Differentiating progressive supranuclear palsy‐parkinsonism (PSP‐P) from Parkinson's disease (PD) is clinically challenging.This study aimed to develop an automated Magnetic Resonance Parkinsonism Index 2.0 (MRPI 2.0) algorithm to distinguish PSP‐P from PD and to validate its diagnostic performance in two large independent cohorts.We enrolled 676 participants: a training cohort  from our center and an independent testing cohort  from an international research group. We developed a new in‐house algorithm for MRPI 2.0 calculation and assessed its performance in distinguishing PSP‐P from PD and control subjects in both cohorts using receiver operating characteristic curves.r = 0.91, P < 0.001) was found between automated and manual MRPI 2.0 values.The automated MRPI 2.0 showed excellent performance in differentiating patients with PSP‐P from patients with PD and control subjects both in the training cohort  and in the international testing cohort , suggesting the generalizability of the results. The automated MRPI 2.0 also accurately distinguished between PSP‐P and PD in the early stage of the diseases (AUC = 0.91 [0.84–0.97]). A strong correlation (Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder SocietyOur study provides an automated, validated, and generalizable magnetic resonance biomarker to distinguish PSP‐P from PD. The use of the automated MRPI 2.0 algorithm rather than manual measurements could be important to standardize measures in patients with PSP‐P across centers, with a positive impact on multicenter studies and clinical trials involving patients from different geographic regions. © 2022 The Authors.  The clinical differential diagnosis between progressive supranuclear palsy‐parkinsonism (PSP‐P) and Parkinson's disease (PD) may be challenging, especially in the first years after the disease onset.To date, several imaging biomarkers have proved to be useful in distinguishing PSP from PD, and the Magnetic Resonance Parkinsonism Index (MRPI) is one of the most powerful and robust ones.Preliminary results have shown that MRPI 2.0 accurately differentiated patients with PSP‐P from patients with PD,The aim of this study was the development of a fully automated algorithm to calculate MRPI 2.0 and the validation of the automated MRPI 2.0 performance in differentiating patients with PSP‐P from patients with PD and control subjects in two large independent cohorts from different geographic regions.A total of 676 participants were enrolled in this study, divided into a training and a testing cohort. The training cohort included 346 participants  from our center, while the independent testing cohort included 330 participants  from an international research group.off state and the Hoehn and Yahr (H‐Y) rating scale.Participants in the training cohort were consecutively recruited between March 2012 and January 2020 at the Movement Disorder Center of Magna Graecia University, Catanzaro, Italy. The PSP‐P, PD, and control subjects included in the testing cohort were enrolled from seven different centers (Supporting Information Table 123I‐ioflupane Single Photon Emission Computed Tomography (DaTscan), and magnetic resonance imaging (MRI) abnormalities such as lacunar infarctions in the basal ganglia and/or subcortical vascular lesions with diffuse periventricular signal alterations. None of the control participants were younger than 40 years or had a history of neurological, psychiatric, or other major medical illnesses. We also excluded subjects from the study who showed Evans Index >0.32 associated with callosal angle <100 degrees, which is a combination of MRI biomarkers strongly suggestive of normal pressure hydrocephalus.Exclusion criteria for patients with PD and PSP consisted of age <40 years, clinical features suggestive of other diseases, normal striatal uptake on All study procedures and ethical aspects were approved by an institutional review board . Each recruitment site received approval from an institutional review board or ethics committee. Written informed consent according to the Declaration of Helsinki for the use of their medical records for research purposes was obtained from all individuals participating in the study.All patients and controls in the training cohort underwent a brain MRI with a 3‐T MR750 General Electric scanner and an eight‐channel head coil, with a recently described MRI protocol.https://mrpi.unicz.it)Patients and control subjects from the international cohort underwent a brain MRI with 3‐T  or 1.5‐T scanners (26 PD and 17 control subjects), with a protocol including a T1‐weighted volumetric image. The 3D T1‐weighted MRIs were uploaded on the web‐based platform for the automated MRPI calculation  width ratio Fig. . The autP < 0.05. All P values were corrected according to Bonferroni. We assessed the diagnostic performance of the automated MRPI and MRPI 2.0 in differentiating patients with PSP‐P from patients with PD and control subjects, in both the training and testing cohorts. Optimal cutoffs, defined as the values with the highest sum of sensitivity and specificity on the receiver operating characteristic (ROC) curves, and 95% confidence intervals (CIs) were calculated using pROC software package with bootstrapping (n = 2000 iterations).Difference in sex distribution was assessed by Fisher's exact test. The Shapiro–Wilk test was used to check for normality to decide whether parametric or nonparametric tests were appropriate for comparisons. Age at examination was compared using Kruskal–Wallis test followed by pairwise Wilcoxon rank sum test; age at disease onset and disease duration were compared using Wilcoxon rank sum test. Differences in MDS‐UPDRS‐III score, H‐Y score, MRPI, 3 V, FHs, 3 V/FHs ratio, and MRPI 2.0 values were investigated using analysis of covariance on generalized linear models with age and sex as covariates, followed by the Tukey test. The scanner field strength (1.5 and 3.0 T) was also included as covariate in the testing cohort; the MDS‐UPDRS‐III score was also included as covariate in the PSP‐P versus PD comparison of imaging data in both cohorts. All tests were 2‐tailed, and the α level was set at We also investigated the MRPI 2.0 diagnostic performances after excluding the possible age effect. To correct for age effect, we fitted a linear regression model on MRPI 2.0 values in control subjects, both in the in‐house and in the external cohort, and computed residuals for PD, PSP‐P, and control subjects in both cohorts. Logistic regression models were evaluated for PSP‐P versus PD and PSP‐P versus controls classification, using both original MRPI 2.0 data and residuals. For each model, a ROC analysis was performed, and the De Long test was used to compare ROC curves evaluated on raw MRPI 2.0 values to ROC curves evaluated on residuals. The effect of sex on MRPI 2.0 classification of PSP‐P was evaluated in male and female subjects separately.Logistic regression models were also used to evaluate the association between MRPI 2.0 values and the probability of having PSP‐P in both cohorts, taking into account the proportion between patients with PD and PSP‐P. The correlation between automated and manual MRPI 2.0 values was evaluated using Pearson's correlation test. The intraclass correlation coefficient was calculated to investigate the reproducibility of automated MRPI 2.0 measurements. Statistical analyses were performed using R statistical software  and the ROCR package for R.In this study, we developed a fully automated algorithm to calculate the MRPI 2.0. This software provided the calculation in 92.5% of cases (625/676 MRI DICOM images), and failure occurred in only 7 patients with PSP‐P, 21 patients with PD, and 23 control subjects, who were excluded from the subsequent analyses. Thirty of the failures were due to errors in the automated MRPI calculation, while the remaining 21 failures were due to errors in the automated measurement of the 3 V or FHs width, probably caused by motion artefacts that did not allow the automatic identification of anatomical landmarks used for the segmenting procedures.The final training cohort included 312 patients from our center of Catanzaro, Italy , while the independent testing cohort included 313 patients from several international research groups . The demographic, clinical, and imaging data of patients and control subjects in the two cohorts are summarized in Table P = 0.638; PSP‐P versus controls: D = 0.92, P = 0.360), demonstrating the generalizability of our findings , while no differences were found between patients with PD and control subjects Table . In the ngs Fig. . The MRPngs Fig. . Logistings Fig. . MRPI 2.ngs Fig. .P < 0.05), and this difference was slightly larger in the PSP‐P group. Thus, the classification accuracy of MRPI 2.0 in distinguishing patients with PSP‐P from patients with PD and control subjects was slightly higher in the male than in the female cohort   Fig. . MoreoveFinally, we evaluated the diagnostic performance of MRPI 2.0 in an intention‐to‐treat analysis, taking into account the failures of the automated algorithm ,In addition, this new automated MR biomarker accurately distinguished PSP‐P from PD in a subcohort of patients with short disease duration (up to 3 years from the disease onset), demonstrating its usefulness also in the early stage of the diseases. Our findings in patients with early‐stage PSP are in accordance with previous studies showing that high MRPI 2.0 values predicted the development of PSP clinical features in patients with clinically unclassifiable parkinsonismOverall, this new automated MR algorithm provides reliable and reproducible results significantly improving standardization of the measurements across centers. This finding together with its generalizability makes automated MRPI 2.0 a valid MR biomarker for PSP‐P classification with a positive impact