The Development of a Risk Assessment Model for Inedible Rendering Plants in Canada: Identifying and Selecting Feed Safety-Related Factors.

The Canadian Food Inspection Agency (CFIA) is developing an establishment-based risk assessment model to categorize rendering plants that produce livestock feed ingredients (ERA-Renderer model) according to animal and human health risks (i.e., feed safety risks) and help in determining the allocation of inspection resources based on risk. The aim of the present study was to identify and select feed-safety-related factors and assessment criteria for inclusion in the ERA-Renderer model. First, a literature review was performed to identify evidence-based factors that impact the feed safety risk of livestock feed during its rendering processes. Secondly, a refinement process was applied to retain only those that met the inclusion conditions, such as data availability, lack of ambiguity, and measurability. Finally, an expert panel helped in selecting factors and assessment criteria based on their knowledge and experience in the rendering industry. A final list of 32 factors was developed, of which 4 pertained to the inherent risk of a rendering plant, 8 were related to risk mitigation strategies, and 20 referred to the regulatory compliance of a rendering plant. A total of 179 criteria were defined to assess factors based on practices in the Canadian rendering industry. The results of this study will be used in the next step of the model development to estimate the relative risks of the assessment criteria considering their impact on feed safety. Once implemented, the CFIA's ERA-Renderer model will provide an evidence-based, standardized, and transparent approach to help manage the feed safety risks in Canada's rendering sector.

Current Understanding of Equine Gut Dysbiosis and Microbiota Manipulation Techniques: Comparison with Current Knowledge in Other Species.

Understanding the importance of intestinal microbiota in horses and the factors influencing its composition have been the focus of many studies over the past few years. Factors such as age, diet, antibiotic administration, and geographic location can affect the gut microbiota. The intra- and inter-individual variability of fecal microbiota in horses complicates its interpretation and has hindered the establishment of a clear definition for dysbiosis. Although a definitive causal relationship between gut dysbiosis in horses and diseases has not been clearly identified, recent research suggests that dysbiosis may play a role in the pathogenesis of various conditions, such as colitis and asthma. Prebiotics, probiotics, and fecal microbiota transplantation to modulate the horse's gastrointestinal tract may eventually be considered a valuable tool for preventing or treating diseases, such as antibiotic-induced colitis. This article aims to summarize the current knowledge on the importance of intestinal microbiota in horses and factors influencing its composition, and also to review the published literature on methods for detecting dysbiosis while discussing the efficacy of gut microbiota manipulation in horses.

The Microbiota and Equine Asthma: An Integrative View of the Gut-Lung Axis.

Both microbe-microbe and host-microbe interactions can have effects beyond the local environment and influence immunological responses in remote organs such as the lungs. The crosstalk between the gut and the lungs, which is supported by complex connections and intricate pathways, is defined as the gut-lung axis. This review aimed to report on the potential role of the gut-lung gut-lung axis in the development and persistence of equine asthma. We summarized significant determinants in the development of asthma in horses and humans. The article discusses the gut-lung axis and proposes an integrative view of the relationship between gut microbiota and asthma. It also explores therapies for modulating the gut microbiota in horses with asthma. Improving our understanding of the horse gut-lung axis could lead to the development of techniques such as fecal microbiota transplants, probiotics, or prebiotics to manipulate the gut microbiota specifically for improving the management of asthma in horses.

Plant age-dependent dynamics of annatto pigment (bixin) biosynthesis in Bixa orellana.

