Performance of Hermetia illucens (Diptera: Stratiomyidae) larvae reared on organic waste diets and pupal chitin and chitosan yield.

Recently, much research has been oriented towards the influence of different food wastes and agricultural by-products on the final larval biomass and chemical composition of the insect species Hermetia illucens L. (Diptera: Stratiomyidae). However, there is a gap in the literature regarding the possible relationship between the feeding substrate of H. illucens larvae and chitin. In this context, in the present study, larvae of H. illucens derived from two populations (i.e., UNIPI and UTH), were reared on different diets composed of fruits, vegetables, and meat. Based on the results, the larval survival was high for all diets tested. Larval growth in terms of weight gain, larval length, and feed conversion ratio (FCR) depended on the composition of each diet. The chitin and chitosan composition of larvae, reared on different substrates, did not reveal significant differences. Given the fact that the feeding substrate represent a significant cost in the industrial production of insects, its correlation with a high value product (i.e. chitosan) is important. On the other hand, as the prepupal stage of H. illucens is currently used as animal feed, the metabolization of chitin by farmed animals when the larvae or prepupae were offered as feed could have adverse effects. Thus, depending on the final product that is to be produced, industries could benefit from the establishment of a suitable diet.

Downregulation of PLIN2 in human dermal fibroblasts impairs mitochondrial function in an age-dependent fashion and induces cell senescence via GDF15.

Perilipin 2 (PLIN2) is a lipid droplet (LD)-coating protein playing important roles in lipid homeostasis and suppression of lipotoxicity in different tissues and cell types. Recently, a role for PLIN2 in supporting mitochondrial function has emerged. PLIN2 dysregulation is involved in many metabolic disorders and age-related diseases. However, the exact consequences of PLIN2 dysregulation are not yet completely understood. In this study, we knocked down (KD) PLIN2 in primary human dermal fibroblasts (hDFs) from young (mean age 29 years) and old (mean age 71 years) healthy donors. We have found that PLIN2 KD caused a decline of mitochondrial function only in hDFs from young donors, while mitochondria of hDFs from old donors (that are already partially impaired) did not significantly worsen upon PLIN2 KD. This mitochondrial impairment is associated with the increased expression of the stress-related mitokine growth differentiation factor 15 (GDF15) and the induction of cell senescence. Interestingly, the simultaneous KD of PLIN2 and GDF15 abrogated the induction of cell senescence, suggesting that the increase in GDF15 is the mediator of this phenomenon. Moreover, GDF15 KD caused a profound alteration of gene expression, as observed by RNA-Seq analysis. After a more stringent analysis, this alteration remained statistically significant only in hDFs from young subjects, further supporting the idea that cells from old and young donors react differently when undergoing manipulation of either PLIN2 or GDF15 genes, with the latter being likely a downstream mediator of the former.

Evaluation of ruminal methane and ammonia formation and microbiota composition as affected by supplements based on mixtures of tannins and essential oils using Rusitec.

BACKGROUND: Dietary supplements based on tannin extracts or essential oil compounds (EOC) have been repeatedly reported as a promising feeding strategy to reduce the environmental impact of ruminant husbandry. A previous batch culture screening of various supplements identified selected mixtures with an enhanced potential to mitigate ruminal methane and ammonia formation. Among these, Q-2 (named after quebracho extract and EOC blend 2, composed of carvacrol, thymol, and eugenol) and C-10 (chestnut extract and EOC blend 10, consisting of oregano and thyme essential oils and limonene) have been investigated in detail in the present study with the semi-continuous rumen simulation technique (Rusitec) in three independent runs. For this purpose, Q-2 and C-10, dosed according to the previous study, were compared with a non-supplemented diet (negative control, NC) and with one supplemented with the commercial EOC-based Agolin® Ruminant (positive control, PC). RESULTS: From d 5 to 10 of fermentation incubation liquid was collected and analysed for pH, ammonia, protozoa count, and gas composition. Feed residues were collected for the determination of ruminal degradability. On d 10, samples of incubation liquid were also characterised for bacterial, archaeal and fungal communities by high-throughput sequencing of 16S rRNA and 26S ribosomal large subunit gene amplicons. Regardless of the duration of the fermentation period, Q-2 and C-10 were similarly efficient as PC in mitigating either ammonia (-37% by Q-2, -34% by PC) or methane formation (-12% by C-10, -12% by PC). The PC was also responsible for lower feed degradability and bacterial and fungal richness, whereas Q-2 and C-10 effects, particularly on microbiome diversities, were limited compared to NC. CONCLUSIONS: All additives showed the potential to mitigate methane or ammonia formation, or both, in vitro over a period of 10 d. However, several differences occurred between PC and Q-2/C-10, indicating different mechanisms of action. The pronounced defaunation caused by PC and its suggested consequences apparently determined at least part of the mitigant effects. Although the depressive effect on NDF degradability caused by Q-2 and C-10 might partially explain their mitigation properties, their mechanisms of action remain mostly to be elucidated.