also on research studies and clinical trials involving patients from different geographic regions. The reproducibility guaranteed by automatic measurements also makes this biomarker suitable for longitudinal studies aiming at evaluating disease progression of PSP through repeated measurements over time. A recent studyhttps://mrpi.unicz.it) on registration.This study has several strengths. First, patients with PSP‐P and PD were from two large and independent cohorts, and the performance of MRPI 2.0 in the training cohort was validated in the international independent cohort, thus ensuring the generalizability of the results. Second, this study involves 99 patients with PSP‐P, representing one of the largest PSP‐P cohorts ever described. Third, we demonstrated that MRPI 2.0 was accurate in distinguishing PSP‐P from PD also in the early stage of the diseases, when the differential diagnosis is much more challenging. Fourth, we developed a reliable fully automated algorithm for MRPI 2.0 calculation to reduce the variability of manual measurements, which need expertise for image reconstruction and slice selection. This software is freely available online  compared with pathological data.In conclusion, this study demonstrates that the automated MRPI 2.0 is a powerful, validated, and generalizable MR biomarker for distinguishing PSP‐P from PD in vivo. The automated algorithm for calculating MRPI 2.0 reduces the variability of the measurements across centers, allowing to obtain reliable results worldwide. These findings provide a strong impetus to use automated MRPI 2.0 for supporting clinical diagnosis of PSP‐P, especially in multicentric studies and clinical trials with potential new disease‐modifying therapies.1. Research project: A. Conception, B. Organization, C. Execution;2. Statistical Analysis: A. Design, B. Execution, C. Review and Critique;3. Manuscript: A. Writing of the first draft, B. Review and Critique;Andrea Quattrone: 1A, 1B, 1C, 3A, 3B; Maria G. Bianco: 1B, 1C, 2C, 3A; 3B; Angelo Antonini: 1B, 1C, 3B; David E. Vaillancourt: 1B, 1C, 3B; Klaus Seppi: 1B, 1C, 3B; Roberto Ceravolo: 1B, 1C, 3B; Antonio P. Strafella: 1B, 1C, 3B; Gioacchino Tedeschi: 1B, 1C, 3B; Alessandro Tessitore: 1B, 1C, 3B; Roberto Cilia: 1B, 1C, 3B; Maurizio Morelli: 1B, 1C; Salvatore Nigro: 1B, 1C, 3B; Basilio Vescio: 2A, 2B, 2C; Pier Paolo Arcuri: 1B, 1C; Rosa De Micco: 1B, 1C; Mario Cirillo: 1B, 1C; Luca Weis: 1B, 1C; Eleonora Fiorenzato: 1B, 1C; Roberta Biundo: 1B, 1C; Roxana G. Burciu: 1B, 1C; Florian Krismer: 1B, 1C; Nikolaus R. McFarland: 1B, 1C; Christoph Mueller: 1B, 1C; Elke R. Gizewski: 1B, 1C; Mirco Cosottini: 1B, 1C; Eleonora Del Prete: 1B, 1C; Sonia Mazzucchi: 1B, 1C; Aldo Quattrone: 1A, 1B, 3B.A. Antonini has received compensation for consultancy and speaker‐related activities from UCB, Boehringer Ingelheim, Ever Pharma, Jazz Pharmaceuticals, General Electric, Britannia, AbbVie, Kyowa Kirin, Zambon, Bial, Neuroderm, Theravance Biopharma, Roche, and Medscape; he receives research support from Bial, Lundbeck, Roche, Angelini Pharmaceuticals, Horizon 2020 (Grants 825785 and 101016902), Ministry of Education University and Research , Cariparo Foundation, and Movement Disorder Society for NMS Scale validation. He serves as consultant for Boehringer–Ingelheim for legal cases on pathological gambling.D.E. Vaillancourt has received grant support from the National Institutes of Health, National Science Foundation, and Tyler's Hope Foundation. He is cofounder and manager of Neuroimaging Solutions, LLC.K. Seppi reports personal fees from Teva, UCB, Lundbeck, AOP Orphan Pharmaceuticals AG, Roche, Grünenthal, Stada, Licher Pharma, Biogen, and AbbVie; honoraria from the International Parkinson and Movement Disorder Society; and research grants from the FWF Austrian Science Fund, The Michael J. Fox Foundation, and AOP Orphan Pharmaceuticals AG.R. Ceravolo has received speaking fees from Zambon, UCB Pharma, AbbVie, and Lusofarmaco.A.P. Strafella is a consultant for Hoffman La Roche and received honoraria from GE Health Care Canada LTD, Hoffman La Roche.G. Tedeschi has received speaker honoraria from Sanofi Aventis, Merck Serono, Bayer Schering Pharma, Novartis, Biogen‐Dompé AG, Teva, and Lilly; has received funding for travel from Bayer Schering Pharma, Biogen‐Dompé AG, Merck Serono, Novartis, and Sanofi Aventis; and serves as an associate editor of Neurological Sciences.A. Tessitore has received speaker honoraria from Novartis, Schwarz Pharma/UCB, Lundbeck, AbbVie, and Glaxo.F. Krismer reports receiving personal fees from Institut de Recherches Internationales Servier, Clarion Healthcare, and the Austrian Society of Neurology and grant support from the MSA Coalition outside of the submitted work.All other authors report no disclosures.Figure S1 Automated MRPI 2.0 pipeline, from the mid‐sagittal plane identification to automatic segmentation of the third ventricle width and frontal horns width.Click here for additional data file.Figure S2 Receiver operating characteristic (ROC) curves for assessing the classification performance of automated MRPI (blue) and MRPI 2.0 (red) in differentiating PSP‐P from PD patients, and PSP‐P from control subjects both in the in the training cohort (A and C) and in the testing cohort (B and D). Abbreviations: MRPI, Magnetic Resonance Parkinsonism Index; PSP‐P, Progressive supranuclear palsy‐parkinsonism; PD, Parkinson's disease; AUC, area under the ROC curve.Click here for additional data file.Figure S3 Receiver operating characteristic (ROC) curves for assessing the classification performance of automated MRPI 2.0 in differentiating PSP‐P from PD patients, and PSP‐P from control subjects both in the in the training cohort (A and C) and in the testing cohort (B and D). The red ROC curves were evaluated on raw automated MRPI 2.0 values, while the blue ROC curves were evaluated on residuals after correcting for age. Abbreviations: MRPI 2.0, Magnetic Resonance Parkinsonism Index 2.0; PSP‐P, Progressive supranuclear palsy‐parkinsonism; PD, Parkinson's disease; AUC, area under the ROC curve.Click here for additional data file.Figure S4 Receiver operating characteristic (ROC) curves for assessing the classification performance of automated MRPI 2.0 in differentiating PSP‐P from PD patients (A) and control subjects (B) in male (red) and female subjects (blue). De Long test showed no significant differences in the diagnostic performance of automated MRPI 2.0 between the male and female cohorts . The male cohort included 66 PSP‐P, 204 PD and 92 controls from both cohorts; the female cohort included 33 PSP‐P, 139 PD and 91 controls from both cohorts. Abbreviations: MRPI 2.0, Magnetic Resonance Parkinsonism Index 2.0; PSP‐P, Progressive supranuclear palsy‐parkinsonism; PD, Parkinson's disease; AUC, area under the ROC curve.Click here for additional data file.Table S1 Number of participants for each center.Click here for additional data file.Table S2 Diagnostic performance of the Magnetic Resonance Parkinsonism Index 2.0 in differentiating patients with progressive supranuclear palsy‐parkinsonism from those with Parkinson's disease and control subjects, in the training and testing cohorts.Click here for additional data file.Table S3 Diagnostic performance of the automated Magnetic Resonance Parkinsonism Index and Magnetic Resonance Parkinsonism Index 2.0 in differentiating between patients with progressive supranuclear palsy‐parkinsonism and those with Parkinson's disease, in the early stage of the diseases.Click here for additional data file.Table S4 Diagnostic performance of the Magnetic Resonance Parkinsonism Index 2.0 in differentiating patients with PSP‐P from those with PD and control subjects, also considering the algorithm failures.Click here for additional data file.Appendix S1 Supporting Information.Click here for additional data file.


To the Editor:Balbach et al.P = 6.7 × 10−16) and 54 G12C mutations versus two in SCLC . The lower prevalence in SCLC is suggestive of reduced oncogenic effect, but the difference could be explainable by differences in underlying KRAS G12 mutation rates between SCLC and NSCLC. Therefore, we calculated neutral mutation rates and quantified selection for KRAS G12C in each cancer type with cancereffectsizeR 2.6.4.4, 23rd of 3272 recurrent variants. In SCLC, it was 1.2 × 103—12-fold lower, 1120th of 1259. Lower effect indicates a lesser role in the growth and proliferation of SCLC than of NSCLC. Treatment with sotorasib may trend less beneficial for patients with KRAS G12C SCLC than for patients with KRAS G12C NSCLC.To quantify the tumorigenic effect of KRAS G12C mutation, we aggregated somatic variant calls from whole-exome sequencing of 1042 NSCLC tumorsB2M was present at 88.4% VAF. A similar B2M loss-of-function mutation in SCLC exhibits sixfold higher effect than KRAS G12C—ranking 382nd at 6.9 × 103. Prospects for immune checkpoint therapy should be tempered by the presence of B2M mutation, which has been linked to mismatch-repair deficiency in colon cancer and to acquired resistance to immune checkpoint inhibitors in melanomaPotential treatments may also be informed by the effect sizes of other variants in the patient which are quantified at higher oncogenic effect than KRAS G12C. For example, a point mutation of Margaret Moore: Conceptualization, Investigation, Writing—original draft, Writing—review and editing, Project administration.Yuyang Zhuo: Software, Formal analysis, Investigation, Data curation, Writing—review and editing.Jeffrey Mandell: Methodology, Software, Validation, Formal analysis, Data curation, Writing—review and editing.Stephen Gaffney: Validation, Data curation, Writing—review and editing, Supervision.Jeffrey Townsend: Conceptualization, Methodology, Resources, Writing—original draft, Writing—review and editing, Supervision, Project administration.


