Sublethal toxicity of carbofuran in cattle egret (Bubulcus ibis coromandus): hematological, biochemical, and histopathological alterations / Toxicidade subletal do carbofurano na garça-vaqueira (Bubulcus ibis coromandus): alterações hematológicas, bioquímicas e histopatológicas

Abstract This study was aimed to investigate Carbofuran (CF)-induced pathological changes in cattle egret. Two hundred cattle egrets were reared and equally divided into four groups and given different CF concentrations (0.03 mg/L, 0.02 mg/L, 0.01 mg/L and 0 mg/L (control group)). Hematology, serum biochemistry, histopathology, and immunological markers were studied. Our results confirm that CF induces anemic conditions, leukocytosis, elevated liver enzymatic activity, and alterations in renal biomarkers. Moreover, specific microscopic lesions such as multifocal necrosis, pyknotic nuclei, hemorrhages, congestion, and inflammatory cell proliferation were observed in the liver, kidney, spleen, and thymus. These findings suggest that CF can induce harmful effects, so the application of this pesticide in the field must be strictly monitored to mitigate the possibility of exposure to non-target species.

Resumo Este estudo teve como objetivo investigar as alterações patológicas induzidas por carbofurano (CF) em garças-vaqueiras. Duzentas dessas garças foram criadas e divididas igualmente em quatro grupos e receberam diferentes concentrações de CF: 0,03 mg/L; 0,02 mg/L; 0,01 mg/L; e 0 mg/L (grupo controle). Foram realizadas análises de hematologia, bioquímica sérica, histopatologia e marcadores imunológicos. Nossos resultados confirmaram que CF induz condições anêmicas, leucocitose, atividade enzimática hepática elevada e alterações nos biomarcadores renais. Além disso, lesões microscópicas específicas, como necrose multifocal, núcleos picnóticos, hemorragias, congestão e proliferação de células inflamatórias, foram observadas no fígado, rim, baço e timo. Esses achados sugerem que o CF pode causar efeitos nocivos, portanto a aplicação desse agrotóxico no campo deve ser rigorosamente monitorada para mitigar a possibilidade de exposição a espécies não alvo.

Effects of Pasturella Multocida B: 2 and its immunogens (LPS and OMP) on reproductive hormones in Nili-Ravi Buffaloes / Efeitos de Pasteurella multocida B: 2 e seus imunógenos (LPS e OMP) em hormônios reprodutivos em búfalos Nili-Ravi

Abstract Livestock is a fundamental part of the agriculture industry in Pakistan and contributes more than 11.53% to GDP. Among livestock species, the buffaloes are regarded as the black gold of Pakistan. Being the highest milk producers globally, Nili-Ravi buffaloes are the most famous ones. Buffaloes are affected by many endemic diseases, and "Hemorrhagic septicemia" (HS) is one of them. This study was designed to ascertain the effects of experimental exposure ofP. multocida B:2 (oral) and its immunogens, i.e., LPS (oral and intravenous) and OMP (oral and subcutaneous) on reproductive hormonal profiles in Nili-Ravi buffaloes. Repeated serum samples were collected from the jugular vein of experimental animals for 21 days (0, 02, 04, 08, 12, 16, 20, 24, 36, 48, 72, 120, 168, 216, 264, 360, 456 and 504 hours). Hormonal assays to determine the serum concentrations of Gonadotropin-releasing hormone (GnRH), Follicle-stimulating hormone (FSH), Luteinizing hormone (LH), Estrogen (E2) and progesterone (P4) were performed using (MyBioSource) commercial Elisa kits. The hormonal profile of all treatment groups of the buffalo heifers exhibited significant (P<0.05) variations as compared to the control group (G-1). These results indicate suppression in Nili-Ravi buffaloes' reproductive hormonal profile on exposure to P. multocida B:2 and its immunogens. This influence warrants that exposure to H.S may be a possible reason for delayed puberty and poor reproduction performance in Nili-Ravi buffaloes.