Age affects the production of secondary metabolites, but how developmental cues regulate secondary metabolism remains poorly understood. The achiote tree (Bixa orellana L.) is a source of bixin, an apocarotenoid used in diverse industries worldwide. Understanding how age-dependent mechanisms control bixin biosynthesis is of great interest for plant biology and for economic reasons. Here we overexpressed miRNA156 (miR156) in B. orellana to comprehensively study the effects of the miR156-SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) module on age-dependent bixin biosynthesis in leaves. Overexpression of miR156 in annatto plants (miR156ox) reduced BoSPL transcript levels, impacted leaf ontogeny, lessened bixin production, and increased abscisic acid levels. Modulation of expression of BoCCD4-4 and BoCCD1, key genes in carotenoid biosynthesis, was associated with diverting the carbon flux from bixin to abscisic acid in miR156ox leaves. Proteomic analyses revealed an overall low accumulation of most secondary metabolite-related enzymes in miR156ox leaves, suggesting that miR156-targeted BoSPLs may be required to activate several secondary metabolic pathways. Our findings suggest that the conserved BomiR156-BoSPL module is deployed to regulate leaf dynamics of bixin biosynthesis, and may create novel opportunities to fine-tune bixin output in B. orellana breeding programs.

Longitudinal effects of oral administration of antimicrobial drugs on fecal microbiota of horses.

BACKGROUND: Antimicrobial drug-associated diarrhea (AAD) is the most common adverse effect in horses receiving antimicrobials. Little information on how oral administration of antimicrobials alters intestinal microbiota in horses is available. OBJECTIVE: Investigate changes of the fecal microbiota in response to oral administration of antimicrobials. ANIMALS: Twenty healthy horses. METHODS: Prospective, longitudinal study. Horses were randomly assigned to 4 groups comprising 4 horses each: group 1 (metronidazole); group 2 (erythromycin); group 3 (doxycycline); group 4 (sulfadiazine/trimethoprim, SMZ-TMP); and group 5 (control). Antimicrobials were administered for 5 days. Fecal samples were obtained before (day 0) and at 1, 2, 3, 4, 5, 6, and 30 days of the study period. Fecal microbiota was characterized by high throughput sequencing of the V4 region of the 16S rRNA. RESULTS: Horses remained healthy throughout the study. Richness and diversity in doxycycline, erythromycin, and metronidazole, but not SMZ-TMP groups, was significantly lower (P < .05) at multiple time points after administration of antimicrobials compared with samples from day 0. Main changes in the microbiota were observed during the time of antimicrobial administration (day 2-5; weighted and unweighted UniFrac PERMANOVA P < .05). Administration of erythromycin, doxycycline and, to a lesser extent, metronidazole produced a pronounced alteration in the microbiota compared with day 0 samples by decreasing the abundance of Treponema, Fibrobacter, and Lachnospiraceae and increasing Fusobacterium and Escherichia-Shigella. CONCLUSIONS AND CLINICAL IMPORTANCE: Oral administration of antimicrobials alters the intestinal microbiota of healthy horses resembling horses with dysbiosis, potentially resulting in intestinal inflammation and predisposition to diarrhea.

Colonization of the Gastrointestinal Tract of Chicks with Different Bacterial Microbiota Profiles.

This study aimed to investigate the consequences of early-life microbiota transplantation using different caecal content sources in broiler chicks. We hypothesized that chicks receiving at-hatch microbiota from organic hens would harbour a distinct microbiota from chicks receiving industry-raised broiler microbiota after six weeks of age. Three hundred Cobb broilers eggs were randomly assigned to one of four groups according to the caecal content received: organic laying hens (Organic); autoclaved caecal content of organic laying hens (Autoclaved); conventionally grown broilers (Conventional); and sterile saline (Control). caecal microbiota transplantation was given by gavage on day 1. Ten birds/group were euthanized on days 2, 7, 14, 28, and 42. The caecal tonsils and contents were collected for cytokines and microbiota analyses. The microbiota from chicks receiving live inocula resembled the donors' microbiota from day seven until day 42. The microbiota composition from the chickens who received the Organic inoculum remained markedly different. Starting on day 7, the Organic group had higher richness. Simpson and Shannon's indices were higher in the Conventional group on days 2 and 7. Chickens in the Conventional group presented higher production of IL-1ß and IL-6 in plasma on days 2 and 28, increased IL-6 expression in the caecal tonsils at days 7 and 42, and increased IL-12 expression on day 7. However, the Conventional group was infected with Eimeria spp., which likely caused inflammation. In conclusion, microbiota transplantation using different microbiota profiles persistently colonized newly hatched broiler chicks. Future studies evaluating the importance of microbiota composition during infections with common enteropathogens are necessary. This study also highlights the need for a strict screening protocol for pathogens in the donors' intestinal content.

