A conserved motif in the immune-subdominant RAP-1 related antigen of Babesia bovis contains a B-cell epitope recognized by antibodies from protected cattle.

Introduction: Babesia bovis, a tick-borne apicomplexan parasite causing bovine babesiosis, remains a significant threat worldwide, and improved and practical vaccines are needed. Previous studies defined the members of the rhoptry associated protein-1 (RAP-1), and the neutralization-sensitive rhoptry associated protein-1 related antigen (RRA) superfamily in B. bovis, as strong candidates for the development of subunit vaccines. Both RAP-1 and RRA share conservation of a group of 4 cysteines and amino acids motifs at the amino terminal end (NT) of these proteins. Methods and results: Sequence comparisons among the RRA sequences of several B. bovis strains and other Babesia spp parasites indicate a high level of conservation of a 15-amino acid (15-mer) motif located at the NT of the protein. BlastP searches indicate that the 15-mer motif is also present in adenylate cyclase, dynein, and other ATP binding proteins. AlphaFold2 structure predictions suggest partial exposure of the 15-mer on the surface of RRA of three distinct Babesia species. Antibodies in protected cattle recognize a synthetic peptide representing the 15-mer motif sequence in iELISA, and rabbit antibodies against the 15-mer react with the surface of free merozoites in immunofluorescence. Discussion and conclusion: The presence of the 15-mer-like regions in dynein and ATP-binding proteins provides a rationale for investigating possible functional roles for RRA. The demonstrated presence of a surface exposed B-cell epitope in the 15-mer motif of the B. bovis RRA, which is recognized by sera from protected bovines, supports its inclusion in future subunit epitope-based vaccines against B. bovis.

Self-emulsifying drug delivery systems (SEDDS) containing Cymbopogon citratus essential oil: Enhancing the stability and acaricide efficacy against Rhipicephalus (Boophilus) microplus.

The objectives of this study were to develop a self-emulsifying drug delivery system (SEDDS) to enhance the stability and efficacy of Cymbopogon citratus essential oil or lemongrass oil (LEO) against cattle tick larvae and engorged females. The system with the highest oil loading in SEDDS was composed of LEO (23.33%w/w), Tween 80: SGKH 4000 in a 2:1 ratio as surfactant (66.67%w/w), and propylene glycol as co-surfactant (10%w/w). The selected SEDDS-LEO has a particle size of 18.78 nm with a narrow size distribution (polydispersity index of 0.27). Notably, the stability of SEDDS was superior to that of the original oil, both during long-term storage and under accelerated conditions. SEDDS-LEO at oil concentrations ranging from 1.458% to 5.833% w/v showed a significantly higher percentage of egg-laying reduction against adult ticks compared with the original oil at the same concentrations (p < 0.05). Furthermore, SEDDS-LEO demonstrated greater larvicidal efficacy than the original oil, with lower LC50 and LC90 values of 0.91 mg/mL and 1.20 mg/mL, respectively, whereas the original oil's LC50 and LC90 values were 1.17 mg/mL and 1.74 mg/mL, respectively. Our findings indicate that SEDDS-LEO is a promising candidate for use as an acaricide in the control of tick populations in dairy cattle.

Evaluation of selected risk factors for different stages of digital dermatitis in Dutch dairy cows.

Digital dermatitis (DD) is a painful infectious disease in dairy cattle that causes ulcerative lesions of the skin just above the coronary band, mainly of the hind legs. Estimates for DD prevalence at cow level in the Netherlands range from 20% to 25%. In this study, risk factors for the various stages of DD were identified and quantified. The hind legs of 6766 cows on 88 farms were scored by trained interns, using the M-scoring system (M0-M4.1). Farms in this study were a convenience sample, based on the prevalence of DD as recorded at the latest herd trim, geographical location and willingness of the farmers to participate. A survey with questions about cow environment and herd management was conducted by the intern at the day of scoring. The data were collected between August 2017 and January 2018. DD was found on 38.6% of the scored legs; 49.8% of the cows had DD on at least one leg and M4 was the most frequent stage (20.9%). Not removing manure on a regular basis resulted in lower odds for M2, M4 and M4.1 compared to cleaning by automatic scrapers ten times a day or more (odds ratio [OR]= 0.16, 0.49 and 0.18, respectively). The odds for M2 and M4 lesions were higher in cows aged 3-5 years than in first-calved cows (OR> 1.5 and > 1.7, respectively). Rubber flooring in the passageways resulted in lower odds for both M1 and M2 (OR, 0.06 and 0.32, respectively). Prophylactic use of footbaths treatment with an alternative active compound resulted in significant higher odds for M4 lesions than formalin and a combination of formalin and copper sulphate (OR= 1.69 and 2.04 respectively). The odds for an M4.1 lesion were lower in cows from smaller herds (n = 50-100) compared to large herds (n >100; OR= 0.67).

