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In dairy manure, a wide array of microorganisms, including many pathogens, survive and grow under suitable conditions. This microbial community offers a tremendous opportunity for studying animal health, the transport of microbes into the soil, air, and water, and consequential impacts on public health. The aim of this study was to assess the impacts of manure management practices on the microbial community of manure. The key novelty of this work is to identify the impacts of various stages of manure management on microbes living in dairy manure. In general, the majority of dairy farms in California use a flush system to manage dairy manure, which involves liquid-solid separations. To separate liquid and solid in manure, Multi-stage Alternate Dairy Effluent Management Systems (ADEMS) that use mechanical separation systems (MSS) or weeping wall separation systems (WWSS) are used. Thus, this study was conducted to understand how these manure management systems affect the microbial community. We studied the microbial communities in the WWSS and MSS separation systems, as well as in the four stages of the ADEMS. The 16S rRNA gene from the extracted genomic DNA of dairy manure was amplified using the NovoSeq Illumina next-generation sequencing platform. The sequencing data were used to perform the analysis of similarity (ANOSIM) and multi-response permutation procedure (MRRP) statistical tests, and the results showed that microbial communities among WWSS and MSS were significantly different ( p  < 0.05). These findings have significant practical implications for the design and implementation of manure management practices in dairy farms.

On a milk-producing dairy farm, milk production is correlated with manure production and the number of cattle, and manure is widely used as a soil fertilizer. However, excessive dairy manure production is linked with greenhouse gas emissions and water quality issues. On-farm planning of manure storage and application to enhance soil nutrients are essential in a circular economy to reduce environmental impact, where manure is not landfilled and incinerated. Instead, it creates a nutrient resource for crops and soil. Dairy manure, which is rich in nutrients, is a valuable fertilizer that contains many nutrients such as nitrogen (N), organic matter (OM), phosphorous (P), Potassium (K) and micronutrients. In this work, a pilot field research was conducted between 2016 and 2018 in various parts of California, USA (San Joaquin Valley, Sacramento Valley, Shasta Cascade, and the North Coast of California) to assess physio-chemical characteristics of solid fractions of dairy manure among various dairy farms. A total of 156 samples were collected from the gut ( n  = 107) and toe ( n  = 49) of the manure piles across California for determining total solid (TS), volatile solid (VS), temperature, moisture content and carbon-nitrogen ratio (C: N). Here, using the observations of field study and analysis, we show that C: N, OM and MC of solid fractions of dairy manure vary significantly among dairy farms. The average C: N ratio of manure (26-32) among various regions was close to an ideal C: N value of 24:1 for soil microbes to stimulate nutrient release to crops. Manure pH ranged between 7.0 and 8.0, which was close to an optimal pH range for common crops (6.0-8.0). Moreover, considering less cost and surplus availability, manure will likely continue providing a cost-effective organic fertilizer resource compared to commercial chemical fertilizers.

An increased understanding of the interaction between manure management and public and environmental health has led to the development of Alternative Dairy Effluent Management Strategies (ADEMS). The efficiency of such ADEMS can be increased using mechanical solid-liquid-separator (SLS) or gravitational Weeping-Wall (WW) solid separation systems. In this research, using pilot study data from 96 samples, the chemical, physical, biological, seasonal, and structural parameters between SLS and WW of ADEM systems were compared. Parameters including sodium, potassium, total salts, volatile solids, pH, and E. coli levels were significantly different between the SLS and WW of ADEMS. The separated solid fraction of the dairy effluents had the lowest E. coli levels, which could have beneficial downstream implications in terms of microbial pollution control. To predict effluent quality and microbial pollution risk, we used Escherichia coli as the indicator organism, and a versatile machine learning, ensemble, stacked, super-learner model called E-C-MAN ( Escherichia coli- Manure) was developed. Using pilot data, the E-C-MAN model was trained, and the trained model was validated with the test dataset. These results demonstrate that the heuristic E-C-MAN ensemble model can provide a pilot framework toward predicting Escherichia coli levels in manure treated by SLS or WW systems., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Shetty, Amaly, Weimer and Pandey.)

The Bcl-2 inhibitor venetoclax has yielded exceptional clinical responses in chronic lymphocytic leukemia (CLL). However, de novo resistance can result in failure to achieve negative minimal residual disease and predicts poor treatment outcomes. Consequently, additional proapoptotic drugs, such as inhibitors of Mcl-1 and Bcl-xL, are in development. By profiling antiapoptotic proteins using flow cytometry, we find that leukemic B cells that recently emigrated from the lymph node (CD69+/CXCR4Low) in vivo are enriched for cell clusters simultaneously overexpressing multiple antiapoptotic proteins (Mcl-1High/Bcl-xLHigh/Bcl-2High) in both treated and treatment-naive CLL patients. These cells exhibited antiapoptotic resistance to multiple BH-domain antagonists, including inhibitors of Bcl-2, Mcl-1, and Bcl-xL, when tested as single agents in a flow cytometry-based functional assay. Antiapoptotic multidrug resistance declines ex vivo, consistent with resistance being generated in vivo by extrinsic microenvironmental interactions. Surviving "persister" cells in patients undergoing venetoclax treatment are enriched for CLL cells displaying the functional and molecular properties of microenvironmentally induced multidrug resistance. Overcoming this resistance required simultaneous inhibition of multiple antiapoptotic proteins, with potential for unwanted toxicities. Using a drug screen performed using patient peripheral blood mononuclear cells cultured in an ex vivo microenvironment model, we identify novel venetoclax drug combinations that induce selective cytotoxicity in multidrug-resistant CLL cells. Thus, we demonstrate that antiapoptotic multidrug-resistant CLL cells exist in patients de novo and show that these cells persist during proapoptotic treatment, such as venetoclax. We validate clinically actionable approaches to selectively deplete this reservoir in patients., (© 2021 by The American Society of Hematology.)

Carnivore protoparvovirus 1 (CP1, earlier called Feline panleukopenia virus ) variants such as canine parvovirus (CPV) and feline parvovirus (FPV) are significant, emerging, multihost pathogens of domestic and wild carnivores. The diversity of CP1 variants was studied between 2008 and 2014 in Wayanad, India, where flagship wildlife species such as tigers ( Panthera tigris ) and leopards ( Panthera pardus ) coexist alongside domestic carnivores, including dogs ( Canis lupus familiaris ) and cats ( Felis catus ). Using polymerase chain reaction, FPV and CPV sequences were obtained from the heart blood of a necropsied leopard individual for the first time in the world and from rectal swabs of three sympatric and clinically ill domestic dogs. CP1 amplicons were also detected in a tiger. Cross-species transmission possibilities were identified, as the closest relatives to the leopard FPV sequence were found in domestic cats from a neighboring state.