To investigate magnetic resonance imaging (MRI) features for differentiating ovarian endometrioid adenocarcinoma (OEC) from high-grade serous adenocarcinoma (HGSC).Twenty-three patients with 25 OECs and 93 patients with 139 HGSCs confirmed by surgery and pathology underwent conventional MRI and diffusion-weighted imaging (DWI). The MRI features of the tumors, including laterality, size, shape, configuration, signal intensity, ADC value of solid component, enhancement, ascites, synchronous primary cancer (SPC) of the ovary and endometrium, and clinical stage, were evaluated and compared between two groups.−3 mm2/s) than in HGSCs (0.820 ± 0.112 × 10−3 mm2/s) (P = 0.002). When a mainly cystic mass with mural nodules or papillary projections was associated with any one of homogeneously iso- or hyperintense cystic component on TIWI, a relatively higher ADC value and SPC, the sensitivity, specificity, accuracy, and positive and negative predictive values for characterizing OEC were 87.0%, 93.5%, 92.2%, 76.9%, and 96.7%, respectively.The following characteristics were significantly more common for OECs than HGSCs: unilateral , larger mass , round or oval shape , mainly cystic with mural nodules or papillary projections , cystic component with homogeneous iso- or hyperintensity on T1WI , moderate enhancement , no or mild ascites , and SPC . The ADC value of the solid component was higher in OECs (0.979 ± 0.197 × 10Conventional MRI combining DWI is helpful for differentiating OECs from HGSCs. When an adnexal mass is suspected to be an epithelial ovarian carcinoma by magnetic resonance imaging (MRI), the primary concern of clinicians is that the patient has a high-grade serous adenocarcinoma (HGSC), which is the most frequent epithelial carcinoma and is typically aggressive clinically . HoweverThe institutional ethical board  approved this retrospective study, and informed consent was waived. Between September, 2010 and October, 2014, we searched for the data of patients with OEC at our hospitals information system and picture achiving and communication system. A total of 25 patients with OEC confirmed by surgery and pathology were found. We excluded 2 patients with metastatic OEC. The remaining 23 patients with 25 OECs were reviewed in this study. The mean age of the patients was 54 ± 9 years . As a comparison, 93 patients with 139 HGSCs were served as a control group from 126 surgically and pathologically confirmed cases of HGSC at the same period and database. We excluded 15 patients who received chemotherapy before MR scanning, 8 patients who were performed on a 3.0 T MR scanner, 6 patients with poor image quality and 4 patients without the administration of intravenous contrast. Their mean age was 55 ± 9 years , which was not significantly different from the mean age of patients with OEC (P = 0.508). The diagnosis of SPC was established according to the pathological criteria proposed by Singh ; axial turbo SE T2-weighted imaging (T2WI) with and without fat saturation ; and sagittal and coronal turbo SE T2WI . Axial DWI (19 patients with 20 OECs and 83 patients with 124 HGSCs) was obtained with echo planar imaging  and b factors of 0 and 1000 s/mm2 was placed at targeted areas with the possibly lowest ADC values in the solid components of the tumor, by referring to conventional MR imagings and avoiding areas such as haemorrhage, necrosis and major vascular structures. At least three measurements were obtained and averaged.The MR images were reviewed independently by two radiologists (H.M.L and J.W.Q) with 7 years and 30 years of experience in abdominal imaging, respectively, and were blinded to the original reports . Any discrepancies were resolved by consensus. The following features of the tumors were evaluated: (a) uni- or bilaterality , size and shape; (b) mass configuration ; (c) signal intensity ; (d) enhancement ; (e) amount of ascites  (f) associated findings ; and (g) apparent diffusion coefficient (ADC) value as measured on ADC maps, a circular region of interest (ROI) of at least 1 cmt-test. A P-value less than 0.05 was considered statistically significant. Receiver operating characteristic (ROC) curve analysis was used to determine a cut-off value for differentiating OECs and HGSCs.SPSS 16.0 for Windows  was used for the statistical analysis. The differences between OECs and HGSCs in laterality, shape, mass configuration, signal intensity, enhancement, ascites, SPC, and clinical stage were compared using a Pearson chi-square test. The differences in age, mass size, and ADC values between the two groups were compared using the two independent-sample Student The maximum diameters of the OECs ranged from 3.7 to 22.5 cm (mean 11.1 ± 4.9 cm), and those of the HGSCs ranged from 2.5 to 16.8 cm (8.1 ± 3.4 cm) (P = 0.005). The median size of the tumors was 9.9 cm in OECs versus 7.6 cm in HGSCs. A bilateral mass was found in 2 OEC patients and 46 HGSC patients (P < 0.001). According to the International Federation of Gynecology and Obstetrics (FIGO) staging system, 19 patients with OECs were at stage I (82.6%), 3 were at stage II (13.0%), and 1 was at stage III (4.4%). Pathologically, 14 OECs were grade 1, 8 were grade 2, and 1 was grade 3. For the HGSC group, 6 patients were at stage I (6.4%), 9 were at stage II (9.7%), 73 were at stage III (78.5%), and 5 was at stage IV (5.4%). There was a statistically significant difference in the number of patients at different stages of disease between the two groups (P < 0.000).The MRI features of OEC and HGSC are summarized in Table −3 mm2/s in 20 of 25 OECs versus (0.820 ± 0.112) × 10−3 mm2/s in 116 of 139 HGSCs (P = 0.002)  OECs and isointensity in the remaining 3 (15.0%), while for HGSCs, the solid component showed hyperintensity in 111 of 116 (95.7%) and isointensity in 5 4.3%) (P = 0.173). The mean ADC value of the solid component was (0.979 ± 0.197) × 10.3%  ,11. OvarThe present MRI study showed that OECs were significantly different from HGSCs in laterality, size, shape, configuration, signal intensity, enhancement, ADC value, SPC, clinical stage and ascites. OEC commonly appeared as a large, unilateral, round or oval cystic mass with mural nodules or papillary projections, homogeneous iso- or hyperintensity on T1WI in the cystic component, and moderate enhancement and relatively higher ADC values in the solid component. OEC was commonly associated with SPC and was diagnosed at an early stage. In contrast, HGSC was typically a moderately sized, irregular solid or mixed cystic-solid mass with marked enhancement and a lower ADC value, more common bilaterality and moderate to severe ascites. Although some features had low diagnostic specificity, four features, a cystic mass with mural nodules or papillary projections, homogeneously iso- or hyperintense cystic component on T1WI, higher ADC value and SPC, have a high specificity for OEC. Those four features yielded a sensitivity, specificity, accuracy, positive and negative predictive values for characterizing OEC of 87.0%, 93.5%, 92.2%, 76.9%, and 96.7%, respectively.Histopathologically, it has been well-established that atypical endometriosis is a precursor lesion for OEC. OEC associated endometriosis is found in 20%-40% of cases . In contA previous study showed that 33.3% of OECs were cystic, and the remaining 66.7% were mainly solid, which was inconsistent with our findings . The cys−3 mm2/s in OECs versus (0.820 ± 0.112) × 10−3 mm2/s in HGSCs, and this difference was statistically significant. An optimal ADC value threshold of 0.971 × 10−3 mm2/s yielded a sensitivity, specificity and accuracy of 55.0%, 94.0% and 88.2%, respectively, for differentiating OECs from HGSCs, In our study, the solid component in OECs was mainly mural nodules or papillary projections, which were reported to have relatively low malignant potential [In our study, almost all OECs and HGSCs demonstrated high signal in the solid component on DWI. The mean ADC value of the solid component was (0.979 ± 0.197) × 10otential . Histopaotential . In contApproximately 1-2% of gynecological malignant tumor cases have two or more synchronous primary genital tract carcinomas . ApproxiOur study had several limitations. First, only imaging features of a limited number of patients were evaluated. Therefore, larger samples are necessary to confirm the value of these features for diagnosing OEC. Second, due to the retrospective design of our study, a selective bias was inevitable. Third, a correlation analysis of MRI features with pathology was not performed.In conclusion, our preliminary study demonstrates that a large, round or oval, mainly cystic mass with moderately enhanced mural nodules or papillary projections, a homogeneously iso- or hyperintense cystic component on TIWI, a relatively higher ADC value in the solid component and, commonly, SPC are features that help to differentiate OEC from HGSC. Although the diagnostic performance of any feature alone is not sufficient for diagnosis, the combination of the feature of a mainly cystic mass with mural nodules or papillary projections with any one of the following features: homogeneously iso- or hyperintense cystic component on TIWI, a relatively higher ADC value and SPC, yields high sensitivity, specificity and accuracy for identifying OEC.