Resumo A pecuária é uma parte fundamental da indústria agrícola no Paquistão e contribui com 11,53% do PIB nacional. Entre as espécies de gado, os búfalos são considerados o ouro negro do Paquistão. Sendo os maiores produtores de leite em todo o mundo, os búfalos Nili-Ravi são os mais famosos. Os búfalos são afetados por muitas doenças endêmicas, entre as quais a "septicemia hemorrágica" (SH). Este estudo busca verificar os efeitos da exposição experimental de P. multocida B:2 (oral) e seus imunógenos, ou seja, LPS (oral e intravenoso) e OMP (oral e subcutâneo), nos perfis hormonais reprodutivos em búfalos Nili-Ravi. Amostras de soro repetidas foram coletadas da veia jugular de animais experimentais por 21 dias (0, 2, 4, 8, 12, 16, 20, 24, 36, 48, 72, 120, 168, 216, 264, 360, 456 e 504 horas). Os ensaios hormonais para determinar as concentrações séricas do hormônio liberador de gonadotrofina (GnRH), hormônio foliculoestimulante (FSH), hormônio luteinizante (LH), estrogênio (E2) e progesterona (P4) foram realizados usando kits comerciais Elisa (MyBioSource). O perfil hormonal de todos os grupos de tratamento das novilhas bubalinas apresentou variações significativas (P < 0,05) em relação ao grupo controle (G-1). Esses resultados indicam supressão no perfil hormonal reprodutivo de búfalos Nili-Ravi na exposição a P. multocida B:2 e seus imunógenos. Essa influência garante que a exposição à SH possa ser uma possível razão para o atraso da puberdade e o baixo desempenho reprodutivo em búfalos Nili-Ravi.

Relationship between peptides and the change in quality characteristics of beef strip loin (M. longissimus lumborum) and tenderloin (M. psoas major).

Peptides in fresh and aged beef strip loin (M. longissimus lumborum) and tenderloin (M. psoas major) were quantified to investigate the relationship between proteolysis-induced peptides and beef quality characteristics. A total of 409 and 450 peptides were quantified from strip loin and tenderloin, respectively, and found to be significantly correlated to beef quality characteristics. Changes in redness and yellowness were significantly correlated to the peptides derived from G3P, ENOB, and KCRM in both muscles during 14 days of storage. The peptides produced from MYG, ENOB, HBA, PGK1, and TPIS were strongly associated with improved tenderness, while those derived from major myofibrillar proteins, such as MYH1, MYH2, ACTS, and DESM, were associated with changes in tenderloin color. These results improve our understanding of the association between peptides and changes in meat quality during cold storage, indicating that proteolysis-induced peptides can be indicators of the quality characteristics of fresh and aged meat.

Sonocatalytic degrading antibiotics over activated carbon in cow milk.

Herein, an efficient, simple and economical approach to remove antibiotics (ABX), i.e. ceftiofur hydrochloride, sulfamonomethoxine sodium (SMM), marbofloxacin and oxytetracycline by sonication with activated carbon (AC) from cow milk has been successfully implemented. The pseudo-first-order kinetics constants for the sonolytic and sonocatalytic degrading SMM are 0.036 and 0.093 min-1, respectively. The synergistic efficiency of removing ABX by using sonocatalysis reached 1.8-4.0. Hydrophobic ABX underwent faster degradation than hydrophilic ABX in sonocatalytic systems. Adding 0.5 mmol L-1 H2O2, the optimal concentration, improved the sonocatalytic degradation rates of ABX by 9.1%-28.5%. Surface area and dose of AC play crucial roles in the sonocatalysis of ABX. By sonicating 50 mL of 5.52 µmol L-1 ABX in milk at 500 kHz and 259 W with 20 mg AC for 20-60 min resulted in residual ABX concentrations ranging from 42.6 to 95.1 µg L-1, which meet the relative maximum residue limits set by European Commission.

Development of innovative antioxidant food packaging systems based on natural extracts from food industry waste and Moringa oleifera leaves.

