Novel Approaches to Evaluate and Enhance Neonatal Calf Gastrointestinal Health and Development

Physiological adaptations of the gastrointestinal tract (GIT) epithelial cells as well as methods to manipulate early life GIT microbiota colonization during the neonatal stage, is of great importance to the dairy industry. The first objective of this research was to optimize a method based on evaluation of bovine transcripts in fecal RNA via RTqPCR using L-selectin (SELL) as a marker for polymorphonuclear leukocytes (PMNL), keratin 8 (KRT8) and fatty acid binding protein 2 (FABP2) for GIT enterocytes, and tight junctions in fecal RNA and GIT sections in dairy calves. To test the reliability of the fecal RNA method, fecal and PMNL samples from neonatal calves were used. The expression of KRT8 was greater in fecal RNA than in PMNL. In contrast, a greater SELL expression was observed in PMNL. In another study, postmortem GIT and feces samples were collected from healthy calves for total RNA isolation. Overall, the expression of FABP2 and KRT8 was similar between fecal RNA and the lower GIT. Taken together, these results provide further evidence that the fecal RNA method can potentially be used as a tool to evaluate molecular adaptations of the GIT in dairy calves. The second objective was to evaluate the effects of early life fecal microbiota transplantation (FMT) from healthy adult donors on health and performance of neonatal dairy calves. To test the effects of FMT, newborn calves were subjected to 1×/d inoculations with 25 g of fecal donor material mixed in the milk replacer from 8 to 12 d of age. Results from this study demonstrated that calves subjected to FMT tended to have greater body weight than control calves. The liver function marker paraoxonase was greater in the blood of FMT calves than control at 3wk of age. These results suggest that FMT in neonatal calves has positive effects not only on growth performance but also in mediating liver function

Role of Peroxisome Proliferator-Activated Receptor Gamma on Prevention/Cure of Mastitis