The aim of this study was to examine the situation for elderly patients with diabetes living in nursing homes with regard to diabetes treatment, clinical variables, and vascular complications associated with diabetes. A second aim was to evaluate if the patients were at risk of hypoglycaemia.1c, hypoglycaemic events, and diabetes complications were registered from the medical records. The patients were stratified into a general group and divided into three groups according to HbA1c .This was a cross-sectional study including diabetes patients from all 30 nursing homes in Uppsala County, Sweden. Current antidiabetic medications, HbA1c concentration 56.0 ± 1.2 mmol/mol. Hypoglycaemic events were reported in 24% of the diabetic individuals, and 43.1% of them had HbA1c <52 mmol/mol . Of these, 36% were taking antidiabetic drugs. Another 35.8% of the patients had HbA1c values between 52–73 mmol/mol , and 82% of these patients were taking antidiabetic drugs. Almost 80% of the diabetic patients had either micro- or macrovascular complications, with diabetes duration as an association for both micro- or macrovascular complications and hypoglycaemic events.Of 1,350 individuals, 218 patients were identified with diabetes mellitus. The diabetes duration was 11.2 ± 9.4 years and their serum HbA1c level should be considered, especially for multimorbid elderly patients with low HbA1c and hypoglycaemia.A reduction of the use of antidiabetic drugs with follow-up of HbA The prevalence of diabetes is increasing worldwide, affecting more than 8% of the adult population. Diabetes is one of the leading causes of cardiovascular disease and death . In gene1c values, diabetes treatment, and micro- and macrovascular complications. Patients were divided into three groups according to HbA1c value . A second aim was to examine differences in diabetes treatment between the groups and the risk for excessive treatment and hypoglycaemia.Due to the complexity of ageing and declining body functions, often associated with multiple chronic illnesses, e.g. dementia and polypharmacy, these patients run a greater risk of severe hypoglycaemic complications ,12. Hypo1c value, diabetes duration, age, gender, weight, height, plasma creatinine, current antidiabetic medications, micro- and macrovascular complications, and hypoglycaemic episodes. Retinopathy, nephropathy, and neuropathy were designated as microvascular complications, while myocardial infarction, unstable angina pectoris, brain haemorrhage, brain infarction, and transient ischemic attack were designated as macrovascular complications.In 2012 we collected a list of 1,350 patients from 30 different nursing homes in Uppsala County, Sweden . Uppsala1c . Statistical analyses were made to examine the differences between the groups in antidiabetic treatment, medical dosages, and hypoglycaemic complications.Hypoglycaemic events were defined as plasma blood glucose <4 mmol/L. We categorized hypoglycaemic events as severe or minor events. Events leading to hospitalization, events that needed treatment in the emergency room, and events emerging during hospital care were categorized as severe events. Hypoglycaemic events only noted in a patient’s medical journal were considered as minor events. The patients were first stratified into a general group and then divided into three groups according to HbAPatients on oral antidiabetic drugs (OADs) as well as rapid-acting insulin when needed were recorded as OADs. Patients on diet treatment with rapid-acting insulin when needed were recorded as diet treatment.The study was approved by the regional ethics review board at Uppsala University.t test. To avoid too small subgroups a cut-off level at HbA1c 52 mmol/mol was chosen. A P value <0.05 was considered statistically significant. Multivariable analyses taking co-morbidities was not performed. Findings of association of HbA1c with risk of hypoglycaemia are crude, and they are not adjusted for the influence of differences in other variables, such as age, diabetes duration, or gender. A subgroup analysis on patients on pharmacological treatment for diabetes was performed. Statistical software JMP 5.0 for PC  was used.Results are given as arithmetic means with their standard deviations. Comparisons were made using chi-square tests with the Yates correction factor for analyses in differences between diabetes treatment and hypoglycaemic episodes within the groups. The differences between medication doses were analysed using independent samples 2, and mean age was 84.6 ± 8.0 years. Seven patients had T1DM, and 211 patients had T2DM. Mean HbA1c was 56.0 ± 1.2 mmol/mol (data missing for 18 patients), and average diabetes duration was 11.2 ± 9.4 years. The diabetes duration in the different HbA1c groups was 8.6 ± 1.4, 13.0 ± 11.0, and 16.8 ± 10.7 years, respectively. Longer diabetes duration was correlated with higher HbA1c, (P = 0.006)  were identified with diabetes mellitus (16.1%) . Their m= 0.006) . The creP < 0.05) in the group with HbA1c <52 mmol/mol compared to patients with HbA1c ≥52 mmol/mol. There were no differences between the groups (<52 and ≥52 mmol/mol), comparing numbers of patients being treated with only OADs (P = 0.80). The number of patients with insulin treatment and HbA1c ≥52 mmol/mol was higher (P < 0.05) compared to patients with HbA1c <52 mmol/mol. Also, the number of patients with combined OADs + insulin treatment was higher (P < 0.05) for patients with HbA1c ≥52 mmol/mol than for patients with HbA1c <52 mmol/mol.Eighty-two patients (37.6%) were on diet treatment, 60 patients (27.5%) were on OADs, 44 patients (20.6%) had insulin treatment, and 25 patients (11.5%) had combined OADs + insulin treatment. The number of patients with diet treatment was higher  (P = 0.365), intermediate-acting insulin (P = 0.271), mixed insulin (P = 0.490), and continuous rapid-acting insulin (P = 0.490).Of a total of 69 patients on insulin treatment, 9 patients had HbAmmol/mol . For thr= 0.319) . Moreove1c <52 mmol/mol compared to groups with HbA1c ≥52 mmol/mol. However, there was no significant difference between the groups <52 and ≥52 mmol/mol regarding hospitalization due to hypoglycaemic events.Hypoglycaemic events were observed in 52 patients . A totaln = 130) showed the same pattern regarding hypoglycaemic events as in the general group  in the three different HbA1c groups. We could not find any differences regarding daily dosage/doses of insulin between the groups with different HbA1c levels. This finding might indicate excessive treatment, especially for the patients in the group with HbA1c <52 mmol/mol.For patients in the group HbAn = 98), and the diabetes prevalence was 15%, with a mean HbA1c value of 57.0 ± 1.2 mmol/mol, and most patients were on antidiabetic drug treatment. Several international studies have evaluated intensive diabetes treatment in T2DM patients regarding complications and that there were negligible effects on preventing micro- and macrovascular complications, particularly after a longer diabetes duration of 8–10 years (The results in this study are in accordance with findings from a previous study in nursing homes in Sweden . It was 10 years . In the 10 years . Studies10 years ,15,16. H10 years ,17. HypoIn this present study we found a total of 106 hypoglycaemic episodes in 52 patients. Compared to observations in previous studies, hypoglycaemic events were less frequent, which may indicate that the method for finding hypoglycaemic events was insufficient. It could not be excluded that episodes of hypoglycaemia were not reported, since hypoglycaemia may mimic several other conditions and, furthermore, many hypoglycaemic episodes are nocturnal.1c, which might reflect a diabetes disease with poor glycaemic control (1c levels, but the patients could still not benefit from intensive glucose-lowering therapy. The rate of death was even higher among patients with more intensive glucose-lowering treatment (The hypoglycaemic events tended to be more severe among patients with higher HbA control . The ACCreatment ,9,18.1c ≥52 mmol/mol. This could possibly, to some extent, explain why hypoglycaemia occurred more frequently with increasing HbA1c. Longer diabetes duration was correlated with higher HbA1c, and these parameters were associated with increased risk for hypoglycaemic episodes and microvascular complications. Almost 80% of the diabetes patients had either micro- or macrovascular complications, and diabetes duration was associated with both hypoglycaemic episodes and micro- or macrovascular complications.Further, in our study, 49 patients received insulin on demand, and they were involved in 50% of all major hypoglycaemic episodes and 50% of all minor hypoglycaemic episodes. The fact that 130 patients received pharmacological treatment shows that the subgroup given insulin on demand more often suffered from hypoglycaemic episodes. Insulin treatment was more common among patients with HbA1c suffering from hypoglycaemic episodes could benefit from a re-evaluation of their pharmacological treatments, including changes of agents used, dose reductions, or even stopping selected treatments (1c should be monitored quite frequently to ensure that serum HbA1c concentrations are kept within reasonable levels.The results from this study suggest that multimorbid elderly patients with low HbAeatments . HbA1c s


Enteric methane from rumen methanogens is responsible for 25.9 % of total methane emissions in the United States. Rumen methanogens also contribute to decreased animal feed efficiency. For methane mitigation strategies to be successful, it is important to establish which factors influence the rumen methanogen community and rumen volatile fatty acids (VFA). In the present study, we used next-generation sequencing to determine if dairy breed and/or days in milk (DIM) (high-fiber periparturient versus high-starch postpartum diets) affect the rumen environment and methanogen community of primiparous Holstein, Jersey, and Holstein-Jersey crossbreeds.Methanobrevibacter (Mbr.) smithii, Mbr. thaueri, Mbr. ruminantium, and Mbr. millerae. The 16S rRNA gene sequence reads clustered at 3 DIM, but not by breed. At 3 DIM, the mean % abundance of Mbr. thaueri was lower in Jerseys (26.9 %) and higher in Holsteins (30.7 %) and Holstein-Jersey crossbreeds (30.3 %) (P < 0.001). The molar concentrations of total VFA were higher at 3 DIM than at 93, 183, and 273 DIM, whereas the molar proportions of propionate were increased at 3 and 93 DIM, relative to 183 and 273 DIM. Rumen methanogen densities, distributions of the Mbr. species, and VFA molar proportions did not differ by breed.When the 16S rRNA gene sequences were processed and assigned to operational taxonomic units (OTU), a core methanogen community was identified, consisting of The data from the present study suggest that a core methanogen community is present among dairy breeds, through out a lactation. Furthermore, the methanogen communities were more influenced by DIM and the breed by DIM interactions than breed differences. In the United States, enteric methane emissions from ruminants are the second largest anthropogenic source of methane, contributing to 25.9 % of all methane emissions and global warming . MethaneThe rumen is an anaerobic environment that houses a microbiome consisting of bacteria, protozoa, fungi, phages, and archaea. The bacteria, protozoa, and fungi  ferment feedstuff consumed by the host, and produce VFA. Acetate, butyrate, and propionate, the predominant VFA in the rumen, are the main energy sources for the host animal. Fermentation byproducts such as carbon dioxide, formate, hydrogen gas, methanol, and methylamines are used by methanogenic archaea for methane production. The majority of the methane is eructated and exhaled out by the ruminant into the environment. For methane mitigation strategies to be successful, it is important to identify factors that may influence the rumen environment and thus, affect the methanogen density and diversity.Methanobrevibacter (Mbr.) are the most abundant species and primarily use hydrogen and carbon dioxide as substrates for methanogenesis .P < 0.01). Propionate proportions were lowest at 273 DIM and highest at 3 and 93 DIM (P < 0.05). Relative to 3 DIM, acetate proportions were higher at 183 and 273 DIM (P <0.001). Isobutyrate and lactate proportions were highest at 273 DIM (P < 0.01). Isovalerate proportions did not differ by DIM  gene/mL whole rumen digesta) were not different by breed (P = 0.93) or DIM (P = 0.25). The mean and standard error (SE) of densities by breed were 6.51 ± 0.04 (Holsteins), 6.53 ± 0.04 (Jerseys), and 6.52 ± 0.04 for (Holstein x Jersey crossbreeds), while the densities by DIM were 6.48 ± 0.05 (3 DIM), 6.49 ± 0.05 (93 DIM), 6.61 ± 0.05 (183 DIM), and 6.49 ± 0.05 (273 DIM), respectively. The rumen methanogen densities  ranged from 357–390 bp with a mean length of 358 bp. The mean and SE of sequence reads per individual by breed were: 19,039 ± 2,667 (Holsteins), 19,096 ± 2,744 (Jerseys), and 19,947 ± 3,034 (Holstein x Jersey crossbreeds). There were 298,689 total unique sequences  with 67,072 chimeras removed. The numbers of unique sequence reads for each time point was: 67,111 (3 DIM), 87,455 (93 DIM), 77,477 (183 DIM), and 66,646 (273 DIM). Principle coordinate analysis (PCoA) did not demonstrate clustering of methanogen communities by breed. However, rumen methanogen communities clustered at 3 DIM, but not at other time points  and higher in Holsteins (30.7 %) and Holstein-Jersey crossbreeds (30.3 %) (P < 0.001). At 93 DIM, a lower abundance of the species Mbr. thaueri was observed in Holsteins  and Holstein-Jersey crossbreeds  when compared to Jerseys (35.0 %). The species, Mbr. ruminantium (P < 0.05) was higher at 93 DIM than at 273 DIM. . The less abundant methanogen species (<5 %) Mbr. gottschalkii and Mbr. woesei also varied by DIM, but not by breed. At 93 DIM (P < 0.05) and 273 DIM (P < 0.001), the abundance of Mbr. gottschalkii was higher than at 3 DIM. Mbr. woesei (P <0.001) were more abundant at 3 DIM, whereas the order Methanosarcinales was more abundant at 183 DIM (P < 0.01). Less than 1 % of total methanogen sequences were distantly related to the following methanogen genera: Methanoculleus, Methanolobus