Design of a multi-epitope-based vaccine candidate against Bovine Genital Campylobacteriosis using a reverse vaccinology approach.

BACKGROUND: Bovine Genital Campylobacteriosis (BGC), a worldwide distributed venereal disease caused by Campylobacter fetus subsp. venerealis (Cfv), has a relevant negative economic impact in cattle herds. The control of BGC is hampered by the inexistence of globally available effective vaccines. The present in silico study aimed to develop a multi-epitope vaccine candidate against Cfv through reverse vaccinology. RESULTS: The analysis of Cfv strain NCTC 10354 proteome allowed the identification of 9 proteins suitable for vaccine development. From these, an outer membrane protein, OmpA, and a flagellar protein, FliK, were selected for prediction of B-cell and T-cell epitopes. The top-ranked epitopes conservancy was assessed in 31 Cfv strains. The selected epitopes were integrated to form a multi-epitope fragment of 241 amino acids, which included 2 epitopes from OmpA and 13 epitopes from FliK linked by GPGPG linkers and connected to the cholera toxin subunit B by an EAAAK linker. The vaccine candidate was predicted to be antigenic, non-toxic, non-allergenic, and soluble upon overexpression. The protein structure was predicted and optimized, and the sequence was successfully cloned in silico into a plasmid vector. Additionally, immunological simulations demonstrated the vaccine candidate's ability to stimulate an immune response. CONCLUSIONS: This study developed a novel vaccine candidate suitable for further in vitro and in vivo experimental validation, which may become a useful tool for the control of BGC.

A comparison of risk factor investigation and experts' opinion elicitation analysis for identifying foot-and-mouth disease (FMD) high-risk areas within the FMD protection zone of South Africa (2007-2016).

Foot-and-mouth disease is a controlled disease in accordance with the South African Animal Diseases Act (Act 35 of 1984). The country was classified by the World Organisation for Animal Health (WOAH) as having a FMD free zone without vaccination in 1996. However, this status was suspended in 2019 due to a FMD outbreak outside the controlled zones. FMD control in South Africa includes animal movement restrictions placed on cloven-hoofed species and products, prophylactic vaccination of cattle, clinical surveillance of susceptible species, and disease control fencing to separate livestock from wildlife reservoirs. The objectives of this study were to evaluate differences in identifying high-risk areas for FMD using risk factor and expert opinion elicitation analysis. Differences in risk between FMD introduction and FMD spread within the FMD protection zone with vaccination (PZV) of South Africa (2007-2016) were also investigated. The study was conducted in the communal farming area of the FMD PZV, which is adjacent to wildlife reserves and characterised by individual faming units. Eleven risk factors that were considered important for FMD occurrence and spread were used to build a weighted linear combination (WLC) score based on risk factor data and expert opinion elicitation. A multivariable conditional logistic regression model was also used to calculate predicted probabilities of a FMD outbreak for all dip-tanks within the study area. Smoothed Bayesian kriged maps were generated for 11 individual risk factors, overall WLC scores for FMD occurrence and spread and for predicted probabilities of a FMD outbreak based on the conditional logistic regression model. Descriptively, vaccine matching was believed to have a great influence on both FMD occurrence and spread. Expert opinion suggested that FMD occurrence was influenced predominantly by proximity to game reserves and cattle density. Cattle populations and vaccination practices were considered most important for FMD spread. Highly effective cattle inspections were observed within areas that previously reported FMD outbreaks, indicating the importance of cattle inspection (surveillance) as a necessary element of FMD outbreak detection. The multivariable conditional logistic regression analysis, which was consistent with expert opinion elicitation; identified three factors including cattle population density (OR 3.87, 95% CI 1.47-10.21) and proximities to game reserve fences (OR 0.82, 95% CI 0.73-0.92) and rivers (OR 1.04, 95% CI 1.01-1.07) as significant factors for reported FMD outbreaks. Regaining and maintaining an FMD-free status without vaccination requires frequent monitoring of high-risk areas and designing targeted surveillance.

Supplementation of rumen-protected lysine during the close-up period improves vaginal discharge clearance in Holstein dairy cows.

