Impact of a spatial repellent intervention on Anopheles kdr insecticide resistance allele in Sumba, Indonesia.

BACKGROUND: The emergence of insecticide resistance and outdoor transmission in malaria-endemic areas underlines the urgent need to develop innovative tools, such as spatial repellents (SR), that may circumvent this residual transmission. With limited options for effective insecticides, regular resistance monitoring is warranted for selecting and using appropriate tools. This study evaluates the pyrethroid knockdown resistance (kdr) allele before and after implementing a transfluthrin-based spatial repellent (SR) intervention in placebo-treated clusters. METHODS: This study looks at the frequency distribution of the kdr allele in Sumba Island from June 2015 to August 2018. Insecticide susceptibility tests were carried out on female Anopheles sp. aged 3-5 days against permethrin 21.5 µg/ml, deltamethrin 12.5 µg/ml, and transfluthrin 10 µg/ml using CDC bottle assay. PCR sequencing of representative samples from adult mosquito collections and insecticide tests revealed the presence of kdr mutations (L1014F and L1014S) in the VGSC gene. RESULTS: A total of 12 Anopheles species, Anopheles tesselatus, Anopheles. aconitus, Anopheles barbirostris, Anopheles kochi, Anopheles annularis, Anopheles maculatus, Anopheles sundaicus, Anopheles flavirostris, Anopheles balabacensis, Anopheles indefinitus, Anopheles subpictus, and Anopheles vagus were analysed. Anopheles vagus and An. sundaicus predominated in the larval populations. Susceptibility assays for all insecticides identified fully susceptible phenotypes in all species examined. Anopheles increasing frequency of kdr mutant alleles during the 3 year SR deployment was observed in both SR-treated and placebo areas, a statistically significant increase occurred in each arm. However, it is unclear how significant SR is in causing the increase in mutant alleles. The L1014S, knockdown resistance east type (kdr-e) allele was detected for the first time among the mosquito samples in this study. The L1014F, knockdown resistance west type (kdr-w) allele and heteroduplex form (wild-type-mutant) were found in almost all Anopheles species examined, including An. vagus, An. aconitus, An. subpictus, An. tesselatus, An. annularis, An. flavirostris and An. sundaicus. CONCLUSION: The presence of fully susceptible phenotypes over time, along with an increase in the frequency distribution of the L1014F/S mutations post-intervention, suggest drivers of resistance external to the study, including pyrethroid use in agriculture and long-lasting insecticidal nets (LLINs). However, this does not negate possible SR impacts that support resistance. More studies that enable the comprehension of possible SR-based drivers of resistance in mosquitoes need to be conducted.

Dietary supplementation with a mixture of Dunaliella salina and Spirulina enhances broiler performance by improving growth, immunity, digestive enzymes and gut microbiota.

The aim of this study is to evaluate the effect of Dunaliella salina and Spirulina (D + S) mixture on performance, carcass yield, kidney and liver markers, lipid profile, and immune responses of fattening chicks. Two hundred broiler chicks at 7 days old were distributed into 5 experimental groups, 5 replicates each with 8 chicks each. Group 1 was fed on only basal diet; group 2 was fed with basal diet and 0.50 g/kg (D + S); group 3 was fed with basal diet and 1.00 g/kg (D + S); group 4 was fed with basal diet and 1.50 g/kg (D + S); and group 5 was fed with basal diet supplemented with 2.00 g/kg (D + S). The additive mixture (D + S) consisted of (1 D. salina: 1 Spirulina). The experiment lasted for 6 wk. The results demonstrated significantly improved better live body weight, body weight gain, and feed conversion ratio (P<0.01) for groups that received (D + S) at levels of 1.0 and 1.5 g/kg diet compared to other groups at 6 wk of age. There was no significant influence of different levels of dietary feed additives on feed intake or carcass traits. The lipid profile was improved through a reduction of total cholesterol and low-density lipoprotein (LDL) values and increasing high-density lipoprotein (HDL) values, as well as the immune response, which was improved through increasing values of complement 3, immunoglobulin M (IgM), and immunoglobulin G (IgG) in the birds treated with (D + S) compared to the control group. The inclusion of all levels of (D + S)/kg decreased triglyceride, while total protein, albumen, and globulin values (P<0.05 or P<0.01) were higher compared to other groups. The inclusion of the different levels of (D + S)/kg improved liver function, whereas aspartate aminotransferase (AST) and alanine aminotransferase (ALT) values were lower than in other groups (P<0.001). The lowest values of creatinine, urea, and uric acid were noted in birds fed a diet supplemented with 1.50 g (D + S)/kg. Antioxidant levels were improved through increasing values of catalase (CAT) and reduced glutathione (GSH) enzymes in the treated birds with (D + S) compared with the control chicks. Furthermore, digestive enzymes and microbial content were improved in broiler checks fed on diet supplemented with (D + S) compared to the control group. In conclusion, supplementing broiler chicks with a dietary D. salina and Spirulina combination increased their productive performance, immunity, kidney and liver function, lipid profile, and digestive enzymes.

