Impact of residual antibiotics on microbial decomposition of livestock manures in Eutric Regosol: Implications for sustainable nutrient recycling and soil carbon sequestration.

The land application of livestock manure has been widely acknowledged as a beneficial approach for nutrient recycling and environmental protection. However, the impact of residual antibiotics, a common contaminant of manure, on the degradation of organic compounds and nutrient release in Eutric Regosol is not well understood. Here, we studied, how oxytetracycline (OTC) and ciprofloxacin (CIP) affect the decomposition, microbial community structure, extracellular enzyme activities and nutrient release from cattle and pig manure using litterbag incubation experiments. Results showed that OTC and CIP greatly inhibited livestock manure decomposition, causing a decreased rate of carbon (28%-87%), nitrogen (15%-44%) and phosphorus (26%-43%) release. The relative abundance of gram-negative (G-) bacteria was reduced by 4.0%-13% while fungi increased by 7.0%-71% during a 28-day incubation period. Co-occurrence network analysis showed that antibiotic exposure disrupted microbial interactions, particularly among G- bacteria, G+ bacteria, and actinomycetes. These changes in microbial community structure and function resulted in decreased activity of urease, ß-1,4-N-acetyl-glucosaminidase, alkaline protease, chitinase, and catalase, causing reduced decomposition and nutrient release in cattle and pig manures. These findings advance our understanding of decomposition and nutrient recycling from manure-contaminated antibiotics, which will help facilitate sustainable agricultural production and soil carbon sequestration.

Identification and prevalence of ixodid ticks of cattle in case of Aleltu district, Oromia regional state, northern Ethiopia.

BACKGROUND: In Ethiopia, ticks are the major threat to cattle productivity and production, leading to considerable economic losses. The current study was designed to estimate the prevalence of ixodid tick infestation, identify species, assess major risk factors associated with tick infestation and assess public awareness. METHODS: A cross-sectional and questionnaire-based study was conducted from January 2022 to June 2022 in the Aleltu district. The study animals were selected using a simple random sampling method. RESULTS: Of the 400 cattle examined, 303 (75.8%) were found to be infested by one or more tick species. Six species of ticks were identified that belonged to three genera: Amblyomma, Hyalomma and Rhipicephalus, and the subgenus Rhipicephalus (Boophilus). The most common tick species identified in terms of their prevalence and dominance were Rh. (Bo) decoloratus, Rh. evertsi, Am. variegatum, Hy. rufipes, Rh. bergeoni and Rh. praetextatus. In the present study, Rh. (Bo) decoloratus was the most prevalent (56.8%) in the study area. Among the risk factors considered, the prevalence of tick species had a statistically significant (p < 0.05) association with the age, production systems and body condition of animals. Out of 110 people interviewed, 107 (97.3%) believed there was a tick infestation in their village, and almost all farmers 103(93.6%) in the study area were unaware that ticks serve as vectors. CONCLUSIONS: The present study provides preliminary information on the prevalence of tick infestation and the composition of ticks in the Aleltu district. Ticks are a major problem for the cattle in the study area. Therefore, the problem observed in the study area alarms the district and calls for a comprehensive control strategy.

Depth detection limit of a fluorescent object in tissue-like medium with background emission in continuous-wave measurements: a phantom study.

Significance: Although the depth detection limit of fluorescence objects in tissue has been studied, reports with a model including noise statistics for designing the optimum measurement configuration are missing. We demonstrate a variance analysis of the depth detection limit toward clinical applications such as noninvasively assessing the risk of aspiration. Aim: It is essential to analyze how the depth detection limit of the fluorescence object in a strong scattering medium depends on the measurement configuration to optimize the configuration. We aim to evaluate the depth detection limit from theoretical analysis and phantom experiments and discuss the source-detector distance that maximizes this limit. Approach: Experiments for detecting a fluorescent object in a biological tissue-mimicking phantom of ground beef with background emission were conducted using continuous wave fluorescence measurements with a point source-detector scheme. The results were analyzed using a model based on the photon diffusion equations. Then, variance analysis of the signal fluctuation was introduced. Results: The model explained the measured fluorescence intensities and their fluctuations well. The variance analysis showed that the depth detection limit in the presence of ambient light increased with the decrease in the source-detector distance, and the optimum distance was in the range of 10 to 15 mm. The depth detection limit was found to be ∼ 30 mm with this optimum distance for the phantom. Conclusions: The presented analysis provides a guide for the optimum design of the measurement configuration for detecting fluorescence objects in clinical applications.