Evaluation of potassium monopersulfate footbath solution for controlling digital dermatitis in lactating dairy cattle. A randomized clinical trial.

Our objective was to assess potassium monopersulfate as a disinfectant used in footbath to control digital dermatitis (DD) in dairy cows. We hypothesized that a potassium monopersulfate solution would control DD. A 180-day randomized negative controlled trial was conducted in a 265-Holstein free-stall facility. Throughout the trial, foot bathing was performed bi-weekly using a split (left vs. right feet) footbath: one tub filled with 1% potassium monopersulfate (treatment), the other with tap water (control). Digital dermatitis lesions were scored during trimming chute examinations of the unwashed hind heels every 90 days using the modified M-scoring system. Digital dermatitis lesions were re-categorized into four variables: 1) inactive; 2) active; 3) any; 4) inactive or absence of DD lesions. Three longitudinal outcomes were characterized: risks of 1) developing a DD lesion; 2) reactivating an inactive DD lesion; 3) development of an inactive or the absence of the DD lesion. A generalized linear model was used to compare the variables and longitudinal outcomes between treated and control groups. Prevalence of active DD lesions increased from 12.5% to 39.9% between days 0 and 90. This significant increase in prevalence justified the discontinuation of the study on day 90 for ethical reasons. There was no statistical difference between treated and control groups for the first outcome (RR: 1.0; 95% CI: 0.62, 1.7), the second outcome (RR: 1.0; 95% CI: 0.62,-1.7); or the third outcome (RR: 0.88; 95% CI: 0.37, 2.1). A 1% potassium monopersulfate footbath solution appears ineffective to control DD in this study.

Interferon-tau protects bovine endometrial epithelial cells against inflammatory injury by regulating the PI3K/AKT/ß-catenin/FoxO1 signaling axis.

Endometritis is one of the most common causes of infertility in dairy cows, and is histopathologically characterized by inflammation and damage of endometrial epithelium. Interferon-tau (IFN-τ) is a novel type I interferon secreted by ruminant trophoblast cells with low cytotoxicity even at high doses. Previous studies suggested that IFN-τ plays an important role in inflammation. However, the mechanisms whereby IFN-τ may modulate the inflammatory responses in the bovine endometrium are unknown. In the present study, primary bovine endometrial epithelial cells (BEEC) isolated from fresh and healthy uterine horns were used for in vitro studies. The integrity of BEEC was assessed by immunofluorescence staining for cytokeratin 18 (CK-18, a known epithelial marker). For the experiments, BEEC were stimulated with different concentrations of lipopolysaccharide (LPS; 0-20 µg/mL) for different times (0-24 h). Cell viability and apoptosis were assessed via CCK-8 and flow cytometry. In a preliminary study, we observed that compared with the control group without LPS, 10 µg/mL of LPS stimulation for 24 h induced apoptosis. In a subsequent study, 20 or 40 ng/mL of IFN-τ alleviated LPS-induced apoptosis. Relative to the LPS group, western blotting further revealed that IFN-τ inhibited the protein abundance of TLR4 and phosphorylated (p-) p65 (p-p65) and Bax/Bcl-2 ratio, suggesting that IFN-τ can protect BEEC against inflammatory injury. Furthermore, the protein abundance of p-phosphoinositide 3-kinase (p-PI3K), p-protein kinase B (p-AKT), p-glycogen synthase kinase-3ß (p-GSK3ß), ß-catenin, and p-forkhead box O1 (p-FoxO1) was lower in the LPS group, whereas IFN-τ upregulated their abundance. The use of LY294002, a specific inhibitor of PI3K/AKT, attenuated the upregulation of p-PI3K, p-AKT p-GSK3ß, ß-catenin, and p-FoxO1 induced by IFN-τ, and also blocked the downregulation of TLR4, p-p65, and Bax/Bcl-2 ratio. This suggested that the inhibition of TLR4 signaling by IFN-τ was mediated by the PI3K/AKT pathway. Furthermore, compared with the LPS group, the ß-catenin agonist SB216763 led to greater p-FoxO1 and lower p-p65 and cell apoptosis. In contrast, knockdown of ß-catenin using small interfering RNA had the opposite effects. To explore the role of FoxO1 on the inhibition of TLR4 by IFN-τ, we employed LY294002 to inhibit the PI3K/AKT while FoxO1 was knocked down. Results revealed that the knockdown of FoxO1 blocked the upregulation of TLR4 and p-p65 induced by LY294002, and enhanced the inhibition of IFN-τ on TLR4, p-p65, and cell apoptosis. Overall, these findings confirmed that IFN-τ can protect endometrial epithelial cells against inflammatory injury via suppressing TLR4 activation through the regulation of the PI3K/AKT/ß-catenin/FoxO1 axis. These represent new insights into the molecular mechanisms underlying the anti-inflammatory function of IFN-τ in BEEC, and also provide a theoretical basis for further studies on the in vivo application of IFN-τ to help prevent negative effects of endometritis.