We aimed to evaluate the effects of rumen-protected lysine (RPL) supplementation during the close-up period on uterine involution and the resumption of ovarian function in dairy cows. Fifty-two multiparous Holstein cows were categorized based on parity and expected calving date and randomly assigned to the RPL or control (CON) groups. The RPL group received 80 g of RPL daily from day 21 before the expected calving date until parturition. Blood samples were obtained twice weekly from pre-supplementation to 6 weeks postpartum. The onset of luteal activity postpartum was determined via ultrasonography twice weekly for up to 6 weeks postpartum. Uterine involution was tracked at 3 and 5 weeks postpartum through the vaginal discharge score, percentage of polymorphonuclear cells (PMN) in endometrial cytology samples, presence of intrauterine fluid, and gravid horn diameter via ultrasonography. Before supplementation, the RPL group showed amino acid imbalance, which was improved by RPL supplementation. There were no significant differences in the onset of luteal activity, percentage of PMN, intrauterine fluid, or the diameter of the uterine horn between the two groups. The vaginal discharge score in the RPL group decreased from 3 to 5 weeks postpartum, whereas that in the CON groups did not decrease. The number of cows with clinical endometritis was lower in the RPL group. Overall, RPL supplementation during the close-up period enhanced vaginal discharge clearance, potentially averting clinical endometritis, but did not affect the first ovulation in dairy cows.

Long-term outcomes of the Italian Mycobacterium avium subspecies paratuberculosis control programme for dairy cattle.

BACKGROUND: The considerable epidemiological and economic implications of paratuberculosis, caused by Mycobacterium avium subspecies paratuberculosis (MAP), have placed importance on control efforts aimed at preventing MAP transmission. In this context, Italy issued national guidelines for the control and status certification of MAP in dairy cattle in 2013. METHODS: We assessed the long-term outcomes of the Italian MAP control programme for 14 dairy farms located in northern Italy by retrospectively reviewing the results of yearly serological tests, presence of clinical cases, MAP faecal shedding in serologically positive animals, farm management and health ranking as indicators of herd health between 2014 and 2021. RESULTS: A significantly higher number of serologically positive animals were observed between 2014 and 2016 than between 2017 and 2021, as well as an improving trend in the paratuberculosis health ranking for nine of the 14 farms. No clinical cases were reported. MAP shedding was detected in 9.4% of serologically positive animals. Discarding colostrum and prioritised culling of seropositive animals assisted by adoption of standardised serological testing were presumed to have a key role in MAP control, despite the reluctance of some farmers to address hygienic issues and improve the separation of calves from adult animals. LIMITATIONS: The small number of farms included in this study and the fact that these were not randomly selected may limit the generalisability of the findings. CONCLUSIONS: The Italian paratuberculosis control plan has provided measures to limit the uncontrolled spread of MAP infection within and between herds by promoting animal trading between farms certified as negative or low risk.

Identification and distribution of Rhipicephalus microplus in selected high-cattle density districts in Uganda: signaling future demand for novel tick control approaches.