Methanoplanus, Methanospirillium, and Methanosarcina.All sequence reads belonged to the phylum Euryarchaeota. The SGMT and RO clades did not differ by breed or DIM Table . The majae Table . BecauseP < 0.01), while Good’s coverage and the Shannon Diversity indices were highest at 3 DIM (P < 0.05). The most and least OTUs shared between all animals were at 93 and 273 DIM, respectively (Table The 16S rRNA gene sequence reads clustered into 403 (3 DIM), 383 (93 DIM), 590 (183 DIM), and 547 OTUs (273 DIM). Breed and breed by DIM did not affect rumen methanogen diversity measures  clustered into OTU 1–4. At 3, 93, and 183 DIM, the mean abundance of OTU 1 was 30.8 ± 2.0 %, 30.9 ± 2.8 %, and 31.7 ± 1.6 %, respectively. At 273 DIM, OTU 2 was most abundant with 36.1 ± 2.3 %. No breed effects were observed for OTUs 1–4 and the least abundant OTUs. However, the abundance of OTU 1 was lowered (P < 0.05) in Holsteins (28.8 ± 2.0 %) when compared to Holstein x Jersey crossbreeds (34.9 ± 2.0 %). DIM did not affect the distribution of OTU 3 or the least abundant OTUs. The abundance of OTU 2 increased at 273 DIM (P < 0.05), while the abundance of OTU 4 increased at 3 DIM (P <0.05).The top four OTUs shared by all breeds and at each stage of lactation were related to the species r = −0.98, P < 0.0001) . The abundance of the order Methanosarcinales was positively correlated with the order Methanoplasmatales . Several weak correlations were observed between most methanogen taxa and VFA  and positively correlated to propionate . Propionate was positively and negatively correlated to Mbr. ruminantium  and Mbr. thaueri , respectively. Several correlations were observed between individual VFA. Acetate was negatively correlated to propionate , butyrate , and valerate  and positively correlated to isobutyrate . Propionate was negatively correlated to isobutyrate  and lactate  and positively correlated to valerate .Notably, the SGMT and RO clades were negatively correlated (01) Fig. . A negatVFA Fig. . The speThe present study is the first to investigate the rumen methanogen community across a lactation period in three dairy cattle breeds. The purpose of this experiment was to provide more knowledge about the rumen methanogen community and rumen parameters at 3, 93, 183, and 273 DIM in Holstein, Jersey, and Holstein-Jersey crossbreeds. This study identified the core methanogen community with NGS technologies, quantified rumen VFA and methanogen densities, and correlated rumen methanogen species to one another and to VFA.VFA are the main energy source provided to lactating dairy cattle and are the by products of carbohydrate fermentation by rumen bacteria, protozoa, and fungi. Generally, propionate is a precursor to glucose and is increased when animals are provided a high-starch diet or provided the ionophore, monensin. Relative to 183 and 273 DIM, proportions of propionate were increased at 3 and 93 DIM, suggesting a greater demand for glucose by the cow during early lactation. Although the animals were provided 0.06 % monensin pre-partum versus 0.02 % post-partum, it is not possible to correlate the increase in propionate at 3 and 93 DIM with this additive. An effect from monensin would be more plausible at 3 DIM, when the cows were transitioning from a pre-partum to a post-partum diet, but this would not explain why propionate was also increased at 93 DIM.. [Furthermore, the increase in total VFA concentrations observed at 3 DIM suggests an increase in carbohydrate fermentation at the start of lactation. Johnson et al.  stated t.  found th10 mcrA gene copies) from bulls on high-fiber (9.02) and starch diets (9.07) [The present study is the first to compare the methanogen densities in three breeds of dairy cattle and by DIM. In agreement with our hypothesis, the methanogen densities did not vary by breed or DIM. Previously reported methanogen densities   were higs (9.07) . Howevers (9.07) . In a sts (9.07) , the useMbr to be the most predominant genus. As methanogens belonging to the genus Mbr use the rumen fermentation byproducts, such as hydrogen and carbon dioxide as substrates for methanogenesis, it is thought that the high levels of these byproducts in the rumen enable these methanogens to thrive over other species that rely on scarce substrates such as methylamines, methanol, or acetate [Previous work in dairy , 7, 10 a acetate , 15.Mbr is the most abundant archaeal genus in ruminants, there are several species that are distributed into two different phylogenetic clades . In the present study, the SGMT clade was the most dominant branch by breed and DIM. Both Mbr. smithii and Mbr. thaueri made up the majority of the SGMT clade, while Mbr. ruminantium made up the majority of the RO clade. Previous research, using the same archaeal forward primer (Met86F), suggested a difference between SGMT-RO clade distributions between Holstein and Jersey cows [Although sey cows . Howeversey cows . FinallyMbr. smithii, Mbr. thaueri, Mbr. ruminantium, and Mbr. millerae were identified in each breed and at each DIM time point investigated, our data showed the presence of a core methanogen community. Jeyanathan et al. [Because the four methanogen species n et al.  identifiMbr. thaueri was not. Recently, Mbr. thaueri was identified with the same primer pair used in the present study, at a high abundance in wild impalas from South Africa [Mbr. thaueri from previous studies could be due to lack of sequencing depth or diet of the animal. The higher abundance of Mbr. thaueri in Jersey cows at 93 DIM  may be a result of a higher dry matter intake (DMI), but future studies are needed to draw a clear link between DMI, milk yield, and the rumen methanogen species identified. It is conceivable that Mbr. thaueri and the other three methanogen species persisted because the rumen environment and the substrates created by bacteria, protozoa, and fungi enabled these methanogens to thrive.Three out of the four methanogen species in the present study were previously identified in both Holstein and Jersey cows, while h Africa . OmissioMsp. was identified in low abundances. Its mean % abundance was highest around peak lactation (93 DIM) and lowest at 3, 183, and 273 DIM. Similarly, Kumar et al. [Mbr or Msp. At 1–5 DIM, the methanol-utilizing genus Msp was more abundant (4.5 %) in primiparous Holsteins than those from the present study (<1 %). Like in the present study, animals were stomach tubed 2–3 h post-feeding and received a diet before calving with the same NDF content  from the present study were highest at 3 DIM, while the 16S rRNA gene sequences reads clustered during this time as well. At 93, 183, and 273 DIM the sequence reads were mixed in one cluster. Previous research also demonstrated that the Shannon diversity indices of methanogens increased in bulls on a high-fiber diet (0.95) and decreased with a high-starch diet (0.79) . These dMbr. smithii, Mbr. thaueri, Mbr. ruminantium, and Mbr. millerae. These methanogens may play a significant role in methanogenesis and in the utilization of substrates from bacterial, protozoal, and fungal fermentation. However future work is required to better delineate these relationships. The SGMT-RO clades did not vary by breed or DIM, instead, the SGMT clade was dominant in all three breeds. Although our results show that breed does not affect the rumen methanogen taxa per se, more studies are needed to clarify if this finding is consistent in other geographic locations and in dairy cattle consuming varying diets.The data presented here are the first to characterize the rumen methanogen communities in three dairy cattle breeds across a lactation period. NGS produced over 1 million sequence reads and demonstrated that diversity was different at 3 DIM. Notably, a core methanogen community persisted and consisted of four species, From May 2013 to May 2014, 22 primiparous lactating dairy cattle (7 Holstein (H), 8 Jersey (J), and 7 first generation Holstein x Jersey crossbreeds (X)) were co-housed at the University of Vermont (UVM) Paul Miller Research Complex in South Burlington, VT. At 3 DIM, one Jersey was excluded from all data analyses because of post-partum health concerns. The Institutional Animal Care and Use Committee at the UVM approved all animal sample collection methods under protocol # 13–031. All animals calved within a 2-month period. At 3, 93, 183, and 273 DIM, whole rumen digesta samples (50 mL) were collected 2-3 h post-feeding at 0900 h via stomach intubation from each animal. To collect rumen samples, a flexible milk hose (2.54 cm diameter) was passed through a speculum to the esophagus and to the rumen. The hose was marked at 200 cm to indicate the approximate location of the rumen. Once the tube contacted the fiber mat and rumen sounds were heard, a 600 cc livestock drench gun  collected the digesta. Whole digesta samples were immediately frozen at −20 °C to minimize microbial activity.Prior to calving, all animals consumed a pre-partum total mixed ration (TMR) diet. Within 24 h post-partum, each cow was transitioned to a diet that was higher in starch and lower in NDF in comparison to the diet fed pre-partum Table . Throughg for 20 min at 4 °C and the resulting supernatant was filtered through a 25 mm hardened ashless filter  to remove debris. Subsequently, the samples were diluted 1:1 with 0.06 M oxalic acid containing 50 μM trimethyl acetic acid  and analyzed for VFA by gas chromatography  coupled with a flame ionization detector and a customized packed column (2 m x 2 mm ID glass) with 4 % carbowax and 80/120 Carbopac B-DA  with nitrogen as carrier gas (15 mL/min flow rate). The other gases were purified air at 300 mL/min and hydrogen makeup gas at 30 mL/min. The column was operated at 175 °C; the total run time was 25 min. Both the injector and detector temperature were kept at 200 °C. The injection volume was 1 μL. The identification of VFA was based on retention times against known standards using software Star Chromatography v5 (Varian) and quantified for their concentrations using respective VFA standards. Results are expressed in mM VFA.The whole rumen digesta samples were spun in a Beckman J2-21 centrifuge at 10,000 x Across 4 time points , 87 individual whole rumen digesta samples were collected. The previously frozen samples were thawed overnight at 4 °C. Each sample was vortexed for 30s to homogenize the sample and break up the solid particles that settled to the bottom of the conical tube. The microbial DNA was extracted using the repeated-bead beating plus column (RBB + C) method  and foll10 of the copy number of the mcrA gene per mL of rumen digesta was determined by real-time PCR, each sample was amplified in triplicate. Each real-time PCR included: 12.5 μL of SYBR® Green Mix, 6.5 μL of double distilled water, 2.5 μL of the methanogen-specific primer pair, mcrA-F and mcrA-R [2 value greater or equal to 0.997. Using the BioRad CFX Manager (v.3.0) a model equation of the standard curve (y = mx + b) was used, where “x” was the log of the starting quantity, “b” the y-intercept of the quantification value (Cq), and “m” the slope of the line for log starting quantity versus Cq values. Individual densities were calculated using previously established methods [The logd mcrA-R , and 1 μd mcrA-R ) or negad mcrA-R . The fiv methods .The archaeal-specific primer pair, Met86F  and Met4The program MOTHUR, version 1.33.3, was used to perform bioinformatics analyses in-house . Each tiTo determine the number of unique sequences in the data set, the command unique.seqs was used. A Needleman-Wunsch pairwise alignment and an aligned reference file of known rumen methanogen 16S rRNA gene sequences were used with the command align.seqs to align the unique sequences. Chimeric sequences were identified with UChime  and remoIn order to detect any bacterial 16S rRNA gene sequences, the sequences were classified with the 16S rRNA reference Ribosomal Database Project (RDP) files provided by MOTHUR. These files contained known bacteria and methanogen sequences from kingdom to genus taxonomic levels. The files were modified to contain species-level names. Any bacteria sequences (<0.01 %) were removed. The online RDP Classifier was used at a 95 % confidence threshold to further quality check the sequences. Sequences with an unknown root were removed. Taxonomy and fasta files containing 765 known archaeal species names and 16S rRNA gene sequences were used to classify the sequence reads into taxonomic species. The command cluster.split, was used with a 2 % cutoff to cluster the 16S rNA gene sequences into OTUs. Once OTUs were formed, they were classified with the command, classify.otu. The command summary.single calculated OTU-based alpha diversities, Shannon Diversity Index, Chao I richness estimator, Inverse Simpson index, and Good’s coverage.The subsample parameter in MOTHUR was used to analyze the same number of sequences per individual sample. Shared OTUs within and between breeds were counted with get.sharedseqs. To correlate OTUs , the otu.association command used a Pearson correlation. The command, merge.file combined the fasta files of unique sequences. A subsample of 100,000 sequences was taken and the distances between sequences with a 2 % cutoff were calculated with dist.seqs. Once a phylip distance file was created, the clearcut command created a phylogenetic tree file. The output from clearcut was used with Fast Unifrac to perform a PCoA . The PCoP < 0.05 and trends were declared at 0.05 ≤ P ≤ 0.10.All data were analyzed with a repeated measures ANOVA model in SAS 9.4  with PROC MIXED. The model included breed, DIM, and breed by DIM interactions as fixed effects and used the Kenward-Roger method to determine the degrees of freedom. A Pearson correlation, to determine the relationship between VFA and methanogen taxa, was performed with the PROC CORR. The online data visualization tool, Plotly, was used to generate a heatmap of the correlation values (r). Statistical significance was declared at The present study was performed in accordance with the Institutional Animal Care and Use Committee at the University of Vermont under protocol # 13–031.The data sets that support the results of the present study are included within the journal article.