Waste cooking oil processing over cobalt aluminate nanoparticles for liquid biofuel hydrocarbons production.

The catalytic conversion of waste cooking oil (WCO) was carried out over a synthetic nano catalyst of cobalt aluminate (CoAl2O4) to produce biofuel range fractions. A precipitation method was used to create a nanoparticle catalyst, which was then examined using field-emission scanning electron microscopy, X-ray diffraction, energy dispersive X-ray, nitrogen adsorption measurements, high-resolution transmission electron Microscopy (HRTEM), infrared spectroscopy, while a gas chromatography-mass spectrometer (GC-MS) was used to analyze the chemical construction of the liquid biofuel. A range of experimental temperatures was looked at including 350, 375, 400, 425, and 450 °C; hydrogen pressure of 50, 2.5, and 5.0 MPa; and liquid hour space velocity (LHSV) of 1, 2.5, and 5 h-1. As temperature, pressure, and liquid hourly space velocity increased, the amount of bio-jet and biodiesel fractional products decreased, while liquid light fraction hydrocarbons increased. 93% optimum conversion of waste cooking oil over CoAl2O4 nano-particles was achieved at 400 °C, 50 bar, and 1 h-1 (LHSV) as 20% yield of bio-jet range,16% gasoline, and 53% biodiesel. According to the product analysis, catalytic hydrocracking of WCO resulted in fuels with chemical and physical characteristics that were on par with those required for fuels derived from petroleum. The study's findings demonstrated the nano cobalt aluminate catalyst's high performance in a catalytic cracking process, which resulted in a WCO to biofuel conversion ratio that was greater than 90%. In this study, we looked at cobalt aluminate nanoparticles as a less complex and expensive alternative to traditional zeolite catalysts for the catalytic cracking process used to produce biofuel and thus can be manufactured locally, which saves the cost of imports for us as a developing country.

The Effect of Varying Dentin Chroma and Enamel Thickness on the Color of A2 Double-layered Resin Composite.

OBJECTIVES: To evaluate chroma (C*) and overall color of double-layered (DL) resin composite (RC) restorations with various dentin shades and enamel thicknesses. METHODS: Enamel specimens were fabricated using custom-made molds to replicate VITA shade tabs with variant enamel thicknesses (0.5, 0.7, and 1.0 mm) (n=7) from two RC: Clearfil-Majesty (CM) shade (A2), and Vit-l-escence (VL), shade (pearl-neutral). Dentin specimens (shades A1, A2, and A3) were fabricated using custom molds corresponding to the enamel molds. Each enamel specimen was paired with three different dentin specimens. L*a*b* parameters were measured with VITA Easyshade-V. Color difference between DL specimens and the A2 VITA shade tab were calculated with the CIEDE2000 formula. Relationships among enamel thickness, ΔE00, C* of dentin layer, C* of DL, and change in chroma were assessed by Spearman rank correlations. ΔE00 was compared among groups using one-way analysis of variance with Tukey post-hoc adjustment for all possible pairwise group comparisons (experiment-wise α=0.05). RESULTS: There was no statistical difference among C* of DL specimens (p=0.65, 0.53) for CM and VL, respectively. Combinations of enamel thickness/ dentin shade had a significant difference in ΔE00 (p>0.05). No significant correlation was observed among enamel thickness and C* of dentin, and C* of the DL (p>0.05). Significant correlations were observed between ΔE00 of the VL DL and C* DL (r=-0.8, p<0.001); and ΔE00 of CM DL and enamel thickness (r=0.5, p<0.001). CONCLUSIONS: Enamel thickness did not affect C* of the dentin layer. Unlike VL RC, variations in dentin shades with CM produced a closer match to the A2 shade tab. Enamel is recommended to be 0.7 mm or less.

Human behavior determinants of exposure to Anopheles vectors of malaria in Sumba, Indonesia.