Investigation of the vaginal microbiota of dairy cows through genetic sequencing of short (Illumina) and long (PacBio) reads and associations with gestational status.

The vaginal microbiota has been shown to be important in local immune regulation and may play a role in reproduction and fertility. Next-generation sequencing (NGS) technologies have been used to characterize the bovine vaginal microbiota, mainly using short-read sequencing (Illumina). However, the main limitation of this technique is its inability to classify bacteria at the species level. The objective of this study was to characterize the bovine vaginal microbiota at the species level using long-read sequencing (PacBio) and to compare it with the results of short-read sequencing. In addition, the vaginal microbiota of cows that became pregnant after artificial insemination (AI) was compared with that of infertile animals. Thirteen Holstein cows had vaginal swabs collected prior to AI. DNA was extracted and subjected to Illumina and PacBio sequencing to characterize the V4 region and the entire 16S rRNA gene, respectively. PacBio sequencing yielded 366,509 reads that were assigned to 476 species from 27 phyla. However, none of the most abundant reads (>1%) could be classified at the species level. Illumina sequencing yielded more reads and consequently was able to detect a more observed species, but PacBio sequencing was able to detect more unique and rare species. The composition of the vaginal microbiota varies according to the sequencing method used, which might complicate the interpretation of results obtained in the majority of the current studies. The present study expands on the current knowledge of bovine microbiota, highlighting the need for further efforts to improve the current databanks.

Effect of time of sample collection after onset of diarrhea on fecal microbiota composition of calves.

BACKGROUND: The effect of time of sample collection after onset of diarrhea on the fecal microbiota composition of calves is unknown. OBJECTIVE: Compare the fecal microbiota of calves with diarrhea onset on the day of sampling (D <24h), and calves having had diarrhea for >24 to 48 hours (D 24-48h). ANIMALS: Thirty-one diarrheic calves (20 D <24h and 11 D 24-48h), 3 to 7 days of age. METHODS: Cross-sectional study. Diarrhea was defined as a calf with loose feces or watery feces. Assessment of the fecal microbiota was performed by sequencing of 16S ribosomal RNA gene amplicons. RESULTS: Richness and diversity were not statistically different between D <24h and D 24-48h (P > .05), but bacterial membership and structure differed significantly (AMOVA, P < .001 for both comparisons). Linear discriminant analysis effect size (LefSe) showed an enrichment of Faecalibacterium, Phocaeicola, Lachnospiracea, and Lactobacillus in the feces of D <24h calves, whereas Escherichia/Shigella, Ligilactobacillus, Clostridium_Sensu_Stricto, Clostridium_Incerta_Sedis, and Enterococcus were enriched in the D 24-48h calves. CONCLUSION AND CLINICAL IMPORTANCE: Rapid changes in fecal microbiota occur during the first 48 hours of diarrhea with an enrichment of lactic acid-producing bacteria in D <24h followed by an enrichment in Escherichia/Shigella and Clostridium spp. in D 24-48h. The time from diarrhea onset to sampling appears to affect the bacterial composition. Researchers should standardize times for fecal collection based on the time of diarrhea.

Health indicators in surplus calves at the time of arrival at auction markets: Associations with distance from farms of origin in Québec, Canada.