Antiviral activity of bovine type III interferon against bovine viral diarrhea virus is greatly reduced in bovine turbinate cells due to limited expression of IFN lambda receptor 1 (IL-28Rα).

Introduction: The antiviral activity of recombinant bovine interferon lambda 3 (bovIFN-λ3) against bovine viral diarrhea virus (BVDV) has been demonstrated in vitro in Madin-Darby bovine kidney cells (MDBK) and in vivo in cattle. However, anti-BVDV activity of bovIFN-λ3 has not been studied in bovine respiratory tract epithelial cells, supposedly a primary target of BVDV infection when entering the host by the oronasal route. Methods: Here we investigated the anti-BVDV activity of bovIFN-λ3 in bovine turbinate-derived primary epithelial cells (BTu) using BVDV infection and immunoperoxidase staining, TCID50, RT-qPCR, DNA and transcriptome sequencing, and transfection with plasmids containing the two subunits, IL-28Rα and IL-10Rß that constitute the bovIFN-λ3 receptor. Results: Our immunoperoxidase staining, RT-qPCR, and TCID50 results show that while BVDV was successfully cleared in MDBK cells treated with bovIFN-λ3 and bovIFN-α, only the latter, bovIFN-α, cleared BVDV in BTu cells. Preincubation of MDBK cells with bovIFN-λ3 before BVDV infection was needed to induce optimal antiviral state. Both cell types displayed intact type I and III IFN signaling pathways and expressed similar levels of IL-10Rß subunit of the type III IFN receptor. Sequencing of PCR amplicon of the IL-28Rα subunit revealed intact transmembrane domain and lack of single nucleotide polymorphisms (SNPs) in BTu cells. However, RT-qPCR and transcriptomic analyses showed a lower expression of IL-28Rα transcripts in BTu cells as compared to MDBK cells. Interestingly, transfection of BTu cells with a plasmid encoding IL-28Rα subunit, but not IL-10Rß subunit, established the bovIFN-λ3 sensitivity showing similar anti-BVDV activity to the response in MDBK cells. Conclusion: Our results demonstrate that the sensitivity of cells to bovIFN-λ3 depends not only on the quality but also of the quantity of the IL-28Rα subunit of the heterodimeric receptor. A reduction in IL-28Rα transcript expression was detected in BTu as compared to MDBK cells, despite the absence of spliced variants or SNPs. The establishment of bovIFN-λ3 induced anti-BVDV activity in BTu cells transfected with an IL-28Rα plasmid suggests that the level of expression of this receptor subunit is crucial for the specific antiviral activity of type III IFN in these cells.

Effects of brewery by-products based silage on productive performance of crossbred dairy cows.

The present study aimed to evaluate the effect of an increasing levels of brewery by-products based silage on productive performances of 3/4 Friesian x Boran mid-lactating cows. Experimental cows had similar in initial milk yield (11.7 ± 1.0), average days in milk (81.7 ± 6.1) and live weight (LW, 430.7 ± 40.3 kg) but different in parities (2-5).The dietary treatments were arranged randomly in 4 × 4 Latin Square Design that included ad libitum natural pasture hay feeding for all treatments as a roughage source plus a commercial dairy concentrate mix supplemented at 0.5 kg DM (dry matter)/liter of milk produced/day for cows in the control group (T1) and 0.3, 0.5 and 0.7 kg DM of brewery by-products based silage per liter of milk yield/cow/day for cows in T2, T3 and T4 groups, respectively. The study revealed that the daily milk yield of experimental cows was influenced by dietary treatments with relatively higher daily milk yield being recorded (P < 0.05) for cows in the T4 (13.9 l) followed by T3 (13.8 l). Milk composition of cows remained unchanged (P > 0.05) except for fat percentage of the milk that showed a declining trend (P < 0.05) with incremental inclusion levels of brewery by-products based silages. The highest net income (NI, 437.9 Eth. Birr) and marginal rate of return (MRR, 800.7%) was obtained for cows receiving brewery by-products based silage at the rate of 0.7 kg/liter of milk yield as compared to cows in the other treatment groups. Further study is required on the long term effect of brewery by-products based silage supplementation on productive, reproductive performance, and milk microbial qualities.

