Charles L. Stoltenow

Risk Factors Associated with Anthrax Outbreak in Animals in North Dakota, 2005 : A Retrospective Case-Control Study

Objective. We identified the risk factors associated with the anthrax outbreak of 2005 in animals in North Dakota. Methods. Medical records of the 2005 anthrax outbreak were obtained from the Veterinary Diagnostic Laboratory at North Dakota State University. Additional data were obtained from the North Dakota state veterinarians office, and supplemental questionnaires were administered to producers. The data obtained included ecological and environmental factors, animal health factors, and management factors. Results. Anthrax occurred from July 1 to October 12, 2005. The cases were located in eastern North Dakota around the Red River Basin. Ransom, LaMoure, and Barnes counties reported most cases (71%). Species affected included cattle, bison, horses, sheep, elk, deer, pigs, and llamas. The predominant symptom was sudden death (38%) followed by bleeding from orifices (17%). Chi-square analysis indicated significant differences between case and control premises on the following variables: death reported on neighboring pasture, vaccination period, dry conditions, wet conditions, antibiotic use, multiple vaccination, and type of predator (coyote). Factors that significantly (p<0.05) predicted anthrax occurrences on the final logistic regression model were vaccination, use of antibiotics during an outbreak, and period of vaccine administration (before or during the outbreak). Conclusions. The characteristics of the anthrax outbreak regarding time and place of occurrence, animals affected, clinical signs reported, and mortality rate were consistent with previous reports of natural anthrax outbreaks in animals. A number of factors that significantly predicted anthrax occurrence in animals in the 2005 outbreak in North Dakota were identified. This information is important in planning appropriate control and prevention measures for anthrax, including recommending the right vaccination and treatment regimens in managing future anthrax outbreaks.

A Review of Management Practices for the Control of Anthrax in Animals: The 2005 Anthrax Epizootic in North Dakota – Case Study

Outbreaks of anthrax have diverse consequences on society. Establishing the appropriate control strategies is very important and crucial in reducing the socio‐economic impact of the disease. Control measures are aimed at breaking the cycle of infection, and their implementation must be adhered to rigorously. The objectives of this paper were: (i) to review the control strategies currently used in management of anthrax in animals and (ii) to describe management strategies used by producers in North Dakota during the 2005 anthrax outbreak in livestock. Anthrax control strategies were divided in to strategies that apply before, during, and after an anthrax outbreak. This paper also highlights the problems or constraints faced by North Dakota producers in controlling anthrax during the outbreak of 2005.

Development of an Assay to Determine Single Nucleotide Polymorphisms in the Prion Gene for the Genetic Diagnosis of Relative Susceptibility to Classical Scrapie in Sheep

The objective of this study was to develop a reliable Taqman® 5′ Nuclease Assay for genotyping sheep for scrapie susceptibility. The sheep prion gene contains 2 single nucleotide olymorphisms (SNPs) that may mediate resistance to classical scrapie, one at codon 136, alanine (A) or valine (V), and another at codon 171, arginine (R) or glutamine (Q). The R allele appears to confer resistance to classical scrapie, with the AA136 RR171 genotype the most resistant to scrapie and QR171 only rarely infected in the US sheep population. The Assays by Design® protocol was used for development of probes and rimers for codon 136 and Primer Express® for codon 171. Commercially available kits were used to isolate genomic DNA from blood or muscle. For validation, 70 SNP determinations for each codon were compared to commercial testing with an error rate of less than 1%. Then, 935 samples from blood (n = 818) and muscle (n = 117) were tested for both codons with 928 successful determinations and only 7 samples (<1% of total samples) that needed repeating. Genotypes were AA QQ (n = 102; 11.0%), AV QQ (n = 28; 3.0%), AA QR (n = 396; 42.7%), AV QR (n = 54; 5.8%), and AA RR (n = 348; 37.5%). Thus, 86% of the sheep tested (n = 798) contained R at codon 171 and were expected to be scrapie-resistant. This new Taqman® 5′ Nuclease SNP genotyping assay is accurate, easy to perform, and useful in the study of classical scrapie in sheep and its prevention through selective breeding programs to eliminate highly susceptible animals.

Rabies in an American Bison from North Dakota

In North Dakota (USA) during April 1998, a ranched female bison (Bison bison) was found dead. At gross necropsy, there was profound hair loss and consolidated lung lobes. Intracytoplasmic neuronal inclusions suggestive of Negri bodies were observed in the brain stem and hippocampus, and a diagnosis of rabies was confirmed by the fluorescent antibody test. Antigenic typing demonstrated the occurrence of a rabies virus variant associated with skunks from the upper mid-western USA. This case of a rabid bison was one of only four such instances recorded from the USA over the past 40 yr, and is the first case report of rabies in a bison that reports clinical, pathologic, and antigenic findings. Although rabies in bison is rare, veterinarians and wildlife managers that work closely with such non-traditional species are reminded of the dangers that zoonoses such as rabies present.

Effects of Nutrition and Genotype on Prion Protein (PrPC) Gene Expression in the Fetal and Maternal Sheep Placenta

For placental transmission of scrapie to occur, the normal cellular prion protein (PrPC) must be converted to an abnormal infectious form known as PrPSc. PrPC genotype influences susceptibility to contracting scrapie, but we still do not understand whether genotype or expression levels of PrPC are important in transmission of scrapie. Some evidence exists that nutrition affects expression levels of PrPC. Thus, we evaluated the effects of genotype and nutrition on PrPC mRNA and protein expression in adolescent ewes fed at control (100% of National Research Council [NRC] requirements) or restricted (60% of NRC) levels of diet intake during two periods of pregnancy (days 50-90 and days 90-130)]. Gravid uteri (n=50) from singleton pregnancies were collected at day 130, and placentomes were either separated into caruncular (CAR; maternal) or cotyledonary (COT; fetal) placenta and snap-frozen for PrPC mRNA expression or perfusion fixed for PrPC protein expression. PrPC genotypes were determined (codons 136 and 171) using SNP assay. There were no genotype effects on PrPC mRNA expression in CAR or on PrPC protein expression in either CAR or COT, but PrPC mRNA expression in COT was greater (P<0.02) when codon 136 was homozygous for alanine. Some PrPC protein-positive cells were found in the epithelium of CAR, but most were found in trophoblast binucleate and mononucleate cells of COT. In CAR, from days 90 to 130, PrPC protein abundance was greater (P=0.003) in diet-restricted ewes than in control ewes, but was less uniformly distributed (P<0.007). Additionally, in COT, from days 90 to 130, PrPC protein was less uniformly distributed (P<0.01) in diet-restricted ewes. The localized increase in PrPC protein expression, found in ewes diet-restricted late in pregnancy, may suggest a protective role for PrPC in placental biology. Further study is needed to evaluate whether nutrition, PrPC genotype, and PrPC expression levels influence placental transmission of scrapie.