Effect of different doses of camelina cake inclusion as a substitute of dietary soyabean meal on growth performance and gut health of weaned pigs.

Camelina cake (CAM) is a co-product proposed as an alternative protein source; however, piglet data are still limited. This study aimed to evaluate the effect of different doses of CAM in substitution of soyabean meal on the growth, health and gut health of weaned pigs. At 14 d post-weaning (d0), sixty-four piglets were assigned either to a standard diet or to a diet with 4 %, 8 % or 12 % of CAM. Piglets were weighed weekly. At d7 and d28, faeces were collected for microbiota and polyamine and blood for reactive oxygen metabolites (ROM) and thyroxine analysis. At d28, pigs were slaughtered, organs were weighed, pH was recorded on gut, colon was analysed for volatile fatty acids (VFA) and jejunum was used for morphological and gene expression analysis. Data analysis was carried out using a mixed model including diet, pen and litter as factors; linear and quadratic contrasts were tested. CAM linearly reduced the average daily gain from d0-d7, d0-d14, d0-d21 and d0-d28 (P ≤ 0·01). From d0-d7 increasing CAM linearly decreased feed intake (P = 0·04) and increased linearly the feed to gain (P = 0·004). CAM increased linearly the liver weight (P < 0·0001) and affected the cadaverine (P < 0·001). The diet did not affect the ROM, thyroxine, intestinal pH, VFA and morphology. All doses of CAM increased the α diversity indices at d28 (P < 0·05). CAM at 4 % promoted the abundance of Butyricicoccaceae_UCG-008. Feeding with CAM enhanced resilience in the gut microbiome and can be evaluated as a potential alternative protein source with dose-dependent limitations on piglet growth performance.

Secular Trend in GFR Decline in Non-Dialysis CKD Based on Observational Data From Standard of Care Arms of Trials.

RATIONALE & OBJECTIVE: The standard of care (SoC) group of randomized controlled trials (RCTs) is a useful setting to explore the secular trends in kidney disease progression because implementation of best clinical practices is pursued for all patients enrolled in trials. This meta-analysis evaluated the secular trend in the change of glomerular filtration rate (GFR) decline in the SoC arm of RCTs in chronic kidney disease (CKD) published in the last 30 years. STUDY DESIGN: Systematic review and meta-analysis of the SoC arms of RCTs analyzed as an observational study. SETTING & STUDY POPULATIONS: Adult patients with CKD enrolled in the SoC arm of RCTs. SELECTION CRITERIA FOR STUDIES: Phase 3 RCTs evaluating GFR decline as an outcome in SoC arms. DATA EXTRACTION: Two independent reviewers evaluated RCTs for eligibility and extracted relevant data. ANALYTICAL APPROACH: The mean of GFR declines extracted in the SoC arm of selected RCTs were pooled by using a random effects model. Meta-regression analyses were performed to identify factors that may explain heterogeneity. RESULTS: The SoC arms from 92 RCTs were included in the meta-analysis with a total of 32,202 patients. The overall mean GFR decline was-4.00 (95% CI, -4.55 to-3.44) mL/min/1.73m2 per year in the SoC arms with a high level of heterogeneity (I2, 98.4% [95% CI, 98.2-98.5], P<0.001). Meta-regression analysis showed an association between publication year (ß estimate, 0.09 [95% CI, 0.032-0.148], P=0.003) and reduction in GFR over time. When evaluating publication decade categorically, GFR decline was-5.44 (95% CI, -7.15 to-3.73), -3.92 (95% CI, -4.82 to-3.02), and -3.20 (95% CI, -3.75 to -2.64) mL/min/1.73m2 per year during 1991-2000, 2001-2010, and 2011-2023, respectively. Using meta-regression, the heterogeneity of GFR decline was mainly explained by age and proteinuria. LIMITATIONS: Different methods assessing GFR in selected trials and observational design of the study. CONCLUSIONS: In the last 3 decades, GFR decline has decreased over time in patients enrolled in RCTs who received the standard of care. TRIAL REGISTRATION: Registered at PROSPERO with record number CRD42022357704. PLAIN-LANGUAGE SUMMARY: This study evaluated the secular trend in the change in glomerular filtration rate (GFR) decline in the placebo arms of randomized controlled trials (RCTs) that were studying approaches to protect the kidneys in the setting of chronic kidney disease. The placebo groups of RCTs are useful for examining whether the rate of progression of kidney disease has changed over time. We found an improvement in the slope of change in GFR over time. These findings suggest that adherence to standards of kidney care as implemented in clinical trials may be associated with improved clinical outcomes, and these data may inform the design of future RCTs in nephrology.