AbstractSantalumaustrocaledonicum Vieill.  is endemic to New Caledonia  and Vanuatu, where several varieties are recognized. The Loyalty Islands sandalwood variety is here reinstated as Santalumaustrocaledonicumvar.glabrum Hürl. emend. Butaud & P.Firmenich, mut. char. It was previously considered a synonym of the type variety; however, new morphological and genetic studies confirmed its distinctiveness. The key for New Caledonian varieties of Santalumaustrocaledonicum has been updated and a short description of its essential oil composition and organoleptic quality is given.Sandalwoods encompass 19 species restricted to southeast Asia and the Pacific. The species  Santalum  comprising 19 species restricted to southeast Asia and the Pacific , Isle of Pines, Loyalty Islands and Vanuatu;Santalumaustrocaledonicumvar.pilosulum N.Hallé in the vicinity of Nouméa on the southwest coast of Grande-Terre;Santalumaustrocaledonicumvar.minutum N.Hallé from the northwest coast of Grande-Terre.var.austrocaledonicum in New Caledonia and Vanuatu which are linked to geographical distribution , short and wide juvenile leaves ;Isle of Pines: medium-sized seeds , long and narrow juvenile leaves ;Grande-Terre: small seeds , long and very narrow juvenile leaves .No comprehensive study of the variation of these characters is available in Vanuatu due to lack of herbarium specimens from most of sandalwood populations there.varietyloyaltensis but his manuscript was never published.For this reason, More recently, molecular studies of New Caledonian sandalwood showed strong genetic differentiation between islands and led to the recognition of two evolutionarily significant units, i.e. Grande-Terre and Isle of Pines for the first, and Loyalty Islands for the second , 2007.The New Caledonian sandalwood is subject to exploitation through harvesting and is grown in plantations to satisfy the increasing international demand of sandalwood essential oil. However, its taxonomy appears to be in need of revision to more precisely describe the variability of this New Caledonian biodiversity hotspot species  and to cA first step of this revision is presented here, with the recognition of an endemic sandalwood variety from the Loyalty Islands based on morphological and molecular studies as well as examination of living plants and herbarium specimens.PageBreakSantalumaustrocaledonicum, was described by E. Vieillard 1090, holotype P00645808). Later, it was confirmed that this species was also present on the Isle of Pines, Loyalty Islands and Vanuatu  represented erroneously by samples gathered around Nouméa  which have villous flowers.In 1964, Hürlimann described a new variety based on a sample collected on the island of Maré in the Loyalty Islands , SantaluSantalum by Hallé recognized the three varieties mentioned in the introduction. Hallé reduced var.glabrum of Hürlimann to synonymy under the type (autonym) variety, considering it superfluous due to confusion by Hürlimann on what was really the type of the species. Indeed, the latter described var.glabrum in comparison with what is now recognized as var.pilosulum, this one differing morphologically from var.austrocaledonicum. The true var.austrocaledonicum and var.glabrum are in fact identical relative to the morphological differences pointed out by Hürlimann in his diagnosis.In 1988, the revision of the New Caledonian glabrum for the endemic Loyalty sandalwood, its description must be amended to distinguish it from the true type specimen.As the native Loyalty Islands sandalwoods are quite homogeneous, only one variety is to be recognized for the entire Loyalty archipelago, which is the one of Hürlimann. Thus, to reinstate the varietal name Taxon classificationPlantaeSantalalesSantalaceaeHürl. emend. Butaud & P.Firmenichmut. char.Santalumaustrocaledonicumvar.glabrum Hürl., Mém. Mus. Hist. Nat., ser. B, Bot. 15(1): 15 (1964).M.G. Baumann-Bodenheim 14762 .New Caledonia, Loyalty Islands, Maré, près de Rawa, arbre, 8 m, en fleurs et en fruits, forêt mésophile, 17 July 1951, Santalumaustrocaledonicumvar.glabrum is most similar to var.austrocaledonicum in its glabrous inflorescence and leaves wider than 1.5 cm, which differentiates them from the other New Caledonian Santalumaustrocaledonicum varieties. Santalumaustrocaledonicumvar.glabrum differs from Santalumaustrocaledonicumvar.austrocaledonicum by the seed size, which is more than 7.5 mm wide for the former and less than 7.5 mm for the latter.PageBreakLeaves glabrous; petiole canaliculate, 7–13 mm long; blades of the mature leaves 3.5–6.0 (–6.6) × (1.6–) 2.0–3.5 (–4.1) cm, usually elliptic or rarely obovate, apex obtuse to acute or apiculate, base acute, secondary veins mostly 7–9 pairs. Inflorescences glabrous, in axillary or terminal panicles, usually trichotomous and several times branched, with 10–40 flowers; peduncles 10–38 mm long. Flowers bisexual with outer surface of petal greenish and glabrous; pedicels 1.5–2 mm long. Petals 4, narrowly triangular, 2.5–3.0 × 1.5–2.0 mm, inner surface white when opening turning brownish later; petal internal margin glabrous. Stamens 4, surrounded by long hairs at the base; the outer ones reaching the anther apex, the inner ones reflexed in the cup-shaped disk; anthers 1.5–2.7 × 0.8–1.0 mm. Disk concave, more than 2 mm deep; disk lobes fleshy and erected between petals, 1.3–1.4 × 0.8–0.9 mm. Ovary unilocular, conic, acute, 1.1–1.7 × 0.6 mm; style free, 4 mm long; stigma 3 or 4 lobed. Fruit a globose fleshy drupe, 15–21 × 12–15 mm when fresh, topped by the petal scars 3–5 mm diameter, green turning red to deep purple and black at maturity. Seed globose, with a hard endocarp (8.5–) 9.0–11.5 × 7.5–10.0 (–10.5) mm.Shrub to small tree 2–10 m tall, trunk up to 30 cm dbh; bark rough, grey to reddish-brown, longitudinally fissured; heartwood fragrant, yellowish to brownish. Flowering and fruiting probably occurring throughout the year but with some peaks; herbarium samples provide the following data: flowers from December to August, fruits from January to August.New Caledonia, endemic to Loyalty Islands; known only from Ouvéa, Lifou and Maré islands Figure . Not recAcaciaspirorbis Labill., Acalypha spp., Acronychialaevis J.R.Forst. & G.Forst., Dodonaeaviscosa (L.) Jacq., Elattostachysapetala (Labill.) Radlk., Glochidionbillardierei Baill., Melochiaodorata L.f., Morindacitrifolia L., Pandanusmacrocarpus (Brongn.) Solms, Podonepheliumhomei (Seem.) Radlk., Polysciasbracteata(R.Vig.)Lowrysubsp.bracteata., Psidiumguajava L., and Schinusterebenthifolius Raddi. Loyalty sandalwood is a hemiparasitic tree like all Santalum species; its pollination is insect-mediated whereas its fleshy fruits are dispersed mainly by doves and pigeons : B (1+2) ab . Its population size is estimated at more than 10,000 mature individuals (excluding criteria C and D) with an extent of occurrence around 8,000 km² and an area of occupancy around 1,000 km². Criteria A can not be used due to PageBreaklack of knowledge of generation length and magnitude of population size reduction. Three locations (one per island) can be distinguished without any fragmentation. A continuing decline is observed and projected in terms of habitat quality and number of mature individuals due to harvest, competition with invasive plant species  Cass., Lantanacamara L.), hybridization with other Santalumaustrocaledonicum varieties used in plantations, and changes in the traditional agricultural system . Indeed, despite provincial regulations establishing quota and exploitability criteria, illegal logging is still occurring  or “tapakai” . Moreover, two new endemic varieties of Santalumaustrocaledonicum are expected based on the recent study of The taxonomy of  studies  showed s but also as a promising new raw material for the fraPageBreakgrance industry , pilosulum (surroundings of Nouméa) and austrocaledonicum (Isle of Pines), leaving unknown the quality of each individual variety.Essential oil of industry . These evar.glabrum), two samples were analyzed for their main sesquiterpenoids but also for their organoleptic properties:To investigate the quality of Loyalty Islands sandalwood essential oil  obtained in 2015 from Distillerie de Boulouparis  with Ouvéa sandalwood,Loyalty Oil 2 (EI2) obtained in 2002 from Michel Point and processed by Koop Cuada distillery with Maré and Lifou sandalwoods.The gas-chromatography was performed on an apolar column  to 280° (2’) at 50°/min., vector gas: hydrogen). Its results are given in Table The organoleptic evaluation showed that the Loyalty essential oil is lighter and less milky than the Indian sandalwood; the former has also a character less woody but more animal and masculine .Santalumaustrocaledonicumvar.glabrum or Loyalty sandalwood constitutes a true substitute for Indian sandalwood in the perfume industry.These findings are in accordance with


Despite the current obesity epidemic, maternal underweight remains a common occurrence with potential adverse perinatal outcomes.We aimed to investigate the relationship between weight gain during pregnancy, and fetal growth in underweight women with low and late fertility. Women body mass index (BMI), defined according to the World Health Organization’s definition, gestational weight gain (GWG), defined by the Institute of Medicine and National Research Council and neonatal birth weight were prospectively collected at maternity ward of Policlinico Abano Terme  in 793 consecutive at term, uncomplicated deliveries.2), 551 (69.5 %) as normal weight, 107 (13.4 %) as overweight, and 39 (4.9 %) as obese, respectively. In all mother groups, GWG was within the range recommended by IOM 2009 guidelines. However, underweight women gained more weight in pregnancy (12.8 ± 3.9 kg) in comparison to normal weight (12.3 ± 6.7 kg) and overweight (11.0 ± 4.7 kg) women and their GWG was significantly higher (p < 0.001) with respect to obese women 5.8 ± 6.1 kg). In addition, offspring of underweight women were comparable in size at birth to offspring of normal weight women, whereas they were significantly lighter to offspring of both overweight and obese women.Among those, 96 (12.1 %) were categorized as underweight (BMI < 18.5 kg/mPre-pregnancy underweight does not impact birth weight of healthy, term neonates in presence of normal GWG. Presumably, medical or personal efforts to reach ‘optimal’ GWG could be a leading choice for many women living in industrialized and in low-income countries. It ho common . For inso common  and 9.0 o common  are undeo common , a populommon [2 .When BMI-specific ranges for “optimal” weight gain were introduced by IOM , the natHence, there is a need to see the contextual relevance of these determinants to our own population, representative of developed, industrialized countries supporting advanced educational levels, good socio-economic status, and low and late fertility. We therefore undertook an analytical study with a reference group of pre-gestational low BMI women to determine the direction and magnitude of the effect of maternal underweight and GWG on fetal growth in singleton, at term, uncomplicated pregnancies.This prospective study was performed at the maternity wards of the Division of Perinatal Medicine of Policlinico Abano Terme, Abano Terme, Italy. The hospital where the study took place is located in an industrialized area supporting advanced educational levels, good socio-economic status, occupation, and low and late fertility.The study group consisted of 793 consecutive mothers of singleton, healthy neonates delivered at term. Women that didn’t speak Italian, underwent general anaesthesia, with psychological treatment and with infants hospitalized in NICU, were excluded from the study.After informed consent was obtained, they answered sociodemographic questions before and after pregnancy, obstetric history, smoking, psychological care, plan to lactate an infant without medical or obstetrical complications precluding routine discharge.It was an analytical study with the use of secondary data . The stuEvery singleton at term (>37 < 42 weeks) pregnancy with known GWG indicators and maternal and neonatal outcomes were eligible. After informed consent was obtained, they answered sociodemographic questions before and after pregnancy, obstetric history, smoking, psychological care, plan to lactate an infant without medical or obstetrical