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\"A sweet and silly buffalo who tries to bluff and fluff his way into being bigger than he really is\"-- Provided by publisher.

Mitochondria play a pivotal role in energy production, metabolism, and cellular signaling, serving as key regulators of cellular functions, including differentiation and tissue-specific adaptation. The interplay between mitochondria and the nucleus is crucial for coordinating these processes, particularly through the supply of metabolites for epigenetic modifications that facilitate nuclear-mitochondrial interactions. To investigate tissue-specific mitochondrial adaptations at the molecular level, we conducted RNA sequencing data analyses of kidney, heart, brain, and ovary tissues of female buffaloes, focusing on variations in mitochondrial gene expression related to amino acid metabolism. Our analysis identified 82 nuclear-encoded mitochondrial transcripts involved in amino acid metabolism, with significant differential expression patterns across all tissues. Notably, the heart, brain, and kidney-tissues with higher energy demands-exhibited elevated expression levels compared to the ovary. The kidney displayed unique gene expression patterns, characterized by up-regulation of genes involved in glyoxylate metabolism and amino acid catabolism. In contrast, comparative analysis of the heart and kidney versus the brain revealed shared up-regulation of genes associated with fatty acid oxidation. Gene ontology and KEGG pathway analyses confirmed the enrichment of genes in pathways related to amino acid degradation and metabolism. These findings highlight the tissue-specific regulation of mitochondrial gene expression linked to amino acid metabolism, reflecting mitochondrial adaptations to the distinct metabolic and energy requirements of different tissues in buffalo. Importantly, our results underscore the relevance of mitochondrial adaptations not only for livestock health but also for understanding metabolic disorders in humans. By elucidating the molecular mechanisms of mitochondrial function and their tissue-specific variations, this study provides insights that could inform breeding strategies for enhanced livestock productivity and contribute to therapeutic approaches for human metabolic diseases. Thus, our findings illustrate how mitochondria are specialized in a tissue-specific manner to optimize amino acid utilization and maintain cellular homeostasis, with implications for both animal welfare and human health.

Smallholder large ruminant production in Lao People's Democratic Republic (Laos) is characterised by low reproductive efficiency. To determine if common abortifacient bovid infectious diseases are involved, a serological investigation was conducted. Sera was collected from stored and fresh cattle (n = 390) and buffalo (n = 130) samples from 2016-18 from, and then examined for associations in a retrospective risk factor study of 71 herds. The sera were assayed for antibodies to Neospora caninum, bovine viral diarrhoea virus (BVDV), Leptospira interrogans serovar Hardjo and Brucella abortus using commercially available enzyme-linked immunosorbent assay kits. These pathogens were detected in buffalo samples at 78.5% (95% CI 71.4-85.6), 0%, 2.3% (95% CI 0-4.9) and 0%, respectively, and in cattle at 4.4% (95% CI 2.4-6.4), 7.7% (95% CI 3.1-12.3), 12.8% (95% CI 9.5-16.1) and 0.26% (95% CI 0-0.8), respectively. Exposure of buffalo to N. caninum was positively associated with buffalo age, with a predicted seropositivity at birth of 52.8%, increasing to 97.2% by 12 years of age (p = 0.037). Exposure of cattle to L. interrogans serovar Hardjo was more prevalent in females compared to males, was associated with higher titres of BVDV, and was more prevalent in the wet season compared to the dry season. Exposure of cattle to BVDV was more prevalent in males compared to females, the wet and dry seasons were comparable, and was associated with rising antibody titres against N. caninum and L. interrogans serovar Hardjo. The risk factor survey identified that the probability of herds being N. caninum positive increased with farmer age, if farmers believed there were rodents on farm, and if farmers weren't aware that canids or rodents could contaminate bovid feed on their farm. The probability of a herd being positive to L. interrogans serovar Hardjo increased on farms where multiple cows shared the same bull, where farmers had lower husbandry knowledge, and on farms that used water troughs. The probability of a herd being BVDV seropositive increased with increasing herd size and increasing titres to N. caninum. The benchmarking of bovid exposure to emerging abortifacient pathogens and identification of their risk factors potentially informs disease prevention strategies, supporting efforts to establish a biosecure beef supply for enhanced smallholder livestock productivity, public health and food security in Laos and surrounding countries.