Malaria vector control interventions in Sumba, Indonesia, have not been able to eliminate malaria. Human drivers of exposure to Anopheles bites were investigated as part of a larger clinical trial evaluating the impact of a spatial repellent product on malaria incidence. Human behavioral observations (HBOs) evaluating temporal and spatial presence, sleeping behaviors, and insecticide treated net (ITN) use, were collected parallel to entomological collections-indoor and outdoor human landing catches (HLCs), and house hold surveys. Data demonstrates that mosquito access to humans, enabled by structurally open houses, is evident by the similar entomological landing rates both inside and outside households. The presence of animals inside houses was associated with increased mosquito entry-however, the number of humans present inside houses was not related to increased mosquito landing. Analyzing mosquito landing rates with human behavior data enables the spatial and temporal estimation of exposure to Anopheles bites, accounting for intervention (ITN) presence and usage. Human behavior adjusted exposure to Anopheles bites was found to be highest in the early in the evening, but continued at lower levels throughout the night. Over the night, most exposure (53%) occurred when people were indoors and not under the protection of nets (asleep or awake) followed by exposure outside (44%). Characterized gaps in protection are outdoor exposure as well as exposure indoors-when awake, and when asleep and not using ITNs. Interestingly, in the primary trial, even though there was not a significant impact of the spatial repellent on vector biting rates by themselves (16%), when factoring in human behavior, there was approximately 28% less exposure in the intervention arm than in the placebo arm. The treated arm had less human behavior adjusted bites in all spaces evaluated though there was proportionally higher exposure indoors. This analysis points to the importance of using HBOs both towards understanding gaps in protection as well as how interventions are evaluated. To mitigate ongoing transmission, understanding context specific spatial and temporal exposure based on the interactions of vectors, humans and interventions would be vital for a directed evidence-based control or elimination strategy.

Assessing performance and stability of yellow rust resistance, heat tolerance, and agronomic performance in diverse bread wheat genotypes for enhancing resilience to climate change under Egyptian conditions.

Yellow rust and heat stress adversatively impact the growth and production of bread wheat in particular under rising adverse environmental conditions. Stability of grain yield is a pivotal purpose of plant breeders to improve wheat production and ensure global food security especially under abrupt climate change. The objective of this study was to assess the performance and stability of diverse bread wheat genotypes for yellow rust resistance, heat stress, and yield traits. The studied genotypes were evaluated in two different locations under two sowing dates (timely and late sowing) during two growing seasons. The obtained results displayed significant differences among the tested locations, sowing dates, and genotypes for most measured traits. The yellow rust measurements evaluated under the field conditions including final rust severity (FRS), the average coefficient of infection (ACI), and area under disease progress curve (AUDPC) revealed that Giza-171, Misr-1, Gemmeiza-12, Shandweel-1, Sids-13, Line-1, Line-2, and Line-55 had better resistance. Based on heat sensitivity measurements, Line-1 and Line-2 followed by Line-35, Shandweel-1 and Line-55 were classified as more tolerant to heat stress compared with the remaining genotypes. The genotypes Line-55, Gemmeiza-12, Giza-171, Line-1, Line-2, and Misr-1 were able to maintain acceptable agronomic performance under timely and late sowing dates in all evaluated environments. Different statistical procedures were employed to explore the adaptability and stability of tested genotypes i.e., joint regression, stratified ranking, Wricke's Ecovalence values, cultivar superiority, additive main effects, and multiplicative interaction (AMMI), AMMI stability value, and genotype plus genotype-by-environment interaction (GGE). The applied stability parameters were quite similar for describing the stability of the evaluated wheat genotypes. The results indicated that Gemmeiza-12, Giza-171, Sids-12, Sids-13, Misr-1 Shandweel-1, Line-1, Line-2, and Line-55 were desirable and stable. The heatmap and hierarchical clustering were exploited for dividing the evaluated bread wheat genotypes into different clusters based on yellow rust resistance measurements, heat tolerance indices, and agronomic performance. Line-1 and Line-2 had the best performance for all rust resistance, heat tolerance, and agronomic performance followed by Giza-171, Line-55, Line-35, Gemmeiza-12, Shandweel-1, Misr-1, and Sids-13. In conclusion, our findings provide evidence of utilizing promising genotypes in rust resistance, heat tolerance, and agronomic performance in breeding programs for improving wheat grain yield stability mainly under climate change.

Impact of a spatial repellent product on Anopheles and non-Anopheles mosquitoes in Sumba, Indonesia.