BACKGROUND: Rhipicephalus (Boophilus) microplus (Canestrini, 1888), the Asian blue tick, is a highly invasive and adaptable ectoparasite. This tick species has successfully established itself in most regions of the world, with movement of cattle being a major driver for its spread. In the recent past, R. microplus ticks have been reported in three districts of Uganda. Information on its spread and distribution are vital in deepening our understanding of the ecological scenarios that lead to tick persistence and in the formulation of control strategies. This is especially important in the cattle-dense districts. METHODS: We randomly collected tick specimens from 1,461cattle spread across seven cattle dense districts located in the Central, Karamoja and West Nile regions of Uganda from January to September 2020. The ticks were identified using standard morpho-taxonomic keys and the R. microplus tick species identities were confirmed by sequencing of the ITS2 region, 12S rRNA and 16S rRNA genes and phylogenetic analyses. RESULTS: Adult ticks (n = 13,019) were collected from 1,461 cattle. Seventeen tick species were identified based on morpho-taxonomic keys and the majority (47.4%; n=6184) of these were R. appendiculatus. In total, 257 R. microplus ticks were found infesting cattle in 18 study sites in the districts of Amudat, Kaabong, Napak (Karamoja region) and Arua (West Nile region). The identity of R. microplus was confirmed using molecular technics. No R. microplus tick was recorded in the districts of Lyantonde and Nakaseke (Central region). Arua district accounted for 82.1% (n=211) of the R. microplus ticks recorded followed by Napak district at 16.3% (n=42), while Amudat and Kaabong districts accounted for 1.5% (n=4). Rhipicephalus microplus and R. decoloratus co-existed in 6 of the 13 study sites in Arua district, while in another 6 study sites, no R. decoloratus was recorded. In the Karamoja region districts R. decoloratus co-existed with R.microplus. Of the total 618 ticks belonging to four species of the subgenus Boophilus recorded in this study, R. decoloratus accounted for 50.04% (n=334), followed by R. microplus at 41.58% (n=257), R. geigyi at 2.75% (n=17) and R. annulatus at 1.61% (n=10). In the districts of Amudat, Kaabong and Napak, R. decoloratus was more dominant (76.1%; n=179) of the three Rhipicephalus (Boophilus) tick species recorded, followed by R. microplus (19.5%; n=46) and R. geigyi (4.2%; n=10). Contrariwise, R. microplus was more dominant (84%; n=211) in Arua district followed by R. decoloratus (10.7%; n=27), R. annulatus (3.9%; n=10) and R. geigyi (1.1%; n=3). Phylogenetic analyses of the ITS2 region, 12S rRNA and 16S rRNA genes revealed subgrouping of the obtained sequences with the previously published R. microplus sequences from other parts of the world. CONCLUSION: Rhipicephalus microplus ticks were found infesting cattle in four districts of Uganda. The inability to find R. decoloratus, an indigenous tick, from six sites in the district of Arua is suggestive of its replacement by R. microplus. Rhipicephalus microplus negatively affects livestock production, and therefore, there is a need to determine its distribution and to deepen the understanding of the ecological factors that lead to its spread and persistence in an area.

Positive cases of bovine genital campylobacteriosis and bovine trichomonosis in the eradication program of La Pampa, Argentina: 2008-2021.

Bovine genital campylobacteriosis, caused by the gram-negative bacteria Campylobacter fetus venerealis, and bovine trichomonosis, caused by the parasite protozoan Tritrichomonas foetus, are venereal diseases that occur with long intercalving periods and abortion. The control of both diseases relies on microbiological testing and culling infected bulls. Vaccination and antibiotic treatment may help in controlling campylobacteriosis but are not recommended for trichomonosis control. Several regions of the world have active control programs for trichomonosis, not campylobacteriosis. In Argentina, the state of La Pampa aims to eradicate trichomonosis and campylobacteriosis by imposing annual diagnostic testing of every bull and slaughtering positive animals. Prior studies indicated a declining trend in the prevalence of campylobacteriosis and trichomonosis in La Pampa. It was also proposed that the prevalence of one disease could be estimated from the prevalence of the other. The purpose of this retrospective analysis of data gathered from 2008 to 2021 was to determine the La Pampa program's efficacy. Descriptive statistics were employed to determine the reason behind the correlation between tricomonosis and campylobacteriosis diagnostic results. The outcomes refute the notion that this program of venereal eradication was a success. Furthermore, an excess of false positives in both diagnoses may have contributed to the correlation between the prevalences of campylobactriosis and trichomonosis. The practice of killing animals without verifying positive results hinders the determination of disease prevalence and results in the death of numerous healthy animals.

Control of Rhipicephalus microplus on taurine cattle with fluralaner in a subtropical region.

BACKGROUND: In Rio Grande do Sul, Brazil, a region with a subtropical climate, Rhipicephalus microplus is present in taurine cattle raised for beef and milk. In addition, ticks resistant to multiple acaricides are present in this region. Recently, fluralaner (isoxazoline) was launched on the market. Thus, there is a need to evaluate the effects of fluralaner for the control of R. microplus on taurine cattle. In addition, occurrence of myiasis by Cochliomyia hominivorax larvae after tick parasitism and weight gain of cattle during the experimental period were evaluated. METHODS: Thirty naturally infested cattle were divided into two experimental groups: T01, treated with fluralaner (2.5 mg/kg) pour-on; T02, control. T01 received fluralaner on Days 0 (early summer in January), 42 and 84 (early autumn), whereas T02, a control group, received palliative treatment with a spray formulation when the group mean was ≥ 30 ticks. Counts of R. microplus females and calculation of the efficacy of fluralaner were performed on Days 3, 7, 14, 28, 35, 42, 56, 70, 84, 98, 112 and 126. The occurrence of myiasis was assessed throughout the study period. In addition, the weight, weight gain and daily weight gain of the animals were evaluated. RESULTS: In the 12 evaluations performed, the parasitic load of T01 was near zero. Fluralaner showed 99.5% efficacy on the 3rd day after the first treatment and 100% efficacy from Day 7 to Day 126. Cochliomyia hominivorax larvae (n = 6; p = 0.0251) were found only in the control group (T02). At the end of the study, the animals subjected to treatments with fluralaner gained 32.8 kg more than the animals in the control group. CONCLUSIONS: Application of fluralaner in summer and autumn, with 42-day intervals between treatments, was effective to control R. microplus on taurine cattle, which also gained more weight than control cattle. Additionally, no cases of myasis were documented in animals treated with fluralaner.