[Antimicrobial in vitro effects of eight essential oils on Pasteurella multocida and Mannheimia haemolytica from nasopharyngeal swab samples of fattening calves]. / Antimikrobielle in-vitro-Wirkung acht ätherischer Öle auf Pasteurella multocida und Mannheimia haemolytica aus Nasenrachentupferproben von Mastkälbern.

INTRODUCTION: Essential oils are secondary metabolites of aromatic plants and are used in phytotherapy to treat various diseases. In the present study, eight selected essential oils - ajwain oil (Trachyspermum ammi L.), fennel oil (Foeniculum vulgare Mill. subsp. vulgare var. vulgare), thyme oil chemotype (ct.) thymol (Thymus vulgaris L.), tea tree oil (Melaleuca alternifolia Cheel.), oregano oil (Origanum vulgare L.), mountain savory oil (Satureja montana L.), lemongrass oil (Cymbopogon citratus (DC.) Stapf.) and eucalyptus oil (Eucalyptus globulus Labill.) -were examined for their antibacterial effect against Pasteurella (P.) multocida and Mannheimia (M.) haemolytica isolates from deep nasopharyngeal swab samples of fattening calves using agar diffusion and microdilution. All eight essential oils were effective against the tested isolates. Lemongrass oil proved to be the most potent of all eight essential oils, while fennel oil was only weakly effective. Different antimicrobial effects were observed between the two research methods. The effectiveness of ajwain, thyme, oregano and mountain savory oils was comparable in agar diffusion. However, this could not be reproduced using the microdilution method. P. multocida was found to be more sensitive to all essential oils tested than M. haemolytica. This study shows that the tested essential oils have antimicrobial in-vitro effects on P. multocida and M. haemolytica isolates and that the examination method is associated with the test result.

INTRODUCTION: Les huiles essentielles sont des métabolites secondaires de plantes aromatiques et sont utilisées en phytothérapie pour le traitement de différentes maladies. Dans la présente étude, huit huiles essentielles sélectionnées ­ huile d'ajowan (Trachyspermum ammi L.), huile de fenouil (Foeniculum vulgare Mill. subsp. vulgare var. vulgare), huile de thym chémotype (ct.) thymol (Thymus vulgaris L.), huile d'arbre à thé (Melaleuca alternifolia Cheel.), huile d'origan (Origanum vulgare L.), huile de sarriette de montagne (Satureja montana L. ), huile de citronnelle (Cymbopogon citratus (DC.) Stapf.) et huile d'eucalyptus (Eucalyptus globulus Labill.) ­ ont été étudiées par diffusion sur gélose et microdilution pour leur effet antibactérien sur des isolats de Pasteurella (P.) multocida et de Mannheimia (M.) haemolytica provenant d'échantillons d'écouvillons nasaux profonds de veaux d'engraissement. Les huit huiles essentielles se sont révélées efficaces sur les isolats testés. L'huile de citronnelle s'est avérée être la plus puissante des huit huiles essentielles, tandis que l'huile de fenouil n'était que faiblement efficace. Des effets différents ont été observés entre les deux méthodes de recherche utilisées. Par exemple, l'efficacité des huiles d'ajowan, de thym, d'origan et de sarriette de montagne était comparable dans la diffusion sur gélose. Cependant, cela n>a pas pu être reproduit avec la méthode de microdilution. P. multocida s'est révélée plus sensible que M. haemolytica à toutes les huiles essentielles testées. Cette étude montre premièrement que les huiles essentielles testées ont une efficacité antimicrobienne in vitro sur des isolats cliniques de P. multocida et de M. haemolytica. Deuxièmement, elle montre que la méthode d'examen est associée au résultat du test.

Realization of reproductive qualities of cows and productivity of young stock.