BACKGROUND: The East Nusa Tenggara province, Indonesia, contributed to 5% of malaria cases nationally in 2020, with other mosquito-borne diseases, such as dengue and filariasis also being endemic. Monitoring of spatial and temporal vector species compositions and bionomic traits is an efficient method for generating evidence towards intervention strategy optimization and meeting disease elimination goals. METHODS: The impact of a spatial repellent (SR) on human biting mosquitoes was evaluated as part of a parent cluster-randomized, double-blinded, placebo-controlled trial, in Sumba, East Nusa Tenggara. A 10-month (June 2015-March 2016) baseline study was followed by a 24-month intervention period (April 2016 to April 2018)-where half the clusters were randomly assigned either a passive transfluthrin emanator or a placebo control. RESULTS: Human-landing mosquito catches documented a reduction in landing rates related to the SR. Overall, there was a 16.4% reduction (21% indoors, and 11.3% outdoors) in human biting rates (HBR) for Anopheles. For Aedes, there was a 44.3% HBR reduction indoors and a 35.6% reduction outdoors. This reduction was 38.3% indoors and 39.1% outdoors for Armigeres, and 36.0% indoors and 32.3% outdoors for Culex species. Intervention impacts on the HBRs were not significant and are attributed to large inter-household and inter cluster variation. Anopheles flavirostris, Anopheles balabacensis and Anopheles maculatus individually impacted the overall malaria infections hazard rate with statistically significance. Though there was SR-based protection against malaria for all Anopheles species (except Anopheles sundaicus), only five (Anopheles aconitus, Anopheles kochi, Anopheles tessellatus, An. maculatus and An. sundaicus) demonstrated statistical significance. The SR numerically reduced Anopheles parity rates indoors and outdoors when compared to the placebo. CONCLUSION: Evidence demonstrating that Anopheles vectors bite both indoors and outdoors indicates that currently implemented indoor-based vector control tools may not be sufficient to eliminate malaria. The documented impact of the SR intervention on Aedes, Armigeres and Culex species points to its importance in combatting other vector borne diseases. Studies to determine the impact of spatial repellents on other mosquito-borne diseases is recommended.

Entry-Level Expectations for Pathologists' Assistants.

OBJECTIVES: The aim of this study was to assess expectations of performance that exist in the marketplace for entry-level pathologists' assistants (PathAs), defined as recent graduates of a pathologists' assistant program on their first day of employment. METHODS: A voluntary, anonymous survey was distributed to pathologist and PathA members of the American Society for Clinical Pathology by email. We assessed 98 professional activities of PathAs using a 5-point scale of expectations based on levels of trust placed in them. We also collected demographic information. RESULTS: A total of 728 participants responded to this survey, including 280 pathologists and 448 PathAs. We classified 98 activities according to expectations: independent performance (20/98), developing independence (48/98), and not expected of PathAs (5/98). Some activities (25/98) were indeterminate yet likely represent areas of developing independence. CONCLUSIONS: This study demonstrates an expectation for entry-level PathAs to perform some activities included in the scope of practice independently but eventually to develop independent proficiency for most professional activities. A minority of activities were identified as responsibilities that are not expected of PathAs. Entry-level PathAs, therefore, remain "works in progress," with an expectation for independent performance of core activities while developing abilities in many areas of professional practice.

Treatment with specific and pan-plasma membrane calcium ATPase (PMCA) inhibitors reduces malaria parasite growth in vitro and in vivo.

BACKGROUND: Rapid emergence of Plasmodium resistance to anti-malarial drug mainstays has driven a continual effort to discover novel drugs that target different biochemical pathway (s) during infection. Plasma membrane Calcium + 2 ATPase (PMCA4), a novel plasma membrane protein that regulates Calcium levels in various cells, namely red blood cell (RBC), endothelial cell and platelets, represents a new biochemical pathway that may interfere with susceptibility to malaria and/or severe malaria. METHODS: This study identified several pharmacological inhibitors of PMCA4, namely ATA and Resveratrol, and tested for their anti-malarial activities in vitro and in vivo using the Plasmodium falciparum 3D7 strain, the Plasmodium berghei ANKA strain, and Plasmodium yoelii 17XL strain as model. RESULTS: In vitro propagation of P. falciparum 3D7 strain in the presence of a wide concentration range of the inhibitors revealed that the parasite growth was inhibited in a dose-dependent manner, with IC50s at 634 and 0.231 µM, respectively. RESULTS: The results confirmed that both compounds exhibit moderate to potent anti-malarial activities with the strongest parasite growth inhibition shown by resveratrol at 0.231 µM. In vivo models using P. berghei ANKA for experimental cerebral malaria and P. yoelii 17XL for the effect on parasite growth, showed that the highest dose of ATA, 30 mg/kg BW, increased survival of the mice. Likewise, resveratrol inhibited the parasite growth following 4 days intraperitoneal injection at the dose of 100 mg/kg BW. CONCLUSION: The findings indicate that the PMCA4 of the human host may be a potential target for novel anti-malarials, either as single drug or in combination with the currently available effective anti-malarials.

A comparative study among dietary supplementations of antibiotic, grape seed and chamomile oils on growth performance and carcass properties of growing rabbits.

