Concurrent treatment with a macrocyclic lactone and benzimidazole provides season long performance advantages in grazing cattle harboring macrocyclic lactone resistant nematodes.

In 2013, a 118-day study was initiated to investigate the efficacy of concurrent treatment at pasture turnout with an injectable macrocyclic lactone with activity up to 28 days and an oral benzimidazole, referred to as "conventional" anthelmintics, when compared to treatment with conventional macrocyclic lactone alone or an injectable macrocyclic lactone with extended activity of 100 days or longer. A group of 210 steers were obtained from a ranch in California and transported to Idaho, USA. A total of 176 steers with the highest fecal egg counts were blocked by pre-treatment body weights and pasture location. A total of 44 pasture paddocks were assigned with 4 steers per paddock with 12 paddocks per therapeutic treatment group and 8 paddocks per controls. The four treatments were injectable doramectin (Dectomax®, Zoetis Inc., 0.2 mg kg-1BW, SC), injectable doramectin concurrently with oral albendazole (Valbazen®, Zoetis Inc., 10 mg kg-1BW, PO), extended release injectable eprinomectin (LongRange™, Merial Limited, 1 mg kg-1BW, SC) or saline. Cattle were individually weighed and sampled for fecal egg count on Days 0, 31/32, 61, 88, and 117/118 with an additional fecal sample on Day 14. At conclusion, one steer per paddock was euthanized for nematode recovery. The results from the first 32 days found evidence of macrocyclic lactone resistance against injectable doramectin and extended release eprinomectin. During this period the concurrent therapy provided nearly 100% efficacy based on fecal egg count reduction and a 19.98% improvement in total weight gain compared to controls (P = 0.039). At the conclusion of the 118-day study and past the approved efficacy for the conventional anthelmintics, the concurrent therapy with conventional anthelmintics provided a 22.98% improvement in total weight gain compared to controls (P = 0.004). The 118-day improvement in weight gain for the extended release eprinomectin group (29.06% compared to control) was not statistically different from the concurrent therapy with conventional anthelmintics. The results indicate that concurrent treatment with a conventional macrocyclic lactone and benzimidazole may provide production benefits early in the grazing period that continue throughout the entire period for cattle harboring macrocyclic lactone resistant nematodes. By using two different anthelmintic classes together, macrocyclic lactone resistant parasites were effectively controlled early in the period. Furthermore, the use of an effective conventional anthelmintic treatment regimen without an extended period of drug release may help to promote refugia and decrease the further selection for anthelmintic resistant parasites.

Feeding performance, carcass characteristics, and tenderness attributes of steers sorted by the Igenity tenderness panel and fed zilpaterol hydrochloride.

Steers (n = 560; initial BW = 420 ± 26 kg) were selected from a pool of 1,040, using the IGENITY Profile DNA test for tenderness, sorted into 1 of 4 tenderness genotype (TG) groups [140 tough (TUF), 140 intermediate (INT), 140 tender (TEND), or 140 mixed (MXD)], and subsequently allocated into 56 pens at random, of which one-half (28 pens, 7 pens from each TG) were supplemented the ß-adrenergic agonist zilpaterol hydrochloride (ZH) and the balance fed a control ration. No TG × ZH interaction (P ≥ 0.15) occurred for any measured trait. Cattle from INT TG had less (P < 0.05) DMI during pretreatment (d 0 to 118) and entire trial (d 0 to 143) periods than other TG. Cattle fed ZH had greater (P < 0.01) ADG and G:F, and decreased (P < 0.01) DMI during the treatment period (d 119 to 143). Cattle from the TEND group had greater (P < 0.01) marbling scores, increased (P < 0.02) calculated USDA yield grades (YG), and more (P < 0.02) calculated empty body fat (EBF) than TUF cattle. Cattle receiving ZH during the treatment period had increased (P < 0.01) HCW, dressed yield, and LM area. Additionally, cattle fed ZH exhibited decreased (P < 0.01) EBF, marbling, KPH, and calculated USDA YG. No difference (P > 0.06) in YG distributions were detected among TG, yet TEND cattle were represented by a greater (P < 0.01) proportion of Prime and premium Choice carcasses. Cattle fed ZH exhibited increased (P < 0.01) frequencies of YG 2 carcasses and fewer (P < 0.01) YG 3, 4, and 5 carcasses concurrent with an increase (P < 0.04) in the percentage of Select carcasses. Longissimus steaks from TUF cattle had greater (P < 0.03) Warner-Bratzler Shear Force (WBSF) values at 7 and 14 d postmortem than steaks from INT or TEND cattle. Furthermore, ZH-fed cattle had increased (P < 0.01) WBSF values for all aging periods compared with control cattle. Frequency of steaks with WBSF values <3.9 kg (certified tender) were less (P < 0.05) for the TUF group. Feeding ZH resulted in fewer longissimus steaks (P < 0.01) with WBSF values <3.0 kg (guaranteed tender) across all aging periods; however, no difference in the frequency of steaks with WBSF values <3.9 kg was found after 21 d of aging. Igenity Profile tenderness scores were correlated (P < 0.05) to carcass finish attributes and WBSF values. Commercially available tenderness panels may have the potential to allow for antemortem sorting of cattle into expected tenderness groupings, which could augment feeding management strategies and ultimately lead to increased marketing value for the beef system.