complications precluding routine discharge. The following data were also collected for the babies: gestational age, birth weight, length, head circumference, and mode of delivery.2). The women were categorized into four categories with respect to their BMI as per the standard of institute of Medicine [2), normal (BMI ≥18.5 and < 25 kg/m2), overweight (BMI ≥25 and <30 kg/m2), and obese (BMI ≥30 kg/m2). Gestational age was recorded by last menstrual period and verified by ultrasound as a routine practice within the institution. Total weight gain was calculated by subtracting the pre-pregnancy weight from the last measured weight before delivery and ‘optimal’ ranges were categorized according to 2009 IOM guidelines: underweight 12.5–18 kg, normal weight 11.5–16 kg, overweight 7–11.5 kg, and obese 5–9 kg, respectively [t test were used at 0.05 level of significance. Statistical analyses were carried out using Statistical Package for Social Sciences (SPSS) version 19 .Pre-pregnancy weight was used as recommended by other studies on gestational weight gain –15. BMI Medicine  as underAnthropometrical and clinical features of the study population are shown in Table This cohort study is characterized by advanced educational levels, good socio-economic status, occupation, and low and late fertility. Women were 33.0 ± 4.7 years old, 56.2 % were nulliparous, most completed high school (52.0 %) or graduated from university (33.5 %), and 82.2 % were employed, and 9 (7.8) 49 (6.1) were unemployed. Smoking behaviour was present in 3.6 %. Only 3 neonates, 1 offspring of a normal weight woman and 2 of overweight women, respectively, were NICU admitted .Anthropometrical and clinical characteristics of the mother-infant dyads categorized according to the Institute of Medicine  BMI and Among 793 consecutive women included in this analysis, 96 (12.1 %) were categorized as underweight , 551 (69.5 %) as normal weight, 107 (13.4 %) overweight, and 39 (4.9 %) obese, respectively.Underweight women were 31.9 ± 5.1 years old, 53.1 % were nulliparous, most completed high school (53.1 %) or graduated from university (32.2 %), and 85.4 % were employed, and 9.3 % were unemployed. Smoking behaviour was 6.2 %.p < 0.001) with respect to obese women 5.8 ± 6.1 kg).In all mother groups, GWG was within the range recommended by IOM 2009 guidelines. However, underweight women gained more weight in pregnancy (12.8 ± 3.9 kg) in comparison to normal weight (12.3 ± 6.7 kg) and overweight (11.0 ± 4.7 kg) women and their GWG was significantly higher  (2.0 vs. 5.9 %) and of operative delivery, caesarean section (16.6 vs 27.7 %), and vacuum extractor (2.5 vs. 4.7 %), respectively, in comparison to normal BMI women. The above-described reduced rates appeared also more pronounced with respect to overweight and obese women.Following IOM BMI-specific ranges for “optimal” weight gain in women categorized by weight group, we explored the impact of the combined effects of pre-pregnancy low BMI and GWG on fetal growth, in an urban setting of women living in an industrialized area of North-Eastern Italy. In this cohort study, inclusive of women with advanced educational levels, good socio-economic status, and low and late fertility, pre-pregnancy low BMI was common and involved more than 1:10 women, whereas mean GWG was within the range recommended by IOM 2009 guidelines in this and in the other weight groups.Our results have also shown that offspring of underweight were comparable in size at birth to offspring of normal weight women, and significantly lighter than offspring of both overweight and obese women. However, occurrence of LGA neonates and operative delivery rate, caesarean section and vacuum extractor, were almost halved in underweight women in comparison to normal weight women and even three-four times lower in comparison to underweight or obese women.One of the most important modifiers of pregnancy weight gain and its impact on a mother’s and her baby’s health is a woman’s weight at the start of pregnancy . HoweverThe underweight prevalence in this study is similar to that reported in a Chilean women population  and consUnderstanding these associations is also complex, because pre-gestational BMI and GWG, considered predictors of the maternal nutritional status, are closely linked to lifestyle, prevalent body image models, and socio-cultural factors in industrialized countries and, in contrast, to endemic malnutrition, diseases, and genetic traits in disadvantaged, low income countries . In addiAfter excluding cases of abnormal GWG, a stronger similitude between fetal growth of offspring of underweight and normal weight women appeared. This is the first time this result has been reported, and it suggests that the known relationship between pre-gestational BMI and offspring birth weight obscures, in part, the association between pre-pregnancy underweight or low GWG and birth weight previously reported in low income women, and this must be taken into account. It is possible that the ‘optimal’ weight gain in women with low BMI may have received less attention from clinicians but may indeed play a strong role in the risk of LBW and adverse pregnancy .Although we adopted a reproducible method to classify maternal BMI, GWG and fetal growth, we were not able to exclude some other limitations of this study, which therefore need to be addressed. Firstly, the relevant correlation between birth weight and gestational age of the newborns is lacking. This information could have some impact on postnatal adverse health outcomes  , 34. AnoOur analysis using net maternal weight gain in underweight women living in an industrialized, high income country does not reinforce the association between low BMI and deleterious consequences for themselves or their infants, differently from that reported in underweight, disadvantaged women living in low income countries. Offspring of underweight and normal weight women were comparable in size at birth to offspring of normal weight women and operative deliveries rates, caesarean section and vacuum extractor, were reduced in comparison to normal weight women and even more so in comparison to overweight or obese women. The findings of this study highlight the need for nutritional education and life style adjustments to improve pregnancy outcomes.BMI, body mass index; GWG, gestational weight gain; IOM, the Institute of Medicine; IUGR, intra uterine growth retardation; LGA, large for gestational age; WHO, World Health Organization


Pleurotus ostreatus in solid-state fermentation. Among eight variables 2SO4, KH2PO4, asparagine, and yeast extract), copper sulfate and ammonium sulfate concentrations were demonstrated to significantly influence laccase production. The replacement of ammonium sulfate by yeast extract and the addition of ferulic acid as inducer provided increases of 5.7- and 2.0-fold, respectively, in laccase activity. Optimization of laccase production as a function of yeast extract, copper sulfate, and ferulic acid concentrations was performed by response surface methodology and optimal concentrations were 6.4 g/L, 172.6 μM, and 1.86 mM, respectively. Experimentally, the maximum laccase activity of 151.6 U/g was produced at the 5th day of solid-state fermentation. Lignin content in sugarcane bagasse was reduced from 31.89% to 26.36% after 5 days and to 20.79% after 15 days by the biological treatment of solid-state fermentation.Laccases are oxidative enzymes related to the degradation of phenolic compounds, including lignin units, with concomitant reduction of oxygen to water. Delignification is a necessary pretreatment step in the process of converting plant biomass into fermentable sugars. The objective of this work was to optimize the production of laccases and to evaluate the delignification of sugarcane bagasse by  Laccases are blue multicopper oxidases able to oxidize a variety of phenolic compounds and, in the presence of a mediator , also nonphenolic compounds –3. It isLaccases are indicated for several applications in different sectors, that is, in the food and beverages industry to remove undesired phenolics which are responsible for browning, haze formation, and turbidity, in the pulp and paper industry, for biopulping and biobleaching processes, and in the textile industry, for dyes decolourization, and there are also applications in the fields of nanotechnology, bioremediation, and synthetic chemistry , 9.The utilization of agroindustrial wastes as substrates for fermentative processes producing high added value products  has been widely explored, since they are easily available and rich in carbon and often present disposal problems –12. SugaOne of the main challenges in the utilization of lignocellulosic biomass in fermentative processes is the transformation of the complex polysaccharides into simple sugars that can be assimilated by microorganisms. This can be achieved by chemical or enzymatic hydrolysis, preceded by appropriate pretreatments that enhance the efficiency of hydrolysis by lignin removal .Delignification can be performed by thermochemical processes or by the biological route, using enzymes or microorganisms. The advantages of biological delignification over the thermochemical methods may include mild reaction conditions, higher product yields, fewer side reactions, less energy demand, and less reactor resistance to pressure and corrosion .Lignin decomposition in nature is primarily attributed to the metabolism of microorganisms. Among all other organisms, white-rot basidiomycetes degrade lignin more rapidly and extensively than other groups  through  Ceriporiopsis subvermispora, Phellinus pini, Phlebia spp., and Pleurotus spp. preferentially attack lignin more readily than hemicellulose and cellulose. Many white-rot fungi, however, such as Trametes versicolor, Heterobasidion annosum, and Irpex lacteus, exhibit a pattern of simultaneous decay characterized by degradation of all cell wall components [Some white-rot fungi such asmponents .Solid-state fermentation is an interesting technology to be applied in the valorization of agroindustrial residues and can be economically feasible for the production of many biotechnological products . It is a Pleurotus ostreatus (coded Pl 22 Em).The objective of this work was to optimize the fermentation conditions for the production of laccases and to evaluate the delignification of sugarcane bagasse, through the process of solid-state fermentation, using a selected strain ofThe sugarcane bagasse, generated by the sugar and alcohol industry, was provided by the private company Ourofino Agronegócio, located in the region of Ribeirão Preto, São Paulo, Brazil. The bagasse was previously washed with water and dried at 60°C and the portion presenting particle sizes greater than 2 mm was grinded in a knife mill. Separation of the fractions presenting different particle sizes was performed by sieving. Contents of lignin, total extractives, ashes, and moisture were determined according to the TAPPI norms T222, T264, T413, and T264, respectively. Holocellulose content was calculated by difference.4·7H2O (0.3 g/L), FeSO4·7H2O (0.005 g/L), MnSO4·H2O (0.00156 g/L), ZnSO4·7H2O (0.0014 g/L), CaCl2 (0.3 g/L), and CoCl2 (0.002 g/L) [4·5H2O as the inducer , (NH4)2SO4 as the nitrogen source , KH2PO4 as the source of potassium and phosphorus , asparagine as the supplementary aminoacid , yeast extract as the source of vitamins and aminoacids , and pH . The pH was adjusted with HCl 1 M or NaOH 1 M. These variables were chosen on the basis of the composition of the basidiomycetes synthetic medium. In order to evaluate the effect of water activity (Aw) and temperature, the bagasse was humidified with different volumes of saline solution  and the cultures were incubated at 25, 29, and 33°C. For the subsequent experiments, the concentration of KH2PO4 and the pH were set at 1.5 g/L and 5.5, respectively; no asparagine was added; initial Aw was fixed at 0.993 or 10 mL/g and temperature at 29°C. The studied variables were nitrogen source 2SO4, 2.5 g/L), inorganic inducer , and organic inducer .Erlenmeyer flasks containing 1 g of sugarcane bagasse (particle size between 0.8 and 2 mm 50% and <0.8 mm 50%) were autoclaved and humidified with a sterilized saline solution presenting the following constant composition: MgSO002 g/L) . For the P. ostreatus (Pl 22 Em), available at the culture collection of the Bioprocess Engineering and Biotechnology Department  was reactivated in PDA dishes and after 7 days of growth, 4 disks of 7 mm diameter were transferred to Czapek liquid medium containing the antibiotic cephalexin (0.08 g/L). After 5 days of growth at 30°C and 120 rpm, the mycelium was separated from the residual medium by a sieve, homogenized with a spatula and resuspended in the residual medium to a lower final volume . 