Elucidating the Mechanisms of Cell-to-Cell Crosstalk in Probiotics Co-culture: A Proteomics Study of Limosilactobacillus reuteri ZJ625 and Ligilactobacillus salivarius ZJ614.

Limosilactobacillus reuteri ZJ625 and Ligilactobacillus salivarius ZJ614 are potential probiotic bacteria with improved benefits when administered to the host as a multi-strain preparation. To elucidate the mechanisms of cell-to-cell crosstalk between these two strains, we studied their intracellular and extracellular proteomes in co-culture by liquid-chromatography mass-spectrometry (LC-MS) using Dionex Nano-RSLC and fusion mass spectrometer. The experiment consisted of five biological replicates, and samples were collected during the mid-exponential growth phase. The quantitative proteomic profiles revealed several differentially expressed proteins (DEPs), which are down- or up-regulated between and within groups for both the intracellular and extracellular proteomes. These DEPs include proteins synthesising autoinducer-2, a sensor compound for cell-to-cell bacterial crosstalk during quorum sensing in mixed culture. Other important DEPs identified include enolase, phosphoglycerate kinase, and l-lactate dehydrogenase, which play roles in carbohydrate metabolism. Proteins associated with transcription, ATP production and transport across the membrane, DNA repair, and those with the potential to bind to the host epithelium were also identified. The post-translational modifications associated with the proteins include oxidation, deamidation, and ammonia loss. Importantly, this study revealed a significant expression of S-ribosylhomocysteine lyase (luxS) involved in synthesising autoinducer-2 that plays important roles in quorum sensing, aiding bacterial cell-to-cell crosstalk in co-cultures. The proteome of L. salivarius ZJ614 was most affected when co-cultured with L. reuteri ZJ625. In contrast, omitting some medium components from the defined medium exerted more effects on L. reuteri ZJ625 than L. salivarius ZJ614.

A Forty-Year Analysis of the Literature on Babesia Infection (1982-2022): A Systematic Bibliometric Approach.

Babesia infection is a tick-borne protozoan disease associated with significant veterinary, economic, and medical importance. This infection affects many hosts, ranging from wild to domestic animals and including man. All vertebrates serve as potential carriers due to the huge diversity of the species. Babesiosis has been associated with severe economic loss in livestock production, especially in cattle farming, and is also a major public health concern in man, which could be fatal. The infection is usually opportunistic, ranging from asymptomatic to symptomatic, usually in immunocompromised subjects or under conditions of stressful management. This study was designed to uncover trends in relation to publication growth and further explore research output regarding babesiosis from data indexed in the WoS. The WoS is the only platform used to map publications on Babesia infection. The search term "babesiosis" or "Babesia infection" was used to extract articles published across the study period from 1982 to 2022. The inclusion criteria were restricted to only articles for the analysis. The results from the search query showed that a total of 3763 articles were published during the study period with an average of 91.70 ± 43.87 articles annually and an average total citation (n = 1874.8). An annual growth rate of 2.5% was recorded during the study period. The year 2021 had the highest number of published articles (n = 193, 5.1%) and citations (n = 7039). The analysis of the most relevant keywords and titles showed that infection (n = 606, 16.1%), babesiosis (n = 444, 11.7%), and Babesia (n = 1302, 16%) were the most relevant keyword plus (ID), author keyword (DE), and title, respectively. The common conceptual framework analysis through K-means clustering showed two clusters comprising 4 and 41 elements, respectively. The United States of America is the top-performing country in terms of article production (n = 707, 20.8%) and the leading funder for babesiosis research, with two of its agencies ranked at the top. These are the Department of Health and Human Services (n = 254, 6.7%) and the National Institute of Health (n= 238,6.3%). Igarashi I. is the top-performing author (n = 231, 6.1%), while Veterinary Parasitology is ranked the top journal (n = 393, 10.4%) in terms of babesiosis publications. Overall, an increase in publications was observed in the study period, with significant output from developed nations.

'Multi-omics' data integration: applications in probiotics studies.