For the first time, on the basis of complex research, scientifically based and experimentally proven zootechnical feasibility of using the Prevention-N-A biologics developed by us based on the polysaccharide complex of yeast cells Saccharomyces cerevisiae and the aminoglycoside group bactericidal preparation in the technology of obtaining and growing calves in comparison with the previously approved PS-2 preparation. It is proved that three-time intramuscular injection of PS-2 and Prevention-N-A biologics to down-calvers 45-40, 25-20 and 15-10 days before calving in a dose of 10 ml prevents gynecological diseases and increases reproductive function. Under the influence of preparations in cows, the time of separation of the fetal membranes was reduced by 6.0 and 6.4 hours, retention of placenta, post-parturient complications and breast diseases were prevented. The risk of subinvolution of the uterus and endometritis in the first case decreased by 3.0 and 2.0 times, respectively, and in the second - was excluded (P<0.05). Against the background of immunocorrection in cows, the time of onset of heat by 11.6 and 14.2 days, the insemination index by 1.6 and 1.8 times, the service period by 22.4 and 28.4 days, and fertilization in 1 oestrus increased by 2.5 and 3.0 times (P<0.05-0.01). It was found that twice intramuscular injection of PS-2 and Prevention-N-A to calves on 2...3rd and 7... 9th day of life in a dose of 3 ml stimulates their growth and development, reduces the incidence. By the end of the growing period, the animals of the 1st and 2nd experimental groups were superior in live weight to control peers by 4.6 and 7.0 kg, rearing - 13.8 and 17.0 kg, and fattening - by 19.4 and 24.2 kg, respectively (P<0.05-0.01). A similar pattern occurred in the nature of changes in exterior measurements and the growth coefficient of animals of the compared groups. In calves of the experimental groups, the incidence of respiratory and digestive organs decreased by 2.3 and 7.0 times, the recovery time - by 1.3 and 4.3 days, respectively, compared to the control (P<0.05).

[Mass transfer of bilirubin and bovine serum albumin in hollow fiber membrane module of artificial liver].

Understanding the mass transfer behaviors in hollow fiber membrane module of artificial liver is important for improving toxin removal efficiency. A three-dimensional numerical model was established to study the mass transfer of small molecule bilirubin and macromolecule bovine serum albumin (BSA) in the hollow fiber membrane module. Effects of tube-side flow rate, shell-side flow rate, and hollow fiber length on the mass transfer of bilirubin and BSA were discussed. The simulation results showed that the clearance of bilirubin was significantly affected by both convective and diffusive solute transport, while the clearance of macromolecule BSA was dominated by convective solute transport. The clearance rates of bilirubin and BSA increasd with the increase of tube-side flow rate and hollow fiber length. With the increase of shell-side flow rate, the clearance rate of bilirubin first rose rapidly, then slowly rose to an asymptotic value, while the clearance rate of BSA gradually decreased. The results can provide help for designing structures of hollow fiber membrane module and operation parameters of clinical treatment.

Effect of onion skin powder on color, lipid, and protein oxidative stability of premade beef patty during cold storage.

The impact of premade beef patty (BBP) with red onion skin powder (OSP) at 0, 1, 2, and 3% levels on color, lipid, and protein oxidative stability, and infection degree of microorganisms during cold storage was investigated. The objective was to determine the effect of color by L*, a*, b*, and the content of MetMb. The inhibitory effect of OSP on the oxidation of lipid and protein was studied based on TBARS and the carbonyl content of protein in samples at different storage times. TVB-N content was used to characterize the degree of infection of microorganisms and their effect on meat quality. The results showed that the addition of OSP reduced the pH, L *, a*, and b * values of BBP, and improved the hardness, springiness, gumminess, and cohesiveness of BBP, but had no significant effect on the chewiness of BBP (p > 0.05). After 12 days of storage, the carbonyl group and TBARS content in the BBP supplemented with 3%OSP was significantly lower than that in the control group (p < 0.05). Furthermore, the addition of OSP significantly inhibited the TVB-N increase during beef patty storage. These results indicated that OSP has a good research prospect as a natural antioxidant or preservative.

Metagenomic analysis reveals high diversity of gut viromes in yaks (Bos grunniens) from the Qinghai-Tibet Plateau.