0.2 mL of the homogenized mycelium (containing 4% of dry biomass) was transferred to the fermentation flasks, which were manually homogenized and incubated for 5 days (optimization studies) or for 3 to 7 days (kinetic study).The strain ofFermentations were prepared as described in Analysis of the results and determination of the mathematical model were performed using the software Statistica 5.0 , and determination of the optimal levels was performed through the Solver Excel tool .2PO4·H2O, 50 mM, pH 7.0) [Enzymes produced by solid-state fermentation were extracted by solid-liquid extraction using sodium phosphate buffer as solvent (NaH pH 7.0) . The ferμL of ABTS 20 mM , sample , and sodium citrate buffer  up to 1 mL. Oxidation of ABTS was followed by absorbance increase at 420 nm . The enzyme activity was expressed in international units (U), where one unit of enzyme activity is defined as the amount of enzyme that oxidizes 1 μmol of substrate in 1 min.The enzymatic activity of laccase was assayed by the oxidation of ABTS . The reaction mixture contained 100 4·7H2O (0.3 g/L), FeSO4·7H2O (0.005 g/L), MnSO4·H2O (0.00156 g/L), ZnSO4·7H2O (0.0014 g/L), CaCl2 (0.3 g/L), CoCl2 (0.002 g/L), KH2PO4 (1.5 g/L), and pH 5.5. Yeast extract  and copper sulfate (sterilized by filtration) were added to the saline solution to reach final concentrations of 6.4 g/L and 173 μM, respectively. Ferulic acid (sterilized by filtration) was added after 48 h of fermentation to a final concentration of 1.86 mM. The inoculum was prepared according to Erlenmeyer flasks containing 3 g of sugarcane bagasse (particle size between 0.8 and 2 mm 50% and <0.8 mm 50%) were autoclaved and humidified with a saline solution  presenting the following constant composition: MgSO P. ostreatus Pl 22 Em.R2 = 0.96873).According to the Pareto analysis, the variables that presented significant effects on laccase production at the confidence level of 90% were copper sulfate and ammonium sulfate concentrations, both presenting positive standardized effects of 11.25 and 5.003, respectively  Kummer (type Florida) liquid culture medium caused a 30-fold increase in total laccase activity, and Hou et al. [ P. ostreatus  when Cu2+ 1 mM was added to the liquid culture medium. Baldrian and Gabriel [2+ not only induces laccase by the expression of laccase genes in P. ostreatus CCBAS-447  but also positively affects activity and stability of the enzyme.As far as the effect of copper sulfate is concerned, the addition of copper as an inducer of laccase production has been already reported in literature. Different studies have shown that laccase production is regulated by metal ions such as Cugulation , 23. Palgulation  demonstru et al.  reported Gabriel  conclude Phanerochaete chrysosporium [ P. ostreatus , however, a higher concentration of nitrogen in the medium did not repress but rather slightly stimulated mineralization of lignin, as reported by Stajić et al. [ P. ostreatus, produce higher ligninolytic enzyme activities in response to a nitrogen-rich medium.As far as the effect of ammonium sulfate is concerned, the production of ligninolytic enzymes has been associated with the secondary metabolism and with conditions of limited nitrogen for many white rot fungi, including the model organism for laccase production and lignin degradationosporium . For P. ć et al. . Kaal etć et al.  also sug P. ostreatus Pl 22 Em when different sources of nitrogen were used and when ferulic acid was added is presented in The comparison between the laccase activities produced by the strain P. ostreatus (strain 32) were peptone and yeast extract, in comparison with urea, ammonium sulfate, and ammonium tartrate. These organic nitrogen sources increased laccase activity in 1.55, 1.99, and 1.46, respectively, (peptone) and 1.40, 1.79, and 1.32, respectively (yeast extract). Mishra and Kumar [ P. ostreatus MTCC1804 , yeast extract was preferred to inorganic nitrogen sources, reaching 23 U/g dry substrate against 2.2 U/g (without nitrogen supplementation), 10.11 U/g (with ammonium sulfate), and 13.0 U/g (with urea). Highest activity, however, was obtained in the presence of cyanobacterial biomass and copper sulfate 1 mM (65 U/g after 10 days). These results may be attributed to the presence of some additives (nutrients/activators) and favorable C : N ratio of the organic nitrogen sources [These results are in accordance with Hou et al. , who demnd Kumar  also dem sources . P. ostreatus Pl 22 Em (89.18 ± 3.95 versus 44.23 ± 2.44 U/g dry substrate). Ferulic acid is a known inducer of laccase production. The structure of this organic acid is similar to that of coniferyl alcohol, the most abundant monolignol of the three lignin precursors [ P. ostreatus IT01 in submerged fermentation with glucose and lactose as substrates. A peak of laccase activity  was observed at the 15th day of fermentation (3-fold increase when compared to control). Ferulic acid was also shown to be the best inducer of laccase activity in Pleurotus sajor-caju [ Pycnoporus cinnabarinus after 10 days, in contrast with around 10 U/g without inducers. It is worth noting that the values achieved in this study in the presence of copper sulfate (44.23 ± 2.44 U/g) and ferulic acid (89.18 ± 3.95 U/g) after 5 days of fermentation were promising when compared to other results reported in literature.Addition of ferulic acid to the copper containing medium further enhanced laccase production (2.0-fold) by the strainecursors . Vanhullecursors  reportedjor-caju . In solijor-caju  reported2+), and ferulic acid (Fer) are given in g/L, μM, and mM, respectively.According to 2+ 172.6 μM, and ferulic acid 1.860 mM.The predicted model indicated that the maximum laccase activity (161.3 U/g dry substrate) would be obtained at the following conditions: yeast extract 6.417 g/L, CuR2) of 0.8963, the most significant differences being obtained at the lowest and highest levels . However, when these points were not considered, the model described the laccase production as a function of yeast extract, CuSO4, and ferulic acid concentrations, within the range of 5.5 to 8.5 g/L, 112.5 to 187.5 μM, and 1.5 to 2.5 mM, respectively, with a correlation coefficient of 0.9798.Experiments for the verification of the predicted model  revealed P. ostreatus in solid-state fermentation are 65.42 U/g with copper as inducer [Other values reported in the literature for laccase production by inducer , 9 U/g w inducer , and 36  inducer . P. ostreatus hybrids without exogenous inducers [ P. ostreatus [ Pycnoporus cinnabarinus after 14 days of solid-state fermentation in columns and activities near 80 U/g after 6 days [ Pycnoporus cinnabarinus after 10 days [ P. ostreatus Pl 22 Em in this study (151.6 U/g) after 5 days of solid-state fermentation was promising in terms of enzyme activity and productivity.inducers , 12.2 U/streatus , 90 U/g r 6 days , and 70  10 days . The max Paecilomyces carneus was inoculated to a culture of the ligninolytic fungus Trametes maxima. Also, studies on gene expression could be performed since the physiological behavior of many laccase producing organisms suggests the presence of elements responsive to metals (MRE), xenobiotics (XRE), heat shock (HSE), and oxidative stress  within the promoter region of laccase genes [ Pleurotus ostreatus laccase POXA1b and one of its variants, 1H6C, using Aspergillus niger as a host, and obtained production levels of 35,000 U/L and 60,000 U/L, respectively. You et al. [ Ganoderma lucidum laccase in Pichia pastoris.Further enhancement in the production of laccase could be achieved by other strategies such as coculturing ligninolytic fungi and filamentous microfungi, as demonstrated by Cupul et al. . These ase genes . Regardise genes  developeu et al.  reached The culture conditions optimized for laccase production were applied in the biotreatment of sugarcane bagasse with the aim of evaluating lignin degradation. The sugarcane bagasse received from the industry presented the following particle size distribution: 21.2% between 0.8 and 2.0 mm, 22.8% < 0.8 mm, and 56% > 2.0 mm. After grinding and classification, the particle size distribution changed to 42.4% between 0.8 and 2.0 mm and 57.6% < 0.8 mm. The physicochemical composition of sugarcane bagasse before and after the biotreatment of solid-state fermentation is presented in  Phanerochaete chrysosporium presented delignification times of around two weeks, the kappa number  being reduced from 33 to less than 10 for the kraft pulp, and the lignin content decreasing from 26.5% to 21.3% for the CTMP. Delignification of sugarcane bagasse by Ceriporiopsis subvermispora during 30 days resulted in a pulp yield of 46–54% [ Dichomitus squalens.The process developed by Pellinen et al.  to deligf 46–54% . Meza etf 46–54%  presentef 46–54%  reported P. ostreatus 22 Em, which demonstrated to be a high producer of laccases on sugarcane bagasse after 5 days of solid-state fermentation, presented a moderate efficiency of delignification with the highest rate achieved between the 5th day and the 10th day. However, delignification is not the only beneficial action of laccases with respect to the use of lignocellulosic biomass. Although this action facilitates the cellulose hydrolysis and glucose release, there is also a contribution related to the degradation of phenolic compounds released during pretreatment, which reduces the toxicity of the broth for a subsequent fermentation [The process of delignification is very complex and involves the synergistic action of laccases and peroxidases. Although laccases are directly involved in delignification through the oxidation of phenolic structures, their production level is not necessarily related to the rate of delignification. Also, lignin degradation can occur until the latest stages of fermentation, even if the peak of enzymatic activity is achieved earlier . The funentation , 44. P. ostreatus Pl 22 Em in solid-state fermentation of sugarcane bagasse was significantly affected by the concentrations of nitrogen source, copper sulfate, and ferulic acid. The use of an organic nitrogen source (yeast extract) provided and increase of 5.7-fold in laccase production, in comparison with the inorganic source (ammonium sulfate). The predicted model indicated that the maximum laccase activity (161.3 U/g of sugarcane bagasse) would be obtained at the following conditions: yeast extract 6.417 g/L, Cu2+ 172.6 μM, and ferulic acid 1.860 mM. Experimentally, the maximum laccase activity of 151.6 U/g was produced, under optimized conditions, at the 5th day of solid-state fermentation, which is higher than that obtained in other solid-state fermentations so far reported on sugarcane bagasse. On the other hand, the process of biological delignification reduced the lignin content of sugarcane bagasse from 31.89% to 26.36% after 5 days and to 20.79% after 15 days of solid-state fermentation, which represents a moderate efficiency of delignification. The highest rate of lignin degradation was achieved between the 5th day and the 10th day.The level of laccase activity produced by


