Draft genome sequence of Leuconostoc falkenbergense isolated from naturally fermented buffalo milk curd.

This study reports the draft genome of Leuconostoc falkenbergense strain BSMRAU-M1L5, isolated from artisanal buffalo milk curd in Bangladesh. The draft genome spans 1,776,471 bp, with 50× coverage and 96 contigs.

Genomic insights into antibiotic resistance genes in Leuconostoc citreum strains isolated from artisanal buffalo milk curd in Bangladesh through whole-genome sequencing.

We sequenced the genome of Leuconostoc citreum strains BSMRAU-M1L6 and BSMRAU-M1L13 isolated from artisanal buffalo milk curd in Bangladesh. The draft genomes of BSMRAU-M1L6 and BSMRAU-M1L13 are 1,869,891 and 1,890,611 bp, respectively, with 50.0× coverage (both) and 65 and 75 contigs, respectively.

Facile synthesis of CuONPs using Citrus limon juice for enhancing antibacterial activity against methicillin-resistant Staphylococcus aureus, beta-lactamase and tetracycline-resistant Escherichia coli.

Antimicrobial resistance (AMR) resulting from indiscriminate use of antibiotics in various fields of agriculture such as livestock farming, aquaculture, and croup fields become an emerging catatroph for the health (human, animal) and environment. Among those, poultry farming has been considered as one of the major contributors of multidrug-resistant (MDR) bacteria. Focusing this, the present research is designed for green synthesis of copper oxide nanoparticles (CuONPs) with the aim of their application in antibiotic-free poultry farming for curving use of antibiotics in that sector. For that, antibacterial CuONPs were nanoformulated to decrease the required doses of bulk CuSO4. We used a CuSO4·5H2O solution as a Cu2+ source and Citrus limon juice as a reducing agent as well as capping agent. Particle yield was initially confirmed by the λmax specific to CuONPs (295 nm) using UV-Vis spectroscopy. The presence of the Cu-O group during particle formation and crystallinity with the purity of yielded NPs was confirmed with Fourier-transform infrared spectroscopy and X-ray diffractometry. The round to spherical CuONPs of 92-155 nm average size was confirmed with atomic force, scanning electron, and transmission electron microscopy. The concentration of yielded NPs was calculated with the dynamic light scattering. The physical characterization tools indicated a maximum CuONPs yield with a 0.001 M ion source with 15% reducing agents after 12 h reduction. Antibacterial effectivity was tested against methicillin-resistant Staphylococcus aureus and tetracycline- and beta-lactamase-resistant Escherichia coli, confirmed by PCR amplicon band at 163 bp, 643 bp, and 577 bp for the mecA, blaTEM-1 and tetA genes, respectively. An antibiogram assay of CuONPs showed a maximum zone of inhibition of 26 ± 0.5 mm for the synthesized particles. The minimum inhibitory and bactericidal concentrations were 1.6 µg ml-1 and 3.1 µg ml-1, respectively, for broad-spectrum application. Finally, the biocompatibility of CuONPs was determined by demonstrating a nonsignificant decrease of BHK-21 cell viability at <2 MIC doses for complying their future in vivo applicability.

Nano-Enabled Antivirals for Overcoming Antibody Escaped Mutations Based SARS-CoV-2 Waves.