Active packaging that prolongs food shelf life, maintaining its quality and safety, is an increasing industrial demand, especially if integrated in a circular economy model. In this study, the fabrication and characterization of sustainable cellulose-based active packaging using food-industry waste and natural extracts as antioxidant agents was assessed. Grape marc, olive pomace and moringa leaf extracts obtained by supercritical fluid, antisolvent and maceration extraction in different solvents were compared for their antioxidant power and phenolic content. Grape and moringa macerates in acetone and methanol, as the most efficient and cost-effective extracts, were incorporated in the packaging as coatings or in-between layers. Both systems showed significant free-radical protection in vitro (antioxidant power 50%) and more than 50% prevention of ground beef lipid peroxidation over 16 days by indirect TBARS and direct in situ Raman microspectroscopy measurements. Therefore, these systems are promising for industrial applications and more sustainable farm-to-fork food production systems.

Water extracts of Tibetan medicine Wuweiganlu attenuates experimental arthritis via inducing macrophage polarization towards the M2 type.

ETHNOPHARMACOLOGICAL RELEVANCE: Wuweiganlu (WGL) is a well-known formulation described in the "Four Medical Scriptures of Tibetan medicine", which is mainly used for the treatment of Rheumatoid Arthritis (RA) and other chronic ailments prescribed by Tibetan medicine. Nonetheless, the active constituents present in the water extracts of Wuweiganlu (WGLWE) specifically targeting arthritis treatment are largely unknown. AIM OF THE STUDY: The aim of this study is to explore the effects and underlying mechanisms of the active components in WGLWE on RA. MATERIALS AND METHODS: We utilized ultra-performance liquid chromatography coupled with Q-TOF mass spectrometry (UPLC-Q-TOF-MS) to identify the main chemical compositions of WGLWE. The polarization effect of WGLWE on bone marrow-derived macrophages (BMDM) was determined. A rat model of collagen-induced arthritis (CIA) was established by injecting an emulsion of bovine type II collagen mixed with an equal volume of incomplete Freund's adjuvant into the tail, paw and back of rats. A WGLWE-based ointment was topically applied to the legs and paws of the rats for 30 days. The rats' ankles were photographed to measure the degree of swelling. Micro-CT was used to image the knee joint and paw of rats, and the bone mineral density (BMD) and bone volume fraction (BV/TV) of knee joint in rats were analyzed. High-frequency ultrasound imaging of the rat knee joint was performed to observe knee joint effusion. Further, the serum levels of tumor necrosis factor (TNF-α), interleukin-6 (IL-6), IL-10, and arginine (Arg-1) in CIA rats were detected by enzyme-linked immunosorbent assay (ELISA). Immunohistochemistry (IHC) and immunofluorescence (IF) co-staining were employed to detect the expression levels of inflammatory factors in synovium. RESULTS: A total of 28 main components were identified in WGLWE, and these compounds can directly bind to the inflammatory pathway proteins such as JAK2, NFκB and STAT3. In vitro experiments demonstrated that WGLWE promoted the transformation of M1 macrophages into M2 macrophages and suppressed the release of proinflammatory cytokines TNF-α and IL-6. In vivo studies showed that WGLWE effectively reduced ankle swelling, alleviated knee joint effusion, and improved BV/TV while also reducing synovial inflammation levels. Furthermore, WGLWE compounds induced the transition of M1-type macrophages to M2-type macrophages in synovial tissue, resulting in decreased secretion of inflammatory factors TNF-α, WGLWE improved the synovial inflammatory state. CONCLUSION: Our results indicated that WGLWE alleviated joint inflammation in CIA rats and the underlying mechanism may be related to inducing the transformation of bone marrow-derived M1 macrophages to M2 macrophages, leading to an increase in the secretion of anti-inflammatory factors and a decrease in pro-inflammatory factors. Therefore, WGLWE may be used as a potential herbal preparation for the treatment of RA.

Molecular insights on the binding of chlortetracycline to bovine casein and its effect on the thermostability of chlortetracycline.