The physiological well-being of buffaloes, encompassing phenotypic traits, reproductive health, and productivity, depends on their energy status. Mitochondria, the architects of energy production, orchestrate a nuanced interplay between nuclear and mitochondrial domains. Oxidative phosphorylation complexes and associated proteins wield significant influence over metabolic functions, energy synthesis, and organelle dynamics, often linked to tissue-specific pathologies. The unexplored role of ATP synthase in buffalo tissues prompted a hypothesis: in-depth exploration of nuclear-derived mitochondrial genes, notably ATP synthase, reveals distinctive tissue-specific diversity. RNA extraction and sequencing of buffalo tissues (kidney, heart, brain, and ovary) enabled precise quantification of nuclear-derived mitochondrial protein gene expression. The analysis unveiled 24 ATP synthase transcript variants, each with unique tissue-specific patterns. Kidney, brain, and heart exhibited elevated gene expression compared to ovaries, with 10, 8, and 19 up-regulated genes, respectively. The kidney showed 3 and 12 down-regulated genes compared to the brain and heart. The heart–brain comparison highlighted ten highly expressed genes in ATP synthase functions. Gene ontology and pathway analyses revealed enriched functions linked to ATP synthesis and oxidative phosphorylation, offering a comprehensive understanding of energy production in buffalo tissues. This analysis enhances understanding of tissue-specific gene expression, emphasizing the influence of energy demands. Revealing intricate links between mitochondrial gene expression and tissue specialization in buffaloes, it provides nuanced insights into tissue-specific expression of nuclear-encoded mitochondrial protein genes, notably ATP synthase, advancing the comprehension of buffalo tissue biology.

The goals of ruminant protein nutrition are to provide sufficient rumen-degradable protein (RDP) for optimizing nitrogen utilization efficiency and achieve the desired level of animal productivity, all while minimizing dietary crude protein (CP) intake. The present study aimed to examine the effects of decreasing dietary protein by reducing RDP on the growth performance, energy, and nitrogen metabolism of dairy buffalo heifers. Three diets were formulated: high RDP (H-RDP, RDP =  80.36 g/kg, C P =  15.42%), medium RDP (M-RDP, RDP =  72.79 g/kg, C P =  14.03%), and low RDP (L-RDP, RDP =  66.48 g/kg, C P =  12.87%), yielding digestible crude protein (DCP) balances of 166.07, 114.89, and 66.59 g/d, respectively. The diets were fed to 36 heifers (259.19 ± 3.93 kg) divided into three groups (12 heifers per treatment) over a 51-day trial. The results showed that when dietary RDP decreased from 80.36 g/kg to 72.79 g/kg, the average daily gain (ADG) and dry matter intake (DMI) remained stable; however, a further decrease to 66.48 g/kg resulted in a 19.12% decrease in ADG and an 6.80% decrease in DMI ( P  < 0.05). The feed-to-gain (F/G) ratio increased linearly with decreasing dietary RDP (P  < 0.01) and was significantly lower in the M-RDP and H-RDP groups than that in the L-RDP group ( P  < 0.05). The apparent digestibility of CP decreased linearly ( P =  0.01) and was significantly higher ( P  < 0.05) in the H-RDP group than in the L-RDP group. There was a linear increase in fecal nitrogen efficiency (FN)/IN, which was significantly lower ( P  < 0.05) in the H-RDP group than in the L-RDP group. Reducing dietary RDP linearly decreased urinary nitrogen excretion (UN) ( P  < 0.01) without affecting the efficiency of the retained nitrogen (RN), intake nitrogen (IN), and energy profiles. Overall, this study revealed that reducing dietary RDP (from 80.36 to 72.79 g/kg) and CP (from 15.42% to 14.03%) effectively reduced UN excretion while maintaining dairy buffalo heifer performance. Collectively, these findings suggest an optimal dietary RDP range of 72.79–80.36 g/kg for 13- to 15-month-old dairy buffalo heifers.