[Selective dry cow therapy on dairy farms in Rhineland-Palatinate, Saarland and Hesse, Germany - a survey among farmers]. / Selektives antibiotisches Trockenstellen bei Milchkühen in Rheinland-Pfalz, dem Saarland und Hessen ­ eine Umfrage unter Landwirten.

OBJECTIVE: With the Regulation (EC) 6/2019, antibiotic drying off of the entire dairy herd is no longer permissible. Hence, it is necessary to establish selective antibiotic drying off (SDCT: Selective Dry Cow Therapy) in dairy herds. With the publication of the PraeRi study in 2020, systematic data for the implementation of SDCT on farms became available for several German states. For Rhineland-Palatinate, Saarland and Hesse this type of information is only available from individual projects. Therefore, the aim of this survey was to increase the knowledge concerning the implementation of SDCT in dairy farms located in these states. MATERIAL AND METHODS: An online questionnaire was sent via newsletters to farmers and was published in the regional farmers' bulletins in the described catchment area. The questionnaire inquired about the saving of antibiotics during drying off, the criteria guiding the farmer's decision (cell count from monthly dairy herd improvement data (DHI), mastitis history, microbiological examination of quarter foremilk samples, California mastitis test), use of teat sealants and the type of dry off procedure (abrupt/gradual). RESULTS: A total of 103 questionnaires were evaluated, making the response rate ~1% for Hesse, ~3% for Saarland, and ~5% for Rhineland-Palatinate based on the number of included farms. Approximately 29% of the farmers dried off one out of four cows, 20% half, 23% three out of four and 13% all cows without using antibiotics. Eighty-nine farm managers based their decision on the somatic cell counts of DHI. Additional criteria influencing the decision were the mastitis history, results of the California Mastitis Test, or a combination of both. In 76 farms cows were dried off abruptly. In 79 farms teat sealers were used. CONCLUSIONS: Application of SDCT is established in most of the farms that participated in the survey, even though the proportion varied between farms. Legal requirements are not the only reason farmers need to increasingly deal with SDCT; sustainability programs of the dairies rely on selective drying off as well. Herd veterinarians should be supportive in implementing these measures to achieve good udder health while reducing the use of antimicrobials to a necessary minimum.

Immunomodulatory effect of Lacticaseibacillus casei CB054 supplementation in calves vaccinated against infectious bovine rhinotracheitis.

Probiotics are live microorganisms that, confer health benefits to the host when supplemented in adequate amounts. They can promote immunomodulation by inducing phagocyte activity, leukocyte proliferation, antibody production, and cytokine expression. Lactic acid bacteria (BAL) are important probiotic specimens with properties that can improves ruminant nutrition, productivity and immunity. The aim of the present study was to evaluate the immunomodulatory effect of the supplementation with Lacticaseibacillus casei CB054 in calve vaccinated against bovine infectious rhinotracheitis (IBR). Calve were vaccinated with a commercial IBR vaccine, on day 0 and received a booster dose on day 21. L. casei CB054 was orally administered (4 ×109 UFC) for 35 days, while a non-supplemented control group received Phosphate Buffer Saline (PBS). Stimulation of bovine splenocytes with L. casei CB054 markedly enhanced mRNA transcription levels of cytokines IL2, IL4, IL10 and IL17 genes. Calves supplemented with L. casei CB054 showed significantly higher (p < 0.05) specific anti-BoHV-1 IgG levels, higher serum neutralization, as well as higher mRNA transcription for IL2, IL4, IL10 and IL17 genes in Peripheral Blood Mononuclear Cells (PBMCs) comparing with control calves. Supplemented calve had an average weight gain of ∼14 kg more than non-supplemented during the experimental period. These results suggest that L. casei CB054 supplementation increase immunogenicity of a commercial IBR vaccine in cattle and improve weight gain.