Bovine casein was selected as a model protein to evaluate the impact of food matrix on the thermal degradation of antibiotics. Fluorescence quenching and isothermal titration calorimetry experiments revealed that chlortetracycline (CTC) could spontaneously bind to casein via hydrogen bonding and hydrophobic interactions. The amino acid residues forming the binding pocket were further identified using molecular docking, while saturation transfer difference NMR deciphered that the binding of CTC engages its -N(CH3)2 group. Moreover, the degradation behavior of free CTC versus that bound in casein-CTC complex was compared during thermal treatment. Compared with free CTC, a lower first-order rate constant was observed in the presence of casein. Removal of casein shortened the half-life of CTC by at least 48.1% at low concentrations. Elucidating that the formation of protein-antibiotic complexes alters the amenability of antibiotics to degradative reactions, which could help eliminate residual antibiotics and guarantee the safety of dairy products.

Characterization and determination of casein glycomacropeptide in dairy products by UHPLC-MS/MS based on its characteristic peptide.

The ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS) method was built to quantify the casein glycomacropeptide (CGMP) in bovine dairy products accurately based on targeted proteomics. Qualitative analysis of theoretical peptides was carried out using high-resolution mass spectrometry (HRMS) and protein software. Isotope-labeled characteristic peptides were acquired via the labeled amino acid condensation method to correct the matrix effects. Peptide MAIPPK was the representative characteristic peptide for distinguishing the CGMP from κ-casein through trypsin digestion. After optimizing the pre-treatment conditions, the final 8% oxidant concentration was selected and the 10% formic acid concentration with 2.5 h oxidation time. Moreover, the results of methodological verification showed that the recovery rate was 103.7%, meanwhile the precision of inter-day and intra-day was less than 5%. In conclusion, the research demonstrated the characteristic peptide MAIPPK could quantitatively applied to detect CGMP in dairy products.

Label-Free quantitation of milk oligosaccharides from different mammal species and heat treatment influence.

Milk oligosaccharides (MOs) exhibit significant variations in concentrations and patterns among different species. However, there is limited knowledge about milk oligosaccharides in domestic animals and the impact of heat treatment on them. Here, we developed an LC-ESI-MS/MS method to analyze 11 milk oligosaccharides in 7 distinct species simultaneously. The results showed that human milk presented a completely different composition pattern of milk oligosaccharides from animals. In detail, animal milk predominantly contained sialylated oligosaccharides, and human milk had high levels of fucosylated neutral oligosaccharides. Notably, sheep milk exhibited similarities to human milk in terms of oligosaccharides composition. Then, the milk samples from dairy cows were treated with two common industrial heat treatments. We found that 65 °C treatment had no significant effect on the concentration of milk oligosaccharides, whereas 135 °C heating was associated with their decline, suggesting that high temperatures should be avoided in the processing of oligosaccharides supplemented/enriched products.

Kestose Increases the Relative Abundance of Faecalibacterium spp. and Nominally Increases Cow Milk Tolerant Dose in Children with Cow's Milk Allergy - Preliminary Results.

A single-arm study was conducted with 10 children aged 2-12 years with severe cow's milk allergy (CMA) requiring complete allergen elimination. Subjects were administered kestose, a prebiotic, at 1 or 2 g/day for 12 weeks. Results of a subsequent oral food challenge (OFC) showed a statistically significant increase in the total dose of cow's milk ingestion (1.6 ml vs. 2.7 ml, p = 0.041). However, the overall evaluation of the OFC results, TS/Pro (total score of Anaphylaxis Scoring Aichi (ASCA)/cumulative dose of protein), showed no statistically significant improvement, although the values were nominally improved in seven out of 10 subjects. The 16S rDNA analysis of fecal samples collected from the subjects revealed a statistically significant increase in the proportion of Faecalibacterium spp. (3.8 % vs. 6.8%, p = 0.013), a type of intestinal bacterium that has been reported to be associated with food allergy. However, no statistically significant correlation was found between Faecalibacterium spp. abundance and the results of the OFC.

Evolutionary origin of genomic structural variations in domestic yaks.