The effect of a ketogenic diet on weight loss in CKD: a randomized controlled trial in obese stage G1-3a CKD patients.

This study describes a multicentre randomized controlled trial comparing the effects of a ketogenic diet with a low-energy standard diet containing 0.8 g/kg/day on weight loss and metabolic alterations in adult patients with mild-to-moderate non-diabetic chronic kidney disease (CKD) and mild-to-severe obesity. The study is being conducted to understand the impact of the ketogenic diet on weight loss in these patients, as the existing evidence on the ketogenic diet's effect in CKD patients is limited and inconclusive. The study will enrol mild-to moderate adult CKD patients (Stages G1-3a) with albumin to creatinine ratio ≥200 mg/g, without diabetes, with obesity (body mass index ≥30 kg/m2), and stable body weight and estimated glomerular filtration rate from at least 3 months. The primary outcome will be weight loss at 6 months, and secondary outcomes will include adherence to prescribed dietary regimens, body composition changes, changes in standardized blood pressure measurements, metabolic parameters, lipid profile, liver profile, mineral bone disease biomarkers, and changes in renal function and albuminuria. The findings of this study will contribute to a better understanding of the potential benefits and risks of the ketogenic diet in CKD patients with obesity. The results will help guide future research on the ketogenic diet and renal health.

Insight into the long-term impact of birth weight on intestinal development, microbial settlement, and the metabolism of weaned piglets.

Infant mortality of low birth body weight (LBBW) piglets can reach 10% and is mainly due to gut and immune system immaturity which can lead to a higher risk in the long term. This study aimed to assess the impact of birth body weight (BBW) on piglet metabolism, gut status, and microbial profile from weaning to 21 d postweaning. At birth, 32 piglets were selected for their BBW and inserted into the normal BBW (NBBW:1.38 ±â€…0.09 g) or the LBBW (0.92 ±â€…0.07 g) group. The piglets were weighed weekly from weaning (d0) to d21. At d9 and d21, 8 piglets/group were slaughtered to obtain the distal jejunum for morphology, immunohistochemistry, and gene expression analysis, colon content for microbiota and short-chain fatty acid (SCFA) analysis, and intestinal content for pH measurement. Blood was collected for metabolomic, haptoglobin (Hp), and reactive oxygen metabolite (ROM) analysis. The LBBW group had a lower body weight (BW) throughout the study (P < 0.01), a lower average daily gain from d9-d21 (P = 0.002), and lower feed intake (P = 0.02). The LBBW piglets had lower Hp at d9 (P = 0.03), higher ROMs at d21 (P = 0.06), and a net alteration of the amino acid (AA) metabolism at d9 and d21. A higher expression of NFKB2 was observed in the LBBW piglets at d9 (P = 0.003) and d21 (P < 0.001). MYD88 expression was enhanced in NBBW piglets at d9 (P < 0.001). The LBBW piglets had a lower villus height, absorptive mucosal surface (P = 0.01), and villus height:crypt depth ratio (P = 0.02), and a greater number of T-lymphocytes in both the epithelium and the crypts (P < 0.001) at d21. At d21, the LBBW piglets had higher lactic acid, acetate, butyrate, and valerate, and also higher SCFA in the colon (P < 0.05). The LBBW piglets had a higher Shannon index (P = 0.01) at d9 and a higher abundance of SCFA-fermenting bacteria. In conclusion, the present study confirmed that LBBW could impact the gut mucosal structure, immunity, and inflammatory and oxidative status, leading to an altered AA metabolism, and delaying the recovery from weaning.