This review discusses receptor-binding domain (RBD) mutations related to the emergence of various SARS-CoV-2 variants, which have been highlighted as a major cause of repetitive clinical waves of COVID-19. Our perusal of the literature reveals that most variants were able to escape neutralizing antibodies developed after immunization or natural exposure, pointing to the need for a sustainable technological solution to overcome this crisis. This review, therefore, focuses on nanotechnology and the development of antiviral nanomaterials with physical antagonistic features of viral replication checkpoints as such a solution. Our detailed discussion of SARS-CoV-2 replication and pathogenesis highlights four distinct checkpoints, the S protein (ACE2 receptor coupling), the RBD motif (ACE2 receptor coupling), ACE2 coupling, and the S protein cleavage site, as targets for the development of nano-enabled solutions that, for example, prevent viral attachment and fusion with the host cell by either blocking viral RBD/spike proteins or cellular ACE2 receptors. As proof of this concept, we highlight applications of several nanomaterials, such as metal and metal oxide nanoparticles, carbon-based nanoparticles, carbon nanotubes, fullerene, carbon dots, quantum dots, polymeric nanoparticles, lipid-based, polymer-based, lipid-polymer hybrid-based, surface-modified nanoparticles that have already been employed to control viral infections. These nanoparticles were developed to inhibit receptor-mediated host-virus attachments and cell fusion, the uncoating of the virus, viral gene expression, protein synthesis, the assembly of progeny viral particles, and the release of the virion. Moreover, nanomaterials have been used as antiviral drug carriers and vaccines, and nano-enabled sensors have already been shown to enable fast, sensitive, and label-free real-time diagnosis of viral infections. Nano-biosensors could, therefore, also be useful in the remote testing and tracking of patients, while nanocarriers probed with target tissue could facilitate the targeted delivery of antiviral drugs to infected cells, tissues, organs, or systems while avoiding unwanted exposure of non-target tissues. Antiviral nanoparticles can also be applied to sanitizers, clothing, facemasks, and other personal protective equipment to minimize horizontal spread. We believe that the nanotechnology-enabled solutions described in this review will enable us to control repeated SAR-CoV-2 waves caused by antibody escape mutations.

FeedMaster: A least-cost feed formulation App for minimizing the cost and maximizing milk yield.

Objective: The study aimed to develop and assess an Android app designed for farmers with a low educational status that can formulate a least-cost ration. Materials and Methods: First, a computer-android-based app named BLRI FeedMaster was developed to guide users in formulating a balanced ration at the least cost. A survey was conducted on 30 livestock officers and 18 farmers with 50 cattle to evaluate its efficacy at the field level. The study outcomes were milk yield, feeding cost, milk composition, time, and cost for management before and after using the BLRI FeedMaster app. Descriptive statistics and paired sample t-tests were used to analyze the data. Results: After adopting the BLRI FeedMaster app, a significant increase was observed in daily average milk yield (9.39 ± 0.32 l from 8.37 ± 0.36 l), while a considerable decrease was observed in daily average feed quantity (4.88 ± 0.15 kg from 5.60 ± 0.17 kg) and feed cost (BDT 28.00 ± 0.50 from BDT 29.75 ± 0.49). Besides, the number of visits, time, and cost for seeking professional services regarding feed, health care, and other information was significantly minimized. The number of visits decreased to 0.36 ± 013 from 3.07 ± 0.38, and the consumed time was reduced from 270 ± 34.30 to 235.71 ± 59.42 min (p < 0.05) after adopting the app. Conclusion: Hence, this app was very beneficial for farmers with a low economic and educational background and may ultimately help farmers with profitable animal farming and sustainable production in the least developed countries like Bangladesh.

Fatty acid and amino acid profiles of cheese, butter, and ghee made from buffalo milk.

Objective: The objective was to assess the chemical composition, cholesterol, fatty acid (FAs), and amino acid (AAs) profiles of buffalo cheese, butter, and ghee. Materials and Methods: Buffalo milk (raw) was collected from the Bangladesh Agricultural University (BAU) Dairy Farm, BAU, Mymensingh-2202, Bangladesh. Cheese, butter, and ghee were prepared at the Dairy Chemistry and Technology Laboratory, Department of Dairy Science, BAU, Mymensingh, Bangladesh, and subjected to subsequent analyses. The gross nutritional composition and AAs profile of milk were analyzed prior to the manufacture of cheese, butter, and ghee. The gross nutritional composition of milk and dairy products was analyzed by applying an automated milk analyzer and the Association of Agricultural Chemists techniques, respectively. The cholesterol, FAs, and AAs contents of cheese, butter, and ghee were determined by the Bangladesh Council for Scientific and Industrial Research, Dhaka, Bangladesh. Furthermore, atherogenic and thrombogenic indices were also calculated using reference equations. Results: The results indicated that the buffalo milk is a good source of first-rate nutrients (dry matter: 16.50%, fat: 7.50%, protein: 3.75%). Findings indicated that the butter was significantly rich with (p < 0.05) total solids and fat where higher (p > 0.05) protein, carbohydrate, and minerals were found in cheese. The saponification, Reichert-Meissl, Polenski, and Kirschner values of buffalo ghee were found to be 225, 30, 1.2, and 25, respectively. A significant (p < 0.05) variation was found in the cholesterol content of buffalo cheese, butter, and ghee. Butter and ghee had 40.14 and 39.57 mg more cholesterol, respectively, than cheese. The results revealed identical FA profiles except for C24:0 among the three dairy products where the major FA compositions were C4:0, C14:0, C16:0, and C18:0 and C18:1 cis-9. The atherogenicity index and thrombogenicity index of cheese, butter, and ghee were statistically similar (p > 0.05). Butter was found with the most conducive anti-atherogenic and anti-thrombogenic characteristics due to lower saturated and higher polyunsaturated FAs. However, all the AAs concentrations were statistically higher (p < 0.05) in cheese than in butter and ghee. Conclusion: To conclude, buffalo cheese is superior to butter and ghee as regards nutrient density, but consumers can choose other foods based on their choice.