Yak has been subject to natural selection, human domestication and interspecific introgression during its evolution. However, genetic variants favored by each of these processes have not been distinguished previously. We constructed a graph-genome for 47 genomes of 7 cross-fertile bovine species. This allowed detection of 57,432 high-resolution structural variants (SVs) within and across the species, which were genotyped in 386 individuals. We distinguished the evolutionary origins of diverse SVs in domestic yaks by phylogenetic analyses. We further identified 334 genes overlapping with SVs in domestic yaks that bore potential signals of selection from wild yaks, plus an additional 686 genes introgressed from cattle. Nearly 90% of the domestic yaks were introgressed by cattle. Introgression of an SV spanning the KIT gene triggered the breeding of white domestic yaks. We validated a significant association of the selected stratified SVs with gene expression, which contributes to phenotypic variations. Our results highlight that SVs of different origins contribute to the phenotypic diversity of domestic yaks.

First report of molecular confirmation and phylogenetic analysis of ocular seteriasis in buffalo in India using 12S rRNA.

An adult Indian buffalo (Bubalus bubalis) presented with corneal opacity, irritation, and excessive lacrimation from the left eye in the Referral Veterinary Polyclinic-Teaching Veterinary Clinical Complex (RVC-TVCC), Indian Veterinary Research Institute, Izatnagar. Clinical examination revealed a whitish thread-like worm in the left eye's anterior chamber. The worm was surgically removed from the eye with supportive nerve blocks. Light microscopy was used for parasite morphological identification, which provided insight into the worm as female Setaria sp. Genomic DNA was isolated, and polymerase chain reaction amplification of 12S rRNA was conducted for molecular confirmation of the parasite. The amplicon was sequenced and analysed by bioinformatics software. Sequence data showed an amplicon size of 243 bp. Phylogenetic analysis with reference data from the NCBI Genbank database revealed the worm was S. digitata, with a similarity of 99.17%. The common predilection site of S. digitata is in the peritoneal cavity of natural hosts like cattle and buffalo and is mostly non-pathogenic. The aberrant migration of the parasite larva to the brain and eye commonly occurs in goats, sheep, and horses, causing clinical conditions like cerebrospinal nematodiasis (lumbar paralysis) and ocular setariasis, respectively. Nevertheless, until now, there have been no reports of ocular setariasis in buffalo. This report is the first unusual occurrence of ocular setariasis in buffalo and its molecular confirmation and phylogenetic analysis using 12S rRNA.

Investigation of virulence factors, phylogenetic grouping, multiple-locus variable number tandem repeat analysis, and antimicrobial susceptibility of E. coli isolated from aborted bovine fetal tissue.

Escherichia coli is an important microorganism for cattle breeding. The aim of this study was to investigate the presence of phylogenetic groups, virulence factors, genotyping with multi-locus variable tandem repeat analysis (MLVA), and susceptibility to commonly used antimicrobial agents in E. coli strains isolated from aborted bovine fetal samples. In this study, phylogrouping and various virulence genes were analyzed by PCR in E. coli strains isolated from 637 bovine fetal tissue samples. Consequently, E. coli was isolated and identified in 24 samples in culture. Of the 24 isolates identified as positive, 12.5% were defined as group A, 83.3% as B1, and 4.2% as group B2. Of the E. coli isolates, virulence factor fimH was identified in eight (33.3%), traT in 15 (62.5%), ompT in five (20.8%), CNF1 in one (4.16%), and CNF2 in six (25%). Seven genotypic groups were determined as a result of the analysis with the MLVA 10 method. According to the antimicrobial susceptibility test results, high resistance was determined against amoxicillin/clavulanic acid and oxytetracycline. In conclusion, strains of E. coli containing CNF1, CNF2, fimH, traT, and ompT virulence factors can be associated with bovine abortions. It is noteworthy that the dominant phylogenetic group B1 has been observed in cases of cattle abortions.

Risk factors associated with enzootic bovine leukosis in Boyacá and Cundinamarca municipalities, Colombia.