The drawback of the high prolificacy selection in the swine industry in the past decades is an increase in the number of piglets born with a low birth body weight (LBBW). This study aimed to assess performance, metabolism, gut status, and microbial profile in piglets born with low (0.92 ±â€…0.07 g) and normal birth body weight (1.38 ±â€…0.09 g). Piglets were weighed weekly from weaning (25 d) until 3 weeks postweaning (end of the trial). At d9 and d21, 8 piglets/group were slaughtered to obtain blood for metabolomic, haptoglobin, reactive oxygen metabolite analyses, colon content for microbiota and short-chain fatty acid, intestinal content for pH measurement, distal jejunum for morphology, immunohistochemistry, and gene expression. The LBBW resulted in lower body weight through the study (P < 0.001), lower average daily gain from d9 to d21 (P = 0.002), and lower feed intake (P = 0.02). The LBBW piglets had a lower villus height, absorptive mucosal surface (P = 0.01), and villus height:crypt depth ratio (P = 0.02), and a greater number of T-lymphocytes in both the epithelium and the crypts (P < 0.001) at d21. In conclusion, the present study confirmed that LBBW could impact the gut mucosal structure, immunity, and inflammatory and oxidative status, leading to an altered AA metabolism, and delaying the recovery from weaning.

Linseed supplementation in the diet of fattening pigs: Effect on the fatty acid profile of different pork cuts.

Linseed supplementation improves the fatty acid (FA) profile of meat, primarily by increasing α-linolenic acid (ALA). Pork is one of the most consumed meats, though the high level of saturated FA, therefore, a change of the FA profile is needed to improve the health properties. The aim of this work was to study the influence of extruded linseed supplementation on the FA profile in 5 different pork cuts, increasing their nutraceutical properties. Sixty pigs were divided into two groups, which were fed two different diets: control (C) and experimental (L, 8% supplementation with extruded flaxseed). Five cuts were sampled: backfat (Bf), bacon (B), Boston shoulder (Bs), ham lean part (Hl) and ham fatty part (Hf). The L diet reduced fat content in Hf (-6%) and B (-11%), while the other cuts did not show differences. Moreover, L group showed a higher level of n-3 PUFA (approx. 9-fold), with a significant reduction in the n-6/n-3 from 20 to 2.5. In cuts rich in fat (Bf, B and Hf) of L group, the level of n-3 PUFA was higher than the thresholds set by EU to obtain the claim "Source of omega-3 fatty acids". In contrast, the lean cuts (Hl and Bs) did not exceed the threshold of n-3 PUFA required to obtain the claim, as consequence of the low level of fat. The results demonstrated that a diet with 8% extruded linseed enhanced the nutraceutical quality of pork meat.

Squamata reptiles as a potential source of helminth infections when preyed on by companion animals.

BACKGROUND: Squamate reptiles cohabiting with companion animals may represent a source of helminth infections, especially through predation by dogs and cats with an outdoor lifestyle. METHODS: In order to assess the role of reptiles as intermediate/paratenic hosts of trophically transmitted helminths, synanthropic reptiles (n = 245) captured from different ecological settings (i.e., households, dog shelters, urban, peri-urban and rural areas or natural parks) of southern Italy were examined for endoparasites. Parasitic cysts (i.e., larval forms of acanthocephalans, cestodes and nematodes) and free helminths (i.e., adult nematodes and digeneans) were morphologically and molecularly identified, and statistical analysis was carried out to evaluate the correlations between reptiles, infections, and ecological settings. RESULTS: Overall, 31% of reptiles were positive for at least one helminth, with Podarcis siculus (18.7%) and Tarentola mauritanica (8.1%) being the most frequently infected species. Among the parasites of medical interest, Joyeuxiella echinorhyncoides showed the highest prevalence (19.7%), followed by Diplopylidium acanthotetra (10.5%), Joyeuxiella pasqualei, Mesocestoides lineatus (5.6%) and Physaloptera sp. (3.9%). Macracanthorhynchus hirudinaceus was detected once. Podarcis siculus and T. mauritanica were associated with cestode infections. CONCLUSIONS: The wide range of helminths detected here in reptiles living in sympatry with pets and the fact that many of these helminth species are parasitic and may infect companion animals (e.g., J. pasqualei, J. echinorhyncoides, D. acanthotetra, Physaloptera sp.) and humans (i.e., Macracanthorhynchus hirudinaceus, Mesocestoides lineatus) indicate the potential health risk associated with pets preying on these small vertebrates. Our results indicate the need for complementary investigations of trophically transmitted parasites in dogs and cats living in sympatry with reptiles.