The concept of probiotics is witnessing increasing attention due to its benefits in influencing the host microbiome and the modulation of host immunity through the strengthening of the gut barrier and stimulation of antibodies. These benefits, combined with the need for improved nutraceuticals, have resulted in the extensive characterization of probiotics leading to an outburst of data generated using several 'omics' technologies. The recent development in system biology approaches to microbial science is paving the way for integrating data generated from different omics techniques for understanding the flow of molecular information from one 'omics' level to the other with clear information on regulatory features and phenotypes. The limitations and tendencies of a 'single omics' application to ignore the influence of other molecular processes justify the need for 'multi-omics' application in probiotics selections and understanding its action on the host. Different omics techniques, including genomics, transcriptomics, proteomics, metabolomics and lipidomics, used for studying probiotics and their influence on the host and the microbiome are discussed in this review. Furthermore, the rationale for 'multi-omics' and multi-omics data integration platforms supporting probiotics and microbiome analyses was also elucidated. This review showed that multi-omics application is useful in selecting probiotics and understanding their functions on the host microbiome. Hence, recommend a multi-omics approach for holistically understanding probiotics and the microbiome.

Accuracy of Machine Learning Classification Models for the Prediction of Type 2 Diabetes Mellitus: A Systematic Survey and Meta-Analysis Approach.

Soft-computing and statistical learning models have gained substantial momentum in predicting type 2 diabetes mellitus (T2DM) disease. This paper reviews recent soft-computing and statistical learning models in T2DM using a meta-analysis approach. We searched for papers using soft-computing and statistical learning models focused on T2DM published between 2010 and 2021 on three different search engines. Of 1215 studies identified, 34 with 136952 patients met our inclusion criteria. The pooled algorithm's performance was able to predict T2DM with an overall accuracy of 0.86 (95% confidence interval [CI] of [0.82, 0.89]). The classification of diabetes prediction was significantly greater in models with a screening and diagnosis (pooled proportion [95% CI] = 0.91 [0.74, 0.97]) when compared to models with nephropathy (pooled proportion = 0.48 [0.76, 0.89] to 0.88 [0.83, 0.91]). For the prediction of T2DM, the decision trees (DT) models had a pooled accuracy of 0.88 [95% CI: 0.82, 0.92], and the neural network (NN) models had a pooled accuracy of 0.85 [95% CI: 0.79, 0.89]. Meta-regression did not provide any statistically significant findings for the heterogeneous accuracy in studies with different diabetes predictions, sample sizes, and impact factors. Additionally, ML models showed high accuracy for the prediction of T2DM. The predictive accuracy of ML algorithms in T2DM is promising, mainly through DT and NN models. However, there is heterogeneity among ML models. We compared the results and models and concluded that this evidence might help clinicians interpret data and implement optimum models for their dataset for T2DM prediction.

Formulation of Chemically Defined Media and Growth Evaluation of Ligilactobacillus salivarius ZJ614 and Limosilactobacillus reuteri ZJ625.

Lactic acid bacteria are increasingly becoming important dietary supplements due to their health benefits when consumed in adequate quantity. The increasing attention on these important microbes has necessitated an in-depth understanding of their physiological processes, such as nutritional requirements and growth patterns, to better harness their probiotic potentials. This study was carried out to determine the nutritional requirements for the growth of L. salivarius ZJ614 and L. reuteri ZJ625 from a chemically defined medium and evaluate growth kinetics by fitting different sigmoidal growth models. The complete CDM contains 49 nutritional ingredients such as glucose, Tween 80®, mineral salts, buffers, amino acids, vitamins, and nucleotides at defined concentrations. In addition, the minimal nutritional requirements of the isolates were determined in a series of single-omission experiments (SOEs) to compose the MDM. Growth curve data were generated by culturing in an automated 96-well micro-plate reader at 37°C for 36 h, and photometric readings (optical density: OD600) were taken. The data were summarized in tables and charts using Microsoft Excel, while growth evaluation was carried out using open-source software (Curveball) on Python. The results revealed that omission of the amino acids, vitamins, and nucleotides groups resulted in 2.0, 20.17, and 60.24% (for L. salivarius ZJ614) and 0.95, 42.7, and 70.5% (for L. reuteri ZJ625) relative growths, respectively. Elimination of the individual CDM components also indicates varying levels of growth by the strains. The growth curve data revealed LogisticLag2 and Baranyi-Roberts models as the best fits for L. reuteri ZJ625 and L. salivarius ZJ614, respectively. All the strains showed appreciable growth on the CDM and MDM as observed in de Man-Rogosa-Sharpe (MRS) broth. We also described the growth kinetics of L. reuteri ZJ625 and L. salivarius ZJ614 in the CDM, and the best models revealed the estimated growth parameters.