Background: Enzootic bovine leukosis (EBL) is a lymphoproliferative disorder caused by the bovine leukemia virus (BLV), a virus of the Retroviridae family. The infection is distributed worldwide, and a high percentage of animals infected by the BLV are asymptomatic and act as carriers of the virus in many cattle populations. Aim: To identify the risk factors associated with EBL in the municipalities of Boyacá and Cundinamarca (Colombia). Methods: A simple descriptive cross-sectional study with random sampling was conducted. A total of 1,140 blood samples were taken from cattle (females and males) from the municipalities of Chiquinquirá, Ubaté, and San Miguel de Sema of different breeds and age groups. The samples were processed using the commercial ELISA SERELISA® BLV Ab Mono Blocking kit (sensitivity 97%, specificity 98%). The data were processed with the statistical programs WinEpi and Epi Info® version 7.2.4.0, estimating the prevalence ratio, implementing the chi-square test (p ≤ 0.05) and logistic regression. Results: A true prevalence (TP) and apparent prevalence (AP) of 23.61% and 22.7% in Ubaté, 19.22% and 18.1% in Chiquinquirá, and 15.61% and 14.3% in San Miguel de Sema, respectively, were established. Bovines 2-4 years old were the most prevalent in Ubaté and Chiquinquirá (37.5% and 21.21%, respectively), while in San Miguel de Sema individuals >4 years had the highest percentage of antibodies (18.3%). The Holstein breed had a higher prevalence in Ubaté and San Miguel de Sema (26.02% and 19.67%), and crossbreeds were more BLV-seroprevalence in Chiquinquirá (20.20%). In Ubaté, re-use of needles was identified as a risk factor, contaminated blood in needles is considered one of the main routes of transmission. On the other hand, manual milking was identified as a risk factor in San Miguel de Sema. Conclusion: The non-implementation of an individual needle per animal in Ubaté; the Holstein breed and manual milking in San Miguel de Sema were identified as risk factors for the presence of antibodies against the disease. EBL prevention and control plans should be established that focus on the implementation of management and sanitary practices based on herd biosecurity.

Role of nuc gene in Staphylococcus aureus to phagocytic activity in different cattle infections.

Background: Staphylococcus aureus is a typical pathogenic agent that causes several morbidities and mortalities which variable largely following the severity of bacteria and activity of host immunity. Aim: Isolation of S. aureus from different cattle infections, molecular detection of nuc gene in positive S. aureus isolates, and identification of the effectiveness of the phagocytic activity. Methods: Totally, 100 cattle with various infections (25 wounds, 25 abscesses, 25 nasal discharges, and 25 ear swaps) were selected and subjected to collection of swabs under controlled conditions. All collected samples were cultured on mannitol salt agar (MSA) and assessed by biochemical tests. Targeting the nuc gene, all study MSA positive isolates were examined molecularly by conventional polymerase chain reaction (PCR), and then subjected to antibiotic susceptibility test. Jugular venous blood was collected from all infected animals in addition to 20 healthy cattle that were selected as a control group to estimate the phagocytic activity of S. aureus isolates. Results: The findings of MSA culture revealed a total of 80 positive samples of S. aureus as 23, 21, 20, and 16 positive isolates for nasal discharge, abscess, wound, and ear swab, respectively; based on its morphology, cultural trait, and biochemical test. Subsequently, PCR assaying of MSA-positive isolates demonstrated an overall 59 positive samples as 14, 16, 12, and 17 positive isolates for nasal discharge, abscess, wound, and ear swabs, respectively. Antibiotic susceptibility testing of S. aureus-positive PCR isolates reported a significantly high sensitivity to chloramphenicol and vancomycin, and a high resistance to penicillin. Finally, there was a considerable decline in phagocytic activity in particular 2 weeks post-infection as a result of bacterial invasion. Conclusion: This study shows a high prevalence of S. aureus in cattle infections, and the protocol includes a regular screening of cattle infection and suitable therapy based on antibacterial susceptibility test is of great importance in the long-term control of the pathogen. However, additional molecular studies targeting other genes of S. aureus and the role of immune markers in different infections should be aimed.