A Comparison of the Efficacy of Two Omeprazole Formulations in the Treatment of Equine Gastric Ulcer Syndrome in Racehorses: A Blinded, Randomized Clinical Trial.

Equine gastric ulcer syndrome (EGUS) is the most common disease of the stomach in horses and treatment is based on the oral administration of omeprazole for at least 28 days. Aim of this study was to compare the efficacy of two formulations of oral omeprazole-powder paste and gastro-enteric resistant granules-in the treatment of naturally occurring gastric ulcers in racehorses. Thirty-two adult racehorses, aged between 2 and 10 years old, with clinical signs of EGUS were included in this blinded, randomized clinical trial. Two gastroscopies were performed to evaluate gastric lesions in the squamous or glandular mucosa before and after 28 days of treatment. After the first gastroscopy, 2/32 horses were excluded because affected by equine squamous gastric disease (ESGD) 1/4. The remaining horses were divided into 4 groups, and were treated with 2 formulations of omeprazole (gastroenteric resistant granules, group 1, or powder paste, group 3) or with 2 placebo formulations (granules, group 2, or paste, group 4). Treatments were performed in the placebo horses affected by ESGD or equine glandular gastric disease after the T28 gastroscopy control. No differences were found between groups at T0. Differences were found between T0 vs. T28 for granular (P = .002) and powder paste (P = .01) formulations. No differences were detected between the two groups receiving omeprazole formulations at T28 (0.34) and between T0 vs. T28 for either of the groups receiving placebo treatments. For all the variables, the size of the effect was greater than 0.5, proving that the treatments had a considerable effect. Gastro-enteric resistant granules and powder paste omeprazole showed similar efficacy in the treatment of ESGD. The glandular mucosa responded poorly to treatment with omeprazole.

Sustained Recovery of Kidney Function in Patients with ESKD under Chronic Dialysis Treatment: Systematic Review and Meta-Analysis.

The prevalence of recovery of kidney function (RKF) in patients under maintenance dialysis is poorly defined mainly because of different definitions of RKF. Therefore, to gain more insights into the epidemiology of RKF, we performed a systematic review and meta-analysis of studies addressing the prevalence of sustained (at least for 30 days) RKF in patients under maintenance dialysis. Acute kidney injury (AKI) and RKF in the first 90 days of dialysis were the main exclusion criteria. Overall, 7 studies (10 cohorts) including 2,444,943 chronic dialysis patients (range: 430-1,900,595 patients) were meta-analyzed. The period of observation ranged from 4 to 43 years. The prevalence of RKF was 1.49% (95% C.I.:1.05-2.11; p < 0.001] with high heterogeneity I2: 99.8%, p < 0.001. The weighted mean dialysis vintage before RKF was 294 ± 165 days; RKF persisted for a weighted mean of 27.5 months. The percentage of RKF was higher in studies from the U.S. (1.96% [95% C.I.: 1.24-3.07]) as compared to other countries (1.04% [95%C.I.: 0.66-1.62]; p = 0.049). In conclusion, sustained RKF unrelated to AKI occurs in about 1.5% of patients under maintenance dialysis. On average, RKF patients discontinue chronic dialysis about ten months after starting treatment and live free of dialysis for more than two years. The higher prevalence of RKF reported in the U.S. versus other countries suggests a major role of country-specific policies for dialysis start.

Breath Alcohol Test Results in Equine Veterinarians after Performing an Abdominal Ultrasound with Ethanol.

Transcutaneous abdominal ultrasonography using alcohol saturation is used in the diagnostic workup of a variety of conditions in horses. The duration of the examination and the amount of alcohol used in each case may vary depending on several factors. The aim of this study is to describe the breath alcohol test results obtained by veterinarians performing abdominal ultrasound on horses. Six volunteers were enrolled, after written consent, and a Standardbred mare was used for the whole study protocol. Each operator performed a total of 6 ultrasounds by pouring the ethanol solution from a jar or by spray application, for a duration of 10, 30, and 60 min. An infrared breath alcohol analyzer was used immediately after completing the ultrasonography and at 5-min intervals until a negative result was obtained. Positive results were obtained for 0-60 min after the procedure. A statistically significant difference was found between the groups that used more than 1000 mL, 300-1000 mL, and less than 300 mL of ethanol. No significant differences were observed between the type of ethanol administration and the time of exposure. Based on this study, equine vets who perform ultrasound on horses can test positive at the breath alcohol test for up to 60 min following ethanol exposure.