Anthelmintic resistance of Ostertagia ostertagi and Cooperia oncophora to macrocyclic lactones in cattle from the western United States.

In June 2008, 122 yearling heifers with a history of anthelmintic resistance were obtained from pastures in northern California and transported to a dry lot facility in southwestern Idaho, USA. Fifty heifers with the highest fecal egg counts were selected for study enrollment. Candidates were equally randomized to treatment with either injectable ivermectin (Ivomec, Merial, 0.2 mg kg(-1) BW), injectable moxidectin (Cydectin), Fort Dodge, 0.2 mg kg(-1) BW), oral fenbendazole (Safe-Guard), Intervet, 5.0 mg kg(-1) BW), oral oxfendazole (Synanthic), Fort Dodge, 4.5 mg kg(-1) BW), or saline. At 14 days post-treatment, nematodes were recovered from the abomasum, small intestine, and large intestine. Parasitism was confirmed in the control group when 10/10 animals were infected with adult Ostertagia ostertagi and 9/10 animals with both developing and early L(4) stages of O. ostertagi. Similarly, 9/10 animals were parasitized with adult Cooperia spp. Fenbendazole and oxfendazole efficacy verses controls were >90% against adult Cooperia spp., while moxidectin caused an 88% parasite reduction post-treatment (P<0.05). Ivermectin treatment resulted in no reduction in adult Cooperia spp. Based on geometric mean percent reduction versus saline controls, all four treatments were >or=90% efficacious against adults of O. ostertagi, while moxidectin and fenbendazole were equally effective against developing and inhibited early L(4) stages (P<0.05). Ivermectin was not efficacious for developing or inhibited early L(4) stages of O. ostertagi. Oxfendazole failed to decrease O. ostertagi developing L(4) larvae by >90% but was efficacious for inhibited early L(4) larvae. Based on the results of this study, a source of multi-species anthelmintic resistance in cattle has been identified in the western United States.

Pathogenicity and protective activity in pregnant goats of a Brucella melitensis Deltaomp25 deletion mutant.

The Brucella melitensis mutant BM 25, which lacks the major 25 kDa outer membrane protein Omp25, has previously been found to be attenuated in the murine brucellosis model. In the present study, the capacity of the Deltaomp25 mutant to colonise and cause abortions in the caprine host was evaluated. The vaccine potential of BM 25 was also investigated in goats. Inoculation of nine pregnant goats in late gestation with the B. melitensis mutant resulted in 0/9 abortions, while the virulent parental strain, B. melitensis 16M, induced 6/6 dams to abort (P<0.001, n=6). BM 25 also colonised fewer adults (P<0.05, n=6) and kids (P<0.01, n=6) than strain 16M. The Deltaomp25 mutant was found capable of transient in vivo colonisation of non-pregnant goats for two weeks post-infection. Owing to the ability of BM 25 to colonise both non-pregnant and pregnant adults without inducing abortions, a vaccine efficacy study was performed. Vaccination of goats prior to breeding with either BM 25 or the current caprine vaccine B. melitensis strain Rev. 1 resulted in 100 per cent protection against abortion following challenge in late gestation with virulent strain 16M (P<0.05, n=7). However, unlike strain Rev. 1, BM 25 does not appear to cause abortions in late gestation based on this study with a small number of animals. The B. melitensis Deltaomp25 mutant, BM 25, may be a safe and efficacious alternative to strain Rev. 1 when dealing with goat herds of mixed age and pregnancy status.

