A simple molecular method to identify and quantify genera of gastrointestinal nematodes of cattle.

Classic approaches for antemortem identification of gastrointestinal nematodes (GIN) require coproculture of eggs and morphological examination. While adequate for diagnosis, many PCR techniques cannot easily quantify mixed infections without controls and/or standard curves. Herein, we developed a simple and rapid test for differentiating and quantifying mixed infections of GIN using PCR products separated by capillary electrophoresis. Among the cattle GIN, the ITS2 region is sufficiently distinct in length to delineate among the most common infecting genera, Ostertagia ostertagi = 373 bases (b), Haemonchus contortus (placei) = 366b, Cooperia punctata (oncophora) = 376b, Trichostrongylus axei = 372b, and Oesophagostomum radiatum = 357b. Conserved primers were synthesized that span the ITS2 where one primer was fluorescently labeled with 6-FAM. DNAs from infective L3 were PCR amplified then loaded onto an ABI 3130 sequencer adapted for size fragment analysis. Resulting peak amplitudes were both diagnostic and quantitative on a relative basis. As proof of principle, quantification was performed on PCR fragments from mixed species pairs of Ostertagia ostertagi, Cooperia punctata, and Haemonchus contortus and analyzed using Gene Marker V1.85 software. In all cases, linear responses were observed where R2 > 0.97 and line slopes ranged between 0.90 and 1.1. When tested on eggs from naturally infected animals, the assay showed superior results on two farms when compared to coproculture and morphological identification. Using wildlife-derived samples, results coincided well with deep amplicon sequencing. The assay is adaptable to large-scale studies, does not require comparative PCR controls, and should be compliant with GIN from small ruminant livestock.

Cooperia punctata: effect on cattle productivity?

Cooperia spp. have become the most prevalent parasites in United States cow/calf operations as observed in the USDA NAHMS (National Animal Health Monitoring System) Beef Cow/Calf survey in 2008. This is at least in part due to the widespread use of macrocyclic lactones that have recently been shown to have a reduced activity against these parasites. The effects of Cooperia spp. on cattle productivity are largely unknown. This study was conducted to assess their effect upon cattle housed under conditions found in American feedlots. Two hundred yearling calves (average weight 460 lb/209 kg) were acquired from northwestern Arkansas and northeastern Oklahoma and were vaccinated and dewormed upon arrival at the feedlot. Animals were comingled and preconditioned for approximately one month, and were fed a standard growing ration throughout the study. Calves were randomly divided into two groups (n=80, infected and control) and each group was further divided into two replicate pens (n=40). Calves from the two infected pens were orally inoculated with a gavage of 1 × 10(5) and 0.825 × 10(5) infective larvae of a recent isolate of Cooperia punctata on day 0 and 14, respectively, with the two control pens receiving a similar volume of tap water. Data collected included biweekly fecal egg counts, daily individual feed consumption and weight gain over the 60-day test period. The presence of C. punctata (>99% of recovered worms) was confirmed by necropsy and recovery from the small intestine on days 35 and 60 post infection (PI) in a subset of animals. Egg counts were positive by day 14 PI and remained at numbers similar to values seen in field studies. The control group gained weight 7.5% more rapidly (p=0.02) than infected animals (3.24 lb/1.47 kg per day vs. 3.0 lb/1.36 kg per day, respectively). The Cooperia-infected calves also consumed 1.5 lb (0.68 kg) less dry feed per day than the control animals (p=0.02). These data suggest that C. punctata has a deleterious effect on both appetite and nutrient uptake or utilization. At necropsy (days 35 and 60), the draining mesenteric lymph nodes of infected animals were increased in size and the small intestinal mucosa was thickened and covered with a thick layer of mucus in the infected animals. The most prominent histological changes in the Cooperia- infected animals included a moderate increase in the number of intraepithelial lymphocytes and globule leukocytes, as well as aggregates of eosinophils within the lower lamina propria. The only significant difference was an increase in the goblet cell density at day 60. Anthelmintic sensitivity/resistance of the Cooperia isolate used was determined by treatment of one pen of infected calves with a macrocyclic lactone and the other pen with a benzimidazole at the completion of the study. The macrocyclic lactone treatment (n=40) did not remove the parasites (FECRT=8.8%), while treatment with a benzimidazole was very effective (FECRT=98.1%). This study demonstrated that C. punctata has a significant effect on cattle productivity, both reduced weight gain and decreased feed intake compared to controls.

Comparison of single vaccination versus revaccination with a modified-live virus vaccine containing bovine herpesvirus-1, bovine viral diarrhea virus (types 1a and 2a), parainfluenza type 3 virus, and bovine respiratory syncytial virus in the prevention of bovine respiratory disease in cattle.

Objective-To compare effects of administration of a modified-live respiratory virus vaccine once with administration of the same vaccine twice on the health and performance of cattle. Design-Randomized, controlled trial. Animals-612 mixed-breed male cattle with unknown health histories. Procedures-Cattle were randomly assigned to 1 of 2 treatment groups (single vaccination treatment group [SVAC group] vs revaccination treatment group [REVAC group]) during the preconditioning phase of production. All cattle were given a modified-live respiratory virus vaccine. Eleven days later, REVAC group cattle received a second injection of the same vaccine. During the finishing phase of production, cattle from each treatment group were either vaccinated a third time with the modified-live respiratory virus vaccine or given no vaccine. Health observations were performed daily. Blood and performance variables were measured throughout the experiment. Results-During preconditioning, no significant differences were observed in performance or antibody production between groups. Morbidity rate from bovine respiratory disease was lower for SVAC group cattle; however, days to first treatment for bovine respiratory disease were not different between groups. No significant differences in body weights, daily gains, or dry-matter intake between groups were observed during the finishing phase. Revaccination treatment group cattle had improved feed efficiency regardless of vaccination protocol in the finishing phase. Conclusions and Clinical Relevance-Vaccination once with a modified-live respiratory virus vaccine was as efficacious as vaccination twice in the prevention of bovine respiratory disease of high-risk cattle, although feed efficiency was improved in REVAC group cattle during the finishing period.

Reproductive safety of vaccination with Vista 5 L5 SQ near breeding time as determined by the effect on conception rates.

Replacement heifers (N=799; 10 to 13 months of age) were vaccinated with Vista 5 L5 SQ (Intervet; a reconstituted vaccine-bacterin product containing modified-live cultures of infectious bovine rhinotracheitis [IBR] virus, bovine viral diarrhea virus [BVDV; types 1 and 2], parainfluenza-3 virus, and bovine respiratory syncytial virus and inactivated cultures of Leptospira serovars canicola, grippotyphosa, hardjo, icterohaemorrhagiae, and pomona with a proprietary adjuvant) at either 40 plus/minus 5 days (control; n=399) or 3 days (test; n=400) before peak breeding day. By 40 plus/minus 5 days before peak breeding day, heifers in both groups had greater average titers to IBR, BVDV types 1 and 2, and four of the five Leptospira antigens assessed as compared with prevaccination titers on day -90 plus/minus 25 days. Conception rates were not affected by treatment. This study suggests that conception rates will not differ between heifers vaccinated with Vista 5 L5 SQ 3 days before breeding and those vaccinated approximately 40 days before breeding.