This study evaluated the associations between estimated distance from farms' locations to auction markets, and health indicators of surplus dairy calves sold during summer 2019 and winter 2020 in Québec, Canada. A total of 3,610 animals from 1,331 different farms were used in this cross-sectional cohort study. Geographic coordinates (latitude and longitude) were obtained for each farm and the 2 participating livestock auction markets. Calves' abnormal physical signs (APS) were noted upon arrival at the auction market as they were examined by trained research staff. The haversine distance between the farm and the auction market was evaluated using geographic coordinates and categorized. Generalized linear mixed models were used for statistical analyses. The main APS observed were ocular discharge (34.9%), abnormal hide cleanliness (21.2%), swollen navel (17.2%), dehydration score 1 (at least one of the 2 following clinical signs: persistent skin tent or sunken eye, 12.9%), and dehydration score 2 (both clinical signs mentioned above, 6.5%). Calves from farms located at greater distances from the auction markets (≥110 km) had a higher risk ratio [RR = 1.08; 95% confidence internal (CI) = 1.03, 1.13] for dehydration than those from lesser distances (0-25 km). During the summertime, a RR of 1.18 (95% CI = 1.15, 1.22) was observed for dehydration compared with wintertime. A 2-way interaction between estimated distance and season showed a higher prevalence of ocular discharge for calves from farms at distances greater than or equal to 110 km during the summer (RR = 1.11; 95% CI: 1.04, 1.20) than for calves from farms located at lesser distances (0-25 km). These results demonstrate that calves from farms located at greater distances from the auction markets had more APS, mainly during the summer. A better understanding of the transport conditions and interaction with management at the farm of origin is determinant to mitigate the impact of the journey on surplus calf health.

Effect of dietary iron supplementation on the equine fecal microbiome.

Iron is an essential element for all living organisms, including bacteria, as several virulence factors and replication components are influenced by iron concentration. The objective of this study was to determine whether the composition and diversity of the fecal microbiota of adult horses are affected by supplemental dietary iron. Ten clinically healthy horses were randomly divided into a control and an iron-supplemented group (n = 5). The treated group was supplemented with oral ferrous sulphate monohydrate (720 ppm of iron), whereas the control group received 320 ppm of iron daily for 15 d. Fecal samples were collected before and 5, 10, 15, and 30 d after supplementation and frozen at -80°C. DNA was sequenced using an Illumina MiSeq platform and data were analyzed using the software Mothur and linear discriminant analysis (LDA) effect size (LEfSe). Iron supplementation caused no change in the overall composition of the fecal microbiota, but some minor changes were observed in the low-abundant bacteria, as well as an increased alpha diversity after 15 d of supplementation. Significant differences in community composition of the fecal microbiota over time were observed in both groups, highlighting the importance of a control group, as there are variables that cannot be controlled in microbiome studies.

Le fer est un élément essentiel pour tous les organismes vivants, y compris les bactéries, car plusieurs facteurs de virulence et composants de réplication sont influencés par la concentration en fer. L'objectif de cette étude était de déterminer si la composition et la diversité du microbiote fécal des chevaux adultes sont affectées par la supplémentation en fer alimentaire. Dix chevaux cliniquement sains ont été divisés au hasard en un groupe témoin et un groupe supplémenté en fer, n = 5 par groupe. Le groupe traité a reçu un supplément oral de sulfate ferreux monohydraté (720 ppm de fer) et le groupe témoin a reçu 320 ppm de fer par jour pendant 15 jours. Des échantillons fécaux ont été prélevés avant la supplémentation et 5, 10, 15 et 30 jours après la supplémentation puis congelés à −80 °C. L'ADN a été séquencé à l'aide de la plateforme Illumina MiSeq et les données ont été analysées à l'aide des logiciels Mothur et analyse de la fonction discriminante linéaire taille de l'effet LefSe. La supplémentation en fer n'a provoqué aucun changement dans la composition du microbiote fécal, mais certains changements ont été observés chez les bactéries peu abondantes, ainsi qu'une augmentation de la diversité alpha après 15 jours de supplémentation. Au fil du temps, des différences significatives dans la composition de la communauté bactérienne ont été observées dans les deux groupes, soulignant l'importance d'un groupe témoin, car il existe des variables qui ne peuvent être contrôlées dans les études sur le microbiome.(Traduit par les auteurs).

Effects of concentrated fecal microbiota transplant on the equine fecal microbiota after antibiotic-induced dysbiosis.