Variations in fatty acid and amino acid profiles of doi and rasomalai made from buffalo milk.

OBJECTIVE: This study investigated and compared the chemical composition, cholesterol content, fatty acid (FA), and amino acid (AA) profiles of doi and rasomalai made from buffalo milk. MATERIALS AND METHODS: Bangladesh Agricultural University Dairy Farm, Mymensingh-2202, Bangladesh was the source of raw buffalo milk. Then, doi and rasomalai were produced and analyzed. Prior to the production of doi and rasomalai, the gross composition and AAs of milk were evaluated. Milk and dairy products were evaluated for gross composition using an automated milk analyzer and the Association of Agricultural Chemists techniques, respectively. At the Bangladesh Council for Scientific and Industrial Research, Dhaka, Bangladesh, the cholesterol, FA, and AA levels of doi and rasomalai were determined. Additionally, atherogenic and thrombogenic indices were determined using established equations. RESULTS: The results indicated that the majority of the proximate components were significantly greater (p < 0.05) in rasomalai than in doi. Rasomalai had 3.64 mg more cholesterol (p > 0.05) than doi. The FA profile was identical across doi and rasomalai with the exception of oleic acid (C18:1cis-9), which was 1.50% greater (p < 0.05) in rasomalai. The atherogenicity index was found to be statistically higher in doi than in rasomalai (p > 0.05). Similarly, the thrombogenic index was found to be significantly higher (p > 0.05) in doi (1.98) when compared to the rasomalai (1.92). The concentrations of all AAs were found to be quantitatively higher in doi than in rasomalai (p > 0.05). CONCLUSION: The conclusion is that buffalo milk rasomalai appears to have a higher nutritional density than buffalo milk doi.

Unraveling the Genetic Diversity and Population Structure of Bangladeshi Indigenous Cattle Populations Using 50K SNP Markers.

Understanding the genetic basis of locally adapted indigenous cattle populations is essential to design appropriate strategies and programs for their genetic improvement and conservation. Here, we report genetic diversity measures, population differentiation, and structure of 218 animals sampled from six indicine cattle populations of Bangladesh. Animals were genotyped with Illumina Bovine SNP50K BeadChip along with genotyped data of 505 individuals included from 19 zebu and taurine breeds worldwide. The principal component analysis (PCA) showed clear geographic separation between taurine and indicine lineages where Bangladeshi indigenous cattle clustered with South Asian zebu populations. However, overlapped clusters in PCA, heterozygosity estimates, and Neighbor-Joining phylogenetic tree analysis revealed weak genetic differentiation among the indigenous cattle populations of Bangladesh. The admixture analysis at K = 5 and 9 suggests distinct genetic structure of the studied populations along with 1 to 4% of taurine ancestry. The effective population size suggested a limited pool of ancestors particularly for Sahiwal and North Bengal Grey cattle. In conclusion, these findings shed insights into the genetic architecture of six indigenous cattle populations of Bangladesh for the first time and suggested as distinct gene pools without potential admixture with zebu or taurine populations.

