Raphaël Vanderstichel

The two-step Fagan's nomogram: ad hoc interpretation of a diagnostic test result without calculation

In 1975, Fagan published a nomogram to help practitioners determine, without the use of a calculator or computer, the probability of a patient truly having a condition of interest given a particular test result. Nomograms are very useful for bedside interpretations of test results, as no test is perfect. However, the practicality of Fagans nomogram is limited by its use of the likelihood ratio (LR), a parameter not commonly reported in the evaluation studies of diagnostic tests. The LR reflects the direction and strength of evidence provided by a test result and can be computed from the conventional diagnostic sensitivity (DSe) and specificity (DSp) of the test. This initial computation is absent in Fagans nomogram, making it impractical for routine use. We have seamlessly integrated the initial step to compute the LR and the resulting two-step nomogram allows the user to quickly interpret the outcome of a test. With the addition of the DSe and DSp, the nomogram, for the purposes of interpreting a dichotomous test result, is now complete. This tool is more accessible and flexible than the original, which will facilitate its use in routine evidence-based practice. The nomogram can be downloaded at: www.adelaide.edu.au/vetsci/research/pub_pop/2step-nomogram/.

Managing marine disease emergencies in an era of rapid change

Infectious marine diseases can decimate populations and are increasing among some taxa due to global change and our increasing reliance on marine environments. Marine diseases become emergencies when significant ecological, economic or social impacts occur. We can prepare for and manage these emergencies through improved surveillance, and the development and iterative refinement of approaches to mitigate disease and its impacts. Improving surveillance requires fast, accurate diagnoses, forecasting disease risk and real-time monitoring of disease-promoting environmental conditions. Diversifying impact mitigation involves increasing host resilience to disease, reducing pathogen abundance and managing environmental factors that facilitate disease. Disease surveillance and mitigation can be adaptive if informed by research advances and catalysed by communication among observers, researchers and decision-makers using information-sharing platforms. Recent increases in the awareness of the threats posed by marine diseases may lead to policy frameworks that facilitate the responses and management that marine disease emergencies require.

Evaluation of anti-Ostertagia ostertagi antibodies in individual milk samples as decision parameter for selective anthelmintic treatment in dairy cows

The aim of this study was to explore whether anti-Ostertagia ostertagi antibody levels measured by ELISA in individual milk samples hold promise as a decision parameter for either herd-level decisions or selective anthelmintic treatments by investigating (1) the relationship between individual and bulk-tank milk ELISA results; (2) the relationships of individual milk ELISA results with non-parasitic cow factors and (3) the relationship between individual milk ELISA results and the milk production response after anthelmintic treatment. Twelve farms were randomly allocated to a whole-herd treatment with eprinomectin or a placebo in October 2004 and individual milk samples and a bulk-tank milk sample were collected 1 month before and 1 month after treatment. Linear mixed models were used to investigate the associations of ELISA results with (a) breed, actual milk production, lactation stage, somatic cell count, age and sampling month and (b) the milk production response after anthelmintic treatment. There was a reasonable correlation between the mean individual and bulk-tank milk ELISA results (r=0.72). Individual cow ELISA results increased with higher lactation number and were lower in November than September. The associations with the other non-parasitic factors were weak and not significant. Milk yield responses to anthelmintic treatment were greater when treatment was given in early lactation and increased with the pre-treatment ELISA result and cow age. However, these latter two interaction terms were not significant when they were put in the model together. We conclude that (1) O. ostertagi ELISA results from individual milk samples may provide more information on the herds parasitic status than a single bulk-tank milk result; (2) lactation number should be taken into account when interpreting ELISA results from individual milk samples and (3) the value of the O. ostertagi antibody level in individual cow milk samples to predict individual production responses after anthelmintic treatment remains equivocal.

A multicentre prevalence study in Europe on Giardia duodenalis in calves, with molecular identification and risk factor analysis.

The present study aimed to obtain data on the occurrence of Giardia duodenalis in calves in four major cattle rearing countries in Europe (Germany, UK, France and Italy), along with genotyping data and risk factors associated with these infections. A total of 2072 calves were sampled on 207 farms. The majority of the animals were Holstein dairy or mixed Holstein calves (n=1565 or 75.5%), and were female (n=1640 or 79.1%). The average age was 7.8 weeks (SD=4.1; median=7; range=2-16 weeks). All fecal samples were tested using a commercially available monoclonal antibody-based ELISA. The overall apparent prevalence of G. duodenalis for the four countries was 45.4% (n=942/2072) and the overall farm prevalence was 89.9% (186/207), with differences in both animal and farm prevalence between the four countries. The prevalence was significantly higher in animals up to 8 weeks (OR=1.88; P<0.001) compared to older calves, and several management factors including contact with the Dam, Frequency of cleaning of the Maternity Pens, and Disinfection of the Calf Housing were found to be associated with infection. Positive samples were withheld for genotyping using the β-giardin and triose phosphate isomerase gene: G. duodenalis assemblage E was most prevalent, although 43% of the isolates were typed as assemblage A, with differences in between countries. Furthermore, 32% of the examined samples was found to be a mixed assemblage A and E infection, which is consistent with previous reports. The results of the present study confirm previous findings in other European countries that G. duodenalis infections are common in calves. The infection especially occurs in animals younger than 2 months, and the proportion of positive animals gradually decreased with increasing age.