Pleurotaenium trabecula of green algae; Chaetoceros lorenzianus, Proboscia alata var. gracillima, Pseudosolenia calcar-avis, and Pseudo-nitzschia pungens of diatoms; Trichodesmium erythraeum and Pseudoanabaena limnetica of cyanophytes. Most of other algal species were fairly distributed at the selected stations of the study area. The total abundance of phytoplankton was relatively low (average of 2989 unit/L) in the eastern coast of Suez Gulf, as compared its western coast and the northern part of the Red Sea. The diversity of phytoplankton species was relatively high (2.35–3.82 nats) with an annual average of 3.22 nats in the present study. The results concluded that most of eastern coast of Suez Gulf is still healthy, relatively unpolluted, and oligotrophic area, which is clearly achieved by the low values of dissolved phosphate (0.025–0.3 μM), nitrate (0.18–1.26 μM), and dissolved ammonium (0.81–5.36 μM). Even if the occurrence of potentially harmful algae species was low, the study area should be monitored continuously. The dissolved oxygen ranged between 1.77 and 8.41 mg/L and pH values between 7.6 and 8.41. The multiple regression analysis showed that the dissolved nitrate and pH values were the most effective factors that controlled the seasonal fluctuations of phytoplankton along the eastern coast of Suez Gulf during 2012–2013.Water samples were seasonally collected from 12 stations of the eastern coast of Suez Gulf during autumn of 2012 and winter, spring, and summer of 2013 in order to investigate phytoplankton community structure in relation to some physicochemical parameters. The study area harbored a diversified phytoplankton community (138 species), belonging to 67 genera. Four algal groups were represented and classified as Bacillariophyceae (90 species), Dinophyceae (28 species), Cyanophyceae (16 species), and Chlorophyceae (4 species). The results indicated a relative high occurrence of some species namely.;  The northern end of the Red Sea bifurcates into the Sinai Peninsula, creating the Gulf of Suez in the west and the Gulf of Aqaba to the east. The gulf is relatively shallow and formed within a relatively young but now inactive Gulf of Suez Rift basin, dating back about 28 million years. It stretches some 300 km north by northwest, terminating at the Egyptian city of Suez and the entrance to the Suez Canal. Along the mid-line of the gulf is the boundary between Africa and Asia. The length of the gulf, from its mouth at the Strait of Jubal to its head at the city of Suez, is 314 km, and it varies in width from 19 to 32 km. The Gulf of Suez is relatively shallow, with a maximum depth of about 64 m; outside its mouth, the depth drops sharply to about 1255 m.However, the Suez Gulf is subjected to sources of pollution such as shipping activities, where transport of oil continues to play a critical role in marine pollution in the northern part of the gulf and Suez Canal. On the other hand, extensive oil production operations are taking place in the gulf, both inshore and offshore. In addition, the gulf is subjected to industrial, agricultural, and domestic sewage; thermal pollution from power, desalination plants; and tourism activities TEAM , b, c. INoticeably, all these pollutants affected the marine ecosystem, which becomes under variable pressure, causing radical changes in marine organisms, including coral reefs, invertebrates, seagrasses, seaweeds, phytoplankton, and others TEAM , b, c.In fact, phytoplankton communities are the basis of many marine and freshwater food webs. Their composition fluctuates depending on hydrological conditions, such as light, temperature, salinity, pH, nutrients, and turbulence  and winter (January), spring (April), and summer (August) of 2013 Fig. . The spe3, NH4 and PO4) were determined spectrophotometrically, and the results were expressed in micromolar according to the methods described by APHA  was calculated according to Shannon and Weaver , while in winter, the lowest ones were reached (18.16 °C).2/L.Seawater pH lied in the alkaline side during all seasons with almost the highest values during summer. The lowest value of pH was recorded during winter at St. 9 (7.60) and the highest during summer at St. 1 (8.41). Whereas, seawater dissolved oxygen (DO) varied between a minimum of 1.77 mg/L during spring at St. 3 and a maximum of 8.41 mg/L during winter at St. 9 with small seasonal differences and an annual average of 4.30 mg O4) was very low during spring and summer at all stations, whereas the maximum value was recorded during autumn at St. 9 (0.30 μM). The dissolved nitrate (NO3) in the gulf ranged between a maximum value of 1.26 μM during autumn at St. 9 and a minimum of 0.18 μM during summer at St. 3. The dissolved ammonium (NH4) varied between a minimum of 0.81 μM during spring at St. 1 and a maximum of 5.36 μM during summer at St. 9, which may be due to the effect of sewage and oil effluents. Generally, nitrate and phosphate exhibited a seasonal cycle with lower concentrations during summer, while dissolved ammonium was the most abundant source of nitrogen during summer , belonging to 67 genera. Four algal groups were represented in the eastern coast of Suez Gulf belonging to Bacillariophyceae (90 species), Dinophyceae 28 species), Cyanophyceae (16 species), and Chlorophyceae (4 species) , while station 1 recorded the lowest diversified one (59 species) Table . On the Guinardia flaccida, Gyrosigma attenuatum, Nitzschia longissima, Nitzschia sigma, Odontella obtusa, Synedra ulna, Thalassiothrix longissima, Phormidium sp., and Ceratium trichoceros. Whereas, 21 species appeared during the three seasons and are considered as semi-perennial. The rest number of species was observed either for one or two seasons. On the other hand, there were few species restricted to one station such as Amphora grevilleana (St. 1); Chaetoceros curvisetus, Synedra crystallina, and Anabaena sp. (St. 2); Asterolampra sp., Paralia sulcata, Dinophysis sp., and Phalacroma sp. (St. 4); Fragillaria pectinalis and Fragillaria construens (St. 7); Protoperidinium divergens (St. 8); Amphora ovalis, Cymbella aspera, Hemiaulus sinensis, Odontella sinensis, Rhizosolenia bergoni, Ceratium egyptiacum, Ceratium karastenii, and Prorocentrum compressum (St. 9), whereas Pediastum clathratum appeared only at St. 11 . These species are The diatoms prevailed during all seasons forming numerically the highest percentage 82 %) during winter, followed by cyanophytes, which formed the highest percentage during summer (24 %). The chlorophytes were absent during winter at all stations and formed almost equal percentages during the three other seasons. The fourth group, dinoflagellates, formed numerically 5–13 %) of the total count during the study period. Like the seasonal pattern, the spatial distribution of phytoplankton revealed the dominance of diatoms at all stations (Table  % of the % duringC. lorenzianus (4.24 %), G. attenuatum (6.16 %), P. pungens (4.05 %), P. alata var. gracillima (4.37 %), P. calcar-avis (7.35 %), T. erythraeum (3.98 %), Pseudanabaena limnetica (3.75 %), and P. trabecula (9.41 %). However, some of these species were also observed with relative high counts during summer and spring, 2013 such as P. alata var. gracillima, P. calcar-avis, and P. trabecula. Most of other algal species were fairly distributed at the different stations in the coastal waters of eastern coast of Suez Gulf during 2012–2013.The total abundance of phytoplankton was relatively low in the present study of the eastern coast of Suez Gulf (average of 2989 unit/L) as compared with the previous studies Table . The totP. pungens (3.06 %), P. alata var. gracillima (3.39 %), P. calcar-avis (3.80 %), and P. trabecula (4.29 %). The species P. trabecula appeared also at St. 10 (9.54 %), with the contribution of Melosira granulata (4.77 %), T. longissima (3.90 %), T. erythraeum (5.78 %), and P. limnetica (3.76 %).On the spatial scale, the average total count was relatively low in all stations. The relatively high abundance was recorded at St. 9 followed by St. 10 with total counts of 4170 and 3840, respectively, whereas St. 1 and St. 3 sustained the lowest average total counts   and dissolved oxygen (r = 0.51) but inversely correlated with pH values (r = −0.63), whereas the groups and dominant species showed varied correlations with physicochemical characteristics as shown in Table 3 – 0.33 pH . The similarity index revealed four clusters  and dissolved oxygen (r = −0.67).Variations in pH can affect algal growth in a number of ways. It can change the distribution of carbon dioxide species and carbon availability, alter the availability of trace metals and essential nutrients, and at extreme pH levels potentially cause direct physiological effects (Chen and Durbin r = 0.51). Generally, the slight increase in dissolved oxygen during winter and spring may be due to the increase of oxygen solubility (Nassar o).Oxygen concentration is an index of the balance between processes of food production and food consumption. This balance is a key descriptor of the changing status of the ecosystem. When the balance is disrupted, the oxygen concentration can fall to low levels Kemker . Accordiy Nassar . Howevery Nassar . This isy Nassar  who repor = 0.69), whereas it was not a limiting factor for other groups.The nutrient enrichment of coastal waters is generally the main factor driving the succession and composition of phytoplankton communities  and the regression analysis (phytoplankton counts = 1379.2341 + 0.662 NO3 – 0.33 pH). However, natural phytoplankton communities typically prefer to take up nitrogen in the reduced form of ammonium rather than the oxidized forms nitrite and nitrate. In the present study, there was no correlation between total phytoplankton abundance and ammonium, whereas there was a negative correlation between diatoms and ammonium (r = −0.46). This may be due to that various phytoplankton groups and taxa exhibit differential abilities to take up and assimilate dissolved organic nitrogen vs. dissolved inorganic nitrogen  and was positively correlated with chlorophytes (r = 0.38), whereas it was not a limiting factor for dinoflagellates and cyanophytes.As all marine coastal areas, diatoms were the dominant group forming high percentage (67.48 %) and prevailed during the four seasons (53.03–82 %) and at all stations (57–80 %). Whereas, the cyanophytes were the second dominant group, indicating the presence of freshwater discharge in the study area. On the other hand, the contribution of three groups Cyanophyceae, Dinogflagellates, and Chlorophyceae increased during summer at high temperature (28.1–31.5 °C). Eker and Kideyş  suggest C. lorenzianus, G. attenuatum, P. calcar-avis in addition to the green alga P. trabecula, which appeared at all stations and during all seasons, except winter, indicating the freshwater discharge. Moreover, there were dominant potentially harmful algae, and they appeared frequently and were P. alata var. gracillima  and nitrate (r = 0.50) and negatively correlated with ammonium (r = −0.51), whereas G. attenuatum was positively correlated with nitrate (r = 0.49). The diatomate species P. calcar-avis was positively correlated with nitrate (r = 0.40) and negatively correlated with ammonium (r = −0.49), and P. pungens was negatively correlated with temperature and pH (r = −0.45 and −0.43), respectively, and positively correlated with phosphate and nitrate (r = 0.63 and r = 0.52), respectively. However, the cyanophyte P. limnetica was influenced only by temperature (r = 0.50). On the other hand, P. trabecula, P. var. gracillima, and T. erythraeum were not affected by any of these physicochemical parameters.However, the dominant species during this study were . Malone , Cyclotem Ismael , N. longP. trabecula (37.79 %), P. alata var. gracillima (11.34 %), and P. limnetica (6.73 %).Marine systems are highly dynamic, with biodiversity changing at seasonal and inter-decadal timescales , cluster 2 (St. 10), cluster 3 , cluster 4 . The lowest similarity level was 41.11 % between St. 1 and St. 10, whereas the highest level was 76.91 % between St. 3 and St. 12. However, cluster 2, which includes only one station, reflects the unique ecological conditions at this station (oil effluents from Petropil Company). In contrast, the other clusters indicated that the included stations in each cluster have more or less similar ecological conditions, depending on the degree of similarity.The study area is considered as oligotrophic and healthy, despite land-base, human, and tourism activities, since the total counts of phytoplankton was low accompanied with low nutrient concentrations and high values of diversity. But due to the appearance of potentially harmful algae species even in low counts, make the region of the eastern coast susceptible to drastic effects at flourishing of these species during favorable conditions. Thus, monitoring continuously of this area is imperative, to follow probable bloom of these species, to predict negative effects resulting from increasing land-based activities in order to protect the eastern coast of Suez Gulf from any undesirable change there.