Bacterial imbalances are observed in intestinal diseases and fecal microbiota transplantation (FMT) has been used to restore the intestinal microbiota of horses. However, there is evidence that the current methods proposed for FMT in horses have limited efficacy. The objective of this study was to concentrate the bacteria present in the donor stool by centrifugation, and to test the effect in horses with antibiotic-induced dysbiosis. One healthy 11-year-old horse was selected as a fecal donor and 9 horses were given trimethoprim sulfadiazine (TMS) for 5 days to induce dysbiosis. Horses received either a concentrated FMT (cFMT, n = 3), fresh unconcentrated FMT (fFMT, n = 3), or 10% glycerol solution (vehicle, VEH, n = 3) by nasogastric tube for 3 days. Fecal samples were collected on Days 0, 4, 9, 11, and 21 for microbiota analysis (Illumina sequencing). The TMS significantly changed the bacterial composition of horses' feces (D0 versus D4). The composition of the cFMT and fFMT recipient horses was significantly different after transplantation compared to after antibiotic-induced dysbiosis (D4 versus D11), whereas the microbiota of the vehicle recipients was not, indicating that both protocols induced transient changes. However, preparation of FMT solutions markedly changed the original composition present in the donor's feces, with significant enrichment of Escherichia genus in the cFMT. Individual susceptibility to restoration of the microbiota was observed in horses, similar to what is known for other species. Our results suggest that concentrating bacteria should not be recommended in preparation of FMT solutions and that further research is required to improve current methods recommended to perform FMT in horses.

Des déséquilibres bactériens sont observés dans les maladies intestinales et la transplantation de microbiote fécal (FMT) a été utilisée pour la restaurer le microbiote intestinal des chevaux. Cependant, que les méthodes actuelles proposées pour FMT chez les chevaux ont une efficacité limitée. L'objectif de cette étude était de concentrer les bactéries présentes dans les selles du donneur par centrifugation, et de tester leur effet chez des chevaux atteints de dysbiose induite par les antibiotiques. Un cheval sain de 11 ans a été sélectionné comme donneur fécal et 9 chevaux ont reçu du triméthoprime sulfadiazine (TMS) pendant cinq jours pour induire une dysbiose. Les chevaux ont reçu soit une FMT concentrée (cFMT, n = 3), une FMT fraîche non concentrée (fFMT, n = 3) ou une solution de glycérol à 10 % (véhicule, VEH, n = 3) par sonde naso-gastrique pendant 3 jours. Des échantillons fécaux ont été prélevés aux jours 0, 4, 9, 11 et 21 pour analyse du microbiote (séquençage Illumina). Le TMS a significativement modifié la composition bactérienne des matières fécales des chevaux (D0 versus D4). La composition des chevaux receveurs cFMT et fFMT était significativement différente après la transplantation par rapport à la dysbiose induite par les antibiotiques (D4 versus D11), alors que le microbiote des receveurs de véhicules ne l'était pas, indiquant que les deux protocoles induisaient des changements transitoires. Cependant, la préparation des solutions FMT a considérablement modifié la composition originale présente dans les matières fécales du donneur, avec un enrichissement significatif du genre Escherichia dans le cFMT. Une susceptibilité individuelle à la restauration du microbiote a été observée chez les chevaux, à l'instar de ce qui est connu chez d'autres espèces. Nos résultats suggèrent que la concentration des bactéries ne devrait pas être recommandée dans la préparation des solutions FMT et que des recherches supplémentaires sont nécessaires pour améliorer les méthodes actuelles recommandées pour effectuer la FMT chez les chevaux.(Traduit par les auteurs).

Connecting Higher-Order Topology with the Orbital Hall Effect in Monolayers of Transition Metal Dichalcogenides.