Attenuation of a Brucella abortus mutant lacking a major 25 kDa outer membrane protein in cattle.

OBJECTIVE: To determine the virulence of a Brucella abortus mutant, BA25, lacking a major 25 kd outer membrane protein (Omp25) in cattle. ANIMALS: 20 mixed-breed heifers in late gestation. PROCEDURE: 10 heifers were inoculated with 1 x 10(7) colony-forming units of the Omp25 mutant via the conjunctival sac, and an equal number were infected with the virulent parental strain B. abortus 2308. The delivery status of the dams was recorded, and colonization was assessed following necropsy. The ability of BA25 to replicate inside bovine phagocytes and chorionic trophoblasts was also evaluated in vitro because of the propensity of virulent brucellae to replicate inside these cells in vivo. RESULTS: The parental strain induced abortions in 5 of 10 inoculated cattle, whereas only 1 of 10 dams exposed to BA25 aborted. Brucella abortus strain 2308 colonized all of the cow-calf pairs and induced Brucella-specific antibodies in 100% of the dams. In contrast, BA25 was isolated by bacteriologic cultural technique from 30% of the calves and 50% of the inoculated dams (n = 10). Of the 10 heifers inoculated with BA25, 4 did not develop Brucella-specific antibodies nor were they colonized by the mutant strain. In bovine macrophages and chorionic trophoblasts, BA25 replicated in significantly lower numbers than the virulent parental strain (n = 3). CONCLUSIONS AND CLINICAL RELEVANCE: The 25 kd outer membrane protein may be an important virulence factor for B. abortus in cattle. The attenuation of the Omp25 mutant in cattle may involve the inability of BA25 to replicate efficiently in bovine phagocytes and chorionic trophoblasts.

Oral vaccination of sexually mature pigs with Brucella abortus vaccine strain RB51.

OBJECTIVE: To develop a novel oral vaccine delivery system for swine, using the rough vaccine strain of Brucella abortus. ANIMALS: 56 crossbred pigs from a brucellosis-free facility. PROCEDURE: In 3 separate experiments, pigs were orally vaccinated with doses of 1 x 10(9) to > 1 x 10(11) CFU of strain RB51 vaccine. The vaccine was placed directly on the normal corn ration, placed inside a whole pecan, or mixed with cracked pecans and corn. RESULTS: Oral vaccination of pigs with vaccine strain RB51 resulted in a humoral immune response to strain RB51 and short-term colonization of the regional lymph nodes. CONCLUSIONS AND CLINICAL RELEVANCE: A viscous liquid such as Karo corn syrup in association with pecans that scarify the oral mucosa are necessary when placing the live vaccine directly onto corn or other food rations. Doses of > 1 x 10(11) CFU of RB51 organisms/pig in this mixture ensures 100% colonization of regional lymph nodes via the oral route. This method may allow an efficient and economical means to vaccinate feral swine for brucellosis.

Re-examination of the role of the Brucella melitensis HtrA stress response protease in virulence in pregnant goats.

Based on previously reported studies describing the experimental infection of pregnant goats with B. melitensis strain RWP5, we proposed that the HtrA protease plays an important role in the virulence of B. melitensis in its natural ruminant host. Subsequent studies, however, have shown that RWP5 is actually an htrA cycL double mutant. In order to definitively evaluate the role of the B. melitensis htrA in virulence, we constructed an authentic htrA mutant and examined this strain in pregnant goats. The findings of these studies indicate that the contribution of the htrA gene product to the virulence of B. melitensis in its natural host is not as great as was previously proposed.