The main objective of this study was to evaluate the effect of chamomile oil (Ch), grape seed oil (GS), their mixture and antibiotic (colistin) (AN) as feed addetives on the productivity of growing rabbits as well as in vitro study to evaluate the antimicrobial activity of both Ch and GS oils. To achive this objective, a total of 96 New Zealand (NZW) weaned rabbits, 5 weeks-old were randomly allotted into eight groups. Rabbits were kept under observation for eight weeks and the trial ended at thirteen weeks-old. The experimental treatments were: 1) Basal diet (BD); 2) BD + antibiotic; 3) BD + 0.5 ml GS/ kg diet; 4) BD + 1.0 ml GS/ kg diet; 5) BD + 1.5 ml GS/ kg diet; 6) BD + 0.5 ml Ch/ kg diet; 7) BD + 1.0 ml Ch/ kg diet and 8) BD + 1.5 Ch/ kg diet. Live body weight (LBW) was markedly elevated (p < 0.05) in groups fed on ration included feed additives compared with the control at weeks 9 and 13 of age. Cumulative body weight gain (BWG) and feed intake (FI) increased (p < 0.05) throughout 5-9 and 5-13 weeks of age in rabbits fed rations plus the studied additives. Feed conversion ratio (FCR) was insignificantly altered by dietary feed additives. Spleen and intestine relative weights reduced (p < 0.05) in groups treated with different studied additives. In view of the experiment finings, it could be concluded that dietary supplementation of GS and Ch have a positive impact on the productivity of growing rabbits than that of the control and antibiotic-treated groups.

(Co/Zn) Al2O4 nano catalyst for waste cooking oil catalytic cracking.

The current work investigated the preparation of Nano-particles of Co/Zn Al2O4 as a catalyst via co-precipitation method. Several analyses, including BET, XRD, HRTEM, EDX, SEM, and FTIR, were used to characterize it. The analysis revealed that the prepared catalyst had an average surface area of 69.20 m2/g, a cross-sectional area of 16.2 m2/molecule, an average particle size of approximately 28 nm, and a pore size of 0.22 cm3/g. The prepared catalyst was used in a bio fuel synthesis process via thermo-catalytic cracking of waste cooking oil (WCO) in a single step batch reactor. Catalyst loading was tested with different weight percentage of 1.5%, 2%, and 2.5%. The pilot study revealed that the best conditions for optimizing bio jet fuel yield were 400 °C, a catalyst loading of 2%, and a reaction time of 30 min.The optimal cut-off from the distillation process of crude liquid bio fuel product which represents a fraction of bio-jet fuel was in the range from 150 to 240 °C.

An inventory of human night-biting mosquitoes and their bionomics in Sumba, Indonesia.

Mosquitoes are important vectors that transmit pathogens to human and other vertebrates. Each mosquito species has specific ecological requirements and bionomic traits that impact human exposure to mosquito bites, and hence disease transmission and vector control. A study of human biting mosquitoes and their bionomic characteristics was conducted in West Sumba and Southwest Sumba Districts, Nusa Tenggara Timur Province, Indonesia from May 2015 to April 2018. Biweekly human landing catches (HLC) of night biting mosquitoes both indoors and outdoors caught a total of 73,507 mosquito specimens (59.7% non-Anopheles, 40.3% Anopheles). A minimum of 22 Culicinae species belonging to four genera (Aedes, Armigeres, Culex, Mansonia), and 13 Anophelinae species were identified. Culex quinquefasciatus was the dominant Culicinae species, Anopheles aconitus was the principal Anopheles species inland, while An. sundaicus was dominant closer to the coast. The overall human biting rate (HBR) was 10.548 bites per person per night (bpn) indoors and 10.551 bpn outdoors. Mosquitoes biting rates were slightly higher indoors for all genera with the exception of Anopheles, where biting rates were slightly higher outdoors. Diurnal and crepuscular Aedes and Armigeres demonstrated declining biting rates throughout the night while Culex and Anopheles biting rates peaked before midnight and then declined. Both anopheline and non-anopheline populations did not have a significant association with temperature (p = 0.3 and 0.88 respectively), or rainfall (p = 0.13 and 0.57 respectively). The point distribution of HBR and seasonal variables did not have a linear correlation. Data demonstrated similar mosquito-human interactions occurring outdoors and indoors and during early parts of the night implying both indoor and outdoor disease transmission potential in the area-pointing to the need for interventions in both spaces. Integrated vector analysis frameworks may enable better surveillance, monitoring and evaluation strategies for multiple diseases.

Efficacy and safety of dihydroartemisinin-piperaquine for the treatment of uncomplicated Plasmodium falciparum and Plasmodium vivax malaria in Papua and Sumatra, Indonesia.