Effect of Cooking with Superheated (SHS) vs. Standard Steam Oven on the Fatty Acids Profile of Different Kinds of Meat and Fish.

We compared the effect of two different kinds of steam oven-a standard (SO) and a superheated steam (SHS) oven-on four different kinds of samples: hamburgers, bovine steaks, pork steaks, and salmon fillets. Ten samples of each meat/fish were divided into three parts. Then samples were analyzed as (i) raw, (ii) cooked with SO, and (iii) cooked with SHS. For each sample, we determined the proximate composition, fatty acid composition and thiobarbituric acid reactive substances (TBARS). The results of fatty acid composition were processed both with a linear model and with a multivariate approach by using three complementary discriminant analysis techniques: canonical (CAN), stepwise (St) and discriminant (DA). SHS was effective in degreasing hamburgers but not the other kinds of samples. Cooking methods selectively affected the fatty acid profile of samples, SHS being higher in MUFA and lower in PUFA n-3 than SO. This result was also confirmed by the discriminant analysis. Finally, samples cooked with SHS showed a lower fatty acid oxidation extent than SO, as the TBARS value was significantly lower in the SHS than in the SO, irrespective of the type of meat/fish cooked.

Equine echocardiography: Can dobutamine infusion correct alterations due to sedation with alpha-2 agonists?

For the echocardiographic examination horses should not be sedated unless absolutely necessary because this alters cardiac dimensions and indices of function. However, some horses do not tolerate the echocardiographic procedure and require sedation to conduct the examination safely and obtain good quality images. The objective of this study was to evaluate whether the concurrent infusion of dobutamine in horses sedated with romifidine counteracts the cardiovascular changes observed with sedation alone. Twelve healthy untrained Standardbred mares were used. Three echocardiographic examinations were performed on the same day for each subject: a) without any treatment under resting conditions (WT); b) under sedation with romifidine administered intravenously (RT); c) under sedation with romifidine and concurrent intravenous infusion with dobutamine (RDT). A three-hour washout period was observed between each examination and the order of the examinations was randomly decided by rolling a dice. The measurements on the images recorded were performed offline at the end of the study protocol and at this point the operator was blinded to the horse and treatment administered. Left ventricular internal diameter (LVID) in diastole, left ventricular free wall (LVFW) in systole, and fractional shortening (FS) were higher in the WT group compared with the other two groups. No differences in the other M-mode and B-mode values were observed. A continuous rate infusion of dobutamine did not counteract the alterations caused by sedation and led to similar echocardiographic measurements to those obtained after romifidine administration.

Novel Board Game versus Active Case-Based Discussion to Teach Final-Year Veterinary Students the Diagnostic Approach to Clinical Cases.

Traditional lectures, where students listen passively, often do not reflect the complexity of real-life decision-making situations. Furthermore, modern students are using online devices for daily activities, and this has a challenging side effect for educators, as many people these days can only maintain their focus if topics are discussed in concise and engaging ways. For these reasons, there is growing interest in the use of games for educational purposes. The aim of this study was to introduce a board game based on the Clue game for final-year veterinary students during their practical activity in large animal medicine. This type of learning process was compared with a classical case-based discussion and evaluated via a survey delivered to the students to both test their acquired knowledge and obtain their evaluation of the activity. A total of 49 students were enrolled in this study. While the board game was evaluated significatively better than the traditional class, no statistically significant differences were observed for the answers given to questions assessing their veterinary skills. The proposed game requires few resources other than a case-based visual materials and analyses from clinical patients, a board, two dice, and some imagination to create cases at the appropriate level for students' knowledge. We conclude that this board game-based activity represents innovative techniques to teach clinical approaches in an interactive way with the same utility as a traditional class but is more enjoyable for the students.

New-onset anemia and associated risk of ESKD and death in non-dialysis CKD patients: a multicohort observational study.