Rhamdia quelen is a commercially important fish in South America. During the survey of myxozoan infections in fishes from Pardo River, Paranapanema River basin, São Paulo State, Brazil, we describe three new species of Henneguya found parasitizing gills of R. quelen: Henneguya bagre n. sp., Henneguya breviscauda n. sp and Henneguya novaerae n. sp. The descriptions were based on myxospores morphology and small subunit ribosomal DNA partial sequences. Phylogenetic analysis showed a clade formed by species that parasitize Siluriformes, with Henneguya jundiai as a sister species of Henneguya bagre n. sp., Henneguya breviscauda n. sp and Henneguya novaerae n. sp. Our study indicates that the parasites infecting R. quelen belong to a lineage of myxozoans infecting Heptapteridae fishes. Using molecular and morphological characterization, the species were identified as new species for the genus Henneguya. Based on our analysis we recommend monitoring the presence of these parasites in farmed fishes, to analyze possible pathologies caused by them. Past works suggest that species compilations from the Paranapanema River basin can reach over 160 fish species4. The Pardo River is one of the main tributaries of the Paranapanema River, composing its basin, and contributing to the region's fish biodiversity. These fishes are potentially exposed to parasitic organisms in their life cycle5. Among the parasites are the myxozoans, which are cnidarians that can infect freshwater and marine fishes all over the world with over 2600 species identified7. In the family Myxobolidae Thélohan, 1892, Henneguya Thélohan, 1892 is one of the most specious genera with more than 200 described species8. Morphological variation of myxospores within a species or morphological similarities between species create challenges for myxosporean species identification11. Some species may have very similar myxospores but are genetically distant from each other, while in other cases, genetic data may show that what appears to be morphologically distinct species are conspecific15. In this context, the use of molecular techniques is essential for genetic differentiation between species.The Paranapanema basin is one of the most important basins in southeast Brazil as evidenced by several fish inventories and ecological studies performedRhamdia quelen  is a species widely distributed in South and Central America, east of the Andes, and between Venezuela and northern Argentina16. It is popularly known as “bagre”, “jundiá” or “mandi-guaru”. This fish species is widely accepted by the consumer market due to its excellent flavored meat, high resistance to cold periods, rapid growth, rusticity regarding handling, easy adaptation to artificial feeding, and lack of intramuscular spines18, representing about 2% of the total fish produced by the Brazilian aquaculture sector19, being very important for fish farming in southern Brazil20. Recently Grigio and Meurer18 proposed R. quelen as an alternative in recirculation systems to the species Oreochromis niloticus , which is currently the most consumed species in Brazil. This catfish has nocturnal habits with a preference for quiet and deep river sites21 with large pebbles or with submerged trunks22. Females reach greater lengths and weights than males, with 66.5 cm total length (TL) being the asymptotic length of females and 52 cm TL of males, reaching more than 3 kg21.The catfish Henneguya spp. have already been described in R. quelen in Brazil: Henneguya jundiai Negrelli, Vieira, Tagliavini, Abdallah and Azevedo, 2019 parasitizing the gills arches of R. quelen from the Jacaré-Pepira river, São Paulo state; Henneguya rhamdia Matos, Tajdari & Azevedo, 2005 parasitizing the gill filaments of R. quelen from Peixe Boi River, Pará state, Brazil; and Henneguya quelen Abrunhosa, Sindeaux-Neto, Hamoy & Matos, 2018 infecting the kidneys of R. quelen from Paracauari River, Marajó Island, Para state25.Three R. quelen, in the Pardo River, Paranapanema River basin, São Paulo state, Brazil.The present study describes the morphology and phylogenetic relationships of three new species of myxozoans parasites found in the gills of the siluriform fish, R. quelen were collected . Although equal efforts were made in all localities, the number of captured hosts was different between the locations. Eleven specimens were collected in locality 1, two specimens in locality 2, and seven specimens in locality 3. Plasmodia of Henneguya spp. were observed in gill filaments of 11 specimens . Specific prevalence can be seen in Table Henneguya based on morphological and ssrDNA sequencing data is presented.In total, 20 specimens of Henneguya novaerae n. sp.  .Type-host: Type-locality: Pardo River, Nova Era Resort, municipality of Botucatu, São Paulo State, Brazil .Other locality: Pardo River, municipality of Salto Grande, São Paulo State, Brazil .Site of infection: histozoic, gill filaments .Type-material: A glass slide with myxospores (hapantotype) was deposited in the collection of the Instituto Nacional de Pesquisa da Amazônia (INPA), Brazil (Num. INPA91). The ssrDNA partial sequences were deposited in GenBank with accession numbers OP070158-OP070160.Etymology: The specific name is derived from the name of the locality (Nova Era Resort) where the host was collected.Zoobank registration code: urn:lsid:zoobank.org:pub:D6824583-AAD4-4FC8-B67F-40233059FDF2.Plasmodia: whitish, round and small, measuring about 0.1 mm. Observed in the primary filaments of the gills, occupying an interlamellar position. Average one plasmodia per parasitized fish.Myxospores: The morphological data are based on the observation of myxospores isolated from a single plasmodium. The myxospores found were elongated and ellipsoidal in frontal view. Two elongate, elliptical shell valves with two long tapering caudal appendages. Average measurements: total length 24.9 ± 0.9 (23.3–26.8) µm, body length 10.7 ± 0.5 (10.1–11.6) µm, body width 3.8 ± 0.3 (3.2–4.2) µm, caudal appendages length 14.7 ± 1.4 (11.5–16.1) µm. Myxospores were biconvex in the lateral view with a thickness of 3.5 ± 0.3 (3.1–4.0) µm. The polar capsules were equal in size and measured 4.8 ± 0.4 (3.8–5.5) µm in length and 1.3 ± 0.2 (1.1–1.7) µm in width, occupying about half of the sporoplasm. Six coils in the polar tubules present inside the polar capsule perpendicular to the longitudinal axis of the polar capsule were observed. Binucleated sporoplasm and symmetrical valves.Henneguya bagre n. sp.  .Type-host: Type-locality: Pardo River, municipality of Salto Grande, São Paulo State, Brazil .Site of infection: histozoic, gill filaments .Type-material: A glass slide with myxospores (hapantotype) was deposited in the collection of the Instituto Nacional de Pesquisa da Amazônia (INPA), Brazil (Num. INPA92). The ssrDNA partial sequence was deposited in GenBank with accession number OP070161.R. quelen (bagre).Etymology: The specific name is derived from the Brazilian popular name of Zoobank registration code: urn:lsid:zoobank.org:pub:D6824583-AAD4-4FC8-B67F-40233059FDF2.Plasmodia: whitish, elongated, occupying the inner region of the primary gill filament , measuring about 3 mm. Average one plasmodia per parasitized fish.Myxospores: The morphological data are based on the observation of myxospores isolated from a single plasmodium. The myxospores found were elongated and ellipsoidal in frontal view. Two elongate, elliptical shell valves with two medium tapering caudal appendages. Average measurements: total length 20.9 ± 0.9 (19.2–21.8) µm, body length 11.2 ± 0.5 (10.5–12.4) µm, body width 5.1 ± 0.3 (4.6–6.1) µm, caudal appendages length 9.8 ± 0.9 (8.2–11.1) µm. Myxospores were biconvex in the lateral view with a thickness of 3.8 ± 0.2 (3.6–4.1) µm. The polar capsules were equal in size and measured 5.9 ± 0.6 (4.9–6.7) µm in length and 1.6 ± 0.1 (1.4–1.8) µm in width, occupying about half of the sporoplasm. Seven coils in the polar tubules present inside the polar capsule perpendicular to the longitudinal axis of the polar capsule were observed. Binucleated sporoplasm and symmetrical valves.Henneguya breviscauda n. sp.  .Type-host: Type-locality: Pardo River, municipality of Avaré, São Paulo State, Brazil .Site of infection: histozoic, gill filaments .Type-material: A glass slide with myxospores (hapantotype) was deposited in the collection of the Instituto Nacional de Pesquisa da Amazônia (INPA), Brazil (Num. INPA93). The ssrDNA partial sequence was deposited in GenBank with accession number OP070162.brevis  and cauda (tail) and referred to the small size of their caudal appendages.Etymology: The specific name is derived from the Latin words: Zoobank registration code: urn:lsid:zoobank.org:pub:D6824583-AAD4-4FC8-B67F-40233059FDF2.Plasmodia: whitish, round and small, measuring about 0.05 mm. Present in the basal region of the gill filaments, occupying an interfilamental position. Average one plasmodia per parasitized fish.Myxospores: The morphological data are based on the observation of myxospores isolated from a single plasmodium. The myxospores found were elongated and ellipsoidal in frontal view. Two elongated, elliptical shell valves with two small tapering caudal appendages. Average measurements: total length 17.1 ± 0.9 (15.9–18.7) µm, body length 10.7 ± 0.4 (9.6–10.8) µm, body width 5.3 ± 0.5 (4.4–6.1) µm, caudal appendages length 7.2 ± 0.8 (5.8–8.2) µm. Myxospores were biconvex in the lateral view with a thickness of 3.7 ± 0.1 (3.6–3.9) µm. The polar capsules were unequal in size. The larger polar capsule measured 5.4 ± 0.2 (5.1–5.6) µm in length and 1.5 ± 0.1 (1.2–1.6) µm in width, while the smaller polar capsule measured 4.6 ± 0.3 (4.2–5.0) µm in length and 1.2 ± 0.1 (1.1–1.3) µm, occupying about half of the sporoplasm. Seven coils in the polar tubules present inside the polar capsule perpendicular to the longitudinal axis of the polar capsule were observed. Symmetrical valves.Henneguya novaerae n. sp. , one partial sequence of Henneguya bagre n. sp. (1873-bp), and one partial sequence of Henneguya breviscauda n. sp. (1898-bp) from ssrDNA were obtained. The partial sequences were obtained from the type-host of the species. Henneguya novaerae n. sp. was 88.2% genetically similar to Henneguya breviscauda n. sp. (195 different nucleotides) and 91.4% genetically similar to Henneguya bagre n. sp. (162 different nucleotides). Henneguya breviscauda n. sp. and Henneguya bagre n. sp. are 91.9% genetically similar to each other (134 different nucleotides). The BLAST search showed that the highest similarity to other myxozoan sequences available in GenBank was no higher than 93.6%. Among the species with partial sequences available on GenBank, the one that most genetically resembled the three new species was H. jundiai. The registered similarity was 93.4% with Henneguya novaerae n. sp. (107 different nucleotides), 90.1% with Henneguya breviscauda n. sp. (182 different nucleotides), and 93.6% with Henneguya bagre n. sp (119 different nucleotides).Three genetically identical partial sequences of the Henneguya/Myxobolus spp. that parasitize Siluriformes fishes of the families Ictaluridae, Pimelodidae, and Heptapteridae, where the three new species are inserted. Henneguya novaerae n. sp. appears as a sister species of H. jundiai, while Henneguya bagre n. sp. and Henneguya breviscauda n. sp. are sister species. All species that integrate the subclade that parasitizes fish of the family Heptapteridae parasitize R. quelen. Clade B is mainly composed of Myxobolus spp. which parasitizes Characiformes fish and Henneguya spp. which parasitizes Esociformes fishes.Phylogenetic analysis of species genetically similar to the new species showed a division into two main polyphyletic clades Fig. . Clade AHenneguya novaerae n. sp., Henneguya bagre n. sp., and Henneguya breviscauda n. sp. were distinguished from the other Henneguya spp. by the morphological or molecular way. The three new species were compared with each other and with species previously described parasitizing Siluriformes fishes from the Americas , also differentiating the two species. Furthermore, the molecular differences reported above support the discrimination of the species.The three new species described in this study are visually similar and the myxospore body is almost identical in morphology and morphometry. However, there is a clear difference concerning the size of the caudal appendage of the species (14.7 ± 1.4 µm vs. 9.8 ± 0.9 µm vs. 7.2 ± 0.8 µm), which also causes a distinction in the size of the total length of the myxospore (24.9 ± 0.9 µm vs. 20.9 ± 0.9 µm vs. 17.1 ± 0.9 µm). Henneguya novaerae sp. was compared with other Henneguya spp. that parasitizes R. quelen. Henneguya jundiai is the species that most resembles Henneguya novaerae sp. However, there are morphometric differences regarding the length and width of the myxospore body. For H. jundiai the maximum reported