Comparison of diagnostics techniques in an outbreak of infectious laryngotracheitis from meat chickens

Various diagnostics techniques were compared for their ability to detect infectious laryngotracheitis (ILT) during an outbreak in chickens aged between 4 and 21 wk. Gross lesions ranged from excess mucus to accumulation of fibrinonecrotic exudate in the larynx and trachea. Syncytial cells with intranuclear inclusion bodies were found in sinus, conjunctiva, larynx, trachea, lung, and air sac. Virus isolation in chicken embryos was attempted in every case. Negative-stain electron microscopy detected herpesvirus in only 6% of the cases. Yet, isolation of ILT virus in the chorioallantoic membrane was presumed by histology in >20% of the samples and confirmed by fluorescent antibody (FA) in 35% of the embryos inoculated with conjunctivas or tracheas from affected birds. Overall, results from histology and FA tests were highly correlated. FA test has the advantage over histology of being diagnostically specific for ILT virus. Polymerase chain reaction was the most sensitive test and detected the viral DNA even in cases where histology and FA were negative. ILT virus DNA was quantified by real-time polymerase chain reaction (Re-Ti ILTV). Histologic and FA results from larynx and trachea were negative if the concentration of the viral DNA was < or =4 of log10. A viral DNA concentration higher than log10 4, as determined by Re-Ti ILTV, was required for clinical ILT to be manifested.     

COMPARISON OF ULTRASONOGRAPHIC AND RADIOGRAPHIC MEASUREMENTS OF CANINE PROSTATE DIMENSIONS

The purposes of this study were to: 1) compare prostatic dimensions measured on radiographic and ultrasonographic images and 2) compare a subjective radiographic assessment of prostate size with a previously-described objective method. Thirty-four male dogs undergoing investigation of prostatic disorders were used. Prostate length and depth were measured from ultrasonographic and radiographic images. A subjective assessment of prostate size ('small', 'normal', or 'enlarged') was made in 29 animals by one of the authors who was unaware of radiographic or ultrasonographic measurements. In addition, the distance from sacral promontory to the pubic brim was also measured. A prostate length or depth of >70% of this distance was defined as 'enlarged' and <70% as 'normal'. After the effects of magnification on radiographic measurements were eliminated, there were no significant differences between prostatic length measured by the two methods. However, a significant difference was obtained between prostatic depth measurements. The subjective assessment agreed with a previously described objective assessment of prostatic size in 21/29 dogs for prostate length but in only 12/29 for depth. Prostatic length varied from 46.6 to 116.4% (mean 75.7%) of the distance from the pubic brim to the sacral promontory. Prostatic depth varied from 33.0% to 94.6% (mean 59.7%) of the same distance. It is recommended that prostate length, rather than depth, be used when evaluating prostate size from lateral abdominal radiographs.     

“Arte et Labore”—A Blackburn Rovers fan's legacy in human complex trait genetics

Through his own research contributions on the modelling and genetic analysis of quantitative traits and through his former students and postdocs, Robin Thompson has indirectly left a major legacy in human genetics. In this short note, we highlight examples of the long‐lasting relevance and impact of Robin's work in human genetics. A lone early study of marker‐assisted selection developed many of the tools and approaches later exploited (often after reinvention) by the human genetics community in GWAS studies and for prediction. Furthermore, a particularly clear example of the pervasive impact of Robin's work is that REML has become the default method to estimate variance components and that genetic predictions exploiting linkage disequilibrium in the population are starting to become used in precision medicine applications.     

Seasonal and Diurnal Changes in Starch Content and Sugar Profiles of Bermudagrass in the Piedmont Region of the United States

Seasonal and diurnal patterns of sugar accumulation in bermudagrass (Cynodon dactylon) pastures were monitored to evaluate risk factors for pasture-associated laminitis of ponies and horses. Bermudagrass was collected from four plots in the morning and afternoon on a weekly basis, from mid-July until late August. Tissue was air-dried to simulate hay, or frozen to retain the sugar profiles of fresh pasture. Samples were analyzed colorimetrically for total water-soluble and ethanol-soluble carbohydrates, and electrochemically for starch. In addition, sugars were separated and quantified by high-performance liquid chromatography coupled to pulsed amperometric detection. The dominant sugars in extracts were glucose, fructose, and sucrose. Some minor peaks, corresponding to tri- and tetrasaccharides, were also detected in some extracts. Starch increased over time in fresh and dried tissue, and concentrations varied diurnally in fresh, but not in dried tissue (P = .021). Sucrose in dried tissue decreased and then increased, with higher concentrations than in fresh tissue on all sampling dates (P = .024). Glucose and fructose exhibited diurnal variation on one and two dates, respectively (P = .034 and .0028, respectively). These results reveal trends in carbohydrate concentrations and profiles that may help to evaluate the likelihood of equine laminitis outbreaks on bermudagrass.     

COLOR DOPPLER AND DOPPLER SPECTRAL ANALYSIS IN THE SPINAL CORD OF NORMAL DOGS

Doppler ultrasonography of the spinal cord was performed in 34 normal, anesthetized dogs following hemilaminectomy. This study was part of an investigation to evaluate the efficacy of a 21-aminosteroid compound and high dose methylprednisolone for the treatment of spinal cord trauma. Grey-scale images of the canine spinal cord were similar to those described for the spinal cord of people. Doppler waveforms of intraparenchymal spinal arteries exhibited high end diastolic blood flow velocities, indicating low resistance to flow. Doppler values (mean ± SD) for arteries immediately ventrolateral to the central canal were: Peak Systolic Velocity = 5.78 ± 2.5 cm/sec, Minimum Diastolic Velocity = 3.5 ± 1.62 cm/sec, Mean Velocity = 4.45 ± 1.96 cm/sec, Minimum Diastolic Velocity = 3.5 ± 1.62 cm/sec, Mean Velocity = 4.45 ± 1.96 cm/sec, Systolic/Diastolic ratio = 169 ± 0.19, Pulsatility Index = 0.53 ± 0.09, and Resistance Index = 0.4 ± 0.06.     