Mastitis is a major endemic disease in dairy cows resulting in significant economic losses for the dairy industry. The peroxisome proliferator-activated receptor gamma (PPARγ) is a nuclear receptor that is able to bind and be activated by natural (e.g., fatty acids) and synthetic (e.g. thiazolidinedione) compounds. PPARγ plays important roles in adipocyte differentiation, inflammation, and re-epithelialization in monogastric. In ruminants, PPARγ may play a role in milk fat synthesis. The aim of this study was to assess the role of PPARγ in host response to mammary infection and milk fat synthesis in ruminants. Our hypothesis is that activation of PPARγ improves the host response to mastitis and increases milk fat yield. By using a synthetic PPARγ agonist in dairy goats in combination with intramammary infection to induce subclinical mastitis, the objectives of the present experiments were to test if activation of PPARγ improves 1) the response to mastitis and 2) milk fat production. To achieve our objectives we performed two in vivo experiments (Experiments 1 and 2). In Experiment 1, 24 Saanen lactating goats with a low body condition score and getting a low-energy diet without vitamin supplementation received a daily intrajugular injection of either 8 mg of 2,4-thiazolidinedione (TZD) per kg of BW or saline (as a control) and, after a week of TZD injection, an intramammary infusion (IMI) of either Streptococcus uberis to induce subclinical mastitis or saline used as a control (6 goats/group). Milk yield and components, body weight, rectal temperature, leukocyte phagocytosis, blood metabolic and inflammation parameters plus insulin, adipocyte size by histology, and expression by RT-qPCR of PPARγ target genes in adipose tissue obtained through biopsy and in mammary epithelial cells (MEC) isolated from milk were assessed. In MEC, expression of CCL2 and IL8 was also measured. Data were analyzed by GLIMMIX of SAS with Mastitis, TZD, and Time and all interactions as main effects and goat as random effect. Statistical significance and tendencies were declared at P 5-fold increase of milk somatic cells count (SCC) in goats receiving Strep. uberis and by 30% decrease of % polymorphonuclear leukocytes in blood. The SCC in milk were overall lower in TZD-treated goats. Mastitis induction but not TZD decreased milk yield and production of milk fat. Goats receiving Strep. uberis had increased concentrations of glucose, triglycerides, and non-esterified fatty acids (NEFA) in blood after IMI. NEFA was not affected in TZD goats, which did not receive Strep. uberis. Inflammatory markers increased in blood of all goats but the increase of haptoglobin was overall lower in TZD treated goats. Indicators of liver activity, including albumin, paraoxonase, and cholesterol, overall decreased after IMI but cholesterol did not decrease in TZD-treated goats. The bactericidal myeloperoxidase was higher in TZD-treated goats after mastitis. Insulin sensitivity was not affected by TZD or mastitis. Adipocytes size increased over time and was higher in TZD goats not receiving Strep. uberis. Subclinical mastitis increased expression of CCL2 and prevented a decrease in expression of IL8. MEC from TZD-treated goats tended to have higher expression of PPARG, FASN and SCD1 after 3 weeks of TZD treatment. Neither mastitis nor TZD affected the expression of genes in adipose tissue. Overall the data of Experiment 1 indicated that the subclinical mastitis model was successfully achieved. The treatment with TZD decreased somatic cells in milk, improved the response of liver, decreased the severity of inflammation, and increased the killing capacity of neutrophils after IMI. The data suggested a more lipogenic adipose tissue in TZD-treated goats but also some active, although minor, nutrigenomic effect of TZD on MEC that may have counteracted the competition of lipid substrates between mammary and adipose tissue. Blood metabolic data suggested that goats responded to Strep. uberis intramammary infusion similar to dairy cows in negative energy balance. Data obtained from Experiment 1 indicated that TZD aids with mastitis response. TZD had some effect on milk fat synthesis but, overall, had a smaller-than-expected nutrigenomic effect probably also due to the low body condition and low energy in the diet of the goats. Thus, the effect of PPARγ on milk fat synthesis is still unclear. The rationale to perform Experiment 2 stemmed from the possibility that the limited nutrigenomic response observed in Experiment 1 was due to a potential dietary deficiency. Subsequent in vitro work demonstrated that TZD is a strong activator of PPAR but only in the presence of 9-cis-retinoic acid, a metabolite of vitamin A and the activation of PPARγ obligate heterodimer Retinoic-X-Receptor (RXR). Therefore, we hypothesized that continuous activation of PPARγ by TZD in dairy goats supplemented with adequate amount of vitamin A improves the inflammatory response to subclinical mastitis. In order to test this hypothesis we used 12 Saanen multiparous goats in early lactation. Goats received a diet that met the NRC requirements, including vitamin A, and a daily injection of 8 mg TZD per kg of BW (n=6) or saline (n=6; CTRL). Following 14 days of treatment, all goats received an IMI of Strep. uberis to induce subclinical mastitis in the right half of the udder with the left half used as control. Metabolic, inflammation, and oxidative-status profiling in blood including 20 parameters was performed. Milk yield, SCC, rectal temperature and leukocytes phagocytosis were measured. Expression of several PPARγ target genes and genes involved in inflammation was measured in MEC, macrophages isolated from milk, and liver tissue. Data were analyzed by GLIMMIX of SAS with treatment (TRT) and Time and TRTxTime interaction as main effects and goat as random effect. For milk and SCC, mammary half was also included in the main effect (including interactions). Statistical significance and tendencies were declared at P < 0.05 and 0.05 ≤ P ≤ 0.10, respectively. Milk yield decreased after IMI but the decrease was larger in TZD-treated goats. SCC increased after IMI but was not affected by TZD administration. Milk fat decreased after IMI in all halves except in the untreated half of TZD-treated goats. In blood within 2 days from IMI, ceruloplasmin, haptoglobin, and glucose were increased while Zn was decreased. These data confirmed successful induction of sub-clinical mastitis and a status of slight inflammation after IMI. None of the parameters in blood was affected by TZD with the exception of a lower bilirubin concentration and a tendency for higher haptoglobin in TZD vs. CTRL after IMI, indicating a more robust response of the liver to inflammation. The stronger inflammation was also supported by a tendency for higher reactive oxygen metabolites in TZD vs. CTRL group after IMI. We also detected a tendency for a higher globulin in TZD vs. CTRL indicating a better adaptive immune system. Leukocyte phagocytosis was strongly reduced by TZD treatment. None of the genes measured were affected by TZD in liver. In milk macrophages and MEC, expression of inflammatory genes was higher compared to control in halves receiving Strep. uberis, whereas no effect of TZD was observed with the exception of a lower SCD1 in TZD-treated goats compared to CTRL. We conclude that, contrary to our hypothesis, in goats receiving NRC recommended amount of vitamin A, TZD had a minor effect on the response to mastitis with a likely better liver response, but a lower phagocytosis and minor effect on expression of genes. Considering both in vivo experiments, we can conclude that TZD has an important effect on inflammatory response in dairy goats receiving low energy diet without vitamins supplementation but the effect disappears for the most part in goats receiving adequate feeding, including vitamin A. Furthermore, the lack of effects on expression of PPARγ target genes does not support TZD being a strong PPARγ agonist in dairy goats