Association between host genetics of sheep and the rumen microbial composition.

A synergy between the rumen microbiota and the host genetics has created a symbiotic relationship, beneficial to the host's health. In this study, the association between the host genetics and rumen microbiome of Damara and Meatmaster sheep was investigated. The composition of rumen microbiota was estimated through the analysis of the V3-V4 region of the 16S rRNA gene, while the sheep blood DNA was genotyped with Illumina OvineSNP50 BeadChip and the genome-wide association (GWA) was analyzed. Sixty significant SNPs dispersed in 21 regions across the Ovis aries genome were found to be associated with the relative abundance of seven genera: Acinetobacter, Bacillus, Clostridium, Flavobacterium, Prevotella, Pseudomonas, and Streptobacillus. A total of eighty-four candidate genes were identified, and their functional annotations were mainly associated with immunity responses and function, metabolism, and signal transduction. Our results propose that those candidate genes identified in the study may be modulating the composition of rumen microbiota and further indicating the significance of comprehending the interactions between the host and rumen microbiota to gain better insight into the health of sheep.

Subtractive Genomics Approach for Identification of Novel Therapeutic Drug Targets in Mycoplasma genitalium.

Mycoplasma genitalium infection is a sexually transmitted infection that causes urethritis, cervicitis, and pelvic inflammatory disease (PID) in men and women. The global rise in antimicrobial resistance against recommended antibiotics for the treatment of M. genitalium infection has triggered the need to explore novel drug targets against this pathogen. The application of a bioinformatics approach through subtractive genomics has proven highly instrumental in predicting novel therapeutic targets against a pathogen. This study aimed to identify essential and non-homologous proteins with unique metabolic pathways in the pathogen that could serve as novel drug targets. Based on this, a manual comparison of the metabolic pathways of M. genitalium and the human host was done, generating nine pathogen-specific metabolic pathways. Additionally, the analysis of the whole proteome of M. genitalium using different bioinformatics databases generated 21 essential, non-homologous, and cytoplasmic proteins involved in nine pathogen-specific metabolic pathways. The further screening of these 21 cytoplasmic proteins in the DrugBank database generated 13 druggable proteins, which showed similarity with FDA-approved and experimental small-molecule drugs. A total of seven proteins that are involved in seven different pathogen-specific metabolic pathways were finally selected as novel putative drug targets after further analysis. Therefore, these proposed drug targets could aid in the design of potent drugs that may inhibit the functionality of these pathogen-specific metabolic pathways and, as such, lead to the eradication of this pathogen.

Chloroquine and Sulfadoxine-Pyrimethamine Resistance in Sub-Saharan Africa-A Review.

Malaria is a great concern for global health and accounts for a large amount of morbidity and mortality, particularly in Africa, with sub-Saharan Africa carrying the greatest burden of the disease. Malaria control tools such as insecticide-treated bed nets, indoor residual spraying, and antimalarial drugs have been relatively successful in reducing the burden of malaria; however, sub-Saharan African countries encounter great challenges, the greatest being antimalarial drug resistance. Chloroquine (CQ) was the first-line drug in the 20th century until it was replaced by sulfadoxine-pyrimethamine (SP) as a consequence of resistance. The extensive use of these antimalarials intensified the spread of resistance throughout sub-Saharan Africa, thus resulting in a loss of efficacy for the treatment of malaria. SP was replaced by artemisinin-based combination therapy (ACT) after the emergence of resistance toward SP; however, the use of ACTs is now threatened by the emergence of resistant parasites. The decreased selective pressure on CQ and SP allowed for the reintroduction of sensitivity toward those antimalarials in regions of sub-Saharan Africa where they were not the primary drug for treatment. Therefore, the emergence and spread of antimalarial drug resistance should be tracked to prevent further spread of the resistant parasites, and the re-emergence of sensitivity should be monitored to detect the possible reappearance of sensitivity in sub-Saharan Africa.

Effects of Lactobacillus rhamnosus and Enterococcus faecalis Supplementation as Direct-Fed Microbials on Rumen Microbiota of Boer and Speckled Goat Breeds.

