCR) analysis and immunoperoxidase staining at 7-day intervals. Serum was obtained on days 0 and 28 for serologic testing by glass slide and tissue culture indirect fluorescent antibody tests. At euthanasia on day 28, gross intestinal lesions were evaluated and ileum samples collected for histologic analyses. Ileal histologic sections from each animal were stained by hematoxylin and eosin, Warthin-Starry silver stain, and immunohistochemistry (IHC). Of the 40 pigs, 36 had gross lesions typical of PE at necropsy. The percentage of agreement between the 2 serologic methods was 94.4%. Immunoperoxidase stain of fecal smears was more sensitive than PCR for detecting fecal shedding, especially on day 21 (89.5% and 60.5%, respectively) and day 28 (59.4% and 37.5%, respectively) post-inoculation. The IHC stain was much more sensitive for detecting infection than the routinely used hematoxylin and eosin and Warthin-Starry silver stains. In conclusion, in experimentally infected pigs, both serologic methods were appropriate techniques for detecting infection. For fecal samples, PCR has low sensitivity. Immunohistochemistry is the best diagnostic tool for formalin-fixed samples.     

Development of a Non-radioactive Dot-blot Hybridisation Assay for the Detection of Pelargonium Flower Break Virus and Pelargonium line Pattern Virus

The ornamental geranium, Pelargonium×hortorum Bailey, is a traditional ornamental plant widely cultivated in Europe and Northern America. Vegetative propagation facilitates rapid spread of viral infections which have detrimental effects on the production and the quality of the crop. A non-radioactive nucleic acid hybridisation method was developed for detection of Pelargonium flower break virus (PFBV) and Pelargonium line pattern virus (PLPV) in infected host plants. This method was significantly more sensitive than the conventional ELISA test when using either purified viral preparations or crude plant extracts. The distribution of the viruses was studied by means of the non-isotopic hybridisation technique. The results indicated that the petioles and the apical blade regions of fully expanded leaves were the best source of test material. The hybridisation procedure enables the detection of PFBV and PLPV in a single assay, and its simplicity allows its application to routine large-scale indexing.     

Porcine myelomonocytic markers: summary of the second international swine CD workshop

Forty five mAbs submitted to the Second International Swine CD workshop were analyzed by six different laboratories for their possible reactivity with porcine myelomonocytic cells using flow cytometry and immunohistochemistry. As a result of these analyses, a new swine workshop cluster, SWC9, composed of two mAbs that recognize an antigen selectively expressed on mature macrophages, was defined. In addition, several mAbs were identified, allowing the differentiation of granulocytes from monocytes/macrophages, or monocytes from macrophages. Further work is required to identify the antigen recognized by these mAbs. Nevertheless, they should already prove useful for the identification of different stages in the macrophage maturation/differentiation, and will certainly aid analyses on the complexity of the mononuclear phagocyte system in the pig. Finally, the cross-reactivity of three anti-human CD14 mAbs with porcine myelomonocytic cells was established in this workshop.     

Mastitis caused by Mycoplasma mycoides subspecies mycoides (large colony type) in goat flocks in Spain

We describe three different outbreaks of mastitis caused by M. mycoides subspecies mycoides LC type (Mmm LC) in three goat flocks from the Extremadura Region of south-west Spain. Thirty-two fast-growing isolates were obtained on Hayflick's and Friis's media with inhibitors from different specimens. All were identified as Mmm LC in spite of their cultural, biochemical and serological features.