The Qinghai-Tibet Plateau (QTP), renowned for its exceptional biological diversity, is home to numerous endemic species. However, research on the virology of vulnerable vertebrates like yaks remains limited. In this study, our objective was to use metagenomics to provide a comprehensive understanding of the diversity and evolution of the gut virome in yak populations across different regions of the QTP. Our findings revealed a remarkably diverse array of viruses in the gut of yaks, including those associated with vertebrates and bacteriophages. Notably, some vertebrate-associated viruses, such as astrovirus and picornavirus, showed significant sequence identity across diverse yak populations. Additionally, we observed differences in the functional profiles of genes carried by the yak gut virome across different regions. Moreover, the virus-bacterium symbiotic network that we discovered holds potential significance in maintaining the health of yaks. Overall, this research expands our understanding of the viral communities in the gut of yaks and highlights the importance of further investigating the interactions between viruses and their hosts. These data will be beneficial for revealing the crucial role that viruses play in the yak gut ecology in future studies.

Optimisation of Early Pregnancy Diagnosis and Resynchronisation in Beef Cows.

The objectives of this study were (1) to investigate the effects of the preovulatory follicle (POF) size on the accuracy of Doppler-based early pregnancy detection, and (2) to determine whether the removal of PGF2α (PGF) treatment during the resynchronisation protocol would affect fertility in beef cows. In Experiment 1, Nelore suckling cows (n = 224) were enrolled in an estradiol-progesterone-based timed artificial insemination (TAI) protocol. At TAI, cows were separated based on the range of POF diameters, as follows: ≤11.0 mm (n = 50), 11.1-12.9 mm (n = 64), 13.0-14.4 mm (n = 62) and ≥14.5 mm (n = 48). On day 22 after TAI, the corpus luteum (CL) blood flow (CLBF) of all cows was examined by colour Doppler ultrasonography to diagnose nonpregnant cows. The cows with the largest POF had the greatest positive predictive value (88.6%; 31 of 35) and diagnostic accuracy (91.7%; 44 of 48). In Experiment 2, Nelore cows (n = 233) were subjected to the same TAI protocol. Fourteen days after TAI, all cows were started on a resynchronisation protocol. Cows diagnosed as nonpregnant based on CLBF, on day 22, received 0.5 mg estradiol cypionate intramuscular (im) and were assigned to receive either 150 µg of PGF (PGF; n = 50) or 2 mL of saline (control; n = 47). Cows treated with PGF had a P/AI of 30.0% compared with a 48.9% P/AI in controls (p = 0.06). Our findings demonstrate that the POF size affects the accuracy of a CLBF-based early pregnancy diagnosis and that the removal of PGF treatment from the resynchronisation protocol tended to increase P/AI of the second TAI.

Pathological, immunological and molecular epidemiological analysis of lumpy skin disease virus in Indian cattle during a high-mortality epidemic.

Lumpy skin disease (LSD) is an economically significant, emerging viral disease of Cattle and Buffaloes. This study aimed to investigate the causes of high mortality in a recent LSD epidemic in India. We examined 1618 animals across seventy outbreaks and conducted post-mortem on 48 cattle out of 513 clinically suspected LSD cases. The morbidity, mortality and case fatality rates recorded were 31.70%, 2.97 and 9.37% respectively. Disease stages were categorized as early (20.81%), mid (42.02%), and late (37.17%) and the distribution of skin lesions was classified as mild (34.14%), moderate (39.39%), and severe (26.47%). Post-mortem findings revealed systemic infection with necrotic and ulcerative nodules on multiple internal organs. Histologically, necrotizing vasculitis and mononuclear cell infiltration with intracytoplasmic inclusions were observed in various organs. The highest viral load was found in skin nodules/scabs, trachea, tongue, and lymph nodes. The viral load was significantly higher in mid- and late-stages of skin nodules and internal organs; whereas, blood from early-stage showed high viral load. The expression of Th1-type and Th2-type cytokines varied significantly across different stages of the disease. The downregulation of the apoptotic intrinsic and upregulation of the extrinsic pathway genes, suggesting that the latter plays a role in LSDV infection. Genetic analysis revealed that the LSD virus (LSDV) isolates were derived from a Kenyan ancestral strain with unique nucleotide changes in RPO30 and P32 gene. In conclusion, the high mortality in the recent Indian LSD epidemic can be attributed to a newly identified, highly virulent strain of LSDV causing systemic infection.