spore body length was 10.0 µm, with an average of 9.5 µm, while for Henneguya novaerae n. sp. the measurements found were 10.7 ± 0.5 (10.1–11.6) µm. Concerning the width of the myxospore body, the opposite occurs, with H. jundiai having larger measurements than those found for Henneguya novaerae n. sp. (4.6 ± 0.4 (4.1–5.5) µm vs. 3.8 ± 0.3 (3.2–4.2) µm, respectively). Furthermore, H. jundiai parasitizes the gill arch of R. quelen, while Henneguya novaerae n. sp. parasitizes the gill filaments. The molecular comparison with 106 different nucleotides  in the partial sequences available for the species reinforces the distinction between these two species. About the other two Henneguya spp. described parasitizing R. quelen, Henneguya novaerae n. sp. presented smaller measures in almost all characteristics, besides the molecular differences that can be observed in the phylogenetic analysis.Henneguya novaerae n. sp. was Henneguya pseudoplatystoma Naldoni, Arana, Maia, Ceccarelli, Tavares, Borges, Pozo & Adriano, 2009. However, H. pseudoplatystoma presents the length of the caudal appendages greater than those observed in Henneguya novaerae n. sp (22.7 ± 1.7 µm vs. 14.7 ± 1.4 (11.5–16.1) µm). Henneguya novaerae n. sp. has also been compared to Henneguya spp. which parasitizes fishes of other orders and other geographic locations. The species that most resembled Henneguya novaerae n. sp. was Henneguya sharifi Molnár, Székely, Mohamed & Shaharom-Harrison, 2006, that parasites Pangasius hypophthalmus  from Malaysia. The two species have similar length and width of the myxospore body but differ in the length of the caudal appendages (14.7 ± 1.4 (11.5–16.1) µm vs. 7.5 (5.9–10.3) µm).In comparison with species that parasitize Siluriformes fishes from the Americas, the species that most resembled Henneguya bagre n. sp. and Henneguya breviscauda n. sp. Henneguya corruscans Eiras, Takemoto and Pavanelli, 2009 is the species that presents the caudal appendages with a size closer (13.7 µm) to that observed in the two new species, but the myxospores body length is greater (14.3 µm), differentiating them.The other species described parasitizing Siluriformes from the Americas have a longer caudal appendage concerning All other species used for comparison did not show morphological or morphometric characteristics resembling the new species. At least one morphometric feature, the number of turns of the polar tubules (varying at least 3 turns), or partial genetic sequences, differed in comparison with the new species.Henneguya novaerae n. sp, Henneguya bagre n. sp. and Henneguya breviscauda n. sp.In the present study, new species of myxozoans were described based on morphometric characteristics, as well as with phylogenetic support, and the sum of the evidence indicated conclusively the existence of three new species, denominated 27. Although H. jundiai has been described in the same main watershed where this study was carried out, geographically close, and morphologically similar, our molecular analyzes showed differences between the referred species and the new species. Furthermore, the preferred location for parasitizing the gills was also different, H. jundiai parasitizing the gill arches and the new species found in the filaments.The sequence similarities and variables sites are important evidence for species identification and discrimination of closely related myxozoans speciesHenneguya spp. with the hosts, with a strong evolutionary signal, as reported by Carriero et al.28, Vieira et al.30, and Úngari et al.31. In the family Myxobolidae, the hosts are the primary evolutionary factors, which contribute to the few monophyletic clades observed in the phylogenies33. In this study, the arrangement of the species was determined primarily by the order of the host, followed by the family of the host. The new species are inserted in a polyphyletic clade composed of species that infect Siluriformes fishes, along with other Henneguya/Myxobolus species that parasitize R. quelen. The phylogenetic tree clearly distinguishes the new species from the other Henneguya spp. deposited in GenBank. This arrangement further corroborates the effectiveness of the ssrDNA gene for analyses of the phylogenetic relationships among myxozoans, given that closely related species can be extremely variable, with their characteristics being determined primarily by the process of coevolution with their respective hosts34.The phylogenetic analysis indicated an affinity of 25 and Negrelli et al.24 reported the presence of species that parasitize Esociformes fishes in their phylogenetic analyses, as observed in this study. These data corroborate the close phylogenetic relationships between myxozoans species that parasitize fishes from the families Esocidae and Heptapteridae, as only genetically similar species were used for this analysis in this study.Abrunhosa et al.H. rhamdia in the GenBank, making it impossible to genetically compare them to the new species or to evaluate the phylogenetic relationships of these species. Future molecular and phylogenetic analysis of H. rhamdia is highly recommended to enable a definitive comparison and study of the relationship between species.Unfortunately, there is no ssrDNA data available for Henneguya novaerae n. sp. was the only species found at two different localities, being close to the source and mouth of the Pardo River. Henneguya bagre n. sp. and Henneguya breviscauda n. sp. were only found at a single locality, and the fish parasitized with Henneguya breviscauda n. sp. were not parasitized by another myxozoan species. There was co-infection of Henneguya novaerae n. sp. and Henneguya bagre n. sp. at locality 3. Although the different number of hosts collected per point may have interfered with the prevalence found, it is possible that different populations of Henneguya spp. inhabit different populations of R. quelen along the Pardo River. The various natural and artificial geographic barriers can contribute to this, but future studies are necessary to clarify this statement.Differences were observed concerning the prevalence of each myxozoan described in this study about the collection localities of the hosts. 35. Unfortunately, because of the small number of infected hosts, priority was given to molecular identifications rather than histological preparations. Despite this, visual observations and macroscopic photos helped us to determine the position and shape of plasmodia in the gills. Future studies should determine which tissue of the gill is the specific site of infection for the new species.The place of formation of plasmodia is an important taxonomic character for the differentiation of histozoic myxozoan speciesHenneguya novaerae n. sp., Henneguya bagre n. sp., and Henneguya breviscauda n. sp., gill parasites of R. quelen from Brazil. This is an important contribution to the understanding of the microparasite fauna of the siluriform species of the Neotropical region. The study indicates too that molecular characteristics and microhabitat preferences in the hosts are important for distinguishing morphologically similar species. Based on our analysis we recommend monitoring the presence of these parasites in farmed fish, to analyze possible pathologies caused by them.The present study describes R. quelen were collected from three localities in Pardo River: a region near the river source , municipality of Botucatu, Nova Era Resort  in June 2021; intermediate region , municipality of Avaré  in January 2022; and a region near the river mouth , municipality of Salto Grande  in May 2022. The collection was authorized by the Instituto Chico Mendes de Conservação da Biodiversidade (SISBIO #60640–1). Fish were collected using casting nets and were euthanazied with sodium thiopental (Thiopentax®), measured, and examined. All procedures followed the recommendations of the Ethical Commission for Animal Experimentation from the São Paulo State University (Unesp), Institute of Biosciences, Botucatu, Brazil (CEUA no 9415260520). The experimental protocol was approved by Ethical Commission for Animal Experimentation from the São Paulo State University (Unesp), (CEUA no 9415260520). The study is reported in accordance with ARRIVE guidelines. Fish were captured using a seine net with a small mesh (mosquito net fishing), cast nets, and a hand net. The voucher specimen of the fish host was fixed in 10% formalin, and deposited in Brazilian fish collection: Laboratory of Fish Biology and Genetics of the São Paulo State University (UNESP), São Paulo State (LBP 32004). Host specimens were examined fresh and the internal and external organs were evaluated in search of plasmodia containing myxospores using a Leica S6 D stereomicroscope  with a 16 × ocular. Samples of gills infected with plasmodia were collected for morphological and molecular analysis. The prevalence of parasites was calculated according to Bush et al.36. The morphological measurements of the fresh myxospores followed the recommendations of Lom and Arthur37. The measurements of 30 myxospores were performed using a computerized image analysis system with differential interference contrast (DIC) . Morphometric data were obtained from myxospores from one plasmodium of the type-host. All measurements are presented in micrometers (μm) and are expressed as means, followed by the standard deviation (SD), and range in parentheses. Illustrations of the myxospores were produced with the aid of a camera lucida mounted on a Leica DMLS microscope with phase-contrast optics. The final images were edited using CorelDRAW X8 software .Specimens of 38 with default settings.Samples containing plasmodia were removed from the gills and fixed in absolute ethanol (100%). Genomic DNA was extracted from the species using the DNeasy® Blood & Tissue Kit , following the manufacturer’s protocol. At least one of the samples used to obtain the partial sequences came from the type-host species. Access to the genetic data was authorized by the Brazilian Ministry of Environment (Sisgen A8A1D2B). Fragments of ssrDNA were amplified using the primers and cycling conditions presented in Table 43 using the Akaike information criterion was GTR + I + G. Phylogenies were reconstructed for each alignment under Bayesian inference (BI) using MrBayes v. 3.244. and Maximum Likelihood (ML) using PhyML45 with 1000 replicates. The BI was run using four Markov chain Monte Carlo searches with 10,000,000 generations and sampling tree topologies every 100 generations. The burning was set to the first 30% of generations; the consensus trees were estimated using the remaining topologies. The nodes with posterior probabilities and bootstrap values greater than 0.95/65 were considered valid. The final trees were edited using CorelDRAW X8. Pairwise genetic distances within and among sequences were calculated using the Kimura-2-parameter (K2P) model and a bootstrap procedure with 1000 replicates in Geneious 7.1.338.The best-fitting models for nucleotide substitution selected by jModelTest software


Vanessa cardui) reaching southern Europe each spring. We address their concerns, and further suggest that the rationale behind their critique is predicated upon unrealistic assumptions.López-Mañas et al.  raise coV. cardui will be absent. Yet, a search of the Global Biodiversity Information Facility (https://www.gbif.org) for botanical records produces >1,000 occurrences of Asteraceae and ∼3,000 occurrences of Malvaceae [families including the principal V. cardui larval hosts  n models –5, 7, 8.n models , 10, is n models .V. cardui migrants across Africa and the Middle East, although we do believe correlative analyses should be based on sound ecological theory and existing observations, such as of the massive spring emergence areas in Morocco (We do not dispute additional populations of  Morocco —the fina