Bovine papular stomatitis virus as a vaccine vector for cattle.

Virus vectored vaccines are not available commercially for cattle even though compelling potential applications exist. Bovine papular stomatitis virus (BPSV), a highly prevalent parapoxvirus, causes self-limited oral lesions in cattle. Ability of virus to accommodate large amounts of foreign DNA, induce low level of antiviral immunity, and circulate and likely persist in cattle populations, make BPSV an attractive candidate viral vector. Here, recombinant BPSV were constructed expressing either Bovine herpesvirus 1 (BoHV-1) glycoprotein gD (BPSVgD), or gD and gB (BPSVgD/gB). Immunization of BPSV serologically-positive calves with BPSVgD or BPSVgD/gB induced BoHV-1 neutralization antibodies and provided protection for three of four animals following a high dose BoHV-1 challenge at day 70 pi. Results indicate BPSV suitability as a candidate virus vector for cattle vaccines.

Routine hoof-trimming data provides insight into the occurrence of claw lesions in Holstein herds in the central region of South Africa.

Claw lesions in dairy cows contribute significantly to lameness, causing distress and discomfort for affected cows and raising welfare concerns. Despite increased awareness, lameness incidence continues to rise. Defining and recording claw traits are particularly problematic. In South Africa (SA), claw data is limited to paper-based records kept by private hoof trimmers. This research analysed claw-trimming data from five dairy farms over 6 years to examine the occurrence and recording of claw lesions in SA Holstein cattle. Lesion identification followed the Claw Lesion Identification in Dairy Cattle brochure. Among the recorded lesions, digital dermatitis (DD) had the highest prevalence (64.02%), followed by sole ulcers (SU; 8.59%), white line disease (WLD; 6.27%), and sole haemorrhage (SH; 4.28%), and most lesions occurred in the rear feet. Chi-square tests and correspondence analysis (CA) were employed to explore the relationships between lesions, feet, and housing. Results indicated that the prevalence of SU and SH showed high similarity for foot and lesion association, and that these were more highly associated with the rear feet. Additionally, the prevalence of DD and interdigital phlegmon were strongly associated, and closely associated with SU, and all these lesions were associated with both dirt lot and free-stall housing systems. CA further confirmed a close association between WLD and SH, and the prevalence of these lesions in the combination housing system. Results of this study highlight the complexity of lesion data and that specific associations between lesions could lead to simplifying the recording thereof. Consolidating the most informative claw lesions into categories will aid in the practical prevention, management, and treatment of lameness on-farm.

Prolonged survival of venereal Tritrichomonas foetus parasite in the gastrointestinal tract, bovine fecal extract, and water.

IMPORTANCE: Nowadays, the routine herd diagnosis is usually performed exclusively on bulls, as they remain permanently infected, and prevention and control of Tritrichomonas foetus transmission are based on identifying infected animals and culling practices. The existence of other forms of transmission and the possible role of pseudocysts or cyst-like structures as resistant forms requires rethinking the current management and control of this parasitic disease in the future in some livestock regions of the world.

Evaluation of a refugia-based strategy for gastrointestinal nematodes on weight gain and fecal egg counts in naturally infected stocker calves administered combination anthelmintics.