Heart and skeletal muscle inflammation (HSMI) disease diagnosed on a British Columbia salmon farm through a longitudinal farm study

Heart and skeletal muscle inflammation (HSMI) is an emerging disease of marine-farmed Atlantic Salmon (Salmo salar), first recognized in 1999 in Norway, and later also reported in Scotland and Chile. We undertook a longitudinal study involving health evaluation over an entire marine production cycle on one salmon farm in British Columbia (Canada). In previous production cycles at this farm site and others in the vicinity, cardiac lesions not linked to a specific infectious agent or disease were identified. Histologic assessments of both live and moribund fish samples collected at the farm during the longitudinal study documented at the population level the development, peak, and recovery phases of HSMI. The fish underwent histopathological evaluation of all tissues, Twort’s Gram staining, immunohistochemistry, and molecular quantification in heart tissue of 44 agents known or suspected to cause disease in salmon. Our analysis showed evidence of HSMI histopathological lesions over an 11-month timespan, with the prevalence of lesions peaking at 80–100% in sampled fish, despite mild clinical signs with no associated elevation in mortalities reported at the farm level. Diffuse mononuclear inflammation and myodegeneration, consistent with HSMI, was the predominant histologic observation in affected heart and skeletal muscle. Infective agent monitoring identified three agents at high prevalence in salmon heart tissue, including Piscine orthoreovirus (PRV), and parasites Paranucleospora theridion and Kudoa thyrsites. However, PRV alone was statistically correlated with the occurrence and severity of histopathological lesions in the heart. Immunohistochemical staining further localized PRV throughout HSMI development, with the virus found mainly within red blood cells in early cases, moving into the cardiomyocytes within or, more often, on the periphery of the inflammatory reaction during the peak disease, and reducing to low or undetectable levels later in the production cycle. This study represents the first longitudinal assessment of HSMI in a salmon farm in British Columbia, providing new insights on the pathogenesis of the disease.

Replication of 2 Subtypes of Low-Pathogenicity Avian Influenza Virus of Duck and Gull Origins in Experimentally Infected Mallard Ducks

Many subtypes of low-pathogenicity avian influenza (LPAI) virus circulate in wild bird reservoirs, but their prevalence may vary among species. We aimed to compare by real-time reverse-transcriptase polymerase chain reaction, virus isolation, histology, and immunohistochemistry the distribution and pathogenicity of 2 such subtypes of markedly different origins in Mallard ducks (Anas platyrhynchos): H2N3 isolated from a Mallard duck and H13N6 isolated from a Ring-billed Gull (Larus delawarensis). Following intratracheal and intraesophageal inoculation, neither virus caused detectable clinical signs, although H2N3 virus infection was associated with a significantly decreased body weight gain during the period of virus shedding. Both viruses replicated in the lungs and air sacs until approximately day 3 after inoculation and were associated with a locally extensive interstitial, exudative, and proliferative pneumonia. Subtype H2N3, but not subtype H13N6, went on to infect the epithelia of the intestinal mucosa and cloacal bursa, where it replicated without causing lesions until approximately day 5 after inoculation. Larger quantities of subtype H2N3 virus were detected in cloacal swabs than in pharyngeal swabs. The possible clinical significance of LPAI virus-associated pulmonary lesions and intestinal tract infection in ducks deserves further evaluation.

Effects of farm management practices and environmental factors on bulk tank milk antibodies against gastrointestinal nematodes in dairy farms across Canada