The assessment of traditional products is important for the sustainability of agricultural systems and the preservation of a unique gastronomic heritage. The present study aims to determine the chemical composition of “Galo de “Barcelos”” (“Barcelos” Rooster) raw meat, used in the preparation of the typical Portuguese dish “Roasted Rooster from “Barcelos””, in order to evaluate and protect this gastronomic and cultural tradition. The influence of the genotype on the final product was verified, concerning total protein, and fat contents, and a favorable ratio of n-6/n-3 fatty acids of the Sasso line was compared to the “Amarela” breed, contributing to the promotion of its gastronomic potential.p ≤ 0.01) were observed, with the breast having a higher protein content (25.1 vs. 19.7%) and less fat (1.9% vs. 6.7%), compared to the drumstick. The fatty acid profile revealed  a similar composition to the roosters reared in the traditional or organic production systems, such as the “Amarela” autochthonous rooster. The “Barcelos” rooster can be regarded as a highly nutritional meat, with an interesting chemical profile ensuring a high-quality traditional product to consumers.Ten roosters produced according to “Barcelos Confraria” rules and ten roosters of autochthonous “Amarela” breed, reared on a similar traditional production system, were analyzed, and the chemical profile of two of the most significant meat portions, breast and drumstick, was determined. The results demonstrated that the “Barcelos” rooster raw meat is rich in proteins (22.3%) and fat (4.31%), particularly in monounsaturated fatty acids (39.1%). Significant differences ( Meat is one of the most nutritious foods, and poultry meat is known for its nutritional quality as it contains a significant amount of high-quality proteins, is highly digestible, has a high content of vitamins and minerals, and has a low portion of saturated fat.Chicken meat is highly recommended for all age groups, and its consumption has been privileged by consumers, causing a significant increase in its production ,2. In PoA precise evaluation of meat physicochemical parameters is based on the determination of its nutritional composition. Meat characteristics, such as pH, moisture, fat, and fat profile or protein, are the most frequently determined parameters with traditional methodologies. However, studies  have beeMeat quality is affected by different parameters, including intrinsic parameters such as age, sex, genetics, nutrition, and physiological state and extrinsic parameters such as the environmental and sanitary conditions, feeding techniques, management, transport, slaughter, and postmortem meat processing. Fat quality or nutrient quantities depend largely on the animal diet or its genetic potential, although the mode of production (organic and free range) influences some aspects of meat composition .Consumers’ concerns about health and nutritional diet quality created a growing interest in poultry products from outdoor production systems ,7. The uWith the considerable growth in consumer concern about animal welfare in recent years, ‘organic’ animal breeding now presents an opportunity for more traditional production systems . The marThe connection of the product to its region and the particularities of its manufacturing processes create new opportunities for farmers and local communities to associate the supply of services with production, which is the traditional products’ main value for their contribution to rural development .Portugal has an extensive variety of traditional food products, strongly associated with the region and the cultural influence of its preparation, constituting a living, unique, and rich gastronomic heritage. Promoting local products and characterizing and guaranteeing authenticity are determining factors for their recognition and sustainability.The “Roasted Rooster from Barcelos” is one of the most typical dishes of local gastronomy, with a strong connection to one of the most emblematic icons of the Portuguese cultural tradition. It is a gastronomic delicacy reminiscent of the Legend of the Rooster and brings to the table the ceremony, history, and symbolism of Santiago Compostela Way. It promotes a journey through the Jacobean tradition and the paths of knowledge and flavors of tradition . The roaBrassica oleracea, var. acephala). These characteristics limit the potential growth of the animals but improve meat quality and organoleptic attributes.The “Confraria Gastronómica o Galo de Barcelos” (gastronomic fraternity) was created in 2016 with the purpose to value and promote the “Galo de Barcelos” (“Barcelos” Rooster) in its cultural and gastronomic aspects. The “Confraria” established a set of production rules, which included feeding, density, housing, and sanitary aspects . To guarThe aim of this study is to provide the chemical characterization of the “Barcelos” rooster, identifying its particularities in relation to an autochthonous breed produced in a traditional system, in order to promote and qualify this important endogenous resource.Brassica oleracea, var. acephala), which were complemented with surplus or by-products from human or animal feeding. The “Barcelos” roosters, at 140 days of age and 4.2 kg of bodyweight (BW), in accordance with “Confraria” production regulations, and the “Amarela” roosters at 3.5 kg average bodyweight, after fasting  of 12 to 16 h, were slaughtered in a local plant, according to legislation rules [pectoralis major and peroneous longus muscles were excised for analysis. All procedures were conducted according to the guidelines of the organism responsible for the Animal Welfare of the Polytechnic Institute of Viana do Castelo—ORBEA-IPVC, in accordance with Decree-Law No. 113/2013 of 7 August.Twenty roosters were used; 10 of “Barcelos” (Sasso broiler genotype) and 10 of the autochthonous “Amarela” breed. “Barcelos” roosters originated from 6 exploitations, associated with the “Confraria”, in the region considered to be the breeding area . These farms are characterized by small effective and traditional production systems, in terms of housing and feed management. “Amarela” roosters originated from 6 exploitations; in the same geographic region and mode of production; with access to pasture; fed on corn and vegetables; and with an average slaughter age of 140 days and an average live weight of 3.5 kg. “Barcelos” roosters, identified with a wing tag, as a serial number, enabling the entire production process to be controlled, originated from slow-growth line roosters (Sasso liner). The animals comprising the sample were raised and kept by backyard producers under extensive conditions and were fed on a specific diet, according to “Confraria” rules, corn, and farm fodder, such as galega-cabbage . The parameters were determined in duplicate according to the AOAC .TM, Hanna Instruments, Eibar, Spain) equipped with a penetration probe, according to the ISO 2917:1974 method [The pH of the samples was measured using a digital portable pH-meter (FC2022/HALO4 method . Moistur4 method . Ash conProtein content was determined according to the Kjeldahl method ISO 937:1978) , with th78 , withThe total fat content was quantified following the Soxhlet (behr ED) method with ether petroleum solvent. The fatty acid profile was determined according to the procedure described by Cruz et al. , after tAn analysis of variance was performed on meat quality data using the IBM SPSS Statistics Base 22.0 for Windows. Descriptive statistics were generated for all the variables in the dataset. The analysis was carried out using a t-test of independent samples with variable grouping of breast and drumstick portions.p ≤ 0.001) in the roosters of the Sasso line than those of the “Amarela” breed, at the same age.The carcass characteristics of the “Barcelos” and “Amarela” roosters are shown in The chemical composition of the “Barcelos” and “Amarela” roosters, expressed in fresh meat weight percentage, is shown in The average values were observed between “Barcelos” and “Amarela” roosters with pH (5.80 vs. 5.90), moisture (72.54 vs. 74.60%), ash (1.13 vs. 1.12%), total fat content (4.31 vs. 3.72%), and protein (22.29 vs. 21.62%) values, as a result of the superior carcass weight of the “Barcelos” rooster.p ≤ 0.001), as well as pH (p ≤ 0.001) and protein (p ≤ 0.001) contents, when compared to the breast meat (p ≤ 0.01) content in comparison with drumstick meat.Results regarding the proximate composition analysis showed that, in both groups of rooster origin, drumstick meat had higher values for total fat (ast meat . “Amarelp ≤ 0.001) and total fat (1.89 vs. 1.90%) (p ≤ 0.001). Related to the drumstick, Barcelos” and “Amarela” roosters differed significantly in terms of moisture (71.81 vs. 75.01%) (p ≤ 0.05), fat (6.72 vs. 5.42%) (p ≤ 0.001), and protein  (p ≤ 0.001).There was also higher variability in the protein (breast) and fat (drumstick) contents, which can be attributed to the interaction between genotype and dietary treatment. When compared by genotype, the breast features significant differences in protein (25.15% “Barcelos” rooster vs. 24.55% “Amarela” breed) (The composition of the fatty acid rooster profile  was exprp ≤ 0.001), C18:0 (p ≤ 0.001), ∑SFA (p ≤ 0.001), C16:1 (p ≤ 0.01), C18:1 (p ≤ 0.01), ∑MUFA (p ≤ 0.001), C18:2n6 (p ≤ 0.001), C20:4n6 (p ≤ 0.001), ∑n-6-PUFA (p ≤ 0.001), C22:5n3 (p ≤ 0.01), C22:6n3 (p ≤ 0.01), ∑n-3-PUFA (p ≤ 0.01), ∑LC-PUFAS (p ≤ 0.001), and ∑PUFA (p ≤ 0.001) fatty acids, were verified.Concerning SFA, palmitic acid (C16:0) and stearic acid (C18:0) were determined. Regarding MUFA, oleic acid (C18:1) was the most abundant, followed by palmitoleic acid (C16:1), and the least abundant acid was eicosenoic acid (C20:1). Significant differences between genotypes in C16:0 (p ≤ 0.001) for C16:1 and C18:1 were verified, with the drumstick portion having the highest values and for eicosenoic acid (C20:1) (p ≤ 0.05), there were higher proportions in the breast portion. For the n-6-PUFA, significant differences (p ≤ 0.001) were found in linoleic (C18:2n6), arachidonic (C20:4n6), and adrenic (C22:4n6) acids. However, the first two had a higher proportion in the drumstick, while adrenic acid was more represented in the breast. For n-3-PUFA, the long-chain docosapentaenoic acid (C22:5n3) and docosahexaenoic acid (C22:6n3) were significantly different and were dominant in the breast portions, while α-linolenic (C18:3n3) was superior in the drumstick sample (p ≤ 0.01). In general, breast was the meat portion that presented higher values in polyunsaturated fatty acids  and saturated (∑SFA) and trans fats, while the drumstick was richer in monounsaturated fatty acids (∑MUFA).C16:0 was the most abundant saturated fatty acid , with sip ≤ 0.01), ∑SFA (p ≤ 0.001), C18:1 (p ≤ 0.01), ∑MUFA (p ≤ 0.001), C18:2n6 (p ≤ 0.001), ∑n-6 PUFA (p ≤ 0.001), C22:5n3 (p ≤ 0.01), C22:6n3 (p ≤ 0.01), ∑n-3-PUFA (p ≤ 0.001), ∑LC-PUFAS (p ≤ 0.001), and ∑PUFA (p ≤ 0.001). Concerning drumstick, they were observed in C16:0 (p ≤ 0.01), C18:0 (p ≤ 0.01), ∑SFA (p ≤ 0.001), C18:2n6 (p ≤ 0.001), ∑n-6-PUFA (p ≤ 0.001), ∑LC-PUFAS (p ≤ 0.05), and ∑PUFA (p ≤ 0.001).Differences between genotypes, in breast, were observed in C18:0  in the pH values found in the drumstick and breast could be explained by the unequal activity of each muscle, with a higher concentration of glycogen and less activity in the muscle with lower pH values [Differences  in moisture content between drumstick and breast were verified, inside the range of values reported by other authors [Statistically significant differences , genotyPoultry meat is known for its low fat because, unlike other meat animals, fat is mainly deposited subcutaneously or in the abdomen, rather than in the meat. In the present study, fat content differences were observed between genotypes, with similar values to those in autochthonous breeds ,33,37,40According to different authors, lipid content in the muscle is influenced by age and genotype, diet composition, and environment ,43. DrumIn the second week of life, approximately 36 and 35% of lean meat is located in the breast and legs, respectively, and as chickens grow, the percentage of lean tissue increases to 44% in breast and reduces to 32% in drumsticks, relative to total carcass lean tissue ,44. A grLipids are important constituents of the diet because, in addition to their high energy values, they provide fat-soluble vitamins and essential fatty acids . In an iAccording to different authors ,41,49, hSimilar results were observed by , concern“Barcelos” and “Amarela” roosters were characterized by high proportions of individual and total n-3 fatty acids and a favorable n-6/n-3 fatty acid ratio. In autochthonous populations, breed and feeding significantly affect fatty acid content , presenting a favorable fatty acid profile ,29, as oThe meat of Blackboned chickens had relatively low contents of saturated fatty acids and, in breast muscle, high contents of polyunsaturated fatty acids . Other aPoultry meat contributes to human nutrition by providing high-quality protein and low levels of fat, with a desirable fatty acid profile, and consumers are increasingly demanding poultry meat products.To guarantee authenticity, and to protect, qualify, and promote the traditional “Barcelos rooster” dish, chemical and sensorial characterization is mandatory. The influence of the genotype must be considered as a relevant factor in the “Barcelos Rooster” qualification, in particular due to the significantly higher total fat and protein contents, and the favorable n-6/n-3 fatty acid ratio of Sasso line, in comparison with “Amarela” authoctonous rooster bred in some traditional production systems.After characterizing the production system and obtaining a product similar to that of native animal genotypes, further studies are needed in the sensorial and in the the confection process characterization, with a view to their qualification.