Background: Anemia is a common complication of chronic kidney disease (CKD), but its incidence in nephrology settings is poorly investigated. Similarly, the risks of adverse outcomes associated with new-onset anemia are not known. Methods: We performed a pooled analysis of three observational cohort studies including 1031 non-anemic CKD patients with eGFR <60 mL/min/1.73 m2 regularly followed in renal clinics. We estimated the incidence of mild anemia (hemoglobin 11-12 g/dL in women and 11-13 g/dL in men) and severe anemia (hemoglobin <11 g/dL or use of erythropoiesis-stimulating agents) during a 3-year follow-up period. Thereafter we estimated the risk of end-stage kidney disease (ESKD) and all-cause death associated with new-onset mild and severe anemia. Results: The mean age was 63 ± 14 years, 60% were men and 20% had diabetes. The mean estimated glomerular filtration rate (eGFR) was 37 ± 13 mL/min/1.73 m2 and the median proteinuria was 0.4 g/day [interquartile range (IQR) 0.1-1.1]. The incidence of mild and severe anemia was 13.7/100 patients-year and 6.2/100 patients-year, respectively. Basal predictors of either mild or severe anemia were diabetes, lower hemoglobin, higher serum phosphate, eGFR <30 mL/min/1.73 m2 and proteinuria >0.50 g/day. Male sex, moderate CKD (eGFR 30-44 mL/min/1.73 m2) and moderate proteinuria (0.15-0.50 g/day) predicted only mild anemia. The incidence of anemia increased progressively with CKD stages (from 8.77 to 76.59/100 patients-year) and the proteinuria category (from 13.99 to 25.02/100 patients-year). During a median follow-up of 3.1 years, 232 patients reached ESKD and 135 died. Compared with non-anemic patients, mild anemia was associated with a higher adjusted risk of ESKD {hazard ratio [HR] 1.42 [95% confidence interval (CI) 1.02-1.98]} and all-cause death [HR 1.55 (95% CI 1.04-2.32)]. Severe anemia was associated with an even higher risk of ESKD [HR 1.73 (95% CI 1.20-2.51)] and death [HR 1.83 (95% CI 1.05-3.19)]. Conclusions: New-onset anemia is frequent, particularly in patients with more severe renal damage and in those with diabetes mellitus. The occurrence of anemia, even of a mild degree, is associated with mortality risk and faster progression towards ESKD.

Assessment of blood and productive parameters in mid-lactation dairy cows fed different diets: replacement of corn silage with triticale silage.

Corn crops require large amounts of resources that affect the environmental sustainability of dairy cow farming systems. The aim of the study was thus to investigate the effects of the replacement of corn silage (CS) with triticale silage (TS) by evaluating blood and productive parameters. The study lasted 7 weeks and involved two groups of 20 Italian Holstein Friesian dairy cows that were homogeneous in terms of parity ( 3 ± 1.5 ), days in milk (DIM) ( 150 ± 85.0 ), and daily milk production ( 26 ± 4.6  kg). Chemical analysis of feeds was carried out weekly. Dry-matter intake was estimated daily. At the beginning and end of the trial, haematological, metabolic, and immunological parameters were analysed. At the same, time body weight and body condition score were measured. Milk characteristics were also analysed weekly. Statistical analysis was performed by ANOVA on data of the second sampling, and a non-parametric test was performed to analyse BCS. Regarding the haematological parameters in the two groups, only lymphocyte values were not in the normal range (2.86 and 2.50 × 10 9  L for CS and TS, respectively). Metabolic parameters were in the normal range except for blood ureic nitrogen (BUN; 13.65 and 14.04  mg dL - 1 ), non-esterified fatty acids (NEFAs; 21.40 and 31.93  µ mol L - 1 ), and Cl (91.99 and 93.50  mmol L - 1 ). Hair cortisol was low (0.94 and 0.91  pg mg - 1 ), indicating the absence of stress signs, as confirmed by the results of other immunological parameters (serum lysozyme (SL), bactericidal activity (SBA), haptoglobin (HP), and oxygen free radicals (OFRs)). Statistical differences were not found either for haematological or biochemical parameters. The total replacement of CS with TS did not affect milk yield and composition. In conclusion, the replacement of CS by TS did not give rise to significant modifications in the parameters investigated and did not alter the health status of the animals, thus suggesting the feasibility of its introduction into the diet of mid-lactation dairy cows.

The Kisspeptin analogue C6 induces ovulation in jennies.