BACKGROUND: Dihydroartemisinin-piperaquine (DHA-PPQ) has been adopted as first-line therapy for uncomplicated falciparum malaria in Indonesia since 2010. The efficacy of DHA-PPQ was evaluated in 2 sentinel sites in Keerom District, Papua and Merangin District, Jambi, Sumatra from April 2017 to April 2018. METHODS: Clinical and parasitological parameters were monitored over a 42-day period following the World Health Organization standard in vivo protocol and subjects meeting the inclusion criteria were treated with DHA-PPQ once daily for 3 days, administered orally. RESULTS: In Papua, 6339 subjects were screened through active and passive cases detection. Of the 114 falciparum and 81 vivax cases enrolled, 102 falciparum and 80 vivax cases completed the 42 day follow up, and 12 falciparum and 1 vivax cases were either lost to follow up or withdrawn. Kaplan-Meier analysis of microscopy readings of 102 falciparum cases revealed 93.1% (95% CI 86.4-97.2) as Adequate Clinical and Parasitological Response (ACPR). No delay in parasite clearance nor severe adverse reaction was observed. Recurrent parasites of Plasmodium falciparum were detected in 7 cases and categorized as late treatment failures (LTF) at days 21, 35, and 42 and one of which was reinfected by Plasmodium vivax at day 42. Two cases were confirmed as recrudescent infection and 4 were re-infection. The PCR-corrected DHA-PPQ efficacy for P. falciparum was 97.9% (95% CI 92.7-99.7). Of the 80 cases of P. vivax that were followed up, 71 cases were completely cured and classified as ACPR (88.8%, 95% CI 79.7-94.7) and 9 cases showed recurrent infection at days 35 and 42, and classified as LTF. In Sumatra, of the 751 subjects screened, 35 vivax subjects enrolled, 34 completed the 42 day follow up. Thirty-three cases were completely cured and classified as ACPR (97.1%, 95% CI 84.7-99.9) and 1 recurrent infection was observed and classified as LTF. No delay in parasite clearance nor severe adverse reaction was observed. Analysis of the Pfk13 gene in P. falciparum cases from Papua revealed no mutations associated with artemisinin resistance in the 20 SNPs previously reported. Analysis of the Pfpm2 gene at day 0 and day of recurrence in recrudescent cases revealed the same single copy number, whereas 3 of the 4 re-infection cases carried 2-3 Pfpm2 gene copy numbers. CONCLUSION: Treatment of falciparum and vivax malaria cases with DHA-PPQ showed a high efficacy and safety.

Use of fumaric acid as a feed additive in quail's nutrition: its effect on growth rate, carcass, nutrient digestibility, digestive enzymes, blood metabolites, and intestinal microbiota.

To investigate the effects of dietary fumaric acid (FUA) on performance, carcasses, nutrient digestibility, blood metabolites, digestive enzymes, and cecal microbiota in Japanese quail chicks. Three hundred unsexed Japanese quail (1-wk-old) were randomly assigned to 5 groups. Supplementation of FUA in the diet of Japanese quail chicks exhibited a significant improvement in growth performance through the different experimental periods studied compared with those receiving unsupplemented one. The digestibility of crude protein (CP) and metabolizable energy (ME) were improved with 10 and 15 g/kg FUA, respectively. Apart from lipase enzyme, birds fed 5 and 15 g/kg FUA recorded higher activity of amylase. There were no significant changes among experimental groups on the relative weights of carcass, gizzard, heart, and dressing. Dietary supplementation of FUA at different levels (P> 0.05) increased total protein (TP) and globulin (GLB) concentrations and A/G % compared with control group. A significant (P < 0.01) decrease in plasma low density lipoprotein (LDL) and total cholesterol (TC) levels and increase in high density lipoprotein (HDL) concentrations were observed in chicks fed with FUA containing diets. Immunoglobulin G (IgG) (P = 0.0026) and M (IgM) (P = 0.0007) levels were greater in groups treated with either 10 or 15 g FUA/kg diet. A significant increase in plasma Ca concentration was noticed in chicks received 15 g FUA/ kg compared with the other groups. Quail chicks received diets containing FUA at different levels exhibited reduced cecal count of coliform, E. coli, and Salmonella as compared with control group. In conclusion, supplementation of fumaric acid (especially 15 g/kg diet) in quail chick diets improved their growth, digestibility of nutrients, immune response, antioxidant status, digestive enzyme, and intestinal health.

Analysis of Genetic Diversity and Population Structure in Bitter Gourd (Momordica charantia L.) Using Morphological and SSR Markers.

