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Ticks and tick-borne diseases cause substantial economic losses to the livestock industry in sub-Saharan Africa. Mazao Tickoff is a novel bioacaricide developed for tick control and is based on the entomopathogenic fungus Metarhizium anisopliae sensu lato (s.l.) isolate ICIPE 7. To date, no randomized controlled study has been undertaken to demonstrate the efficacy of this bioacaricide in reducing natural tick infestation on cattle. To this end, this field trial is designed to evaluate the anti-tick efficacy of Mazao Tickoff on cattle in coastal Kenya compared to a standard chemical tick control protocol. In this prospective, multi-center randomized controlled trial, eligible herds will be randomized by the herd size to the intervention arm in a 1:1:1 ratio to either Triatix ® (active ingredient: amitraz); Mazao Tickoff (active ingredient: M . anisopliae ICIPE 7); or placebo (excipients of the Mazao Tickoff), with a total enrollment target of 1,077 cattle. Treatments will be dispensed on Day 0 (defined individually as the day each animal receives the first treatment) and thereafter every two weeks until Day 182. Ticks will be counted on every animal in each herd (herds to be included have at least one animal bearing at least one tick on Day 0), and thereafter on bi-weekly intervals until Day 182. The primary efficacy assessments of Mazao Tickoff will be based on the mean percentage reduction in tick counts at each post-treatment follow-up visit compared to the placebo group and the Triatix ® arm. Further, the effect of Mazao Tickoff on the prevalence of common cattle pathogens, Anaplasma marginale and Theileria parva , will be determined by assessing incidence and seroprevalence at four different time points. This protocol describes the first rigorous evaluation of the efficacy of Mazao Tickoff and its potential as a viable alternative non-chemical acaricide tool for tick control in Kenya and elsewhere.

In veterinary practice pain alleviation plays a part in managing lameness. The aim of this randomized and placebo-controlled clinical study was to evaluate the effect of a single administration of ketoprofen on locomotion characteristics and weight distribution in cattle with foot (located up to and including the fetlock; n = 31) and (proximal to the fetlock; n = 10) pathologies. Cattle were randomly allocated to either the ketoprofen (group K; intravenous 3 mg/kg of body weight; n = 21) or an equivalent volume of isotonic sterile saline solution (group P; n = 20). Two accelerometers (400 Hz; kinematic outcome = stance phase duration; kinetic outcome = foot load and toe-off), a 4-scale weighing platform (weight distribution and SD of the weight) and a subjective locomotion score were measured before (baseline) and after 1 h and 18 h of treatment. All variables were expressed as differences across contralateral limbs, and the measurements at 1 h and 18 h were compared to the baseline. A repeated measures ANOVA was used to determine the differences between groups K and P. A logistic regression model with a binary outcome (0 = no improvement and 1 = improvement of the differences across the contralateral limbs over time) was calculated. Mean (± SD) of locomotion scores at baseline were not significantly different (P = 0.102) in group K (3.10 ± 0.80) as compared to group P (3.48 ± 0.64). Cattle of group K showed significantly lower differences across contralateral limbs at 1 h as compared to group P for the relative stance phase and the weight distribution. Only the treatment (P versus K) remained a significant factor in the model for relative stance phase (odds ratio (OR) = 6.5; 95% CI = 1.38-30.68) and weight distribution (OR = 6.36; 95% CI = 1.30-31.07). The effects of ketoprofen were evident in improving the differences across contralateral limbs-both for stance phase during walking and weight bearing during standing-after 1 h but not after 18 h of administration.

More than 80% of schistosomiasis patients in China live in the lake and marshland regions. The purpose of our study is to assess the effect of a comprehensive strategy to control transmission of Schistosoma japonicum in marshland regions. In a cluster randomized controlled trial, we implemented an integrated control strategy in twelve villages from 2009 through 2011 in Gong'an County, Hubei Province. The routine interventions included praziquantel chemotherapy and controlling snails, and were implemented in all villages. New interventions, mainly consisting of building fences to limit the grazing area for bovines, building safe pastures for grazing, improving the residents' health conditions and facilities, were only implemented in six intervention villages. Results showed that the rate of S. japonicum infection in humans, bovines, snails, cow dung and mice in the intervention group decreased from 3.41% in 2008 to 0.81% in 2011, 3.3% to none, 11 of 6,219 to none, 3.9% to none and 31.7% to 1.7%, respectively (P<0.001 for all comparisons). In contrast, there were no statistically significant reductions of S. japonicum infection in humans, bovines and snails from 2008 to 2011 in the control group (P>0.05 for all comparisons). Moreover, a generalized linear model showed that there was a higher infection risk in humans in the control group than in the intervention group (OR = 1.250, P = 0.001) and an overall significant downward trend in infection risk during the study period. The integrated control strategy, designed to reduce the role of bovines and humans as sources of S. japonicum infection, was highly effective in controlling the transmission of S. japonicum in marshland regions in China. Chinese Clinical Trial Registry ChiCTR-PRC-12002405.