Management of bovine tuberculosis in Michigan wildlife: current status and near term prospects

Surveillance and control activities for bovine tuberculosis (bTB) in free-ranging Michigan white-tailed deer (Odocoileus virginianus) have now been underway for over a decade. Significant progress has been made, lowering apparent prevalence in deer in the core area by >60%, primarily via reduction of deer densities through hunting, and restrictions on public feeding and baiting of deer. These broad strategies of the Michigan Department of Natural Resources (MDNR), implemented with the cooperation of Michigan deer hunters, halved the deer population in the bTB endemic area. However, as hunters see fewer deer, their willingness to sustain aggressive harvests has waned, and public resentment of control measures has grown. During the past four years, apparent prevalence in core area deer has held approximately steady just below 2%. After bottoming out in 2004 at an estimated 10-12 deer/km(2), deer numbers have since rebounded by ～ 30%. Public compliance with baiting and feeding restrictions has been variable. In general, hunters in the core area do not perceive bTB as a problem, in spite of 13 years of MDNR outreach. To date, MDNR has expended more than US$23 million on TB-related activities. Of late, a substantial portion of that funding has been diverted to support other programs which have suffered from budget shortfalls. Livestock herd breakdowns continue to occur sporadically, averaging 3-4 per year 2005 to present. In total, 46 cattle and 4 captive deer herds have been diagnosed bTB positive statewide, the majority yielding only 1 positive animal. Five cattle herds were twice infected, one thrice. Michigan Department of Agriculture (MDA) policy emphasis has shifted towards obtaining producer support for wildlife risk mitigation and farm biosecurity. Funding has proven a limiting factor, with the majority of the US$63 million spent to date devoted to whole herd testing. Nevertheless, some initiatives justify cautious optimism. Promising research to support eventual vaccination of wild deer continues. Some hunters and landowners have begun to recognize the costs of high deer densities and supplemental feeding. A peninsula-wide ban on baiting and feeding was enacted. Some cattle producers, recognizing their precarious circumstances, have begun work to change long-held prevailing opinions among their peers about farm biosecurity. Yet formidable challenges remain, and evidence suggests that eradication of bTB, if it can be achieved, will take decades, and will require greater public and political resolve than has been demonstrated thus far.     

The role of phagocytic cells in enhanced susceptibility of broilers to colibacillosis after Infectious Bronchitis Virus infection

Colibacillosis results from infection with avian pathogenic Escherichia coli bacteria. Healthy broilers are resistant to inhaled E. coli, but previous infection with vaccine or virulent strains of Infectious Bronchitis Virus (IBV) predisposes birds for severe colibacillosis. We investigated whether IBV affects recruitment and function of phagocytic cells and examined NO production, phagocytic and bactericidal activity, and kinetics of peripheral blood mononuclear cells (PBMC) and splenocytes. Moreover, we measured cytokine mRNA expression in lung and spleen samples. Broilers were inoculated with IBV H120 vaccine or virulent M41 and challenged 5 days later with E. coli 506. A PBS control and E. coli group without previous virus inoculation were also included. Birds were sacrificed at various time points after inoculation (h/dpi). Inoculation with IBV induced extended and more severe colibacillosis than with E. coli alone. At 4dpi, the number of KUL-01(+) PBMC in all E. coli-inoculated groups was significantly higher than in PBS-inoculated birds, which correlated with lesion scores. From 1 to 4dpi, NO production by PBMC from all E. coli-inoculated animals was elevated compared to PBS birds. Bactericidal activity of PBMC in IBV-inoculated animals at 7dpi was lower than in PBS- and E. coli-inoculated birds, but phagocytic capacity and recruitment were not severely impaired. In spleen samples of IBV-infected animals reduced expression of IL-1beta, IL-6, IL-8, IL-10, IL-18 and IFN-gamma mRNA was found 1dpi. Our results suggest that enhanced colibacillosis after IBV infection or vaccination is caused at least by altered innate immunity and less by impairment of phagocytic cell function.     

Differential susceptibility to tetracycline,oxytetracycline and doxycycline of the calf pathogens Mannheimia haemolytica and Pasteurella multocida in three growth media

For clinical isolates of bovine Mannheimia haemolytica and Pasteurella multocida, this study reports minimum inhibitory concentration (MIC) differences for tetracycline, oxytetracycline and doxycycline between cation‐adjusted Mueller‐Hinton broth (CAMHB), foetal bovine serum (FBS) and Roswell Park Memorial Institute (RPMI) medium. MICs were determined according to CLSI standards and additionally using five overlapping sets of twofold dilutions. Matrix effect: (a) free drug MICs and minimum bactericidal concentrations (MBC) for all drugs were significantly higher in FBS than in CAMHB for both pathogens (p < 0.001); (b) MICs and MBCs were higher for CAMHB and FBS compared to RPMI for P. multocida only. Net growth rate for P. multocida in CAMHB was significantly slower than in FBS and higher than in RPMI, correlating to MIC and MBC ranking. Drug effect: doxycycline MICs and MBCs were significantly lower (p < 0.001) in both CAMHB and FBS than tetracycline and oxytetracycline for both pathogens. Only for M. haemolytica were oxytetracycline MIC and MBC significantly lower than tetracycline, precluding the use of tetracycline to predict oxytetracycline susceptibility in this species. Determining potencies of tetracyclines in a physiological medium, such as FBS, is proposed, when the objective is correlation with pharmacokinetic data for dosage determination.