Enzyme-linked immunosorbent assays (ELISAs) have been used as a diagnostic tool to quantify levels of gastrointestinal nematodes in dairy cattle by measuring Ostertagia ostertagi antibodies in milk. Higher levels of O. ostertagi antibodies measured by ELISA methods, referred to as optical density ratios (ODRs), are associated with decreased milk production in dairy cattle. On-farm management practices (e.g. pasturing techniques and anthelmintic usage) can influence the exposure of cattle to nematode infections and the magnitude of acquired worm burdens. Additionally, environmental and climatic factors, such as land elevation and precipitation, may also influence the levels of gastrointestinal parasitism. This repeated cross-sectional study investigated the effect of farm management practices and surrounding environmental factors on bulk tank (BT) ODRs in herds from provinces across Canada, and further examined the potential effects of various anthelmintic treatment protocols on BT ODRs. A total of 195 herds contributed an average of 3.5 BT samples between December 2003 and April 2005. The farm management practices were recorded from a questionnaire asking producers about their pasturing methods (confined, pastured, etc.), pasture sharing practices (e.g. mixing heifers with milking cows) and anthelmintic treatments. Environmental data were downloaded online from various governmental databases (e.g. Natural Resources Canada, Statistics Canada, Environment Canada, etc.). Statistical models, accounting for repeated measures (multiple BT ODRs for each farm) and for clustering of farms within a region (province or ecoregion), were used to analyze environmental and farm management data. Overall, the greater the exposure that heifers and milking cows had to pasture, the higher the levels of anti-parasite antibodies detected in BT samples. Treating the entire herd or treating milking cows at calving reduced BT ODR values. Farms in areas with higher number of rainy days, higher normalized difference vegetation index (NDVI) values, and lower land surface temperatures (LSTs), were also likely to have higher BT ODRs. Seasonal variation was such that late summer and early fall, when parasite load was at its highest, yielded larger BT ODRs. Due to the high clustering effect at the herd level, factors at the herd level (e.g. pasturing methods, anthelmintic administration) had a higher potential impact on bulk-tank measurements than the herds surrounding environmental factors.

Effects of surgical and chemical sterilization on the behavior of free-roaming male dogs in Puerto Natales, Chile.

Population management of free-roaming domestic dogs (Canis lupus familiaris) is of interest due to the threat these animals pose to people, other animals and the environment. Current sterilization procedures for male dogs include surgical and chemical methods. However, little is known about how these procedures affect their behavior. The primary objective of this study was to investigate changes in selected behaviors following chemical and surgical sterilization in a male free-roaming dog (FRD) population in southern Chile. We also examined the association between serum testosterone levels and behaviors thought to be influenced by circulating androgens. A total of 174 dogs were randomly assigned to either a surgical or chemical sterilization group, or a control group. At the onset of the intervention period, 119 dogs remained and 102 dogs successfully completed the study. Each dog was monitored pre- and post-intervention using video recordings, GPS collars, and blood samples for the measurement of testosterone. Analysis of behavior revealed that surgically castrated dogs showed no reduction of sexual activity or aggression when compared to their pre-intervention behavior. Chemically sterilized dogs showed a statistically significant increase in dog-directed aggression, but no change in sexual activity. There was no change in home range size in any groups between the pre- and post-intervention measurement. We found no consistent association between levels of serum testosterone concentration and behavioral changes in any of the groups. This study presents the first detailed behavioral observations following surgical and chemical sterilization in male FRDs. The information generated is highly relevant to communities struggling with the control of FRDs. Complementary studies to further our understanding of the effects of male sterilization on the behavioral and reproductive dynamics of FRD populations are needed.

Necrosuppurative Orchitis and Scrotal Necrotizing Dermatitis Following Intratesticular Administration of Zinc Gluconate Neutralized With Arginine (EsterilSol) in 2 Mixed-Breed Dogs

Intratesticular injection of EsterilSol (zinc gluconate neutralized with arginine) is a chemical sterilant for male dogs sometimes used in population control campaigns. Adverse reactions have been reported in 1% to 4% of treated dogs, but detailed histomorphologic descriptions are lacking. During a behavioral study conducted in the Chilean Patagonia in 2012, severe necrosuppurative orchitis and ulcerative dermatitis were observed in 2 of 36 (6%) dogs sterilized with EsterilSol according to the manufacturer’s instructions. Reactions were noted on days 8 and 7 postinjection and required scrotal ablation on days 8 and 13, respectively; neither reaction was associated with the injection site. Although self-trauma following administration may have contributed, the cause of the adverse reactions is uncertain. EsterilSol is a relatively uncomplicated method to sterilize male dogs, but the occurrence of severe adverse reactions several days after administration emphasizes the need for the provision of long-term monitoring and veterinary care during sterilization campaigns using this product.

Blood Collection from the Facial (Maxillary)/Musculo-Cutaneous Vein in True Frogs (Family Ranidae)

Collection of blood from amphibians, as in other classes of vertebrate animals, is essential to evaluate parameters of health, diagnose hemoparasitism, identify viral and bacterial pathogens, and measure antibodies. Various methods of blood collection have been described for amphibians. Most can be cumbersome (venipucture of femoral vein, ventral abdominal vein or lingual venus plexus) or result in pain or deleterious health consequences (cardiac puncture and toe-clipping). We describe an easy and practical technique to collect blood from frogs and toads that can be used in multiple species and is minimally invasive. The technique consists of puncturing either the facial or, less commonly, the musculo-cutaneous vein and collecting the blood with a capillary tube. These veins run dorsal and parallel to the maxillary bone and can be accessed by quick insertion and withdrawal of a needle through the skin between the upper jawline and the rostral or caudal side of the tympanum. The needle should be of 27 or 30 gauge for anurans weighing more or less than 25 g, respectively. Although the technique has been used by some amphibian researchers for years, it is little known by others and has never been fully described in a peer-reviewed publication.