Refugia-based strategies associated with a combination of anthelmintic drugs belonging to different drug classes are becoming more common management practices to mitigate anthelmintic resistance (AR) in gastrointestinal nematodes (GIN) in small ruminants. Though refugia-based strategies have been largely demonstrated in small ruminants, cattle veterinarians and producers are considering such management strategies in grazing cattle production systems. Implementing refugia-based strategies lowers the amount of anthelmintics used in the herd and therefore slows the progression of AR by allowing a proportion of worms to escape drug selection pressure. The objective of this study was to observe the effect of a refugia-based strategy on body weight (BW), average daily gain (ADG) and fecal egg counts (FEC) of trichostongyle-type nematodes in naturally infected beef calves over a 131-day grazing season when compared with a whole herd treatment strategy, using the same combination of drugs. Stocker calves (n = 160) were ranked by body weight within sex then allocated to 16 paddocks, which were randomly assigned to one of two treatment groups. All calves in Group 1 (n = 80) were administered treatment, while in Group 2 (n = 80) the steer with the highest FEC in eggs per gram (EPG) within the paddock was left untreated. Treated calves received an extended release injectable 5 % eprinomectin (LongRange®, Boehringer Ingelheim Animal Health USA Inc.; 1 mL/50 kg of BW) and a 22.5 % oxfendazole oral suspension (Synanthic®, Boehringer Ingelheim Animal Health USA Inc.; 1 mL/50 kg of BW). Fecal egg counts and BW were recorded on days (D) -35, 0, 21, 131, and 148 to calculate the average fecal egg count reduction (FECR) and ADG for both groups. Linear mixed models, with paddock as the experimental unit, were used for analyses. The EPG differed on D21 (p < 0.01) and D131 (p = 0.057) with Group 2 having a higher average FEC (15.2 EPG D21; 57 EPG D131) compared with Group 1 (0.4 EPG D21; 37.25 EPG D131). However, there was no significant difference in average BW or ADG between treatment groups throughout the study. Results suggest refugia-based strategies could be implemented without significant negative impacts on average BW and ADG across other calves in the herd.

Increased dietary vitamin D3 and calcium partially alleviate heat stress symptoms and inflammation in lactating Holstein cows independent of dietary concentrations of vitamin E and selenium.

Twelve multiparous Holstein cows (42.2 ± 5.6 kg of milk/d; 83 ± 27 d in milk) were used in a split-plot design testing the effects of mineral and vitamin supplementation on the time course of animal performance, metabolism, and inflammation markers during heat stress. The main plot was the average concentrations of dietary vitamin E and Se (adequate: 11.1 IU/kg of vitamin E and 0.55 mg/kg of Se, and high: 223 IU/kg of vitamin E and 1.8 mg/kg of Se, respectively). Within each plot, cows were randomly assigned to (1) heat stress (HS) with adequate concentrations of vitamin D3 and Ca (1,012 IU/kg and 0.73%, respectively), (2) HS with high concentrations of vitamin D3 and Ca (HS+D3/Ca; 3,764 IU/kg and 0.97%, respectively), or (3) pair-feeding (PF) in thermoneutrality with adequate concentrations of vitamin D3 and Ca (1,012 IU/kg and 0.73% Ca) in a Latin square design with 14-d periods and 7-d washouts. The highest rectal temperature was recorded at 1700 h for HS (39.4°C; mean of d 1 to 14), being 1.2 and 0.8°C greater than for PF and HS+D3/Ca, respectively. Respiratory rate and water intake were higher in HS (73 breaths/min and 115 L/d, respectively) relative to PF (28 breaths/min and 76 L/d). Heat stress decreased dry matter intake progressively, reaching a nadir on d 5 to 7 (33% reduction) and was not different between treatments. Milk yield decreased progressively in all treatments, but remained greater in PF relative to HS from d 3 to 14 (10%), whereas HS and HS+D3/Ca were not different. Milk fat, protein, and lactose concentrations and yields were lower in HS relative to PF from d 3 to 14, but not different between HS and HS+D3/Ca. Relative to PF, preprandial insulin concentrations were increased in HS, whereas plasma nonesterified fatty acids were decreased on d 7 and 14. Plasma lipopolysaccharide-binding protein concentrations increased in HS cows on d 7 and 14, respectively, relative to PF, whereas they were reduced in HS + D3/Ca on d 14. Plasma C-reactive protein, tumor necrosis factor-α, and fecal calprotectin were increased in HS relative to both PF and HS+D3/Ca on d 7 and 14. Rectal temperature was positively associated with plasma lipopolysaccharide-binding protein (r = 0.72), tumor necrosis factor-α (r = 0.74), C-reactive protein (r = 0.87), and with milk somatic cells (r = 0.75). Plasma 8-hydroxy-2-deoxyguanosine concentrations presented a 3-way interaction, where 8-hydroxy-2-deoxyguanosine was lower in HS than in PF on d 7 and 14, and lower in HS+D3/Ca relative to HS on d 14 in the adequate vitamin E and Se treatment, but no effects were observed in the high vitamin E and Se group. Plasma superoxide dismutase concentrations increased over time, and were higher in HS relative to PF on d 14, whereas HS+D3/Ca was similar to HS. Heat stress markedly reduced milk production and milk components while increasing markers of leaky gut and inflammation. In contrast, vitamin D3 and Ca supplementation reduced hyperthermia (d 7-14), markers of leaky gut, and inflammation independent of dietary concentrations of vitamin E and Se.