Temporal stability of the rumen microbiome and its longitudinal associations with performance traits in beef cattle.

The rumen microbiome is the focus of a growing body of research, mostly based on investigation of rumen fluid samples collected once from each animal. Exploring the temporal stability of rumen microbiome profiles is imperative, as it enables evaluating the reliability of findings obtained through single-timepoint sampling. We explored the temporal stability of rumen microbiomes considering taxonomic and functional aspects across the 7-month growing-finishing phase spanning 6 timepoints. We identified a temporally stable core microbiome, encompassing 515 microbial genera (e.g., Methanobacterium) and 417 microbial KEGG genes (e.g., K00856-adenosine kinase). The temporally stable core microbiome profiles collected from all timepoints were strongly associated with production traits with substantial economic and environmental impact (e.g., average daily gain, daily feed intake, and methane emissions); 515 microbial genera explained 45-83%, and 417 microbial genes explained 44-83% of their phenotypic variation. Microbiome profiles influenced by the bovine genome explained 54-87% of the genetic variation of bovine traits. Overall, our results provide evidence that the temporally stable core microbiome identified can accurately predict host performance traits at phenotypic and genetic level based on a single timepoint sample taken as early as 7 months prior to slaughter.

Diagnostic utility of acute phase proteins and their ability to guide antibiotic usage in pigs, horses, and cattle: a mapping review.

To mitigate the use of antibiotics for many of the multifactorial diseases seen in pigs, horses and cattle, new diagnostic tools are needed. Acute phase protein (APP) measurements can, in humans, be used to guide antibiotic treatment initiation, evaluate treatment efficacy, and make a prognosis. The aim of this review is to collect evidence on the clinical functionality of APP measurements as a tool to guide antibiotic treatment in pigs, horses, and cattle. Literature was retrieved using Medline, CAB Abstracts and Google Scholar. The acute phase response has been investigated for a plethora of diseases and clinical signs and the major acute phase proteins are elevated in diseased compared to healthy animals. Few studies correlated acute phase response with aetiology, antibiotic treatment efficacy, prognosis, or severity of disease. The existing research does not support that APP can be used to guide antibiotic treatment, but the reported studies indicate that C-reactive protein (CRP) might be able to differentiate between bacterial and non-bacterial causes of disease in pigs. Serum amyloid A (SAA) might reflect underlying aetiology in horses and infectious or non-infectious cases of mastitis in cows.

Milk yield prediction in Friesian cows using linear and flexible discriminant analysis under assumptions violations.

BACKGROUND: The application of novel technologies is now widely used to assist in making optimal decisions. This study aimed to evaluate the performance of linear discriminant analysis (LDA) and flexible discriminant analysis (FDA) in classifying and predicting Friesian cattle's milk production into low ([Formula: see text]4500 kg), medium (4500-7500 kg), and high ([Formula: see text]7500 kg) categories. A total of 3793 lactation records from cows calved between 2009 and 2020 were collected to examine some predictors such as age at first calving (AFC), lactation order (LO), days open (DO), days in milk (DIM), dry period (DP), calving season (CFS), 305-day milk yield (305-MY), calving interval (CI), and total breeding per conception (TBRD). RESULTS: The comparison between LDA and FDA models was based on the significance of coefficients, total accuracy, sensitivity, precision, and F1-score. The LDA results revealed that DIM and 305-MY were the significant (P < 0.001) contributors for data classification, while the FDA was a lactation order. Classification accuracy results showed that the FDA model performed better than the LDA model in expressing accuracies of correctly classified cases as well as overall classification accuracy of milk yield. The FDA model outperformed LDA in both accuracy and F1-score. It achieved an accuracy of 82% compared to LDA's 71%. Similarly, the F1-score improved from a range of 0.667 to 0.79 for LDA to a higher range of 0.81 to 0.83 for FDA. CONCLUSION: The findings of this study demonstrated that FDA was more resistant than LDA in case of assumption violations. Furthermore, the current study showed the feasibility and efficacy of LDA and FDA in interpreting and predicting livestock datasets.