Cymbopogon jwarancusa (C. jwarncusa) on physiological and biochemical features of wheat under arsenic (As) and mercury (Hg) stress. The results showed that higher concentrations of As and Hg pose inhibitory effects on wheat seed germination and seedling growth, including shoot/root length and shoot/root fresh weight. Photosynthetic pigments, such as chlorophyll a and b and carotenoids, were significantly decreased under As and Hg stress. Importantly, the levels of H2O2, lipid peroxidation, and TBARS were increased in wheat seedlings. The activity of antioxidant enzymes, such as CAT, was decreased by As and Hg stress, while the levels of SOD, POD, and APX antioxidant enzymes were increased in root and shoot. Interestingly, the application of PDS (2000 ppm), individually or in combination with either As or Hg stress, enhanced wheat seed germination rate, shoot/root length, and shoot/root fresh weight. However, the levels of H2O2, lipid peroxidation, and TBARS were decreased. Similarly, the levels of SOD, POD, and APX were decreased by PDS under As and Hg stress, while the level of CAT was enhanced by PDS under As and Hg stress. Interestingly, the levels of chlorophyll a and b, and total carotenoids were increased with the application of PDS under As and Hg stress. It is concluded that PDS has the capability to alleviate the phytotoxic effects of As and Hg stress in wheat by modulating the antioxidative defense system and could be an economical solution to reduce the heavy metal stress in crops. Heavy metal stress is a significant factor in diminishing crop yield. Plant-derived smoke (PDS) has been used as a growth promoter and abiotic stress alleviator for the last two decades. Although the roles of PDS have been determined in various plants, its role in ameliorating heavy metal stress in wheat has not been reported so far. Therefore, the present work was conducted to investigate the effect of smoke solution extracted from a wild lemongrass  Heavy metal contamination is one of the environmental threats diminishing plant productivity. Most of the anthropogenic activities and rapid industrialization are the sources of heavy metals in the natural environment . Heavy mAllium cepa, due to which, the absorption of water uptake was disturbed [Hg is a significant contaminant in soils . It is eisturbed . Hg has isturbed ,22,23.Seed germination and development of plants is an effective process that can be affected by various abiotic factors, such as water, light intensity, temperature, oxygen, and heavy metals .4 [Sapium sabiforum has shown improved tolerance to salt and osmotic stresses by increasing germination and post-germination growth in PDS [Eragrostis tef (zucc.) via regulating GA biosynthesis-related genes, enhancing ROS-scavenging antioxidant enzymes and sugar uptake in seeds [Plant-derived smoke (PDS) is a complex chemical mixture of plant-active chemicals that can stimulate a wide range of plant species in a given ecosystem. PDS has been reported to play an important role in the seed germination of various endangered plant seeds . PDS has4 . It has 4 ,29,30. Sh in PDS . Under hh in PDS , karrikiin seeds ,34,35. Ain seeds .Wheat is one of the most frequently farmed cereal crops in the world, with 40 percent of the planet’s population dependent on it for nourishment . The glo2O2, O−2, and OH, resulting in severe cell damage [As and Hg stress affect several physiological processes in wheat plants, particularly crop growth and seed germination. Heavy metals cause an increase in reactive oxygen (ROS) molecules, such as Hl damage ,41.The promontory effect of PDS occurs at various growth and developmental levels in plants, and the present study investigated how PDS affects the growth of wheat under As and Hg stress. Certain morphological, physiological, and biochemical activities of wheat were measured. The data showed that PDS can enhance wheat growth under As and Hg stress conditions by scavenging ROS, and modulating antioxidant enzyme activity. PDS was used at 2000 ppm concentration, which enhanced the germination and post-germination growth of wheat seedlings. It alleviated the negative effects of As and Hg on germination and post-germination growth. Our research demonstrates that PDS could be an economical plant product to enhance wheat physiological and biochemical processes under As and Hg stress, which are essential for the induction of tolerance in wheat.Cymbopogon jwarancusa L. (C. jwarancusa), also locally known as Sargara, was collected from the premises of Kohat University of Science and Technology. C. jwarancusa plants were thoroughly washed with water and shade dried for 15 days. PDS solution was prepared by weighing 333 gm of the semi-shade-dried leaves of C. jwarancusa which were burned in a locally constructed furnace on an electric heater [2O3 and HgCl2.c heater ,43. The The seed variety Paseena, 2017 was used in the experiments. Healthy and uniform-sized seeds were selected and surface sterilized with 0.1% sodium hypochlorite. Next, the seeds were washed with 70% ethanol for 3 min and rinsed with distilled water five times each for 1 min. Seed germination test was conducted thrice which revealed 100% germination, indicating that the wheat seeds were good in health and had the ability to germinate. Seeds of similar size were then chosen and germinated on Petri plates (150 × 60 × 100 mm) containing double-layered Whatman No. 1 filter paper. Before the experiment, the Petri plates and filter paper were pre-sterilized. Each petri dish had twelve seeds placed on it, and three Petri dishes were used for each treatment. Seeds were treated with PDS 2000 ppm, As and Hg 2 mM solution and distilled water, which were considered as controls. All Petri plates were kept at 25 degrees Celsius for 16 h of light and 7 h of darkness at 60% humidity. After 24 h, the rate of germination was measured. Germinated seeds are those that have a protruding radical. When the clear differences were shown on day 7 of germination, the morphological parameters such as shoot/root length and fresh weight of the seedling were measured. All experiments were performed independently three times.2O2 in the cell, under treatment of PDS, As, Hg, As + PDS and Hg + PDS, a reaction mixture of 1 mL of enzyme extract, 1 mL of 10 mM potassium phosphate buffer (PH 7.0), and 2 mL of 1 M sodium phosphate buffer (pH 7.0) was prepared and the absorbance was measured at 390 nm following the method described previously [g before being immediately chilled in an ice bath. The absorbance was measured at 532 nm, and non-specific turbidity was determined at 600 nm.To measure the levels of Heviously . Spectroeviously . After b2HPO4.2H2O) buffer (pH 7.8) (10 mL out of 50 mM). The samples were centrifuged at 4 °C for 20 min at 12,000 rpm after chilling. The supernatant was collected after centrifugation for additional enzymatic examination. After being heated for 30 min at 95 °C and immediately cooled in an ice bath, the homogenate was centrifuged for 10 min at 10,000× g for 10 min.Seedlings were harvested at day 7 following germination to assess antioxidant enzyme activity. As such, 500 mg of fresh wheat seedlings was grounded in a pre-cold mortar and pestle and homogenized in a potassium phosphate  exposed to light (4000 lux for 20 min).To determine the SOD, 250 mL of the total reactive homogeneous mixture was used . After being prepared and wrapped in aluminium foil, the reaction substrate was maintained in the refrigerator to prevent degradation. Next, 2.725 mL of the homogeneous mixture, 25 mL enzyme extract, and 25 mL HInstead of enzyme extract, pure water (25 mL) was used as a control sample. When the combination was exposed to lux, the tubes were wrapped in black cloth for a steady response. Glass cuvettes were used to quantify the samples at 560 nm .2O2 in quantities of 100 mL each. PBS (2.8 mL), guaiacol (0.1 mL), and H2O2 were included in the control sample. The POD activity was determined using a spectrophotometer with an absorbance wavelength of 470 nm. Potassium phosphate, EDTA-Na2, H2O2, ascorbic acid, and enzyme extract were used to make the buffer. The utilized wavelength was 290 nm.Using the procedures described previously , the actp ≥ 0.05) being the least significant variation. The samples were compared using the LSD test.The findings from each test were examined after three repetitions. All of the data were displayed as an average standard deviation using the statistics 9 applications . One-way analysis of variance (ANOVA) was used to establish the significance of variations between the treatment and control groups, with  showed 63.33% germination at 24 h, 75.55% at 48 h, and 100% at 72 h. These results indicate that both As and Hg have inhibitory effects while PDS has a promoting effect on seed germination. Next, we wanted to know whether PDS can alleviate the inhibitory effects of As and Hg. We treated wheat seeds with As or Hg solutions supplemented with PDS. The result showed an increase in seed germination by 53.33%, 73.33%, and 86.66% under As stress, and 56.66%, 76.66%, and 86.66% under Hg stress at 24, 48, and 72 h, respectively . The resSoil pollution from heavy metals is a major environmental problem that limits crop productivity. As and Hg are toxic chemicals that injure plants. As and Hg have adverse effects on plant growth and development. Likewise, the majority of biochemical and physiological processes also get disturbed because of toxicity ,49,50,51Next, the effect of As and Hg was evaluated on wheat seedlings, and the treatments resulted in reduced wheat shoot length by 4.8 and 4.26 cm under As and Hg stress, as compared to control, which was 9.2 cm. On the contrary, individual PDS treatment enhanced shoot length by 14.8 cm. Interestingly, As and Hg treatment followed by PDS treatment enhanced the shoot length to 7. 