The effect of neonatal vaccination for bovine respiratory disease in the face of a dual challenge with bovine viral diarrhea virus and Mannheimia hemolytica.

Bovine respiratory disease is the greatest threat to calf health. In this study, colostrum-fed dairy X beef calves were vaccinated at ∼30 days of age with an adjuvanted parenteral vaccine containing modified live bovine viral diarrhea virus (BVDV) type 1 and type 2, bovine herpesvirus 1 (BHV-1), bovine parainfluenza type 3 virus (PI3V) and bovine respiratory syncytial virus (BRSV) andM. haemolyticatoxoid (Group 1), or intranasal temperature-sensitive BHV-1, BRSV and PI3V concurrently witha parenteral vaccine containing modified live BVDV type 1 and type 2 andM. haemolyticatoxoid (Group 2) or a placebo (Group 3). The calves were challenged ∼150 days post vaccination intranasally with BVDV 1b and then 7 days later intratracheally withM. haemolytica. The calves wereeuthanized 6 days after theM. haemolyticachallenge. Clinical signs following BVDV infection were similar in all groups. There was increased rectal temperatures in the Groups 2 and 3 on day 3 and in Group 3 on days 8-13. Group 1 animals had a slight leukopenia following BVDV infection while Groups 2 and 3 had greater leukopenia. BVDV type 1 and 2 serum titers increased in Group 1 following vaccination while these titers waned in Groups 2 and 3. There were higher levels of BVDV in the buffy coats and nasal samples in Group 2 and Group 3 versus Group 1 (p < 0.01). Interferon-gamma response was higher (p < 0.01) in Group 1 animals than Groups 2 and 3. Group 1 had the lowest percent pneumonic tissue (1.6%) while Group 2 vaccinates had 3.7% and the control Group 3 was 5.3%. Vaccination in the face of maternal antibody with a parenteral adjuvanted vaccine resulted in better protection than the regimen of an intranasal vaccine anda parenteral adjuvanted BVDV andM haemolyticacombination vaccine in a BVDV-M. haemolyticadual challenge.

Immunization with a small fragment of the Schmallenberg virus nucleoprotein highly conserved across the Orthobunyaviruses of the Simbu serogroup reduces viremia in SBV challenged IFNAR-/- mice.

Schmallenberg Virus (SBV), an arbovirus from the Peribunyaviridae family and Orthobunyavirus genus, was discovered in late 2011 in Germany and has been circulating in Europe, Asia and Africa ever since. The virus causes a disease associated with ruminants that includes fever, fetal malformation, drop in milk production, diarrhoea and stillbirths, becoming a burden for small and large farms. Building on previous studies on SBV nucleoprotein (SBV-N) as a promising vaccine candidate, we have investigated the possible protein regions responsible for protection. Based on selective truncation of domains designed from the available crystal structure of the SBV-N, we identified both the N-terminal domain (N-term; Met1 - Thr133) and a smaller fragment within (C4; Met1 - Ala58) as vaccine prototypes. Two injections of the N-term and C4 polypeptides protected mice knockout for type I interferon (IFN) receptors (IFNAR-/-) challenged with virulent SBV, opposite to control groups that presented severe signs of morbidity and weight loss. Viremia analyses along with the presence of IFN-γ secreted from splenocytes re-stimulated with the N-terminal region of the protein corroborate that these two portions of SBV-N can be employed as subunit vaccines. Apart from both proteinaceous fragments being easily produced in bacterial cells, the C4 polypeptide shares a high sequence homology (∼87.1 %) with the corresponding region of nucleoproteins of several viruses of the Simbu serogroup, a group of Orthobunyaviruses that comprises SBV and veterinary pathogens like Akabane virus and human infecting viruses like Oropouche. Thus, we propose that this smaller fragment is better suited for vaccine nanoparticle formulation, and it paves the way to further research with other related Orthobunyaviruses.