Coculturing cumulus oocyte complexes with denuded oocytes alters zona pellucida ultrastructure in in vitro matured bovine oocytes.

Oocyte quality is a key factor affecting success of in vitro embryo production in cattle. Improving the microenvironment of oocytes during in vitro maturation (IVM) can increase developmental rate and embryo quality. Therefore, the objective was to determine whether denuded oocytes (DO) affect embryo development and ultrastructure of the zona pellucida (ZP) in in vitro matured bovine oocytes. Intact immature cumulus-oocytes complexes (COC) obtained from a local abattoir or by ovum pick-up (OPU) were cocultured with and without abattoir-obtained DO at a COC:DO ratio of 1:5. After IVM, DO were removed and intact DO were either fertilized or observed by scanning electron microscopy. Blastocyst quality was evaluated using a TUNEL assay. The ZP pore size decreased after IVM in COC + DO coculture, regardless of their origin (OPU, 310.5 ± 92.5 vs. 428.9 ± 148.5 nm; abattoir, 317.5 ± 68.5 vs. 358.9 ± 128.5 nm; P < 0.05; mean values ± standard deviation). Moreover, the number of ZP pores in OPU COC + DO and COC + DO was greater than those in OPU COC and COC (control) groups (56 ± 4 and 55 ± 7 vs. 50 ± 6 and 42 ± 4; P < 0.05). The rate of blastocyst development in COC + DO and OPU COC + DO groups was greater those in control and OPU COC groups (36.6% and 55.5% vs. 28.1% and 40.0%; P < 0.05). Moreover, the total cell numbers of blastocysts in COC + DO group exceeded that of control (132.91 ± 30.90 vs. 115.44 ± 24.95; P < 0.05), with no significant between OPU COC + DO and OPU COC groups (139.31 ± 42.51 vs. 137.00 ± 61.34). In conclusion, in vitro embryo development competence and quality improved when oocytes were cocultured with DO. Furthermore, there more, but smaller, ZP pores.

Proteomic identification of abnormally expressed proteins in early-stage placenta derived from cloned cat embryos.

It is unknown whether gene expression in cloned placenta during pre- and postimplantation is associated with early pregnancy failure in the cat. In this study, protein expression patterns were examined in early-stage (21-day-old) domestic cat placentas of fetuses derived from AI (CP; N = 4) and cloned embryo transfer (CEP; N = 2). Differentially expressed proteins were analyzed by two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight (TOF) mass spectrometry (MS). A total of 21 proteins were aberrantly expressed (P < 0.05) by >1.5-fold in CEP compared with CP. Compared with CP, 12 proteins were upregulated in CEP (peptidyl-prolyl cis-trans isomerase A, annexin A2, protein DJ-1, adenylate kinase isoenzyme 1, protein disulfide-isomerase A3, actin cytoplasmic 1, serum albumin, protein disulfide-isomerase A6, and triosephosphate isomerase), and nine proteins were downregulated (triosephosphate isomerase; heterogeneous nuclear ribonucleoprotein H; tropomyosin alpha-4; triosephosphate isomerase 1; 60 kDa heat shock protein, mitochondrial; serum albumin; calumenin; keratin type 1; and prohibitin). The identities of the differentially expressed proteins were validated by peptide mass fingerprinting using matrix-assisted laser desorption/ionization-TOF/TOF MS/MS. The abnormally expressed proteins identified in this study might be associated with impaired development and dysfunction of CEP during early pregnancy. Abnormal protein expression might also induce fetal loss and contribute to failure to maintain pregnancy to term.

Improved blastocyst development of single cow OPU-derived presumptive zygotes by group culture with agarose-embedded helper embryos.