Buffalo physiology intricately balances energy, profoundly influencing health, productivity, and reproduction. This study explores nuclear-mitochondrial crosstalk, revealing OXPHOS Complex I gene expression variations in buffalo tissues through high-throughput RNA sequencing. Unveiling tissue-specific disparities, the research elucidates the genomic landscape of crucial energy production genes, with broader implications for veterinary and agricultural progress. Post-slaughter, tissues from post-pubertal female buffaloes underwent meticulous processing and RNA extraction using the TRIzol method. RNA-Seq library preparation and IlluminaHiSeq 2500 sequencing were performed on QC-passed samples. Data underwent stringent filtration, mapping to the Bubalus bubalis genome using HISAT2. DESeq2 facilitated differential expression gene (DEG) analysis focusing on 57 Mitocarta 3-derived genes associated with OXPHOS complex I. Nuclear-encoded mitochondrial protein transcripts of OXPHOS complex 1 exhibited tissue-specific variations, with 51 genes expressing significantly across tissues. DEG analysis emphasized tissue-specific expression patterns, highlighting a balanced OXPHOS complex I subunit expression in the kidney vs. brain. Gene Ontology (GO) enrichment showcased mitochondria-centric terms, revealing distinct proton motive force-driven mitochondrial ATP synthesis regulation. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses emphasized Thermogenesis and OXPHOS pathways, enriching our understanding of tissue-specific energy metabolism. Noteworthy up-regulation of NDUFB10 in the heart and kidney aligned with heightened metabolic activity. Brain-specific up-regulation of NDUFAF6 indicated a focus on mitochondrial function, while variations in NDUFA11 and ACAD9 underscored pivotal roles in the heart and kidney. GO and KEGG analyses highlighted tissue-specific mitochondrial ATP synthesis and NADH dehydrogenase processes, providing molecular insights into organ-specific metabolic demands and regulatory mechanisms. Our study unveils conserved and tissue-specific nuances in nuclear-encoded mitochondrial OXPHOS complex I genes, laying a foundation for understanding diverse energy demands and potential health implications.

Background Cellular metabolism is most invariant process, occurring in all living organisms, which involves mitochondrial proteins from both nuclear and mitochondrial genomes. The mitochondrial DNA (mtDNA) copy number, protein-coding genes (mtPCGs) expression, and activity vary between various tissues to fulfill specific energy demands across the tissues. Methods and Results In present study, we investigated the OXPHOS complexes and citrate synthase activity in isolated mitochondria from various tissues of freshly slaughtered buffaloes ( n  = 3). Further, the evaluation of tissue-specific diversity based on the quantification of mtDNA copy numbers was performed and also comprised an expression study of 13 mtPCGs. We found that the functional activity of individual OXPHOS complex I was significantly higher in the liver compared to muscle and brain. Additionally, OXPHOS complex III and V activities was observed significantly higher levels in liver compared to heart, ovary, and brain. Similarly, CS-specific activity differs between tissues, with the ovary, kidney, and liver having significantly greater. Furthermore, we revealed the mtDNA copy number was strictly tissue-specific, with muscle and brain tissues exhibiting the highest levels. Among 13 PCGs expression analyses, mRNA abundances in all genes were differentially expressed among the different tissue. Conclusions Overall, our results indicate the existence of a tissue-specific variation in mitochondrial activity, bioenergetics, and mtPCGs expression among various types of buffalo tissues. This study serves as a critical first stage in gathering vital comparable data about the physiological function of mitochondria in energy metabolism in distinct tissues, laying the groundwork for future mitochondrial based diagnosis and research.

Studies in humans and laboratory animals link stable gut microbiome “enterotypes” with long-term diet and host health. Understanding how this paradigm manifests in wild herbivores could provide a mechanistic explanation of the relationships between microbiome dynamics, changes in dietary resources, and outcomes for host health. We identify two putative enterotypes in the African buffalo gut microbiome. The enterotype prevalent under resource-abundant dietary regimes, regardless of environmental conditions, has high richness, low between- and within-host beta diversity, and enrichment of genus Ruminococcaceae-UCG-005 . The second enterotype, prevalent under restricted dietary conditions, has reduced richness, elevated beta diversity, and enrichment of genus Solibacillus . Population-level gamma diversity is maintained during resource restriction by increased beta diversity between individuals, suggesting a mechanism for population-level microbiome resilience. We identify three pathogens associated with microbiome variation depending on host diet, indicating that nutritional background may impact microbiome-pathogen dynamics. Overall, this study reveals diet-driven enterotype plasticity, illustrates ecological processes that maintain microbiome diversity, and identifies potential associations between diet, enterotype, and disease. There are stable relationships between diet and microbiome in humans and lab animals. A study on African buffalo finds that diet influences microbiome variation and enterotype formation. Three pathogens may associate with microbiome depending on host diet, suggesting nutrition impacts relationships between gut microbiome and host health.