In Vitro Studies of Nanoparticles as a Potentially New Antimicrobial Agent for the Prevention and Treatment of Lameness and Digital Dermatitis in Cattle.

Digital dermatitis (DD) is the second most prevalent disease in dairy cattle. It causes significant losses for dairy breeders and negatively impacts cows' welfare and milk yield. Despite this, its etiology has not been entirely identified, and available data are limited. Antibiotic therapy is a practical method for managing animal health, but overuse has caused the evolution of antibiotic-resistant bacteria, leading to a loss in antimicrobial efficacy. The antimicrobial properties of metal nanoparticles (NPs) may be a potential alternative to antibiotics. The aim of this study was to determine the biocidal properties of AgNPs, CuNPs, AuNPs, PtNPs, FeNPs, and their nanocomposites against pathogens isolated from cows suffering from hoof diseases, especially DD. The isolated pathogens included Sphingomonas paucimobilis, Ochrobactrum intermedium I, Ochrobactrum intermedium II, Ochrobactrum gallinifaecis, and Actinomyces odontolyticus. Cultures were prepared in aerobic and anaerobic environments. The viability of the pathogens was then determined after applying nanoparticles at various concentrations. The in vitro experiment showed that AgNPs and CuNPs, and their complexes, had the highest biocidal effect on pathogens. The NPs' biocidal properties and their synergistic effects were confirmed, which may forecast their use in the future treatment and the prevention of lameness in cows, especially DD.

Efficiency of immunocastration with an anti-gonadotropin-releasing hormone vaccine on cryptorchid bulls.

Cryptorchid bulls have low economic value owing to the effects of masculinization. Moreover, surgical removal of an ectopic testis is difficult in certain clinical cases. Recently, immunocastration has garnered popularity as a nonsurgical castration method in pig farming; however, the effects of immunocastration on cryptorchid bulls are yet to be yet. Herein, we investigated endocrine changes due to immunocastration in cryptorchid bulls and studied its effectiveness. This study included 13 Holstein bulls diagnosed with cryptorchidism and classified into two groups based on pubertal period: <8 months of age (pregroup) and ≥8 months of age (postgroup). Antigonadotropin-releasing hormone (GnRH) vaccine was used for immunocastration, and two vaccine doses were administered. Blood testosterone and anti-Müllerian hormone (AMH) levels were measured and analyzed for endocrine evaluation. The testosterone levels significantly decreased following the start of immunocastration in both groups, thereby confirming the efficacy of antiGnRH vaccination in cryptorchid bulls. The AMH levels significantly increased in the pregroup with two antiGnRH vaccination, suggesting a compensatory response via the neutralization of GnRH antibodies. The AMH levels did not significantly change in the postgroup, indicating the partial suppression of AMH secretion in Sertoli cells during sexual maturation and failure of Sertoli cell maturation. Thus, we successfully restrained the serum testosterone levels in cryptorchid bulls using antiGnRH vaccine. The testosterone levels are a useful indicator of the immunocastration effect on cryptorchid bulls. Hereafter, a vaccine program that can sustain the castration effect on cryptorchid bulls is necessary.

Field efficacy of Metarhizium anisopliae oil formulations against Rhipicephalus microplus ticks using a cattle spray race.