Comparison of coronary arteries morphometry and distribution in bovines with humans and other animal species.

Background: The left coronary artery (LCA) in the bovines is more developed than the right. Aim: The objective of the study is to describe the bovine coronary system from a morphological point of view, including the morphometry and its distribution. Methods: Arciform suture with 2.0 silk was applied around the sinus orifice and coronary ostium and a number 14 catheter was installed, to perfuse semi-synthetic polyester resin, consisting of a mixture of 85% GP40L palatal with 15% styrene with red color mineral. Results: The average weight of 28 bovine hearts used in our study was 1.534.1 kg. The right coronary artery had a proximal caliber of 5 +/- 0.9 mm. The LCA caliber and length were 9.4 +/- 1.2 and 18.3 +/- 4.8 mm, respectively. This artery was divided into two branches in 85.7% of the cases and trifurcated in 14.3%. The paraconal interventricular branch (PIB) ended more frequently in the apex (46.4%), and its proximal caliber was 6.4 +/- 1.4 mm. The left circumflex branch ended in 82.1% in the subsinusal interventricular sulcus, and its proximal caliber was 5.9 +/- 1.2 mm. The proximal calibers of the PIB and the left circumflex branch did not present statistically significant differences (p = 0.137). The sinoatrial branch presented a dual origin (right and LCA) in 46.4% of the cases and a single origin from the LCA in 53.6% of the samples. In the evaluated hearts, left coronary dominance was observed in most cases (96.4%). Their presence of anastomosis between the branches of the coronary arteries was observed in 57.1% of cases. Conclusion: The presence of a myocardial bridge was found in six hearts (21.4%). In bovines, a high percentage of anastomosis was found, a protective factor in obstruction of the coronary arterial branches.

The use of a homeopathic preparation in the treatment of subclinical form of mastitis in cows.

Background: Mastitis is a disease of productive cows, widespread throughout the world, and characterized by significant economic damage to the dairy industry. The subclinical form of this disease is aggravated by additional difficulties with its diagnosis and the lack of clear treatment protocols. Aim: Therefore, the study of the effectiveness of diagnostic studies and the search for new methods of treatment of latent forms of mastitis is an important direction in scientific research in countries with developed dairy cattle breeding. Methods: Studies conducted on the number of dairy cows of the production cooperative "Izhevsky" of the Akmola region of the Republic of Kazakhstan showed that when using rapid tests Kenotest, Somatest, Mastidine test, and Wideside test, the same results were obtained when the disease was detected in cows. The effectiveness of the tests was at the level of 60%-62% when using the settling sample as a control. Medical procedures were carried out using the Aquaton-2 microwave radiation apparatus and a homeopathic preparation. When using physiotherapy with microwave radiation, a decrease in the level of microbial contamination of milk from the treated part of the udder by 1.5-5 times was observed. Results: Biologically active substances of plant origin in the homeopathic preparation, due to the immunostimulating effect, made it possible to increase the level of γ-globulins in the blood serum of sick animals during the application. Conclusion: The complex use of both methods in the treatment of animals with a subclinical form of mastitis made it possible to reduce the level of somatic cells in the milk of the affected udder lobe to a level that cannot be determined using Kenotest in 4-6 days, which is 2-4 days faster than using these methods separately.

A fully integrated duplex RT-LAMP device for the detection of viral infections.

Respiratory viruses can cause epidemics or pandemics, which are worldwide outbreaks of disease. The severity of these events varies depending on the virus, its characteristics, along with environmental factors. The frequency of epidemics and pandemics caused by respiratory viruses is difficult to predict, but the potential severity of such events underlines the importance of continued monitoring, research, and preparation for emerging infectious diseases. To help improve pandemic preparedness, we created a fully integrated duplex reverse transcription loop-mediated isothermal amplification (RT-LAMP) device targeting two respiratory viruses, influenza A/X-31 virus and bovine coronavirus, as a replacement for SARS-CoV-2. This device can be adapted to any other respiratory virus. In this study, we showed and evaluated a prototype of a microfluidic system, and showed that duplex RT-LAMP can detect and distinguish between the two viruses, with LoDs of 2,000 copies/ml for bovine coronavirus and 200 copies/ml for influenza A/X-31 virus.