Monolayers of transition metal dichalcogenides (TMDs) in the 2H structural phase have been recently classified as higher-order topological insulators (HOTIs), protected by C_{3} rotation symmetry. In addition, theoretical calculations show an orbital Hall plateau in the insulating gap of TMDs, characterized by an orbital Chern number. We explore the correlation between these two phenomena in TMD monolayers in two structural phases: the noncentrosymmetric 2H and the centrosymmetric 1T. Using density functional theory, we confirm the characteristics of 2H TMDs and reveal that 1T TMDs are identified by a Z_{4} topological invariant. As a result, when cut along appropriate directions, they host conducting edge states, which cross their bulk energy-band gaps and can transport orbital angular momentum. Our linear response calculations thus indicate that the HOTI phase is accompanied by an orbital Hall effect. Using general symmetry arguments, we establish a connection between the two phenomena with potential implications for orbitronics and spin orbitronics.

Dynamics of the sucrose metabolism and related gene expression in tomato fruits under water deficit.

The impact of water deficit on sucrose metabolism in sink organs like the fruit remains poorly known despite the need to improve fruit crops resilience to drought in the face of climate change. The present study investigated the effects of water deficit on sucrose metabolism and related gene expression in tomato fruits, aiming to identify candidate genes for improving fruit quality upon low water availability. Tomato plants were subjected to irrigated control and water deficit (-60% water supply compared to control) treatments, which were applied from the first fruit set to first fruit maturity stages. The results have shown that water deficit significantly reduced fruit dry biomass and number, among other plant physiological and growth variables, but substantially increased the total soluble solids content. The determination of soluble sugars on the basis of fruit dry weight revealed an active accumulation of sucrose and concomitant reduction in glucose and fructose levels in response to water deficit. The complete repertoire of genes encoding sucrose synthase (SUSY1-7), sucrose-phosphate synthase (SPS1-4), and cytosolic (CIN1-8), vacuolar (VIN1-2) and cell wall invertases (WIN1-4) was identified and characterized, of which SlSUSY4, SlSPS1, SlCIN3, SlVIN2, and SlCWIN2 were shown to be positively regulated by water deficit. Collectively, these results show that water deficit regulates positively the expression of certain genes from different gene families related to sucrose metabolism in fruits, favoring the active accumulation of sucrose in this organ under water-limiting conditions. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-023-01288-7.

Mobile app for targeted selective treatment of haemonchosis in sheep.

Livestock is an important part of many countries gross domestic product, and sanitary control impacts herd management costs. To contribute to incorporating new technologies into this economic chain, this work presents a mobile application for decision assistance to treatment against parasitic infection by Haemonchus contortus in small ruminants. Based on the Android system, the proposed software is a semi-automated computer-aided procedure to assist Famacha© pre-trained farmers in applying anthelmintic treatment. It mimics the two-class decision procedure performed by the veterinarian with the help of the Famacha© card. The embedded cell phone camera was employed to acquire an image from the ocular conjunctival mucosa, classifying the animal as healthy or anemic. Two machine-learning strategies were assessed, resulting in an accuracy of 83 % for a neural network and 87 % for a support vector machine (SVM). The SVM classifier was embedded into the app and made available for evaluation. This work is particularly interesting to small property owners from regions with difficult access or restrictions on obtaining continuous post-training technical guidance to use the Famacha© method effectively.

Challenges in the surgical treatment of a 8-year old child living with Prune Belly syndrome: case report / Desafios no tratamento tardio da síndrome de Prune Belly: relato de caso

Prune Belly syndrome, also known as Eagle-Barret syndrome, is a rare disease, with a prevalence of 3.8 live births per 100,000 births. Its main characteristic is the hypoplasia of the abdominal muscles, giving rise to the name "prune belly syndrome". The gold standard treatment is surgery, ideally with correction of cryptorchidism and phimosis between 6 to 18 months of life. Correction of urinary malformations and abdominoplasty should be performed up to 4 years of age. Little evidence exists in the literature about late treatment and its implications for prognosis. The context mentioned above led us to present an uncommon case of a six years old child in which a surgical approach was performed later than usual.