Rhipicephalus microplus tick is the main ectoparasite of cattle in Brazil. The exhaustive use of chemical acaricides to control this tick has favored the selection of resistant tick populations. Entomopathogenic fungi, as Metarhizium anisopliae, has been described as a potential biocontroller of ticks. Therefore, the aim of this study was to evaluate the in vivo efficacy of two oil based formulations of M. anisopliae for the control of the cattle tick R. microplus under field conditions using a cattle spray race as a method of treatment. Initially, in vitro assays were carried out with an aqueous suspension of M. anisopliae, using mineral oil and/or silicon oil. A potential synergism between oils and fungus conidia for tick control was demonstrated. Additionally, the usefulness of silicon oil in order to reduce mineral oil concentration, while improving formulation efficacy was illustrated. Based on the in vitro results, two formulations were selected for use in the field trial: MaO1 (107 conidia/mL plus 5% mineral oil) and MaO2 (107 conidia/mL plus 2.5% mineral oil and 0.01% silicon oil). The adjuvants concentrations (mineral and silicon oils) were chosen since preliminary data indicate that higher concentrations caused significant mortality in adult ticks. For this, 30 naturally infested heifers were divided into three groups based on previous tick counts. The control group did not receive treatment. The selected formulations were applied on animals using a cattle spray race. Subsequently, tick load was evaluated weekly by counting. The MaO1 treatment significantly reduced the tick count only on day +21, reaching approximately 55% efficacy. On the other hand, MaO2 showed significantly lower tick counts on days +7, +14, and +21 after treatment, with weekly efficacy achieving 66%. The results showed a substantial reduction of tick infestation, up to day +28, using a novel formulation of M. anisopliae based in the mixture of two oils. Moreover, we have shown, for the first time, the feasibility of employing formulations of M. anisopliae for large-scale treatment methods, such as a cattle spray race, which in turn, may increase the use and adhesion to biological control tools among farmers.

Effect of oestrus synchronisation through ovulation delay by vaccination against foot-and-mouth disease in Hanwoo (Bos taurus coreanae) cows.

BACKGROUND: In Korean cattle, after foot-and-mouth disease (FMD) vaccination, anovulation increases, acute immune response is stimulated. OBJECTIVE: Here, we aimed to improve the fertility rate by ovulation delay caused by the foot-and-mouth disease vaccine. METHODS: 160 cows (control, FMD, FMD+Gn250 and FMD+Gn500 groups, with 40 cows each) were used. We analysed the ovulation delay, ovulation rate, conception rate and acute-phase immune responses. RESULTS: In the group vaccinated only with FMD, the average follicle size was maintained at 12 mm and ovulation was delayed. The ovulation rate of the FMD+Gn500 group (500 µg gonadotropin-releasing hormone (GnRH) injections 3 days after the FMD vaccination) was the highest at 81.8%. The ovulation rate of the FMD+Gn250 group (250 µg GnRH injections 3 days after FMD vaccination) was 54.5%, and that of the control group (not FMD vaccinated) was 53.3%. The conception rate was 52.5% (19/40) in the control group, 37.5% (15/40) in the FMD+Gn250 group, and 67.5% (27/40) in the FMD+Gn500 group. Analysis of acute-phase immune response revealed that the plasma contents of haptoglobin and serum amyloid A increased up to 7 days after vaccination against FMD in all the experimental groups, except the control group. CONCLUSIONS: We concluded that ovulation delay can be employed to improve conception rate after FMD vaccination through a modified ovulation synchronisation method with GnRH.

Farm-level risk factors for digital dermatitis in dairy cows in mountainous regions.

Reduction of risk factors for bovine digital dermatitis (BDD) is crucial in current disease control. However, risk factors that might arise especially in mountainous regions are unknown until now, and an adapted BDD control program is consequently missing. The objective of this observational case-control study was to identify farm-level risk factors for BDD in dairy herds in mountainous regions. To investigate predictors for the occurrence of BDD, 100 farms were visited and information about herd characteristics and management practices, potentially relevant explanatory variables for either introduction or establishment of BDD, were gathered by completing a questionnaire with the farmer or herd manager. Within-herd prevalences of BDD assessed during 3 routine claw trimmings with an interval of 6 mo before the survey were used to define cases (BDD within-herd prevalence of ≥26% during each claw trimming) and controls (no BDD cases in each of the 3 routine claw trimmings before the survey). Data were analyzed using 2 separate binomial generalized linear models according to either establishment or introduction of BDD. After prescreening, 15 of 23 explanatory variables were included in the final analysis, which showed 3 variables related to introduction and establishment, each being significantly associated with the occurrence of BDD within a farm. Results of model 1 (i.e., aspects related to BDD introduction) revealed that access to mountain pastures during the summer season (odds ratio, 95% confidence interval: 0.12, 0.04-0.35), participation in dairy shows (0.32, 0.11-0.94), and the number of new animals introduced into the farm during the last 2 yr (1.28, 1.12-1.52) were significantly associated with the occurrence of BDD. Model 2 (i.e., aspects related to BDD establishment) showed that cows kept in freestalls were at higher risk for BDD compared with those kept in tiestalls (20.65, 1.59-649.37). Furthermore, number of days between diagnosis and treatment of a BDD lesion (10.31, 3.55-81.21) and the amount of concentrate feeding (median 5 kg) per cow and day (7.72, 2.46-6.47) were positively associated with BDD occurrence. In conclusion, the findings of this study provide a set of risk factors that are associated with BDD status within herds in mountainous regions. These results may help in development of adapted control programs for BDD in dairy cows.