Kisspeptins (KPs) are the most potent stimulating neurotransmitters of GnRH release, and consequently KP administration triggers LH and/or FSH release. In small ruminants, KP or its analogs induced an LH surge followed by ovulation in both cyclic and acyclic animals, while in the mare KP only increased LH plasma levels but failed to induce ovulation. This study in jennies compares the endocrinological effects, ovulatory and pregnancy rates of the KP analog C6 and the GnRH analog buserelin acetate. The ovarian activity of nine Amiata jennies was monitored daily by transrectal ultrasound for three complete estrous cycles. Jennies in estrus were assigned, to one of three treatment groups: 50 nmol of the KP analog C6 (injected twice, 24 h apart, C6 group); 0.4 mg buserelin acetate (injected once, Bu group); and 2 mL of saline (injected once, CTRL group). Blood samples were collected at Day-1 (-24 h) Day0 (h0, before treatment), h2, h4, h6, h8, h10, h24 (before second treatment with C6), h26, h28, h30, h32, h34, h48 and every 24 h until ovulation. Jennies were inseminated once at h24 with fresh extended semen from a donkey stallion. Pregnancy diagnoses were performed 14 days after ovulation. On days 5, 10, and 14 after ovulation, for every CL the cross-sectional area (CSA) and the vascularized area (VA) were recorded by color doppler ultrasound and measured. Significantly higher plasma LH levels were found after induction between the Bu and CTRL groups at h6 and h8 (P < 0.05), while tendentially higher differences were found between the Bu/C6 groups and CTRL at h10. Five/9, 4/9, and 2/9 jennies ovulated between 24 and 48 h after induction from the Bu, C6, and CTRL groups respectively, (P > 0.05). Correlations between corpora lutea CSA and VA with serum progesterone concentration were r = 0.31, P = 0.01, r = 0.38, P = 0.01, respectively. Pregnancy rates after artificial insemination did not differ among groups (CTRL: 6/9, 66.7%; C6: 7/9, 77.8%; Bu: 6/9, 66.7%; P > 0.05). Ovulation rates after C6 treatment were comparable to that of Bu, although not different from the CTRL. Pregnancy rates were comparable to the literature in terms of fresh extended donkey semen in every group. This study suggests that stimulation of the Kp system in jennies, in contrast to findings observed in mares, induces ovulation. Further studies using higher doses and/or more animals are needed to better characterize the efficacy of C6 in jennies.

Characterization of the microbial community in ripened Pecorino Toscano cheese affected by pink discoloration.

Pink discoloration defect can cause economic losses for cheese producers due to the impossibility to sell the defected cheese, but few knowledge is currently available on the causes of this defect. To gain more insight on the causes that lead to the formation of pink discoloration in Pecorino Toscano cheese with the Protected Designation of Origin (PDO) status, the bacterial community in defected and not defected cheese was characterized by high-throughput sequencing of bacterial 16S rRNA gene. The bacterial community in the defected cheese significantly differed compared to the control. The relative abundance of the genera Acidipropionibacterium, Enterococcus, Escherichia/Shigella, Lactobacillus, Lentilactobacillus and Propionibacterium was higher in the cheese with pink discoloration defect. The concentration of short chain fatty acids and of lactic acid in cheese was measured and a shift towards the production of propionate in the cheese with pink discoloration defect was observed. Furthermore, the possible involvement of microbially produced vitamin B12 in the formation of pink discoloration was not supported by the data, since a tendency to a lower concentration of vitamin B12 was measured in the defected cheese compared to the control.

Study of the Fatty Acid Profile of Milk in Different Sheep Breeds: Evaluation by Multivariate Factorial Analysis.

A multivariate analysis was used to investigate the fatty acid (FA) profile in three different Italian sheep breeds: Comisana, Massese, and Sarda. A sample of 852 animals was considered: 118 Massese, 303 Comisana, 431 Sarda. Sarda sheep were divided into two groups, based on their breeding origin (298 and 133 reared in Sardinia and Tuscany, respectively). Sarda sheep, bred both in Sardinia and in Tuscany, were considered in different groups, both because in these two regions most of the sheep of this breed are reared, and because they differ in geographical characteristics and in the farming system. The individual milk FA composition of dairy ewes was analyzed with multivariate factor analysis. The extracted factors were representative of the following eight groups of fatty acids or functions: factor 1 (odd branched fatty acids and long-chain fatty acids), factor 2 (sn3_position), factor 3 (alternative biohydrogenation), factor 4 (SCD_1), factor 5 (SCD_2), factor 6 (SCD_3), factor 7 (fat secretion) and factor 8 (omega-3). A factor analysis suggested the presence of different metabolic pathways for de novo short- and medium-chain fatty acids and Δ9-desaturase products. The ANOVA of factor scores highlighted the significant effects of the breed. The results of the present study showed that breed is an important factor in defining the fatty acid profile of milk, combined with the effect of the diet. Breeds reared in the same farming system (Comisana, Massese and Sarda reared in Tuscany) showed significant differences for all the factors extracted. At the same time, we found differences between the Sarda sheep reared in Sardinia and Tuscany, two different regions of Italy.