RACK1 mediates NLRP3 inflammasome activation during Pasteurella multocida infection.

Pasteurella multocida is a gram-negative bacterium that causes serious diseases in a wide range of animal species. Inflammasomes are intracellular multimolecular protein complexes that play a critical role in host defence against microbial infection. Our previous study showed that bovine P. multocida type A (PmCQ2) infection induces NLRP3 inflammasome activation. However, the exact mechanism underlying PmCQ2-induced NLRP3 inflammasome activation is not clear. Here, we show that NLRP3 inflammasome activation is positively regulated by a scaffold protein called receptor for activated C kinase 1 (RACK1). This study shows that RACK1 expression was downregulated by PmCQ2 infection in primary mouse peritoneal macrophages and mouse tissues, and overexpression of RACK1 prevented PmCQ2-induced cell death and reduced the numbers of adherent and invasive PmCQ2, indicating a modulatory role of RACK1 in the cell death that is induced by P. multocida infection. Next, RACK1 knockdown by siRNA significantly attenuated PmCQ2-induced NLRP3 inflammasome activation, which was accompanied by a reduction in the protein expression of interleukin (IL)-1ß, pro-IL-1ß, caspase-1 and NLRP3 as well as the formation of ASC specks, while RACK1 overexpression by pcDNA3.1-RACK1 plasmid transfection significantly promoted PmCQ2-induced NLRP3 inflammasome activation; these results showed that RACK1 is essential for NLRP3 inflammasome activation. Furthermore, RACK1 knockdown decreased PmCQ2-induced NF-κB activation, but RACK1 overexpression had the opposite effect. In addition, the immunofluorescence staining and immunoprecipitation results showed that RACK1 colocalized with NLRP3 and that NEK7 and interacted with these proteins. However, inhibition of potassium efflux significantly attenuated the RACK1-NLRP3-NEK7 interaction. Our study demonstrated that RACK1 plays an important role in promoting NLRP3 inflammasome activation by regulating NF-κB and promoting NLRP3 inflammasome assembly.

Changes in Choline Metabolites and Ceramides in Response to a DASH-Style Diet in Older Adults.

This feeding trial evaluated the impact of the Dietary Approaches to Stop Hypertension diet on changes in plasma choline, choline metabolites, and ceramides in obese older adults; 28 adults consumed 3oz (n = 15) or 6oz (n = 13) of beef within a standardized DASH diet for 12 weeks. Plasma choline, betaine, methionine, dimethylglycine (DMG), phosphatidylcholine (PC), lysophosphotidylcholine (LPC), sphingomyelin, trimethylamine-N-oxide (TMAO), L-carnitine, ceramide, and triglycerides were measured in fasted blood samples. Plasma LPC, sphingomyelin, and ceramide species were also quantified. In response to the study diet, with beef intake groups combined, plasma choline decreased by 9.6% (p = 0.012); DMG decreased by 10% (p = 0.042); PC decreased by 51% (p < 0.001); total LPC increased by 281% (p < 0.001); TMAO increased by 26.5% (p < 0.001); total ceramide decreased by 22.1% (p < 0.001); and triglycerides decreased by 18% (p = 0.021). All 20 LPC species measured increased (p < 0.01) with LPC 16:0 having the greatest response. Sphingomyelin 16:0, 18:0, and 18:1 increased (all p < 0.001) by 10.4%, 22.5%, and 24%, respectively. In contrast, we observed that sphingomyelin 24:0 significantly decreased by 10%. Ceramide 22:0 and 24:0 decreased by 27.6% and 10.9% (p < 0.001), respectively, and ceramide 24:1 increased by 36.8% (p = 0.013). Changes in choline and choline metabolites were in association with anthropometric and cardiometabolic outcomes. These findings show the impact of the DASH diet on choline metabolism in older adults and demonstrate the influence of diet to modify circulating LPC, sphingomyelin, and ceramide species.