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).

Epidrugs: alternative chemotherapy targeting Theileria annulata schizont stage parasites.

The growing emergence of resistance to current anti-theilerial agents necessitates the exploration of alternative approaches to drug discovery. This study evaluated the antiparasitic efficacy of 148 compounds derived from an epigenetic inhibitor library against the schizont stage of a Theileria annulata-infected cell line. Initial screening at a concentration of 10 µM identified 27 compounds exhibiting promising anti-theilerial activity. Further investigation, including determination of the 50% inhibitory concentration (IC50) and host cell cytotoxicity assay, highlighted seven highly effective compounds (SAHA, BVT-948, Trichostatin A, Methylstat, Plumbagin, Ryuvidine, and TCE-5003) against T. annulata-infected cells. Analysis of the active compounds revealed their inhibitory action against various human targets, such as HDAC (SAHA and Trichostatin A), SET domain (Ryuvidine), PRMT (BVT-948 and TCE-5003), histone demethylase (Methylstat), and ROS/apoptosis inducer (Plumbagin). We identified gene orthologs of these targets in Theileria and conducted molecular docking studies, demonstrating effective binding of the compounds with their respective targets in the parasite, supported by in vitro data. Additionally, we performed in silico ADME/T predictions, which indicated potential mutagenic and hepatotoxic effects of Plumbagin, Methylstat, and TCE-5003, rendering them unsuitable for drug development. Conversely, SAHA, Trichostatin A, and BVT-948 showed promising characteristics and may represent potential candidates for future development as chemotherapeutic agents against tropical theileriosis. These findings provide valuable insights into the search for novel anti-theilerial drugs and offer a basis for further research in this area.IMPORTANCETheileria annulata is a protozoan parasite responsible for tropical theileriosis, a devastating disease affecting cattle. Traditional chemotherapy has limitations, and the study explores the potential of epidrugs as an alternative treatment approach. Epidrugs are compounds that modify gene expression without altering the underlying DNA sequence, offering a novel way to combat parasitic infections. This research is pivotal as it addresses the urgent need for innovative therapies against T. annulata, contributing to the development of more effective and targeted treatments for infected livestock. Successful implementation of epidrugs could not only enhance the well-being of cattle but also have broader implications for the control of parasitic diseases, showcasing the paper's significance in advancing veterinary science and improving livestock health globally.

Contribution of herd management, biosecurity, and environmental factors to the risk of bovine tuberculosis in a historically low prevalence region.

Eradication of bovine tuberculosis (bTB) in certain historically low-prevalence regions remains elusive. A complete characterisation of the husbandry practices, biosecurity, and environment where farms are located is crucial to implement targeted in-farm risk mitigation protocols. Here, a detailed survey performed in 94 dairy cattle farms located in Navarra, a low-prevalence region of Spain between 2016 and 2020 was carried out. Data on 73 biosecurity, farm-, and environmental-level factors potentially associated with the risk of bTB occurrence were evaluated using an ordinal logistic regression model: farms were classified based on their prevalence index, a score linked to each farm to account for the severity and recurrence of bTB cases: 22.3% of the farms had a score of 1, 21.3% a score of 2, 26.6% a score of ≥ 3, and 29.8% were negative herds. A statistically significant association between a higher prevalence index and the frequency of badger sightings along with the lease of pastures to sheep during Winter was identified. Farms that detected badgers on a monthly to daily basis in the surroundings and those that leased pastures for sheep flocks during Winter were four [odds ratio, 95% CI (4.3; 1.1-17.5)] and three (3.1; 1.0-9.9) times more likely to have the highest prevalence index, respectively (predicted probabilityprevalence index≥3 = 0.7; 95% CI 0.3-0.9). Conversely, farms that used a vehicle to transport animals from holdings to pastures were less likely (0.1; <0.1-0.3) to present higher levels of prevalence index compared with farms that used none (on foot). Results suggested that the combined effect of farm- and environmental-level risk factors identified here may be hampering disease eradication in Navarra, highlighting the need to implement targeted protocols on farms and grazing plots. An increased awareness of monitoring sheep and wildlife in direct or indirect contact with cattle herds in historically low bTB prevalence areas should be raised.