Background Buffaloes are crucial to agriculture, yet mitochondrial biology in these animals is less studied compared to humans and laboratory animals. This research examines tissue-specific variations in mitochondrial succinate dehydrogenase (SDH) gene expression across buffalo kidneys, hearts, brains, and ovaries. Understanding these variations sheds light on mitochondrial energy metabolism and its impact on buffalo health and productivity, revealing insights into enzyme regulation and potential improvements in livestock management. Materials and methods RNA-seq data from buffalo kidney, heart, brain, and ovary tissues were reanalyzed to explore mitochondrial SDH gene expression. The expression of SDH subunits ( SDHA , SDHB , SDHC , SDHD ) and assembly factors ( SDHAF1 , SDHAF2 , SDHAF3 , SDHAF4 ) was assessed using a log2 fold-change threshold of + 1 for up-regulated and − 1 for down-regulated transcripts, with significance set at p  < 0.05. Hierarchical clustering and differential expression analyses were performed to identify tissue-specific expression patterns and regulatory mechanisms, while Gene Ontology and KEGG pathway analyses were conducted to uncover functional attributes and pathway enrichments across different tissues. Results Reanalysis of RNA-seq data from different tissues of healthy female buffaloes revealed distinct expression patterns for SDH subunits and assembly factors. While SDHA , SDHB , and SDHC showed variable expression across tissues, SDHAF2 , SDHAF3 , and SDHAF4 exhibited tissue-specific profiles. Significant up-regulation of SDHA , SDHB , and several assembly factors was observed in specific tissue comparisons, with fewer down-regulated transcripts. Gene ontology and KEGG pathway analyses linked the up-regulated transcripts to mitochondrial ATP synthesis and the respiratory electron transport chain. Notably, tissue-specific variations in mitochondrial function were particularly evident in the ovary. Conclusion This study identifies distinct SDH gene expression patterns in buffalo tissues, highlighting significant down-regulation of SDHA , SDHB , SDHC , and assembly factors in the ovary. These findings underscore the critical role of mitochondria in tissue-specific energy production and metabolic regulation, suggest potential metabolic adaptations, and emphasize the importance of mitochondrial complex II. The insights gained offer valuable implications for improving feed efficiency and guiding future research and therapies for energy metabolism disorders.

Abstract The objective of the present study was to characterize non-classical class I major histocompatibility complex (MHC) genes in buffaloes and evaluate the expression of these genes in different tissue components of the placenta of buffaloes during pregnancy and in trophoblastic cells after stimulation using lipopolysaccharide (LPS). To do this, DNA was extracted from the blood of buffaloes and was subjected to PCR testing and sequencing of the genes NC3 and MICB. The RNA extracted from the placentome and intercotyledonary region of buffaloes in their first (n = 6), second (n = 6) and third (n = 6) trimesters of gestation was subjected to real-time PCR. Explants were created using the chorioallantoic membrane and two experimental groups were established: control and stimulated with LPS for four hours to evaluate the gene expression profile. Analysis on the sequences obtained showed that the genes NC3 and MICB of buffaloes were homologous with those of cattle, with high similarity in the analysis on the sequence variation pattern. The gene expression analysis showed that the genes assessed were transcribed at stages and in placental tissue that differed from what was seen in cattle. The transcription of these genes varied in the tissues studied, with greater transcription of MICB in the intercotyledonary region over the first third of gestation, while the genes studied in the placentome presented low rates of transcription. The trophoblastic cells of the chorioallantoic membrane stimulated with LPS for six hours did not present non-classic MFC-I transcription alterations. The present study therefore provides additional knowledge regarding the immune regulation of placental tissues of buffaloes. Resumo O objetivo do presente estudo foi caracterizar genes não clássicos do complexo principal de histocompatibilidade (MHC) de classe I em búfalas e avaliar a expressão desses genes em diferentes componentes teciduais da placenta de búfalas durante a gestação e em células trofoblásticas após estimulação com lipopolissacarídeo (LPS). Para isso, o DNA foi extraído do sangue de búfalos e submetido a testes de PCR e sequenciamento dos genes NC3 e MICB. O RNA extraído do placentônio e região intercotiledonar de búfalas no primeiro (n = 6), segundo (n = 6) e terceiro (n = 6) trimestres de gestação foi submetido à PCR em tempo real. Explantes foram criados usando a membrana corioalantóide e dois grupos experimentais foram estabelecidos: controle e estimulado com LPS por quatro horas para avaliar o perfil de expressão gênica. A análise das sequências obtidas mostrou que os genes NC3 e MICB de búfalos foram homólogos aos de bovinos, com alta similaridade na análise do padrão de variação de sequência. A análise da expressão gênica mostrou que os genes avaliados foram transcritos em estágios e em tecido placentário diferentes do observado em bovinos. A transcrição desses genes variou nos tecidos estudados, com maior transcrição de MICB na região intercotiledonar ao longo do primeiro terço da gestação, enquanto os genes estudados no placentoma apresentaram baixas taxas de transcrição. As células trofoblásticas da membrana corioalantóide estimuladas com LPS por seis horas não apresentaram alterações não clássicas na transcrição da MFC-I. O presente estudo, portanto, fornece conhecimento adicional sobre a regulação imune de tecidos placentários de búfalos.