Introdução: A síndrome de Prune Belly é uma doença rara, com prevalência de 3,8 nascidos vivos a cada 100 mil nascimentos. Tem como principal caraterística a hipoplasia da musculatura abdominal gerando a origem do nome "síndrome da barriga de ameixa". O tratamento padrão ouro é cirúrgico, idealmente com correção da criptorquidia e fimose entre 6 a 18 meses e a correção das malformações urinárias e abdominoplastia devem ser realizadas até os 4 anos. Aqui descreve-se um caso ainda menos comum, em que foi realizada abordagem cirúrgica de forma mais tardia que o habitual. Há pouca evidência na literatura acerca do tratamento tardio e suas implicações em relação ao prognóstico.

Dynamics of annatto pigment synthesis and accumulation in seeds of Bixa orellana L. revealed by integrated chemical, anatomical, and RNA-Seq analyses.

Bixin is a commercially valuable apocarotenoid pigment found in the seed aril of Bixa orellana. The dynamics and regulation of its biosynthesis and accumulation during seed development remain largely unknown. Here, we combined chemical, anatomical, and transcriptomic data to provide stage-specific resolution of the cellular and molecular events occurring during B. orellana seed development. Seeds at five developmental stages (S1-S5) were used for analysis of bixin content and seed anatomy, and three of them (S1, S3, and S4) were selected for Illumina HiSeq sequencing. Bixin accumulated in large quantities in seeds compared with other tissues analyzed, particularly during the S2 stage, peaking at the S4 stage, and then decreasing slightly in the S5 stage. Anatomical analysis revealed that bixin accumulated in the large central vacuole of specialized cells, which were scattered throughout the developing mesotesta at the S2 stage, but enlarged progressively at later stages, until they occupied most of the parenchyma in the aril. A total of 13 million reads were generated and assembled into 73,381 protein-encoding contigs, from which 312 were identified as containing 1-deoxy-D-xylulose-5-phosphate/2-C-methyl-D-erythritol-4-phosphate (DOXP/MEP), carotenoid, and bixin pathways genes. Differential transcriptome expression analysis of these genes revealed that 50 of them were sequentially and differentially expressed through the seed developmental stages analyzed, including seven carotenoid cleavage dioxygenases, eight aldehyde dehydrogenases, and 22 methyltransferases. Taken together, these results show that bixin synthesis and accumulation in seeds of B. orellana are a developmentally regulated process involving the coordinated expression of DOXP/MEP, carotenoid, and bixin biosynthesis genes.

Bovine anaplasmosis as a risk factor for retained placenta, mastitis, and abomasal displacement in dairy cattle.

This study aimed to evaluate the frequency of IgG antibodies against A. marginale, the occurrence of this bacterium by qPCR, and the effect of bovine anaplasmosis as a risk factor for clinical cases of retained placenta, mastitis, and abomasal displacement in dairy cattle. For that 179 Holstein cows out of three dairy herds, in the municipality of Sertão, Rio Grande do Sul, Brazil. These cows were on farms that were vulnerable to risk factors that are crucial to susceptibility among these animals to this intracellular hemoparasite. The mean seropositivity for A. marginale from the periods evaluated was 54% on farm A, 69.4% on farm B, and 27.3% on farm C. Molecular diagnosis was performed with qPCR and the mean positivity for A. marginale among the cows on farms A, B, and C in December 2017 was 34.6% (67/179). Infected animals showed clinical cases of retained placenta (6.1%), mastitis (6.1%), and abomasal displacement (0.5%). The association between positivity for anaplasmosis and these clinical cases was assessed through the odds ratio. Our results show that females with a positive qPCR assay for A. marginale had 52.48 times increased probability (OR) to develop clinical cases of retained placenta and mastitis (P < 0.001). These clinical cases negatively impact the productivity of positive females. Thus, implementing preventive and prophylactic control measures to ensure the sanitary quality of the herds is needed to avoid losses due to morbidity and mortality and diminish the economic losses suffered by farmers.

Diagnostic performance of convolutional neural networks for dental sexual dimorphism.