The present investigation was carried out using 51 diverse bitter gourd accessions as material for studying genetic diversity and relatedness using morphological and SSR markers. A wide variation was observed for morphological traits like the number of days to the first female flower anthesis (37.33-60.67), the number of days to the first fruit harvest (47.67-72.00), the number of fruits/plant (12.00-46.67), fruit length (5.00-22.23 cm), fruit diameter (1.05-6.38 cm), average fruit weight (20.71-77.67 g) and yield per plant (513.3-1976 g). Cluster analysis for 10 quantitative traits grouped the 51 accessions into 6 clusters. Out of 61 SSR primers screened, 30 were polymorphic and highly informative as a means to differentiate these accessions. Based on genotyping, a high level of genetic diversity was observed, with a total of 99 alleles. The polymorphic information content (PIC) values ranged from 0.038 for marker BG_SSR-8 to 0.721 for S-24, with an average of 0.429. The numbers of alleles ranged from 2 to 5, with an average of 3.3 alleles per locus. Gene diversity ranged from 0.04 for BG_SSR-8 to 0.76 for S-24, showing a wide variation among 51 accessions. The UPGMA cluster analysis grouped these accessions into 3 major clusters. Cluster I comprised 4 small, fruited accessions that are commercially cultivated in central and eastern India. Cluster II comprised 35 medium- to long-sized fruited accessions, which made up an abundant and diverse group. Cluster III comprised 11 long and extra-long fruited accessions. The polymorphic SSR markers of the study will be highly useful in genetic fingerprinting and mapping, and for association analysis in Momordica regarding several economic traits.

Dietary citric acid enhances growth performance, nutrient digestibility, intestinal microbiota, antioxidant status, and immunity of Japanese quails.

A total of 300 un-sexed Japanese quail chicks (1-wk-old) were randomly allotted to 5 experimental groups to study the effect of citric acid (CA) on performance development, carcass estimates, blood measurements, antioxidant and immune measurements, digestive enzymes, and cecum microbiology traits of growing Japanese quail. The chicks were fed a basal diet supplemented with CA (5, 10, 15, and 20 g/kg in diet) had significantly (P < 0.05) greater live body weights at 3 and 5 wk of age and increased weight gain across all experimental periods (1-3, 3-5 and 1-5 wks of age) compared to the control group. No significant difference was found in any of the measured carcass traits. The digestion coefficients of crude protein, ether extract, crude fiber, and nitrogen free extract as well as metabolizable energy significantly improved (P < 0.05) in all treatment groups compared to the control. CA supplementation from 10 to 20 g/kg had increased digestive enzyme activities (amylase and lipase). All treatment groups had higher (P < 0.05) albumin and globulin concentrations than the control group. A significant (P < 0.05) decrease in phosphorus (P) concentrations in the plasma was observed in all treatment groups. The IgG levels were greater (P < 0.05) in the 5 or 10 g/kg groups than the control group. Chicks fed CA at different levels had significantly decreased caecal content of TBC, coliform, E. coli, and Salmonella. We concluded that the inclusion of CA (especially 10 g/kg diet) in growing Japanese quail diets improved growth performance, immune response, and health.

Correction: Anopheles sundaicus complex and the presence of Anopheles epiroticus in Indonesia.

[This corrects the article DOI: 10.1371/journal.pntd.0008385.].

Anopheles sundaicus complex and the presence of Anopheles epiroticus in Indonesia.

Anopheles sundaicus s.l. is an important malaria vector primarily found in coastal landscapes of western and central Indonesia. The species complex has a wide geographical distribution in South and Southeast Asia and exhibits ecological and behavioural variability over its range. Studies on understanding the distribution of different members in the complex and their bionomics related to malaria transmission might be important guiding more effective vector intervention strategies. Female An. sundaicus s.l. were collected from seven provinces, 12 locations in Indonesia representing Sumatra: North Sumatra, Bangka-Belitung, South Lampung, and Bengkulu; in Java: West Java; and the Lesser Sunda Islands: West Nusa Tenggara and East Nusa Tenggara provinces. Sequencing of ribosomal DNA ITS2 gene fragments and two mitochondrial DNA gene markers, COI and cytb, enabled molecular identification of morphologically indistinguishable members of the complex. Findings allowed inference on the distribution of the An. sundaicus s.l. present in Indonesia and further illustrate the phylogenetic relationships of An. epiroticus within the complex. A total of 370 An. sundaicus s.l specimens were analysed for the ITS2 fragment. The ITS2 sequence alignment revealed two consistent species-specific point mutations, a T>C transition at base 479 and a G>T transversion at base 538 that differentiated five haplotypes: TG, CG, TT, CT, and TY. The TG haplotype matched published An. epiroticus-indicative sequences from Thailand, Vietnam and peninsular Malaysia. The previously described insertion event (base 603) was observed in all identified specimens. Analysis of the COI and cytb genes revealed no consistent nucleotide variations that could definitively distinguish An. epiroticus from other members in the Sundaicus Complex. The findings indicate and support the existence of An. epiroticus in North Sumatra and Bangka-Belitung archipelago. Further studies are recommended to determine the full distributional extent of the Sundaicus complex in Indonesia and investigate the role of these species in malaria transmission.