Comparison of chlorhexidine and alcohol-based antisepsis on the paralumbar fossa in cattle.

OBJECTIVE: To determine skin reaction, post-treatment reduction (immediate effect), and 1 hour post-treatment reduction (sustained effect) of aerobic bacterial colony forming units (CFU) following three antiseptic protocols in cattle. STUDY DESIGN: Prospective, randomized experimental study. ANIMALS: Eighteen cows. METHODS: Three sites in each paralumbar fossa were clipped and randomly assigned to one of three treatment groups: 5 minute 4% chlorhexidine gluconate scrub (CHG); 90 second 80% ethanol scrub (ET); 90 second 70% isopropyl alcohol scrub (IPA). All sites were monitored at all sampling time points and at 24 hours following treatment for adverse skin reaction. Samples were collected pre-, immediately post-, and 1 hour post-treatment and plated in duplicate. Bacterial counts were shifted to eliminate zeroes, log10 transformed, and averaged. ANOVA was used to compare differences in mean reduction in log10 CFU/ml between groups. RESULTS: Reduction in log10CFU/ml was more pronounced immediately after application of IPA (p = .001) and ET (p = .001) than CHG. This reduction was better sustained after preparation with CHG than ET (p = .005) but not IPA. Immediate and sustained reductions in bacterial loads did not differ after application of IPA or ET. No adverse skin reactions were noted. CONCLUSIONS: Skin preparation with alcohol-based antiseptics was well tolerated and improved immediate bacterial reduction compared to CHG. This reduction was better sustained 1 hour after application of CHG than ET, but no difference was detected between CHG and IPA. CLINICAL RELEVANCE: Lack of adverse skin reaction and performance provide evidence to support skin preparation with alcohol-based antiseptics in cattle.

Efficacy of essential oil of Lippia sidoides (Verbenaceae) for controlling the cattle tick Rhipicephalus (Boophilus) microplus on naturally parasitized animals under field conditions.

Efficacy of the essential oil of Lippia sidoides to control the bovine tick Rhipicephalus (Boophilus) microplus was evaluated in tests on naturally infested cattle under field conditions. Twenty cattle were allocated to one of two groups (Control and Treated). Animals in the treated group were sprayed with three liters of 1 % L. sidoides solution (10,000 ppm) and efficacy was calculated from teleogynous counts between 4.5 and 8 mm in length before (days -3, - 2 and - 1) and after treatment (days +3, +7, +14, +21 and +28). The essential oil of L. sidoides showed an average efficacy of 50.7 % between days + 3 and + 21, with greater efficacy in the larvae (63.2 % - p = 0.0001).

The effect of pre-calving injection of trace mineral supplements on periparturient disease incidence in pasture based dairy cows.

Trace minerals, have a role in immune function and a trace mineral supplement (TMS) can improve animal health in dairy herds. This prospective randomised clinical study assessed whether subcutaneous injection of 5.5 mL of TMS (40 mg zinc, 10 mg manganese, 5 mg selenium, 15 mg copper per mL), 14-28 days before planned start of calving (PSC) reduced clinical mastitis (CM), subclinical mastitis (SCM) and purulent vaginal discharge (PVD). From four farms, half of 1700 cows stratified on somatic cell count, age and breed were randomly allocated to treatment or no treatment. Occurrence of CM from - 7 to PSC + 100 days, SCM at PSC + 60 days and PVD at PSC + 24 days was analysed using survival analysis and Bayesian generalised mixed multivariable models. From -7 to PSC +30 days, TMS reduced the adjusted hazard ratio (HR) for CM at quarter and cow level (P < 0.001), with no evidence for an effect beyond 30 days. The adjusted OR (and 95% highest density interval, HDI) for the effect of TMS on CM from -7 to PSC +30 days was 0.40 (95% HDI, 0.26-0.63) at quarter level, 0.51 (95% HDI, 0.38-0.69) at cow level and for SCM, 0.72 (95% HDI, 0.54-0.95). The difference in CM incidence from TMS at the cow level was -2.0% (95% HDI, -3.4 to -1.1%) and -1.2% (95% HDI, -3.2 to - 0.6%) at quarter level. No clear effect was identified of TMS on cumulative incidence of PVD.