Convolutional neural networks (CNN) led to important solutions in the field of Computer Vision. More recently, forensic sciences benefited from the resources of artificial intelligence, especially in procedures that normally require operator-dependent steps. Forensic tools for sexual dimorphism based on morphological dental traits are available but have limited performance. This study aimed to test the application of a machine learning setup to distinguish females and males using dentomaxillofacial features from a radiographic dataset. The sample consisted of panoramic radiographs (n = 4003) of individuals in the age interval of 6 and 22.9 years. Image annotation was performed with V7 software (V7labs, London, UK). From Scratch (FS) and Transfer Learning (TL) CNN architectures were compared, and diagnostic accuracy tests were used. TL (82%) performed better than FS (71%). The correct classifications of females and males aged ≥ 15 years were 87% and 84%, respectively. For females and males < 15 years, the correct classifications were 80% and 83%, respectively. The Area Under the Curve (AUC) from Receiver-operating Characteristic (ROC) curves showed high classification accuracy between 0.87 and 0.91. The radio-diagnostic use of CNN for sexual dimorphism showed positive outcomes and promising forensic applications to the field of dental human identification.

Serum IgM antibody response to Clostridioides difficile polysaccharide PS-II vaccination in pony foals.

OBJECTIVES: Clostridioides difficile (formerly Clostridium difficile) is associated with colitis in foals and mature horses. C. difficile exposes specific phosphorylated polysaccharides (PSs), named PS-I, PS-II and PS-III. These cell-surface PSs are potential vaccine targets, especially the hexasaccharide phosphate PS-II, that has been found in all C. difficile ribotypes examined. Since we previously identified anti-PS-II circulating antibodies in horses, we postulated that vaccinating foals with PS-II may prevent colonization by C. difficile. In this study, we aim to evaluate the IgM antibody responses in foals to PS-II. METHODS: To evaluate the reactogenicity and immunogenicity of C. difficile PS-II in foals, three-to four-month-old foals were vaccinated intramuscularly three times at intervals of three weeks with 100 µg/dose (3 foals) or 500 µg/dose (3 foals) of purified PS-II antigen with aluminum hydroxide adjuvant, or with a placebo preparation (2 foals) containing adjuvant alone. RESULTS: No injection site swelling, pain or fever was observed after vaccination. Two of the three foals receiving 100 µg/dose, and three out of three foals receiving 500 µg/dose of PS-II responded with increases in serum IgM antibodies. No control foals that received the placebo had IgM responses to PS-II. There was a trend towards a higher response rate in foals receiving 500 µg PS-II one week after second vaccination when compared to control foals and towards higher concentrations of serum IgM antibodies in foals receiving 500 µg PS-II. CONCLUSIONS: No adverse reactions were observed following vaccination with PS-II in foals; Serum IgM immune responses were induced by vaccination. A polysaccharide-based vaccine for C. difficile in horses deserves further investigation.

Single-base resolution methylomes of somatic embryogenesis in Theobroma cacao L. reveal epigenome modifications associated with somatic embryo abnormalities.

Propagation by somatic embryogenesis in Theobroma cacao has some issues to be solved, as many morphologically abnormal somatic embryos that do not germinate into plants are frequently observed, thus hampering plant production on a commercial scale. For the first time the methylome landscape of T. cacao somatic embryogenesis was examined, using whole-genome bisulfite sequencing technique, with the aim to understand the epigenetic basis of somatic embryo abnormalities. We identified 873 differentially methylated genes (DMGs) in the CpG context between zygotic embryos, normal and abnormal somatic embryos, with important roles in development, programmed cell death, oxidative stress, and hypoxia induction, which can help to explain the morphological abnormalities of somatic embryos. We also identified the role of ethylene and its precursor 1-aminocyclopropane-1-carboxylate in several biological processes, such as hypoxia induction, cell differentiation and cell polarity, that could be associated to the development of abnormal somatic embryos. The biological processes and the hypothesis of ethylene and its precursor involvement in the somatic embryo abnormalities in cacao are discussed.