Highly Efficient Site-Specific Mutagenesis in Malaria Mosquitoes Using CRISPR.

Anopheles mosquitoes transmit at least 200 million annual malaria infections worldwide. Despite considerable genomic resources, mechanistic understanding of biological processes in Anopheles has been hampered by a lack of tools for reverse genetics. Here, we report successful application of the CRISPR/Cas9 system for highly efficient, site-specific mutagenesis in the diverse malaria vectors Anopheles albimanus, A. coluzzii, and A. funestus When guide RNAs (gRNAs) and Cas9 protein are injected at high concentration, germline mutations are common and usually biallelic, allowing for the rapid creation of stable mutant lines for reverse genetic analysis. Our protocol should enable researchers to dissect the molecular and cellular basis of anopheline traits critical to successful disease transmission, potentially exposing new targets for malaria control

Genome-wide patterns of polymorphism in an inbred line of the African malaria mosquito Anopheles gambiae.

Anopheles gambiae is a major mosquito vector of malaria in Africa. Although increased use of insecticide-based vector control tools has decreased malaria transmission, elimination is likely to require novel genetic control strategies. It can be argued that the absence of an A. gambiae inbred line has slowed progress toward genetic vector control. In order to empower genetic studies and enable precise and reproducible experimentation, we set out to create an inbred line of this species. We found that amenability to inbreeding varied between populations of A. gambiae. After full-sib inbreeding for ten generations, we genotyped 112 individuals--56 saved prior to inbreeding and 56 collected after inbreeding--at a genome-wide panel of single nucleotide polymorphisms (SNPs). Although inbreeding dramatically reduced diversity across much of the genome, we discovered numerous, discrete genomic blocks that maintained high heterozygosity. For one large genomic region, we were able to definitively show that high diversity is due to the persistent polymorphism of a chromosomal inversion. Inbred lines in other eukaryotes often exhibit a qualitatively similar retention of polymorphism when typed at a small number of markers. Our whole-genome SNP data provide the first strong, empirical evidence supporting associative overdominance as the mechanism maintaining higher than expected diversity in inbred lines. Although creation of A. gambiae lines devoid of nearly all polymorphism may not be feasible, our results provide critical insights into how more fully isogenic lines can be created

Recruitment of Veterinarians for Rural Communities through Education: The Veterinary Training Program for Rural Kansas

Veterinarians are important members of rural communities and provide valuable services to both community members and the food production systems in these areas. The population of new veterinary graduates entering rural, mixed animal practice is decreasing and several studies have concluded that a shortage of veterinarians in this area is apparent. The Veterinary Training Program for Rural Kansas (VTPRK) was created to address the educational and financial needs of students entering practice in rural Kansas. The program provides a forgivable loan for a select group of students during veterinary school and specific courses to help prepare these students for success in rural practice

Germline Cas9 expression yields highly efficient genome engineering in a major worldwide disease vector, Aedes aegypti.

The development of CRISPR/Cas9 technologies has dramatically increased the accessibility and efficiency of genome editing in many organisms. In general, in vivo germline expression of Cas9 results in substantially higher activity than embryonic injection. However, no transgenic lines expressing Cas9 have been developed for the major mosquito disease vector Aedes aegypti Here, we describe the generation of multiple stable, transgenic Ae. aegypti strains expressing Cas9 in the germline, resulting in dramatic improvements in both the consistency and efficiency of genome modifications using CRISPR. Using these strains, we disrupted numerous genes important for normal morphological development, and even generated triple mutants from a single injection. We have also managed to increase the rates of homology-directed repair by more than an order of magnitude. Given the exceptional mutagenic efficiency and specificity of the Cas9 strains we engineered, they can be used for high-throughput reverse genetic screens to help functionally annotate the Ae. aegypti genome. Additionally, these strains represent a step toward the development of novel population control technologies targeting Ae. aegypti that rely on Cas9-based gene drives

Professional Services Offered in Rural, Mixed-Animal Veterinary Practices

Rural, mixed animal veterinary practices offer a variety of professional services to their clientele. Practitioners must decide which services to offer and how much time to dedicate to each offering. These decisions are influenced by the perceived need for each service in the area and the potential income that can be generated from the specific service