BACKGROUND: The in vitro culture of presumed zygotes derived from single cow ovum pick-up (OPU) is important for the production of quality blastocysts maintaining pedigree. The aim of the present study was to evaluate the agar chip-embedded helper embryo coculture system for single cow OPU-derived zygotes by assessing embryo quality. METHODS: Cumulus oocyte complexes (COCs) were collected from Hanwoo cows with high genetic merit twice a week using the ultra-sound guided OPU technique and from slaughterhouse ovaries. The Hanwoo cow COCs and slaughterhouse ovaries were matured in vitro, fertilized in vitro with thawed Hanwoo sperm and cultured for 24 h. The presumed zygotes were subsequently placed in three different culture systems: (1) control OPU (controlOPU) with single cow OPU-derived presumed zygotes (2~8); (2) agar chip-embedded slaughterhouse helper embryo coculture (agarOPU) with ten presumed zygotes including all presumed zygotes from a cow (2~8) and the rest from agar chip-embedded slaughterhouse presumed zygotes (8~2); and (3) slaughterhouse in vitro embryo production (sIVP) with ten slaughterhouse ovary-derived presumed zygotes, each in 50 µL droplets. Day 8 blastocysts were assayed for apoptosis and gene expression using real time PCR. RESULTS: The coculture system promoted higher blastocyst development in OPU zygotes compared to control OPU zygotes cultured alone (35.2 vs. 13.9%; P < 0.01). Genes predicted to be involved in implantation failure and/or embryo resorption were down-regulated (P < 0.05) in control OPU zygotes (CD9, 0.4-fold; AKRAB1, 0.3-fold) and in cocultured zygotes (CD9, 0.3-fold; AKRAB1, 0.3-fold) compared to sIVP blastocysts (1.0-fold). Moreover, genes involved in implantation and/or normal calf delivery were up-regulated (P < 0.05 to P < 0.01) in control OPU zygotes (PGSH2, 5.0-fold; TXN, 4.3-fold; PLAU, 1.7-fold) and cocultured zygotes (PGSH2, 14.5-fold; TXN, 3.2-fold; PLAU, 6.8-fold) compared to sIVP (1.0-fold) blastocysts. However, the expression of PLAC8, TGF-ß1, ODC1, ATP5A1 and CASP3 did not differ between the three culture groups. CONCLUSIONS: Results show that the agar chip-embedded helper embryo coculture system enhances developmental competence and embryo quality in cultures of limited numbers of high pedigree single cow OPU presumed zygotes.

Enhanced histone acetylation in somatic cells induced by a histone deacetylase inhibitor improved inter-generic cloned leopard cat blastocysts.

The objective was to determine whether alterations of histone acetylation status in donor cells affected inter-generic SCNT (igSCNT)-cloned embryo development. Leopard cat cells were treated with trichostatin A (TSA; a histone deacetylase inhibitor) for 48 h, and then donor cells were transferred into enucleated oocytes from domestic cats. Compared to non-treated cells, the acetylated histone 3 at lysine 9 (AcH3K9) and histone 4 at lysine 5 (AcH4K5) in the TSA group increased for up to 48 h (P < 0.05). The AcH3K9 signal ratios of igSCNT group was higher than control group 3 h after activation (P < 0.05). Treatment with TSA significantly increased total cell number of blastocysts (109.1 ± 6.9 vs. 71.8 ± 2.9, mean ± SEM), with no significant effects on rates of cleavage or blastocyst development (71.1 ± 2.8 vs. 67.6 ± 2.9 and 12.2 ± 2.6 vs. 11.0 ± 2.6, respectively). When igSCNT cloned embryos were transferred into a domestic cat oviduct and recovered after 8 d, blastocyst development rates and total cell numbers were greater in the TSA-igSCNT group (20.7 ± 3.0% and 2847.6 ± 37.2) than in the control igSCNT group (5.7 ± 2.2% and 652.1 ± 17.6, P < 0.05). Average total cell numbers of blastocysts were approximately 4.4-fold higher in the TSA-igSCNT group (2847.6 ± 37.2, n = 10) than in the control group (652.1 ± 17.6, n = 8; P < 0.05), but were ∼2.9-fold lower than in vivo cat blastocysts produced by intrauterine insemination (8203.8 ± 29.6, n = 5; P < 0.001). Enhanced histone acetylation levels of donor cells improved in vivo developmental competence and quality of inter-generic cloned embryos; however, fewer cells in blastocysts derived from igSCNT than blastocysts produced by insemination may reduce development potential following intergeneric cloning (none of the cloned embryos were maintained to term).