In this multicentric, randomised, blinded and placebo-controlled field study, the effect of treatment with toltrazuril (Baycox® Bovis, Bayer) on oocyst excretion, diarrhoea score and weight gain was studied in Danish dairy herds with confirmed history of eimeriosis (coccidiosis) and prevalence of Eimeria bovis and Eimeria zuernii. Three commercial herds and a total of 71 calves, aged 48 – 135 days, were included. Treatment with a single oral dose of toltrazuril (15 mg/kg) was given after relocation to common pens and one week before expected outbreak of eimeriosis. The effect of treatment was followed by weekly faecal sampling and weighing initially and at the end of a study period of 8 weeks. In Herd 2 and 3 toltrazuril treated calves gained on average 7.95 kg more than placebo treated calves (p = 0.007), and both oocyst excretion and prevalence of Eimeria spp. were significantly reduced the first weeks post treatment. In Herd 1, by contrast, the farmer made some unforeseen changes in the management which entailed relocation to large deep-litter pens 3 – 6 weeks post treatment. In addition, many calves were not treated metaphylactically while few calves excreted oocysts when the trial was initiated. Thus, no significant difference in weight gain was found between toltrazuril and placebo treated calves (p = 0.523), and the oocyst excretion of toltrazuril treated calves was significantly higher during week 7 and 8. Significant differences in faecal scores were observed between the herds (p<0.002) but not between treatment groups in any of the herds. In conclusion, timing of treatment is crucial for optimal effect of metaphylactic toltrazuril treatment on weight gain and oocyst excretion.

This study investigated differences in uterine and serum metabolome associated with clinical cure failure of metritis in dairy cows. Metritis was diagnosed in lactating Holstein cows from two Florida dairies and defined by the presence of fetid, watery, reddish-brown vaginal discharge from 4 to 12 days postpartum (dpp). Cows with metritis (n = 24) were paired with cows without metritis of similar parity and dpp (n = 24). On the day of metritis diagnosis (day 0), all cows with metritis received antimicrobial therapy. The continued presence of the fetid, watery, reddish-brown discharge on day 5 (n = 16) was defined as clinical cure failure, whereas clinical cure was defined by its absence (n = 8). Metabolome analyses of uterine lavage (days 0 and 5) and serum samples (day 0) were conducted using untargeted gas chromatography time-of-flight mass spectrometry. Normalized data were analyzed using partial least squares–discriminant analysis and ANOVA, adjusting P-values for multiple comparisons. Differences in the uterine metabolome on day 0 associated with clinical cure failure were linked to carbohydrate, amino acid, and lipid metabolism. Greater concentrations of arachidonic acid, ribose, and glutaric acid were associated with clinical cure failure, suggesting a greater degree of tissue lesion and inflammation. No differences in the serum metabolome were associated with cure failure. No differences in uterine metabolome were associated with clinical cure failure on day 5. The findings suggest that clinical cure failure is associated with a greater uterine inflammatory process that did not persist until cure assessment day. Summary Sentence Clinical cure failure in dairy cows with metritis is linked to distinct uterine metabolome profiles, inflammatory processes, and energy metabolism present on the day of metritis diagnosis but not on cure assessment day. Graphical Abstract

Buparvaquone (BPQ) is used for the treatment of bovine theileriosis, a tickborne disease caused by parasites of the Theileria genus. Studies on T. annulata have linked the mechanism of BPQ resistance predominantly to genetic variations in the parasite cytochrome b ( cytb ) gene. In addition, cryptic mechanisms of resistance involving the parasite peptidyl-prolyl isomerase ( pin1 ) and dihydroorotate dehydrogenase ( dhodh ) genes require assessment. In India, where bovine theileriosis is endemic, and BPQ is widely used for treatment, it is necessary to establish the prevalence of genetic variations linked to BPQ resistance. In this study, multiplexed PCR amplification and nanopore sequencing method was used for genotyping the complete gene loci of the three target genes. Analysis of 454 T. annulata field samples collected from seven different states of India revealed the presence of previously reported BPQ resistance associated variations S129G, A146T and P253S in cytb gene and A53P in pin1 gene. The A146T and I203V variations in cytb were found to be prevalent and mostly co-occurring, and their role in BPQ resistance needs further evaluation. This study has revealed the presence of previously reported BPQ resistance-linked mutations in cytb and pin1 genes in T. annulata infecting dairy cattle in India and establishes an Oxford nanopore sequencing method suitable for large-scale surveillance of genetic variation in Theileria parasites from field samples.

Mastitis is the most prevalent disease in dairy cattle worldwide and not only causes huge economic losses in the dairy industry but also threatens public health. To evaluate the therapeutic potential of melatonin in mastitis, we examined the ability of melatonin to protect bovine mammary epithelial cells (bMECs) from the harmful effects of lipopolysaccharide (LPS). We found that melatonin inhibited the LPS-binding protein-CD14-TLR4 signaling pathway in bMECs, which had opposing effects on pro-inflammatory and anti-inflammatory mediators. Melatonin decreased LPS-induced expression of pro-inflammatory cytokines, chemokines, and positive acute-phase proteins (APPs), including tumor necrosis factor-α, interleukin (IL)-1β, IL-6, granulocyte-monocyte colony-stimulating factor, chemokine CC motif ligand (CCL)2, CCL5, serum amyloid A, haptoglobin, C-reactive protein, ceruloplasmin, and α-1 antitrypsin, and increased expression of the anti-inflammatory cytokine IL-1Ra and the negative APP fibrinogen. In addition, melatonin increased dityrosine levels but suppressed nitrite levels by upregulating the expression of Nrf2 and heme oxygenase-1 in the Nrf2 antioxidant defense pathway. Finally, melatonin administration increased the viability of LPS-stimulated bMECs. These results suggest that melatonin protects bMECs from LPS-induced inflammatory and oxidant stress damage and provide evidence that melatonin might have therapeutic utility in mastitis.