85 and 7.2 cm, respectively A. In casEragrostis tef (zucc.) under osmotic stresses and heat stress [The principal secondary organs of a plant are its shoots and roots. Increased seedling biomass shows that the seedlings are growing and developing rapidly . In the t stress . Karrikit stress . CollectAs and Hg stress primarily affect plant physiological parameters, such as shoot and root vigor, while PDS alleviates As and Hg stress. Smoke water dramatically increased seedling growth, including shoot and root lengths, in all plants. This could be due to butenolide’s ability to stimulate root growth ,60. LighTo understand whether As and Hg PDS affect photosynthetic pigments in wheat seedlings, 12-day-old leaves of wheat seedlings from each treatment were analyzed for chlorophyll a and b contents. The results indicated that chlorophyll a and b content in wheat seedlings, exposed to PDS (2000 ppm) and As and Hg alone, showed significant variations. The obtained results showed that seedlings treated with PDS (2000 ppm) possessed high chlorophyll a (7.284 µg/mg FW) and b (5.547 µg/mg FW) compared to a (6.43 µg/mg FW) and b (4.506 µg/mg FW) in control seedlings. On the contrary, exposure to As showed a significant decrease in chlorophyll a (3.873 µg/mg FW) and b (3.196 µg/mg FW), while Hg stress caused a reduction in a and b to 2.659 and 2.442 µg/mg FW, respectively. To alleviate the phytotoxic effect of As and Hg through the use of PDS (2000 ppm), the levels of chlorophyll a and b were recorded. Notably, treatment of As + PDS and Hg + PDS had high chlorophyll a and b of 5.4567 and 3.974 µg/mg FW and 5.387 and 3.984 µg/mg FW, respectively, compared to seedlings treated with only As and Hg . These rBrassica napus as a result of Hg and As exposure [Photosynthetic pigments are the most fundamental and basic ingredients in the photosynthesis process and, hence, play a crucial part in the creation of plant food. It is created by plants throughout the production cycle to meet their nutritional requirements. In the current study, As and Hg toxicity resulted in a considerable reduction in chlorophyll and carotenoids production in wheat leaves. These findings support prior studies that revealed a reduction in chlorophyll concentration in exposure . Heavy-mexposure ,68. 2O2 in the roots and shoots of wheat seedlings were measured. Treatment of wheat seedlings with As and Hg stress caused high levels of H2O2 in both root and shoot, as compared to control plants. To find the level of oxidative stress in different seedling parts, levels of H2O2 were measured in shoots and roots individually. The results indicated that the content of H2O2 in the root was 0.709 and 0.691 µM/g FW under As and Hg-treated seedlings, respectively, while the shoot was found to have 0.614 and 0.590 µM/g FW levels of H2O2 in As and Hg-treated seedlings, respectively , while in control, it was 0.403 and 0.403 µM/g FW, respectively. When the PDS was used in combination with As and Hg, the content of H2O2 in roots reduced to 0.462 and 0.453 µM/g FW, while in the shoots, it decreased to 0.515 and 0.504 µM/g FW, respectively. The results indicate that PDS 2000 ppm has the capability to reduce H2O2 concentration under As and Hg stress.The levels of Hectively A,B. ThesTo determine how As, Hg, and PDS affect TBARS levels in wheat, we treated wheat seeds with As, Hg, or PDS alone. The results showed that individual treatment of As and Hg enhanced the levels of TBARS in the roots and shoots, as compared to control. Quantitative data for As-treated roots and shoots showed 0.515 and 0.526 µM/g FW levels, while 0.497 and 0.521 µM/g FW for Hg-treated roots and shoots, respectively. The levels of TBARS in both roots and shoots under As and Hg for control were 0.376 and 0.269 µM/g FW, respectively. Plants treated with PDS 2000 ppm had low levels (0.329 and 0.247 µM/g FW) compared to control. When seedlings were treated with PDS 2000 ppm in a combined solution with As and Hg, the levels of TBARS were significantly reduced in roots/shoots to 0.413 and 0.368 µM/g FW for As and 0.401 and 0.356 µM/g FW for Hg, respectively C,D. The 2O2 and TBARS. Seedlings exposed to As and Hg stress had significantly higher levels of H2O2 and TBARS, while those treated with PDS showed a drop in H2O2 and TBARS contents. These results are in line with those that found that As, Hg, and Ni toxicity increased H2O2 and MDA contents in Glycine max, maize, and Allium species [2O2 and TBARS under salt stress, while the application of PDS reduced the levels of H2O2 and TBARS under salt stress [Heavy metals, such as metalloids, have been shown to contribute to oxidative stress by promoting the generation of free radicals and reactive oxygen species . In the  species ,71,72. At stress . These fThe level of lipid peroxidation was determined and the results indicated the highest level of lipid peroxidation when wheat seedlings were treated with As and Hg. Quantitative analysis showed that the level of lipid peroxidation in roots was 25.28 µM/g FW and 24.81 µM/g FW under As and Hg stress, while in shoots, it was 25.65 µM/g FW and 25.09 µM/g FW, respectively. When the plant was treated with PDS, the level of lipid peroxidation in the root and shoot was 12.96 µM/g FW and 13.68 µM/g FW, and in control, it was 18.37 µM/g FW and 18.47 µM/g FW, respectively. These results showed that PDS has the ability to decrease the level of lipid peroxidation in wheat seedlings. To alleviate the deleterious effect of As and Hg at lipid peroxidation level, PDS was used. The results showed that the lipid peroxidation in the root of wheat treated with As and Hg was 21.41 µM/g FW and 20.99 µM/g FW, while in the shoot it was 22.08 µM/g FW and 21.34 µM/g FW, respectively E,F. ThesThe activity of POD enzymes was increased both in the roots and shoots of the wheat seedlings by 0.520 µM/g FW and 0.693 µM/g FW, respectively, when treated with As, while in Hg stress, it was found to be 0.514 µM/g FW and 0.635 µM/g FW for roots and shoots, respectively. The level of POD was recorded as 0.39 and 0.2.1 µM/g FW indicating lower levels of POD under PDS treatment compared to control. Combined application of PDS with As decreased POD level in roots and shoots by 0.319 µM/g FW, 0.470 µM/g FW, respectively, while in Hg + PDS, the activity of POD enzymes decreased to 0.315 and 0.479 µM/g FW in roots and shoots, respectively, as compared to control, which was 0.237 µM/g FW and 0.473 µM/g FW, respectively A,B.2O2, auxin catabolism, pectin cross linking, and growth regulation [Zea mays) was exposed to arsenite and arsenate, similar outcomes were observed [POD is involved in a variety of mechanisms, such as lignin polymerization in the presence of Hgulation . Under Agulation . When maobserved , which t2O2 is a systemic intracellular signal that activates APX in response to abiotic stress [In our findings, As and Hg increased the activity of APX in wheat seedlings. Briefly, the highest APX activity was found in the roots/shoots of seedlings treated with As and Hg, as compared to control. Quantification of roots and shoots treated with As resulted in 0.749 µM/g FW and 0.805 µM/g FW, while 0.732 µM/g FW and 0.805 µM/g FW with Hg. The lowest activity was recorded in roots (0.349 µM/g FW) and shoots (0.367 µM/g FW) treated with PDS compared to control . Wheat seedlings treated with PDS in combination with As and Hg stress showed a decrease in APX activity compared to the seedlings treated with As and Hg treatment alone. In roots/shoots, the APX activity in As + PDS was 0.522 µM/g FW, and 0.573 µM/g FW, while in Hg + PDS it was 0.492 µM/g FW and 0.539 µM/g FW, respectively C,D. The c stress . Shri etc stress  observedThe SOD enzyme was found to be increased as the concentrations of As and Hg increased. As compared to control, the activity of SOD was increased in the roots and shoots of the wheat seedlings under individual As and Hg stress. The wheat seedlings treated with PDS 2000 ppm had the lowest SOD enzyme in roots and shoots, recorded as 0.147 µM/g FW and 0.147 µM/g FW, while in control, it was 0.196 µM/g FW and 0.273 µM/g FW, respectively. When the plants were treated with As and Hg, the level of SOD enzyme in the roots and shoots was higher compared to control. The level of SOD enzyme in roots and shoots treated with As was 0.406 µM/g FW and 0.679 µM/g FW, while in Hg, it was 0.392 µM/g FW and 0.645 µM/g FW, respectively. The results indicate that PDS 2000 ppm has the ability to decrease the SOD enzyme. To alleviate the adverse effects of As and Hg on roots and shoots, the seedlings were treated with PDS 2000 ppm. It was observed that the level of SOD enzyme in root and shoot seedlings treated with As, in combination with PDS 2000 ppm, was decreased to 0.271 µM/g FW and 0.376 µM/g FW, while in Hg it was 0.264 µM/g FW and 0.363 µM/g FW, respectively E,F. The 2 to O−2 and H2O2. According to the current study, SOD activity was shown to be higher in wheat shoots and roots under As or Hg stress. Previously, under Hg stress, a similar SOD response was seen in Chlamydomonas reinhardtii [The first enzyme of defense against oxidative stress is SOD, which catalyzes the dismutation of Onhardtii . Anothernhardtii . On the Wheat seedlings treated with As and Hg showed the lowest CAT activity in roots/shoots, as compared to control and PDS treatment. It was recorded that the CAT activity in the root was 0.181 µM/g FW and the in shoot was 0.61 (treated with As). The CAT activity in the root was 0.185 µM/g FW and in shoot it was 0.149 µM/g FW (treated with Hg). However, the CAT activity in the root was 0.411 µM/g FW, and in shoot was 0.356 µM/g FW when treated with PDS. The CAT activity in the root was 0.372 µM/g FW and in the shoot was 0.310 µM/g FW in the control seedlings. In combination with Hg, it was found that PDS resulted in 0.237 µM/g FW, while in shoots, it showed 0.223 µM/g FW (treated with PDA + As). It was recorded that the CAT activity in the root was 0.226 µM/g FW and the shoot was 0.226 µM/g FW when treated with PDS + Hg G,H. Over2O2 formation [2O2 produced in cells when SOD activity is high [2O2 [2O2 and lipid peroxidation during salt stress acclimation highlights the importance of maintaining redox homeostasis and preventing oxidative damage.CAT is an antioxidant enzyme that has a high turnover rate and can prevent Hormation . Peroxid is high . There wigh [2O2 . Lately,igh [2O2 . This mi2O2, TBARS, and lipid peroxidation were significantly increased in wheat seedlings. On the other hand, PDS (2000 ppm) enhanced seed germination and the seedling growth of wheat. PDS, in combination with either As or Hg, alleviated the negative effects caused by As and Hg on wheat seedlings. Moreover, PDS significantly modulated the activity of antioxidants, such as POD, APX, SOD, and CAT, which resulted in a significant reduction in ROS (H2O2) and lipid peroxidation under As and Hg stress. This study concludes that PDS (2000 ppm) has the potential to alleviate the phytotoxic effects of As and Hg stress at the morphological and physiological level, which will lay a foundation for further understanding of the role of PDS in the regulation of different abiotic stress tolerance in plants. This study also provides a novel clue for farmers and nursery growers on how to promote the crop seedling’s growth under stressed conditions. It would be even more beneficial to analyze the effect of PDS and heavy metal stresses on the flowering and yield of wheat and other crops, in terms of the number and size of grain. It is also necessary to understand the molecular mechanism of how PDS alleviates the adverse effects of As and Hg stress in wheat.According to the outcomes of this study, heavy metal stress, such as As and Hg, brought about a significant decrease in the growth and biochemical activities of wheat plants. Particularly, the levels of H