Distribution and molecular analysis of Subtilase cytotoxin gene (subAB) variants in Shiga toxin-producing Escherichia coli (STEC) isolated from different sources in Iran.

Subtilase exhibits strong cytotoxicity that was first described in O113:H21 strain in Australia as a plasmid- encoded cytotoxin (subAB1). Subsequently, chromosomal variants including subAB2-1, subAB2-2, and subAB2-3 were described. We aimed to investigate the presence of subAB genes in a collection of Shiga toxin-producing Escherichia coli (STEC) strains (n=101) isolated from different sources in Iran. A collection of 101 archived STEC strains isolated from cattle (n=50), goats (n=25), sheep (n=15), wild captive animals (n=8: persian fallow deer, n=3; caspian pony, n=1; Macaca mulatta, n=4), and humans (n=3) during 2007-2016 were analyzed for the detection of different genes encoding the Subtilase variants, plasmidic and chromosomal virulence genes, phylogroups and serogroups. Overall, 57 isolates (56.4%) carried at least one variant of subAB. Most strains from small ruminants including 93% of sheep and 96% of caprine isolates carried at least one chromosomally encoded variant (subAB-2-1 and/or subAb2-2). In contrast, 12 cattle isolates (24%) only harbored the plasmid encoded variant (subAB1). STEC strains from other sources, including deer, pony and humans were positive for subAB-2-1 and/or subAb2-2. Our results reveal the presence of potentially pathogenic genotypes among locus of enterocyte effacement (LEE)-negative isolates, and some host specificity related to Subtilase variants and other virulence markers that may aid in source tracking of STEC during outbreak investigations.

Estimates of Sequences with Ultralong and Short CDR3s in the Bovine IgM B Cell Receptor Repertoire Using the Long-read Oxford Nanopore MinION Platform.

Cattle produce Abs with an H chain ultralong CDR3 (40-70 aa). These Abs have been shown to have features such as broad neutralization of viruses and are investigated as human therapeutics. A common issue in sequencing the bovine BCR repertoire is the sequence length required to capture variable (V) and isotype gene information. This study aimed to assess the use of Oxford Nanopore Technologies' MinION platform to perform IgM BCR repertoire sequencing to assess variation in the percentage of ultralong CDR3s among dairy cattle. Blood was collected from nine Holstein heifers. B cells were isolated using magnetic bead-based separation, RNA was extracted, and IgM+ transcripts were amplified using PCR and sequenced using a MinION R10.4 flow cell. The distribution of CDR3 lengths was trimodal, and the percentage of ultralong CDR3s ranged among animals from 2.32 to 20.13% in DNA sequences and 1.56% to 17.02% in productive protein sequences. V segment usage varied significantly among heifers. Segment IGHV1-7, associated with ultralong CDR3s, was used in 5.8-24.2% of sequences; usage was positively correlated with ultralong CDR3 production (r = 0.99, p < 0.01). To our knowledge, this is the first study to sequence the bovine BCR repertoire using Oxford Nanopore Technologies and demonstrates the potential for cost-efficient long-read repertoire sequencing in cattle without assembly. Findings from this study support literature describing the distribution of length and percentage of ultralong CDR3s. Future studies will investigate changes in the bovine BCR repertoire associated with age, antigenic exposure, and genetics.

Expression profile of microRNAs in bovine lymphocytes infected with Theileria annulata and treated with buparvaquone.