Salmonella Dublin is a serovar that causes severe infections and cattle. Despite the importance of this agent, research on achieving its elimination from dairy farms is limited, which complicates risk mitigation and control efforts. This study thus aimed to assess the prevalence of S . Dublin on a farm with a history of outbreaks, to understand the dynamics of the infection, characterize the antimicrobial resistance of the isolates, and evaluate their genetic similarity. Multiparous cows in the postpartum phase are nearly five times more likely to shed Salmonella sp. A total of 39 cases of fatal septicemic salmonellosis caused by S . Dublin were confirmed in calves aged 3–5 months. Antimicrobial susceptibility was evaluated in 45 strains of S . Dublin, with 48.9% of the isolates classified as multidrug resistant, including resistance to penicillin (48.9%), tetracyclines (42.2%), and fluoroquinolones (33.3%). Seven multidrug-resistant isolates were selected for genomic sequencing. Among the resistance determinants identified, a mutation in the gyrA gene, present in all sequenced isolates, was notable. Analyses of cgMLST and SNPs revealed that the isolates from healthy animals were closely related to those found in animals with confirmed cases of S . Dublin, confirming that the agent was circulating among healthy animals across various categories. A high similarity was also found between the isolates in this study and strains causing salmonellosis in humans in Brazil, thus reinforcing the zoonotic nature and possible epidemiological link between cattle, and the occurrence of this disease in humans.

Bovine mastitis (BM) is a disease with high incidence worldwide and one of the most relevant bovine pathologies and the most costly to the dairy industry. BM is an inflammation of the udder and represents one of the most difficult veterinary diseases to control. Biofilm formation is considered a selective advantage for pathogens causing mastitis, facilitating bacterial persistence in the udder. In fact, recently some authors drew attention to the biofilm formation ability presented by several mastitis causing pathogens and to its possible relation with recurrent mastitis infections and with the increased resistance to antimicrobial agents and host immune defence system. Actually, up to now, several researchers reported the potential role of cells in this mode of growth in the previous facts mentioned. As a consequence of the presence of biofilms, the infection here focused is more difficult to treat and eradicate, making this problem a more relevant pressing issue. Thus, we believe that a deeper knowledge of these structures in mastitis can help to determine the best control strategy to be used in veterinary practice in order to reduce losses in the dairy industry and to ensure milk safety and quality. The aim of this paper was to review the existing research and consequently to provide an overview of the role of biofilms in BM infections. This review provides an important input of knowledge on the relevance of biofilm formation in bovine mastitis infections and their role in antimicrobial resistance. Graphical Abstract Figure. This review provides an important input of knowledge on the relevance of biofilm formation in bovine mastitis infections and their role in antimicrobial resistance.

Abstract Background Acute infectious bronchopneumonia is widely reported in adult cattle, yet most descriptions focus on postmortem findings. Hypothesis/Objectives To describe signalment, clinical, hematological, and ultrasonographic findings in affected cows, and to identify pathogens and clinical findings associated with outcome. Animals Twenty-one cows from 11 farms. Methods Prospective case series. All cows underwent clinical examination, thoracic ultrasonography, bronchoalveolar lavage for culture and PCR, and CBC. Thoracic ultrasonography was repeated to assess lesion dynamics after antibiotic treatment. Outcomes were classified as favorable (clinical recovery and maintained milk production) or unfavorable (death or reduced milk production leading to premature drying-off and early culling). Descriptive statistics were computed, and univariate analyses were performed to identify variables associated with outcome. Results Mean age at inclusion was 47 months (range, 24-104). Cases occurred mainly in winter (10/21) and during the early postpartum period (10/21); 17/21 were dairy cows. Typical clinical signs associated with acute respiratory disease (coughing, wheezes, and crackles) were rarely observed. Thoracic ultrasonography revealed frequent consolidations (17/21) and pleural effusion (13/21), with lesion size decreasing over time. Mannheimia haemolytica and Pasteurella multocida were identified by PCR in bronchoalveolar lavage samples in 13/21 and 18/21 cases, respectively. Unfavorable outcomes (11/21) were associated with nasal discharge, left shift on CBC, reduced ruminal contractions, higher respiratory rate on day 0, and deeper consolidation lesions on days 0 and 5. Conclusions and clinical importance Ultrasonography is a valuable cow-side tool that supports diagnosis and provides key information on the clinical course of acute infectious bronchopneumonia in adult cows.