The effects on rumen microbial communities of direct-fed probiotics, Lactobacillus rhamnosus and Enterococcus faecalis, singly and in combination as feed supplements to both the Boer and Speckled goats were studied using the Illumina Miseq platform targeting the V3-V4 region of the 16S rRNA microbial genes from sampled rumen fluid. Thirty-six goats of both the Boer and Speckled were divided into five experimental groups: (T1) = diet + Lactobacillus rhamnosus; (T2) = diet + Enterococcus faecalis; (T3) = diet + Lactobacillus rhamnosus + Enterococcus faecalis; (T4, positive control) = diet + antibiotic and (T5, negative control) = diet without antibiotics and without probiotics. Our results revealed that Bacteroidetes, Firmicutes, TM7, Proteobacteria, and Euryarchaeota dominate the bacterial communities. In our observations, Lactobacillus rhamnosus and Enterococcus faecalis supplements reduced the archaeal population of Methanomassiliicocca in the T1, T2 and T3 groups, and caused an increase in the T4 group. Chlamydiae were present only in the T5 group, suggesting that probiotic and antibiotic inhibit the growth of pathogens in the rumen. We inferred, based on our results, that Lactobacillus rhamnosus and Enterococcus faecalis favour the survival of beneficial microbial communities in the goats' rumen. This may lead to an overall improved feed efficacy and growth rate.

Understanding the interactions between Eimeria infection and gut microbiota, towards the control of chicken coccidiosis: a review.

The gastrointestinal tract in poultry harbours a diverse microbial community that serves a crucial role in digestion and protection. Disruption of the gut environment due to Eimeria spp. parasite infection causes an imbalance in intestinal homeostasis, driving the increment of pathogens such as Clostridium species. Coccidiosis infection affects the composition and integrity of gut microbiota, resulting in elevated susceptibility to diseases that pose a serious threat to the overall health and productivity of chickens. Anticoccidial drugs have proven effective in curbing coccidiosis but with concerning drawbacks like drug resistance and drug residues in meat. The exploration of natural alternative strategies such as probiotics and phytochemicals is significant in controlling coccidiosis through modification and restoration of gut microbiota, without inducing drug resistance. Understanding the interaction between Eimeria parasites and gut microbiota is crucial for the control and prevention of coccidiosis, and the development of novel alternative treatments.

TITLE: Comprendre l'interaction entre l'infection à Eimeria et le microbiote intestinal pour lutter contre la coccidiose du poulet : une synthèse. ABSTRACT: Le tractus gastro-intestinal de la volaille abrite une communauté microbienne diversifiée qui joue un rôle crucial dans la digestion et la protection. La perturbation de l'environnement intestinal due à l'infection par le parasite Eimeria spp. provoque un déséquilibre de l'homéostasie intestinale, entraînant une prolifération des agents pathogènes tels que les espèces de Clostridium. L'infection par la coccidiose affecte la composition et l'intégrité du microbiote intestinal, ce qui entraîne une sensibilité élevée aux maladies qui ont un impact sérieux sur la santé globale et la productivité des poulets. Les médicaments anticoccidiens se sont avérés efficaces pour freiner la coccidiose, mais avec des effets négatifs inquiétants comme la résistance aux médicaments et la présence de résidus de médicaments dans la viande. L'exploration de stratégies alternatives naturelles telles que les probiotiques et les composés phytochimiques est importante pour le contrôle de la coccidiose par la modification et la restauration du microbiote intestinal, sans induire de résistance aux médicaments. Comprendre l'interaction entre les parasites Eimeria et le microbiote intestinal est crucial pour le contrôle et la prévention de la coccidiose et le développement de nouveaux traitements alternatifs.

Transgenic Eimeria parasite: A potential control strategy for chicken coccidiosis.