Several miRNA-based studies on Theileria-transformed bovine cells have been conducted; however, the mechanism by which transformed cells exhibit uncontrolled proliferation is not yet fully understood. Therefore, it is necessary to screen more microRNAs that may play a role in the transformation process of host cells infected with Theileria annulata to better understand the transformation mechanisms of Theileria-infected cells. RNA sequencing was used to analyze miRNAs expression in the host bovine lymphocytes infected with T. annulata at different time points after buparvaquone (BW720) treatment and DMSO treatment (control groups). Differential miRNAs related to cell proliferation and apoptosis were identified through comparison with gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) databases, and a regulatory network of miRNA-mRNA was constructed. In total, 272 differentially expressed miRNAs were found at 36, 60 and 72 h. The miRNAs change of bta-miR-2285t, novel-miR-622, bta-miR-2478, and novel-miR-584 were significant. Analysis of 27 of these co-differential expressed miRNAs revealed that 15 miRNAs were down-regulated and 12 miRNAs were up-regulated. A further analysis of the changes in the expression of each of these 27 miRNAs in the three datasets suggested that bta-miR-2285t, bta-miR-345-5p, bta-miR-34a, bta-miR-150, and the novel-miR-1372 had significantly changed. Predicted target genes for these 27 miRNAs were analyzed by KEGG and the results demonstrated that EZR, RASSF, SOCS1 were mainly enriched in the signaling pathway microRNAs in cancer. MAPKAPK2, RELB, FLT3LG, and GADD45B were mainly enriched in the MAPK signaling pathway, and some genes were enriched in Axon guidance. This study has provided valuable information to further the understanding of the regulatory function of miRNAs in the host microenvironment and host-parasite interaction mechanisms.

Effect of temperature on rehydration and fracture resistance of reattached tooth fragments after crown fracture: An in vitro study.

BACKGROUND: Uncomplicated crown fractures of maxillary anterior teeth are common dental injuries, and the bonding of fractured fragments is recommended for management. Rehydration of fragments improves bonding and fracture resistance. Therefore, the aim of this study was to compare the fracture resistance of the reattached tooth fragment of the incisor crown after rehydration using either a steamer or a humidifier. MATERIALS AND METHODS: Bovine teeth were used as a model. Fractured fragments were divided into three groups: Group I (no rehydration), Group II (rehydration by humidifier), and Group III (rehydration by steamer). Fragments were reattached using a standard bonding protocol. Fracture resistance was tested using a universal testing machine. Statistical analysis was performed using analysis of variance and Chi-square tests. RESULTS: The force required to fracture the reattached fragments was significantly higher in Group II (humidifier) compared to Group I (no rehydration) (P = 0.005). Group III (steamer) had a force value similar to Group I, indicating no significant improvement in fracture resistance with steamer rehydration. The temperature inside the steam-based chamber reached an average of 95.7°C with 95% humidity, whereas the conventional humidification chamber had an average temperature of 39.2°C and 84% humidity. CONCLUSION: Within the limitations of the present study, the rehydration protocol using a steam-based chamber was found to have a significantly lower force required to fracture the reattached fragments as compared to humidifier-based chamber. This was not significantly different from the fragments which were reattached without rehydration.

Overexpression of miRNA29a gene inhibits proliferation and promotes apoptosis of jejunal epithelial cells in yak.

MicroRNAs (miRNAs) were identified to be involved in various biological functions by regulating the degradation or suppressing the translation of their downstream target genes. Recent studies have identified miR-29a play a key role in functions of mammal cell differentiation, proliferation, apoptosis, and signal transduction. However, the underlying functions for miR-29a in jejunal epithelial cells biological function still to be investigated. In order to explore the yak jejunal epithelial cells proliferation and barrier dysfunction with over expression of miR-29a gene, three 0-day-old Pamir male yaks were randomly selected and slaughtered in present study, and the jejunal epithelial cells were isolated and cultured to determine yak jejunal epithelial cells proliferation and protein composition on differential expression of miR-29a gene in Pamir plateau. Here, we demonstrated that the overexpression of miR-29a gene could inhibit the proliferation of Pamir yaks jejunum epithelial cells, and contribute to the apoptosis of Pamir yaks jejunal epithelial cells with some extent. A total of 133 differentially expressed proteins were identified in different expression of miR-29a groups by label-free Mass Spectrometry (MS), which could be concluded to two predominant themes: cell proliferation and inflammatory response. Interestingly, GPR41, as a bridge protein, was contacted two predominant themes to involved in Pamir Yaks jejunal mechanical barrier PPI network, and the target proteins displayed strong mutual interactions in the complex PPI network. Overall, our study suggested that the over-expression miR-29a inhibited the jejunal epithelial cells proliferation and the expressions of specific proteins, which damaged jejunal barrier function to slow down the intestine structure and function advanced mature development during young livestock period for influence the enhanced performance of production efficiency.