Tenebrio molitor, has recently been used as a substitute for conventional meat, as it has several advantages, e.g., a lower environmental impact. Knowledge of the effects of temperature and photoperiod on mealworm development is crucial to increase farming efficiency, and to contribute to a sustainable production process. As such data is scarce, we tested the effects of three different temperatures, in combination with three photoperiods, on mealworm survival, development, and growth. We found that temperature strongly affects the survival rate, developmental time, and growth rate of T. molitor larvae. Furthermore, photoperiod influences the developmental time and growth rate. The highest survival rates and growth rates, and shortest developmental times, were observed at 25 and 30 °C at constant darkness. These results are important to improve the mass rearing of mealworms for a sustainable production of food and feed.The mealworm, i.e., larvae of Tenebrio molitor. Knowledge of the effects of temperature, and particularly photoperiod, on mealworm development is scarce, but crucial for the improvement of rearing. Therefore, the effects of three temperatures , in combination with three photoperiods  on mealworm survival, developmental time, and growth rate were tested. We describe a significant effect of temperature on survival rate, developmental time, and growth rate. Furthermore, significant effects of photoperiod on developmental time and growth rate were found. At 25 and 30 °C and constant darkness, the highest survival and growth rate, along with the shortest developmental time, were observed. Our data can be used to improve the mass rearing of mealworms for an efficient production of food and feed.Insects are a potential substitute for conventional meat and can be part of a sustainable human diet due to their valuable nutrients and relatively low environmental production impact. One species that is already produced for human consumption and livestock feed is the mealworm, i.e., larvae of  Tenebrio molitor L. , is widely used for food and feed production [2 production [Insect farming for human consumption is a growing economic sector . The meaoduction ,3. Rearioduction ,5, or a oduction ,7. Moreooduction .Tenebrio molitor has a worldwide distribution. Under natural conditions, it inhabits, for example, rotten wood and other dead organic matter, or sheltered environments like bird nests or burrows. Furthermore, it is a synanthropic, omnivorous species, feeding on stored products. Generally, it prefers dark environments and is negative phototactic [totactic ,10,11.T. molitor is essential for leveraging mealworms for future meat consumption. For example, photoperiod influences mealworm development and growth [Knowledge of abiotic parameters on the development of d growth , and thed growth  described growth .T. molitor is described between 22 and 28 °C [T. molitor egg, larval, pupal and adult survival at different temperatures, with 25 °C as the optimal temperature for high survival rates. Temperatures of 10 °C and 35 °C, however, were described as unfavorable, since survival rates decreased at these temperatures [In addition,  found shnd 28 °C . The autnd 28 °C  tested Teratures . Furthereratures . The auteratures ,17 founderatures  recordedT. molitor is widely used for food and feed production [T. molitor and can contribute to a sustainable meat production.Although oduction , certainTenebrio molitor used in this study originated from a mass rearing from LarveMe . Subsequently, a stock culture with freshly hatched adult beetles was maintained at 22 °C and daylight in a plastic box with aeration slits and filled with about 2 cm of feed .T. molitor adult, randomly selected from the stock culture, were put in a plastic box (9 × 6 × 5 cm) (with a lid containing aeration holes) to initiate mating and oviposition. One cm of feed was filled into the box, and ten boxes each were put in different incubators under experimental conditions: three temperature regimes, i.e., 20 °C, 25 °C, and 30 °C, and three different photoperiods, i.e., long day LD , short day SD , and constant darkness 24D . After three weeks, i.e., when oviposition was completed, adults were removed from the plastic boxes. Subsequently, the presence of young larvae was monitored by screening the boxes for instars four to six . Dead larvae and pupae were removed from experimental boxes weekly to record survival rates. Furthermore, larval weight was assessed once per week to calculate growth rates and head capsule width, in order to monitor instar development, until pupation occurred. Survival rates and growth rates were recorded from fourth to sixth instar larvae until 95% of the larvae per box pupated. Developmental time was recorded from the larval eclosion from the eggs until pupation.To compare mean survival rates, mean developmental times, and mean growth rates among experimental conditions, a two-way ANOVA was performed, with Tukey tests as post-hoc tests (α = 0.05). The normal distribution of data was assessed by conducting a Shapiro–Wilk normality test. All analyses were conducted with SPSS Statistics v. 27. Details on the calculations of survival rate, developmental time, and growth rate are provided in p = 0.001). In general, survival rates among all treatments were high, with mortality rates of less than 10% over a period of 37 weeks (p = 0.003), as well as between mean values at 20 °C and those at 30 °C, with 96.7% (p = 0.006) (p = 0.978). The lowest survival rate was recorded at 20 °C/24D, with a mean survival rate of 90.4%. The highest survival rate was recorded at 25 °C/24D and 30 °C/24D, both with a mean survival rate of 98.5%. We found a significant effect of temperature on the survival of mealworms (37 weeks . Across = 0.006) . There wp = 0.237). The influence of temperature on survival was further corroborated, as we found significant differences among temperature conditions within the various photoperiodic treatments . Across all photoperiods tested, the developmental time of mealworms at 20 °C, with a mean of 184.8 days, was significantly higher than at 25 °C and 30 °C, respectively (both p < 0.001) (p = 0.558). We found a significant influence of temperature on larval developmental time, i.e., from the larval eclosion from eggs until pupation (< 0.001) . There wp = 0.001), as well as a significant interaction between temperature and photoperiod (p < 0.001). Across all temperatures, the mean developmental time under LD with 156.7 days was significantly higher than under SD (p = 0.016) and 24D (p = 0.001), respectively (p = 0.632). In general, mealworm developmental time was shorter under SD or 24D . Across all photoperiods, growth rates among the three temperature regimes were significantly different from each other (p < 0.001 each). The mean growth rate at 20 °C was the lowest with 25.1%, followed by 25 °C with 36.2%, and 30 °C being the highest with 39.2% (Temperature had a significant effect on the growth of th 39.2% .p < 0.001), as well as a significant interaction between temperature and photoperiod (p < 0.001). Across all temperatures, the growth rate under 24D (mean 35.7%) was significantly higher than the growth rate under LD (32.5%) or SD (32.3%), (both p < 0.001). However, there was no significant difference between growth rates under LD and SD (p = 0.962). Mealworms at 25 °C or 30 °C gained more weight in the same time period under 24D as compared to LD or SD. The highest growth rates were recorded at 25 °C and 30 °C and 24D, with means of 41.2% and 41.1%, respectively. The lowest growth rate, i.e., 23.7%, was observed at 20 °C and LD (There was also a significant effect of photoperiod on the growth rate (C and LD . At 20 °C, the highest mealworm growth rate occurred under SD. At 25 °C and 30 °C, the highest growth rate occurred under 24D. At LD and SD, there was an increase in growth rates with rising temperature. At 24D, there was a significant difference between 20 °C and the two other temperature regimes, but the mealworm growth rate at 25 °C and 30 °C was similar .There was a significant influence of temperature on the survival rates of mealworms. Mealworms died more frequently at 20 °C, with a mean survival rate of about 92%, compared to the two other temperature regimes, with a survival rate of about 97%. Generally, more mealworms died at the beginning of the experiment, and older larvae had a higher survival rate. The authors of  describeT. molitor is between 22–28 °C, and that temperatures above 35 °C and below 20 °C are associated with decreasing survival rates and increasing stress.Results from the experiments presented here confirm that higher survival rates are observed at higher temperatures, with an optimal rearing temperature between 25 °C and 30 °C. Moreover, ,17 stateHere, a significant influence of temperature and photoperiod on the developmental time of mealworms was found. In general, we describe shorter developmental times at higher temperatures, with no significant difference between 25 °C and 30 °C. The lowest developmental time of about 125 days was recorded at 25 °C and 24D. Rearing under LD resulted in significantly longer developmental times than under SD and 24D. At all three temperature regimes, the developmental time was similar to those observed in previous studies . MoreoveOur study showed a significantly higher developmental time under LD conditions compared to the other photoperiods. At 25 °C and 30 °C, developmental times under 24D were significantly lower, compared to LD or SD. However, the differences in developmental time between different photoperiods were small, i.e., the influence of photoperiod was low compared to temperature. The authors of ,16 stateWe report a significant influence of temperature and photoperiod on the growth rates of mealworms. Results show higher growth rates at higher temperatures. Rearing under 24D resulted in a significantly higher growth rate than under LD and SD conditions. Weekly growth rates of mealworms were similar to , who recT. molitor larvae at different temperature and photoperiodic conditions. In general, at higher temperature regimes and constant darkness, higher survival and growth rates, as well as shorter developmental times, were observed. These data can be used to increase farming efficiency and contribute to a sustainable production of mealworms for food and feed.Taken together, our study provides important data on the development, growth, and survival of