Backgrounding health associated with area of the truck where cattle were housed during transport

Cattle are commonly moved between geographic regions by using commercial transport carriers, and the vast majority of cattle are transported at least one time during their lives. Both handling and travel associated with moving cattle between locations have been identified as potentially stressful events. The objective of this research was to identify potential associations between calf location within the transport carrier and subsequent calf wellness in the short term (40 to 60 days) following shipment. Health outcomes and average daily gain (ADG) were used to measure calf wellness during the backgrounding period. Although some research has described the overall effect of hauling cattle, we are aware of no recent literature describing the effects of location within the vehicle on subsequent animal wellness and performance

Backgrounding health associated with area of the truck where cattle were housed during transport

Cattle are commonly moved between geographic regions by using commercial transport carriers, and the vast majority of cattle are transported at least one time during their lives. Both handling and travel associated with moving cattle between locations have been identified as potentially stressful events. The objective of this research was to identify potential associations between calf location within the transport carrier and subsequent calf wellness in the short term (40 to 60 days) following shipment. Health outcomes and average daily gain (ADG) were used to measure calf wellness during the backgrounding period. Although some research has described the overall effect of hauling cattle, we are aware of no recent literature describing the effects of location within the vehicle on subsequent animal wellness and performance

Connecting Supply Chain Through Sustainability: Initiating a Multi-Disciplinary, Multi-Industry Approach Using the Case of Beef Cattle

Disciplinary perspectives of supply chains are influenced by the “silo” nature of academia. For example, educators typically train students about appropriate supply chain management starting with the manufacturer of products used in their particular industry, as opposed to investigating the entire supply chain from raw material producer to finished consumer product through disposal or recycling. When educating sustainability, understanding the extent and interrelated nature of the entire supply chain is essential. Without foundational knowledge and systems thinking, students may not understand the impact of business decisions on sustainability, and motivation to make sustainable choices may be lacking. This project aims to develop a Collaborative Grant Type 2 to infuse sustainability education into a variety of baccalaureate programs (e.g., animal sciences, hospitality and textile programs) using the study of beef cattle. Beef cattle present complex issues that involve a global view of the challenge to feed, clothe and fuel 9.6 billion people by the year 2050 while conserving finite resources

Determination of value of bovine respiratory disease control using a remote early disease identification system compared with conventional methods of metaphylaxis and visual observations

Citation: White, B. J., Amrine, D. E., & Goehl, D. R. (2015). Determination of value of bovine respiratory disease control using a remote early disease identification system compared with conventional methods of metaphylaxis and visual observations. Journal of Animal Science, 93(8), 4115-4122. doi:10.2527/jas2015-9079Mitigation of the deleterious effects of bovine respiratory disease (BRD) is an important issue in the cattle industry. Conventional management of calves at high risk for BRD often includes mass treatment with antimicrobials at arrival followed by visual observation for individual clinical cases. These methods have proven effective; however, control program efficacy is influenced by the accuracy of visual observation. A remote early disease identification (REDI) system has been described that monitors cattle behavior to identify potential BRD cases. The objective of this research was to compare health and performance outcomes using either traditional BRD control (visual observation and metaphylaxis) or REDI during a 60-d postarrival phase in high-risk beef calves. The randomized controlled clinical trial was performed in 8 replicates at 3 different facilities over a 19-mo period. In each replicate, a single load of calves was randomly allocated to receive either conventional management (CONV; total n = 8) or REDI (total n = 8) as the method for BRD control. Cattle were monitored with each diagnostic method for the first 30 d on feed and performance variables were collected until approximately 60 d after arrival. Statistical differences (P 0.10) in risk of BRD treatment and REDI calves were not administered antimicrobials at arrival; therefore, REDI calves had a lower (P < 0.01) average number of doses of antimicrobials/calf (0.75 +/- 0.1 doses) compared with CONV calves (1.67 +/- 0.1 doses). In this trial, the REDI system was comparable to conventional management with the potential advantages of earlier BRD diagnosis and decreased use of antimicrobials. Further research should be performed to evaluate the longer-term impacts of the 2 systems