Poultry industry has been very instrumental in curtailing malnutrition and poverty and as such contributing to economic growth. However, production loss in poultry industry due to parasitic disease such as coccidiosis has become a global challenge. Chicken coccidiosis is an enteric disease that is associated with morbidity and mortality. The control of this parasite through anticoccidial live vaccines and drugs has been very successful though with some limitations such as the cost of production of live vaccines, and drugs resistance which is a public health concern. The discovery of Eimeria vaccine antigens such as Apical membrane antigens (AMA)-1 and Immune mapped protein (IMP)-1 have introduced the use of recombinant vaccines as alternative control measures against chicken coccidiosis. Although some protections have been reported among recombinant vaccines, improving their protective efficacy has triggered the search for a novel and efficient delivery vehicle. Transgenic Eimeria, which is constructed either through stable or transient transfection is currently being explored as novel delivery vehicle of Eimeria vaccine antigens. Due to partial protections reported in chickens vaccinated with transgenic Eimeria lines expressing different Eimeria antigens, improving protective efficacy becomes imperative. Recent trends in the design of transgenic Eimeria for potential application in the control of chicken coccidiosis are summarized in this review. We conclude that, with improved protective efficacy using multiple vaccine antigens, transgenic Eimeria parasite could fill the gap in the control of chicken coccidiosis as an efficient anticoccidial vaccine.

Diagnosis and control of chicken coccidiosis: a recent update.

Coccidiosis is a deadly disease that hampers chicken's productivity and welfare. Thus, the disease is a major menace to the global poultry industry. Coccidiosis which is caused by the apicomplexan parasite of the genus Eimeria has seven known species which affect the different parts of the intestinal tract of chickens. The disease which occurs by ingestion of sporulated oocyst has been associated with poor poultry management system. Mixed infection among the species of this parasite contributes to both pathogenicity and misdiagnosis of the disease. A progress in identification and diagnosis approach which cuts across pathological, morphological and molecular has been reported for this parasite. Control measures which include anticoccidial drugs, vaccines and natural products have dominated literature for this disease. However, the emergence of genetic and antigenic diversity with implication on resistance to anticoccidials among different strains of Eimeria parasite has generated concerns on the effectiveness of the current anticoccidial vaccines. A new look on the control strategy therefore becomes imperative. This study reviews the current trends on the identification and control of chicken coccidiosis with focus on (1) Avian coccidiosis (2) Epidemiology of chicken coccidiosis (3) Eimeria parasite and distribution in poultry (4) Diagnosis of Eimeria parasite (5) Control measures of coccidiosis (6) Threats posed by genetic and antigenic diversity of Eimeria parasite on coccidiosis control. Genomic study on diversity of Eimeria parasite becomes imperative for effective vaccine design against coccidiosis.

Genotype effect on distribution pattern of maternally derived antibody against Newcastle disease in Nigerian local chickens.

This study was conducted to evaluate the variation in antibody from Newcastle disease (ND) vaccinated breeder birds and their progenies. The chicks were evaluated for maternally derived antibody (MDA) against ND virus and also the MDA transfer rate for each genotype. The local parent stocks were Frizzle-feathered, Normal-feathered, and Naked neck while an exotic parent stock (Anak Titan) was used for the purpose of comparison. Sera samples were collected from chicks generated at day 3 post-hatch, and sera were also collected from the parent stocks. The sera samples were stored at -20 °C until analysed. Enzyme linked immuno-sorbent assay (ELISA) test was used for the detection and quantification of antibodies to ND in the parent stocks and the chicks using IDEXX Newcastle Disease ELISA test kit. The least squares means of antibody titers for parent stocks were 413.333 ± 68.853, 426.333 ± 121.949, 481.667 ± 145.454, and 1148.333 ± 532.843 for Frizzle-feathered, Normal-feathered, Naked Neck, and Anak Titan, and the calculated MDA transfer rate was 96.4, 93.8, 88.7, and 34.7 % for Frizzle-feathered, Normal-feathered, Naked neck, and Anak Titan, respectively. The mean antibody titers for chicks were 398.6 ± 43.871, 400.0 ± 53.952, 427.0 ± 89.353, and 398.8 ± 57.593 for Frizzle-feathered, Normal-feathered, Naked neck, and Anak Titan, respectively. The antibody titers from parent stocks were higher than those observed in chicks for all the genotypes. The values obtained in this study indicated that parent stocks with their progenies were below protective level. The local parent stocks had higher transfer rate when compared to their exotic counterpart. Hence, there is an indication from this finding that the local chickens had a better transfer rate in relation to Anak Titan which is an exotic breed. Therefore, there is the presence of genetic variability for antibody transfer in the Nigerian local chickens and can be incorporated in designing a breeding program for better adaptive potential.