P = 0.948) and low root mean square error of prediction (RMSEP = 0.108) of partial least squares (PLS) model. Then 18 characteristic wavenumbers were selected based on a hybrid of backward interval partial least squares (biPLS) regression, competitive adaptive reweighted sampling (CARS) algorithm and successive projections algorithm (SPA). These characteristic wavenumbers only accounted for 0.64% of the full wavenumbers. Following that, 18 characteristic wavenumbers were used to build linear and nonlinear determination models by PLS regression and extreme learning machine (ELM), respectively. The optimal model with RP = 0.963 and RMSEP = 0.137 was achieved by ELM algorithm. These results demonstrated that FT-IR spectroscopy with chemometrics could be used successfully to detect talcum powder in tea.This paper investigated the feasibility of Fourier transform infrared transmission (FT-IR) spectroscopy to detect talcum powder illegally added in tea based on chemometric methods. Firstly, 210 samples of tea powder with 13 dose levels of talcum powder were prepared for FT-IR spectra acquirement. In order to highlight the slight variations in FT-IR spectra, smoothing, normalize and standard normal variate (SNV) were employed to preprocess the raw spectra. Among them, SNV preprocessing had the best performance with high correlation of prediction (R Among all the organoleptic evaluation indexes of tea, color plays an especially important role in the quality and the market price of tea13SiO10(OH)2,et al.et al.7Unfortunately, research on talcum powder in tea is uncommon. Chen et al. first used near infrared (NIR) spectroscopy to study the protein in hard red winter wheat1112et al. analyzed dry matter content of tea by near infrared (NIR) and mid-infrared (MIR) spectroscopyet al. applied MIR spectroscopy to detect five adulterants in raw milkInfrared (IR) spectroscopy has been known as a powerful tool for analysis of chemical constituents with specific frequency absorbance of functional groups. More complex structure leads to more absorption bands and more complex spectra. Rapid detection is the biggest advantage of IR spectroscopy. In 1977, Norris The IR spectra generally comprises a wide range of wavenumbers, some of which are useless or irrelevant information for modelingThe aim of this study is to discriminate the tea samples added with talcum powder or not based on the IR spectra. Moreover, a hybrid method of biPLS, CARS and SPA was applied to select the most relevant wavenumbers and to build a model for quantitative detection of talcum powder in tea.−1 (see enlarged peaks in −1 could be found in −1 (related to C-H and O-H)25The average spectra of pure tea sample, tea sample with 1.50 mg/g talcum powder and pure talcum powder are demonstrated in P = 0.948) and a lowest root-mean square error of prediction (RMSEP = 0.108). Therefore, the following chemometric analysis was based on the spectra after SNV preprocessing.In order to obtain a reliable, accurate and stable model, it is necessary to preprocess raw spectra before modeling, as shown in Before establishing quantitative model, principal component analysis (PCA) was used as a classification tool for analyzing tea sample with talcum powder or without. PCA is an unsupervised pattern recognition method, which can indicate the data trend of samples in visualizing dimension spaces−1 were divided into 20 intervals with the same width manually. Then, leaving one subinterval out at a time, and the remaining subintervals were combined to build a PLS model. Repeating the previous step, several models with different number of intervals were achieved. Comparing the root-mean square error of cross validation (RMSECV) of each model, the one with lowest RMSECV was chosen as the best model. Thus, the intervals in the best model are the optimal ones. In this research, the final selected intervals were: 881–1080, 1182–1381, 1583–1682, 1783–1983, 2285–2384, 2686–2785 cm−1. Then these 936 wavenumbers were used to build a PLS model (Model 5), and the results were shown in P = 0.949 and lower RMSEP = 0.107. However, the wavenumbers of Model 5 for modeling were still too much, which accounted for 33.46% of the full wavenumbers. So, CARS was used to reduce wavenumbers after biPLS processing. The process of CARS was shown in In order to simplify the model and improve the performance of model, biPLS was first used to select the most relevant wavenumbers in this research. During the process of biPLS, firstly, the full spectrum from 881 to 3581 cm−1) were selected as the most useful variables through SPA, which was only 0.64% of the full wavenumbers shown in −1 were the characteristic wavenumbers of talcum powder. Subsequently, these 18 wavenumbers were treated as input variables for establishing PLS model (Model 7), and the results were shown in P = 0.931 and RMSEP = 0.124 of Model 7 were still acceptable. The most exciting thing was that the dimension of spectral variables reduced from 2800 to 18 characteristic wavenumbers, which resulted in a pretty simple linear model for the quantitative detection of talcum powder in tea. Meanwhile, these characteristic wavenumbers laid a solid foundation for revealing the detection mechanism of talcum powder in tea by IR spectroscopy. In conclusion, all these results indicated that a hybrid wavenumbers selection method, which combined biPLS regression, CARS algorithm and SPA, was effective for extracting the optimal wavenumbers from full spectrum. Meanwhile, it had good performance of the quantitative detection model of talcum powder in tea. More details of results of different wavenumbers selection methods were shown in However, 111 wavenumbers were also a little more for establishing a simple and convenient model. Therefore, SPA was utilized on the basis of these 111 wavenumbers. It could be seen in P = 0.963 and RMSEP = 0.137 shown in In order to further improve the accuracy of the detection model, ELM was used to build a nonlinear model based on the above 18 characteristic wavenumbers. In the modeling, the activation function of ELM was set to “sigmoidal”. The number of hidden nodes, which was the only parameter needed to be determined manually, was varied from 1 to 80. The best model was achieved using 36 hidden nodes. The optimal results were obtained with R−1 were the characteristic wavenumbers of talcum powder. And the linear and nonlinear detection models were built with these 18 characteristic wavenumbers by PLS and ELM, respectively. The optimal model was achieved with RP = 0.963 and RMSEP = 0.137 based on ELM. All these results sufficiently indicated that FT-IR spectroscopy could be considered as a useful tool for rapid detection of talcum powder in tea.In this research, the feasibility of FT-IR spectroscopy for the detection of talcum powder in tea was investigated. Comparing with previous studies, a hybrid of biPLS regression, CARS algorithm and SPA were first used to select optimal wavenumbers, which effectively extracted 18 characteristic wavenumbers from the full spectrum (3581 wavenumbers). Among these 18 wavenumbers, 1016(Si-O-Si), 1182 and 1249(CO-O), 1340 . Firstly, Tea samples were ground into powder and filtered through a 60-mesh sieve. Then, 4.00 g pure sample was blended with 0.00 mg, 0.50 mg, 1.00 mg, 1.40 mg, 2.00 mg, 2.60 mg, 3.00 mg, 3.40 mg, 4.00 mg, 4.40 mg, 5.00 mg, 5.60 mg and 6.00 mg talcum powder. So the dose levels were 0.00 mg/g, 0.15 mg/g, 0.25 mg/g, 0.35 mg/g, 0.50 mg/g, 0.65 mg/g, 0.75 mg/g, 0.85 mg/g, 1.00 mg/g, 1.10 mg/g, 1.25 mg/g, 1.40 mg/g and 1.50 mg/g. Successively, 1.00 g blended sample was added into 49.00 g KBr medium, and mixed adequately for the following IR spectroscopy scanning. The 140 samples with concentrations of 0.00 mg/g, 0.25 mg/g, 0.50 mg/g, 0.75 mg/g, 1.00 mg/g, 1.25 mg/g and 1.50 mg/g were chosen as the calibration samples and the left 70 samples with concentrations of 0.15 mg/g, 0.35 mg/g, 0.65 mg/g, 0.85 mg/g, 1.10 mg/g and 1.40 mg/g were subsumed into the prediction set. Meanwhile, the calibration set was validated by full cross validation method. So the calibration, validation and prediction sets obtained 140, 140 and 70 samples respectively.−1 by a Jasco FT-IR-4100 spectrometer  coupled with a TGS detector and a ZnO crystal sampling accessory with transmission mode. The spectral resolution was 4 cm−1. Each spectrum was the average of 100 scans. During the whole experiment, the temperature was kept at about 25 °C and the humidity was kept at a stable level in the laboratory.The IR spectra of samples were measured in the wavenumber range of 400–4000 cm−1.Raw spectra frequently contained noises besides sample information, so the first 500 and last 400 spectral data were deleted to remove noises, and the following analysis was based on the spectra in range of 881–3581 cmIn this research, a hybrid wavenumbers selection method, which combined backward interval partial least squares (biPLS) regression, competitive adaptive reweighted sampling (CARS) algorithm and successive projections algorithm (SPA), was applied for IR spectral wavenumbers selection. Backward interval partial least squares (biPLS) regression can extract the spectral wavenumbers highly related to the chemical structureC, RCV, RP, RMSEC, RMSECV, RMSEP)38Partial least squares (PLS) regression is a traditional technique for building linear models, which is able to not only extract principal component from both input and output data, but also determine the direction on which input and output data have the largest covariance323335How to cite this article: Li, X. et al. Rapid detection of talcum powder in tea using FT-IR spectroscopy coupled with chemometrics. Sci. Rep.6, 30313; doi: 10.1038/srep30313 (2016).