Fatores de risco e recomendações atuais para prevenção de infecção associada a cateteres venosos centrais: uma revisão de literatura

Backgound and Objectives: Infections related to central venous catheter (CVC) use constitute an important a problem. It is estimated that approximately 90% of bloodstream infections (BSI) are caused by CVC use. This study aims at reviewing the risk factors and current recommendations for prevention of infections associated with central venous catheter use. Methods: A total of 12 articles published in the last 5 years and indexed in the databases of the Latin American and Caribbean Literature on Health Sciences (LILACS), Nursing Database (BDENF), International Literature on Health Sciences (Medline/Pubmed) were selected, as well as publications related to the recommendations for BSI prevention, such as: Institute for Healthcare Improvement (IHI), Centers for Disease Control and Prevention (CDC) and the National Health Surveillance Agency (ANVISA). Results: Two categories were identified: prevention and control measures and risk factors for BSI associated with central venous catheter use. Conclusions: Some recommendations that were well-defined over the years have been questioned by some authors and continuing training and education of the multidisciplinary team are the most important factors for the prevention of bloodstream infections associated with CVC use.Justificativa e Objetivos: As infecções relacionadas ao uso cateter venoso central (CVC) constituem-se em um problema de grande magnitude. Estima-se que aproximadamente 90% das infecções de corrente sanguínea (ICS) são causadas pelo uso de cateter venoso central. Este estudo objetiva levantar os fatores de risco e recomendações atuais para prevenção de infecção associada a cateteres venosos centrais. Método: Foram selecionados 12 artigos, publicados nos últimos 5 anos e indexados nas bases de dados Lilacs (Literatura Latino-Americana e do Caribe em Ciências da Saúde), Bdenf (Base de dados de enfermagem), Pubmed (Literatura Internacional em Ciências da Saúde), Journal of Infusion Nursing, além de publicações referentes às recomendações para prevenção de infecção de corrente sanguínea (ICS), como: Institute for Healthcare Improvement (IHI), Centers for Disease Control and Prevention (CDC) e Agência Nacional de Vigilância Sanitária (ANVISA). Resultados: Foram evidenciadas duas categorias: medidas de prevenção e controle e fatores de risco para infecção em corrente sanguínea associada ao uso de cateter venoso central. Conclusões: Algumas recomendações bem definidas ao longo dos anos vêm sendo questionadas por alguns autores e o treinamento e educação continuada da equipe multidisciplinar são os fatores mais importantes para prevenção de infecção da corrente sanguínea por cateter central

Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear un derstanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5–7 vast areas of the tropics remain understudied.8–11 In the American tropics, Amazonia stands out as the world’s most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepre sented in biodiversity databases.13–15 To worsen this situation, human-induced modifications16,17 may elim inate pieces of the Amazon’s biodiversity puzzle before we can use them to understand how ecological com munities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple or ganism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region’s vulnerability to environmental change. 15%–18% of the most ne glected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lostinfo:eu-repo/semantics/publishedVersio

Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear understanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5,6,7 vast areas of the tropics remain understudied.8,9,10,11 In the American tropics, Amazonia stands out as the world's most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepresented in biodiversity databases.13,14,15 To worsen this situation, human-induced modifications16,17 may eliminate pieces of the Amazon's biodiversity puzzle before we can use them to understand how ecological communities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple organism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region's vulnerability to environmental change. 15%–18% of the most neglected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lost

Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear understanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5,6,7 vast areas of the tropics remain understudied.8,9,10,11 In the American tropics, Amazonia stands out as the world's most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepresented in biodiversity databases.13,14,15 To worsen this situation, human-induced modifications16,17 may eliminate pieces of the Amazon's biodiversity puzzle before we can use them to understand how ecological communities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple organism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region's vulnerability to environmental change. 15%–18% of the most neglected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lost

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030