Efficacy of some feed additives to attenuate the hepato-renal damage induced by aflatoxin B1 in rabbits.

The present trial was conducted to evaluate the beneficial role of some feed additives (potassium sorbate; Sor, hydrated sodium calcium almuniosilicate; Hsc and l-methionine; L-M) in attenuating the hepatic and renal toxicity of aflatoxin B1 (AFB1) in rabbits. A total number of 72 NZW growing rabbits (5 week-old) were divided into six experimental groups (four replicates with three rabbits each) as follows: control group, AFB1 group (supplemented with AFB1 0.3 mg/kg diet), AFB1 + Sor group (AFB1 0.3 mg/kg diet + Sor 2 g/kg diet), AFB1 + Hsc group (AFB1 0.3 mg/kg diet + Hsc 5 g/kg diet), AFB1 + L-M group (AFB1 0.3 mg/kg diet + L-M 8 g/kg diet) and AFB1 + Mix group (AFB1 0.3 mg/kg diet + 2 Sor + 5 Hsc + 8 L-M g/kg diet). Serum levels of total protein, albumin and globulin were significantly reduced by AF. AF increased the serum activity of aspartate aminotransferase (AST), alanine amino transferase (ALT) and alkaline phosphatase (ALP) enzymes. While, they were reduced in AF + Sor and AF + Mix groups compared with AF group. AF increased the level of cystatin C and Beta-2 microglobulin (BMG). All other supplements significantly reduced the level of cystatin C than AF; however, this reduction was more pronounced in AF-L-M group. AF + Sor, AF + L-M and AF + Mix equally reduced the BMG level than AF and AF + HSc, however, still higher than control. AF elevated the total cholesterol (TC) and LDL-cholesterol levels. A significant reduction in HDL cholesterol was seen in AF group. Additionally, AF induced pathological alterations in the liver and kidney of exposed rabbits on the other hands, the three additives separately or in mix attenuated the Af-induced alterations. In conclusion, the dietary supplementation of Sor, L-M, Hsc or their mixture was effective in ameliorating the negative effects of AFB1 on liver and kidney function and structure in rabbits with more better improvement obtained by Sor or L-M separately.

Early administration of phosphodiesterase 5 inhibitors after induction of diabetes in a rat model may prevent erectile dysfunction.

BACKGROUND: The possible role of phosphodiesterase 5 inhibitors (PDE5Is) in prevention of negative effect of diabetes mellitus (DM) on erectile function is not well settled. OBJECTIVES: To investigate the effect of early administration of vardenafil on erectile function, cavernosal structure, and genes expression in a rat model of DM. MATERIALS AND METHODS: This experimental study was carried out at Suez Canal University's research laboratory. This study was conducted on a total of 60 adult male Albino Wistar rats, aged 60-80 days and weighing an average of 200 g. Rats were equally divided into six groups of 10 rats each: Group I (sham); Group II (DM with no treatment); Groups III, IV, V, and VI received vardenafil started at day 1, week 4, week 8, and week 12 after induction of DM, respectively. Functional study assessment of all groups was performed before euthanization, and then tissues were harvested for histopathological, ultrastructural, and molecular examinations. RESULTS: There was a significant difference of intracavernosal pressure between early (94 ± 2.18) and late (40.5 ± 1.94) treatment groups (p = 0.011). Histopathological and ultrastructural changes of DM with no treatment and late treatment groups showed distorted cavernous architecture and extensive fibrosis. There was significant difference of smooth muscle to collagen ratio between early and late treatment groups (p = 0.035). There was significant upregulation of nNOS(p = 0.021) and iNOS (p = 0.047) in early vs. late treatment group. The difference was insignificant in eNOS (p = 0.386) or TGF-ß1(p = 0.149). DISCUSSION AND CONCLUSION: Early treated rats with vardenafil had preserved erection and normal cavernosal structure, ultrastructure and gene expression of iNOS, nNOS, eNOS, and TGF-ß1. Quantification of gene expression would improve our knowledge regarding cytokines expression and molecular background of DM-associated ED. Clinical application of this result may encourage early administration of PDE5I to prevent deleterious effects of DM on erectile function in newly diagnosed DM patients with probable uncontrolled blood glucose.