What is the optimal timing to initiate strategic control of Rhipicephalus microplus in taurine cattle in a tropical region?

Cattle tick control poses a significant challenge for livestock in tropical and subtropical regions. The objective of this study was to determine the most suitable timing to initiate a strategic tick control program and to identify the ideal number of acaricide treatments for adult taurine cattle (Bos taurus taurus) in a tropical region throughout the year. Three groups with 10 bovines each were performed: T01 (strategic treatment in late autumn/winter/late spring, every 28 days), T02 (strategic treatment to act in the "first tick generation" - early spring/summer/early autumn, every 28 days) and T03 (control). Tick counts (females 4-8 mm) were conducted every 14 days. If the tick burden in any group reached 30 or more during these counts, we applied an additional treatment. Over the course of a year, T02 required significantly fewer (p < 0.05) acaricide treatments than T01, with nine treatments for T02 and eleven for T01. Furthermore, during the tick counts, animals in T02 showed a lower tick burden compared to those in T01. Initiating the strategic tick control program in early spring, corresponding to the first tick generation, proved more effective than starting in autumn. This approach not only required fewer acaricide treatments but also resulted in a reduced tick burden. These benefits are particularly valuable in terms of animal welfare and managing acaricide resistance issues.

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.

Biosecurity, health and disease management practices among the dairy farms in five districts of Bangladesh.

Herd health management is a planned program to optimize health, welfare, and production of dairy cows. A cross-sectional study focused on herd health management of dairy farms, was carried out to assess the biosecurity, health, and disease management practices with associated socio-demographic factors. A structured questionnaire with closed- and open-ended questions was employed for farmers' interviews to acquire information on dairy farms. Data were collected from randomly selected 205 dairy farms in five districts (Dhaka-Savar, Mymensingh, Gazipur, Rangpur, and Sirajganj) of Bangladesh. Overall, 1.5% farms had "good", 17% had "moderate" and 81.5% had "poor" levels of biosecurity, health and disease management practices (BHDMP). Analysis of variance revealed that mean BHDMP scores were significantly (p < 0.05) higher in farms of Rangpur (27.6 ± 0.8) and Gazipur (26.7 ± 0.6) than those of the other three districts. Educational status had a significant effect on BHDMP scores. The farms owned by graduated farmers had the highest mean BHDMP score (30.4 ± 1.1) while the farms operated by illiterate farmers had the lowest BHDMP score (20.4 ± 0.4). Multiple linear regression analysis also revealed that educational status (secondary to post-graduation), training experience, area of study (Rangpur district), and herd size were significantly associated with BHDMP score of the farms. The findings lay a basis for improving dairy farms' biosecurity, health and disease management practices, which further envisage the adoption and implementation of herd health management programs.

Dairy farmers' knowledge, awareness and practices regarding bovine lameness in Malaysian dairy farms.

Given the data paucity on dairy farmers' perspectives regarding bovine lameness and hoof diseases, particularly in South East Asian countries, this study was conducted to assess the knowledge, attitude and practices toward lameness and hoof health among dairy cattle farmers in Malaysia. An online-based and face-to-face survey was conducted among 114 dairy farmers from four states in Peninsular Malaysia. Data were analysed using descriptive statistics, principal component analysis and an independent sample t-test. Overall, farmers demonstrated satisfactory knowledge and attitude regarding lameness and its impact on dairy cattle welfare and production. Lameness was ranked the second most important health issue in dairy farms after mastitis. Notably, 90% reported the presence of at least one lame cow on their farms, and 55% stated lameness as the reason for culling their cows. While sole ulcer was the hoof lesion mostly identified by farmers, 75% of them underestimated lameness prevalence on their farms and rarely implemented management strategies such as preventive hoof trimming and footbath. Farmers' educational qualification influenced their understanding of the impact of lameness on dairy cattle production. Despite reflecting satisfactory knowledge and attitude towards lameness in dairy cows, farmers in this study need to improve their current management practices to address lameness problem in their herds. Educating farmers on the importance of early detection and prompt treatment, and preventive measures are crucial for lameness control and improving hoof health in these dairy farms.

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.

Inferring transmission routes for foot-and-mouth disease virus within a cattle herd using approximate Bayesian computation.