Multivariate characterisation of the phenotypic traits of Djallonke and Sahel sheep in Northern Ghana.

The characterisation of the small ruminant populations in developing countries will play a major role in the maintenance of the genetic resources as the basis for future improvement in livestock production. The present study aimed at morphological characterisation of the two main breeds of sheep in Ghana by assessing variation within and between breed populations using principal component and discriminant analyses. The two breeds were the Sahel and the Djallonke sheep of both sexes and of two groups namely, young (1 year old, consisting of 74 animals) and mature sheep (≥ 2 years old, comprising 219 animals). The analysis of variance revealed significant (P < 0.05) differences in the morphological traits of the Sahel and the Djallonke sheep breeds with higher values recorded for the former. Sexual dimorphism was in favour of male animals in all the morphological traits examined. Mature animals also had comparative advantage over the young. Two principal components were extracted to discern the structure of the two genetic groups. The most discriminating traits between the two sheep breeds were rump height, height at withers, neck girth and pin-bone width. Mahalanobis distance between the two genetic groups was 5.723 (P < 0.0001). The developed discriminant functions clearly discriminated and classified the Sahel and the Djallonke sheep into their breeds of origin, thus yielding 100, 93.4 and 90.4 % accurate classification for the rams, ewes and the overall sheep population, respectively. The present approach would greatly help in establishing management and conservation policies for the sustainable production of the two Ghanaian sheep breeds.

Application of principal component and discriminant analyses to morpho-structural indices of indigenous and exotic chickens raised under intensive management system.

The objectives of this study were to investigate the relationships between body weight and morpho-structural indices to predict body weight from their orthogonal body shape characters using principal component analysis and to morphologically classify the chicken genotypes using multivariate discriminant analysis. Data used were from 273 randomly selected 12-weeks-old indigenous chickens of normal-feathered (NF), frizzle-feathered (FF), naked-neck (NN) and Anak Titan (AT) genotypes. Phenotypic correlation among body weight and most biometric traits ranged from 0.227-0.876, -0.7-0.901, 0.034-0.968 and -0.207-0.849 for NF, NN and AT chickens, respectively. Factor analysis with varimax rotation of interrelated traits revealed three principal components which accounted for 83.1%, 74.4%, 78.8% and 76.5% of the total variance in NF, FF, NN and AT chickens in the order listed. Breast girth, keel length, thigh length, shank length and wing length were found to be the most discriminating variables to separate the chicken genotypes. The longest distance (72.54) occurred between AT and NF genotypes while the shortest distance (4.27) was recorded for FF and NN genotypes. Classification results showed that 85.2% of AT genotype was correctly classified into their source population. However, 22.7% of NF was misclassified as NN, while 33.3% of NN was misclassified as NF chickens. These results suggest that there is high rate of gene flow between these two indigenous chicken genotypes. Information obtained from this study may be considered useful in breed improvement programmes for selection, characterization, conservation and better management of Nigerian indigenous chickens.

Effect of crossbreeding on fertility, hatchability and embryonic mortality of Nigerian local chickens.

A total of 970 eggs were collected from matings involving three genotypes of Nigerian local chickens (Normal-feathered, Frizzle-feathered and Naked neck) and one exotic broiler breeder strain (Anak Titan) to evaluate the effect of crossbreeding on fertility, hatchability and embryonic mortality. Mating was achieved through artificial insemination. Sire genotype significantly (P < 0.05) affected percent fertility and percent dead-in-shell. Naked neck sire genotype had the highest dead-in-shell (19.5%) with the least being 7.5% for Frizzle-feathered chickens. Frizzle-feathered sire genotype had the highest fertility (90.5%) and hatchability (91.4%). Dam genotype had a significant effect (P < 0.01) on fertility and hatchability, with Anak Titan dam having the highest fertility and hatchability of 88.2% and 94.6%, respectively. Fertility and hatchability were significantly (P < 0.05) influenced by the interactive effect of sire and dam genotypes with Frizzle-feathered × Anak Titan having 98.5% fertility and 96.8% hatchability. Both straight and reciprocal crosses involving Frizzle-feathered genotype and Anak Titan resulted in higher mean values for fertility and hatchability when compared to crosses involving the other two local chicken genotypes. This result suggests that the use of the Frizzle-feathered genotype appears to be more desirable in fertility and hatchability trials.