Use of western immunoblot analysis for testing moose serum for Brucella suis biovar 4 specific antibodies.

To determine if 12 moose (Alces alces) from northern Alaska with agglutinating antibodies specific for Brucella spp. had been exposed to either B. suis biovar 4 or B. abortus biovar 1, western immnnoblot serologic analysis was performed. Differential serologic responses to strain specific A and M antigenic variances of the lipopolysaccharide O-polysaccharide sugar allowed strain identification. Prior to examination, test sera were absorbed with killed whole cells from either B. abortus biovar 1, containing predominately A antigen (A+ M-); B. melitensis biovar 1, containing essentially M antigen (A- M+); or B. suis biovar 4, containing both antigenic tyes (A+ M+). The resulting sera were then examined by western immunoblot for recognition of either B. abortus biovar 1, B. melitensis biovar 1, or B. suis biovar 4 cell lysates. The results of this study indicate that these moose were exposed to B. suis biovar 4, a known pathogen of caribou (Rangifer tarandus) from arctic Alaska.

Biosafety of Brucella abortus strain RB51 for vaccination of mature bulls and pregnant heifers.

OBJECTIVE: To determine shedding and colonization profiles in mature sexually intact bulls and pregnant heifers after vaccination with a standard calfhood dose of Brucella abortus strain RB51 (SRB51). ANIMALS: 6 sexually mature 3-year-old Jersey bulls and 7 mixed-breed heifers in midgestation. PROCEDURE: Bulls and pregnant heifers were vaccinated IM with the standard calfhood dose of 3x10(10) colony-forming units of SRB51. After vaccination, selected body fluids were monitored weekly for vaccine organism shedding. Pathogenesis was monitored in bulls by weekly breeding soundness examination and, in heifers, by delivery status of the calf. Vaccine organism colonization was assessed by obtaining select tissues at necropsy for bacterial culture. Serologic analysis was performed by use of numerous tests, including complement fixation, an SRB51-based ELISA, and immunoblot analysis. RESULTS: After vaccination, none of the vaccinated bulls or heifers shed SRB51 in their secretions. Results of breeding soundness examination for bulls were normal as was delivery status of the pregnant heifers (6 live births, 1 dystocia). At necropsy, SRB51 was not recovered from any of the selected tissues obtained from bulls, heifers, or calves; however, serologic analysis did detect SRB51-specific antibodies in all cattle. CONCLUSIONS AND CLINICAL RELEVANCE: Vaccination with the standard calfhood dose of SRB51 administered IM was not associated with shedding or colonization in sexually mature bulls or pregnant heifers. Also, under conditions of this study with small numbers of animals, IM vaccination with SRB51 does not appear to cause any reproductive problems when administered to sexually mature cattle.

Safety of Brucella abortus strain RB51 in Bison.

To determine the safety of Brucella abortus strain RB51 (SRB51) vaccine in American bison (Bison bison), 31 animals from a herd with brucellosis were used. In October 1996, 10 adult bison males and seven calves were vaccinated with the standard calfhood cattle dose of 1.8 x 10(10) colony forming units (CFU) of SRB51 subcutaneously while the adult females received the standard adult cattle dose of 1 x 10(9) CFU. Western immunoblot indicated the presence of SRB51 antibodies following vaccination. To evaluate prolonged bacterial colonization of tissues, the adult males, calves, and three adult females were divided into two groups which were slaughtered at either 13 or 16 wk post-vaccination. At necropsy, tissue samples were obtained for B. abortus culture from the liver, spleen, lymph nodes, and reproductive tract of each animal. While B. abortus field strain was cultured from one adult bull, no SRB51 was isolated from any of the animals. Seven pregnant females were monitored until parturition for signs of abortions and fetal lesions. Six cows delivered healthy calves and one delivered a dead full-term calf that was brucellae negative. Based on these results, administration of SRB51 to bison did not cause prolonged bacterial colonization of tissues in calves, adult males, or adult females. Furthermore, SRB51 did not induce abortions following vaccination in the second month of gestation.