To control an outbreak of an infectious disease it is essential to understand the different routes of transmission and how they contribute to the overall spread of the pathogen. With this information, policy makers can choose the most efficient methods of detection and control during an outbreak. Here we assess the contributions of direct contact and environmental contamination to the transmission of foot-and-mouth disease virus (FMDV) in a cattle herd using an individual-based model that includes both routes. Model parameters are inferred using approximate Bayesian computation with sequential Monte Carlo sampling (ABC-SMC) applied to data from transmission experiments and the 2007 epidemic in Great Britain. This demonstrates that the parameters derived from transmission experiments are applicable to outbreaks in the field, at least for closely related strains. Under the assumptions made in the model we show that environmental transmission likely contributes a majority of infections within a herd during an outbreak, although there is a lot of variation between simulated outbreaks. The accumulation of environmental contamination not only causes infections within a farm, but also has the potential to spread between farms via fomites. We also demonstrate the importance and effectiveness of rapid detection of infected farms in reducing transmission between farms, whether via direct contact or the environment.

Rehydration post-transport: duration of oral fluid therapy on behavior, biochemical measures of hydration, and health of neonatal dairy calves.

The objective of this study was to determine the impact of 0, 1, 2, or 3 d of oral electrolyte solutions (OES) following transport on calf behavior, biochemical measures of hydration, and subsequent health. Two cohorts of 60 Holstein bull calves ~3 to 7 d of age were included in this experiment. Calves (n = 30/treatment) were randomized to 1 of 4 treatments on arrival at a calf-raising facility: (1) 1-d OES, (2) 2-d OES, (3) 3-d OES, or (4) no OES (control). Calves were evaluated for signs of arthritis, depression, dehydration, diarrhea, fever, navel inflammation, and respiratory disease on arrival (day 0) and 1, 2, 3, and 7 d after arrival. Blood was obtained following each examination to assess biochemical measures of hydration, including serum electrolytes, glucose, hematocrit, lactate, pH, and serum total protein (STP). Calves were fitted with 3D accelerometers attached to the right hind leg along the metatarsus bone to continuously measure lying time. Linear regression models with Gaussian or Poisson link functions were used to test differences between treatments in blood parameters, as well as disease frequency and behavior, respectively. Calves spent the most time lying immediately post-transport (day 0); however, there was no effect of OES treatment on lying time. On arrival, 90% (95% CI: 84.6% to 95.4%) of calves had mild to moderate dehydration, and there was no significant difference in hydration status across treatments. Mildly dehydrated calves that received 1- and 2-d OES had higher STP concentration compared to the control. Moderately dehydrated calves that received 3- vs. 1-d OES had higher (101.4 vs. 93.7 mmol/L, respectively; P = 0.01) glucose concentration, and 3- vs. 2-d OES (101.4 vs. 96.2, respectively; P = 0.07) calves tended to have higher glucose concentration. The prevalence of failed transfer of passive immunity (STP < 5.1 g/dL) and hypoglycemia (glucose < 4.95 mmol/L) was 26.7% (18.6% to 34.7%) and 76.5% (68.7% to 84.2%), respectively. The frequency of moderate dehydration was lower for calves that received 2-d OES compared to the control (count ratio: 0.4; 0.2% to 0.8%; P = 0.009). These results underline the need for more research on rehydration strategies to help inform calf processing protocols. Future research should investigate preconditioning strategies to minimize the impact of marketing and transport on dairy calves. Continued nutritional-based research is also needed to better support calves' recovery post-transport.

Most male calves are sold and transported from the dairy farm soon after birth. Typically, calves are transported without access to milk and/or water, and they often arrive at calf-raising facilities with varying degrees of dehydration. This study provided calves with 0 (control), 1, 2, or 3 consecutive days of oral electrolyte solutions following transportation and assessed calf behavior, biomarkers of hydration, and subsequent health post-transport. Most calves were dehydrated and hypoglycemic (low blood glucose levels) on arrival at the calf-raising facility. Calves spent the most time lying immediately post-transport; however, electrolytes did not impact total lying time, the number of lying bouts, or lying bout duration. Providing calves with electrolytes for 2 d improved moderate dehydration (assessed via skin tent test) compared to the control. These results suggest that providing calves with 2 consecutive days of electrolytes following transportation can improve hydration status; however, more research is needed to mitigate dehydration prior to arrival at calf-raising facilities.