Rapid assessment of the blood-feeding histories of wild-caught malaria mosquitoes using mid-infrared spectroscopy and machine learning.

BACKGROUND: The degree to which Anopheles mosquitoes prefer biting humans over other vertebrate hosts, i.e. the human blood index (HBI), is a crucial parameter for assessing malaria transmission risk. However, existing techniques for identifying mosquito blood meals are demanding in terms of time and effort, involve costly reagents, and are prone to inaccuracies due to factors such as cross-reactivity with other antigens or partially digested blood meals in the mosquito gut. This study demonstrates the first field application of mid-infrared spectroscopy and machine learning (MIRS-ML), to rapidly assess the blood-feeding histories of malaria vectors, with direct comparison to PCR assays. METHODS AND RESULTS: Female Anopheles funestus mosquitoes (N = 1854) were collected from rural Tanzania and desiccated then scanned with an attenuated total reflectance Fourier-transform Infrared (ATR-FTIR) spectrometer. Blood meals were confirmed by PCR, establishing the 'ground truth' for machine learning algorithms. Logistic regression and multi-layer perceptron classifiers were employed to identify blood meal sources, achieving accuracies of 88%-90%, respectively, as well as HBI estimates aligning well with the PCR-based standard HBI. CONCLUSIONS: This research provides evidence of MIRS-ML effectiveness in classifying blood meals in wild Anopheles funestus, as a potential complementary surveillance tool in settings where conventional molecular techniques are impractical. The cost-effectiveness, simplicity, and scalability of MIRS-ML, along with its generalizability, outweigh minor gaps in HBI estimation. Since this approach has already been demonstrated for measuring other entomological and parasitological indicators of malaria, the validation in this study broadens its range of use cases, positioning it as an integrated system for estimating pathogen transmission risk and evaluating the impact of interventions.

Effects of acute normovolemic hemodilution and allogeneic blood transfusion on postoperative complications of oral and maxillofacial flap reconstruction: a retrospective study.

OBJECTIVE: Patients undergoing oral and maxillofacial flap reconstruction often need blood transfusions due to massive blood loss. With the increasing limitations of allogeneic blood transfusion (ABT), doctors are considering acute normovolemic hemodilution (ANH) because of its advantages. By comparing the differences in the (Δ) blood indices and postoperative complications of patients receiving ABT or ANH during the reconstruction and repair of oral and maxillofacial tumor flaps, this study's purpose was to provide a reference for the clinical application of ANH. METHODS: The clinical data of 276 patients who underwent oral and maxillofacial flap reconstruction from September 25, 2017, to October 11, 2021, in the Department of Oral and Maxillofacial Surgery, Sun Yat-sen Memorial Hospital, were retrospectively analyzed. According to the intraoperative blood transfusion mode, the patients were divided into two groups: ABT and ANH. The differences in the (Δ) blood indices and the incidence of postoperative complications between the groups were analyzed. RESULTS: Among the 276 patients who had ANH (124/276) and ABT (152/276), there were no differences in (Δ) Hb, (Δ) PT, or (Δ) FIB (P > 0.05), while (Δ) WBC, (Δ) PLT, (Δ) APTT and (Δ) D-dimer were significantly different (P < 0.05). The blood transfusion method was not an independent factor for flap crisis (P > 0.05). The wound infection probability in patients with high post-PTs was 1.953 times greater than that in patients with low post-PTs (OR = 1.953, 95% CI: 1.232 ∼ 3.095, P = 0.004). A normal or overweight BMI was a protective factor for pulmonary infection, and the incidence of pulmonary infection in these patients was only 0.089 times that of patients with a low BMI (OR = 0.089, 95% CI: 0.017 ∼ 0.462). Moreover, a high ASA grade promoted the occurrence of pulmonary infection (OR = 6.373, 95% CI: 1.681 ∼ 24.163). The blood transfusion mode (B = 0.310, ß = 0.360, P < 0.001; ANH: ln hospital stay = 2.20 ± 0.37; ABT: ln hospital stay = 2.54 ± 0.42) improved the length of hospital stay. CONCLUSION: Preoperative and postoperative blood transfusion (Δ) Hb, (Δ) PT, and (Δ) FIB did not differ; (Δ) WBC, (Δ) PLT, (Δ) APTT, and (Δ) D-dimer did differ. There was no difference in the effects of the two blood transfusion methods on flap crisis, incision infection or lung infection after flap reconstruction, but ANH resulted in a 3.65 day shorter average hospital stay than did ABT.

Host preference and human blood index of Phlebotomus orientalis, an exophilic sand fly vector of visceral leishmaniasis in eastern Sudan.

Visceral leishmaniasis (VL, kala azar), caused by Leishmania donovani, transmitted by Phlebotomus orientalis, is a serious systemic disease that causes high morbidity and mortality rates in Sudan and other parts of East Africa and the world. Despite progress in understanding the epidemiology of the disease in East Africa, little is known about the host preference of P. orientalis in kala azar endemic villages of Sudan, which have some of the highest VL incidence rates in the world. The present study used host choice experiments and blood-meal identification approaches to determine the host preference of P. orientalis in kala azar endemic villages in Gedarif state, eastern Sudan. In the host choice experiment, tent traps were used to compare the attractiveness of cows, donkeys, sheep and goats for host-seeking P. orientalis. In the blood-meal identification study, blood-fed P. orientalis females, captured inside houses and peri-domestic habitats, were subjected to molecular typing using cytochrome b gene (cyt b) amplification and sequence analysis. Cows and donkeys were the most attractive to blood-seeking P. orientalis, followed by goats. Similarly, the blood-meal analysis of P. orientalis showed that the vector preferentially feeds on cows, followed by donkeys, humans and goats. The human blood index of P. orientalis was 19.4% (42/216), indicating a high zoophilic habit of the vector, both inside and outside the houses. Although the order of host preference varied by location, it was clear that cows are the most preferred host of P. orientalis in the area. Results are discussed in relation to the role of domestic/livestock animals in VL zoopotentiation and zooprophylaxis. Inference is made on the potential impact of insecticide treatment of cows in control of the vector and the transmission of VL in Sudan and other parts of East Africa.

[Incidence of silicosis among excavation workers].

Objective: To assess the incidence of silicosis among excavation workers in Suzhou city. Methods: In 2011-2016, we investigated the incidence of silicosis among 3190 excavation workers who participated in the mining operations from 1971 to 1974 in Suzhou city. To compare the general condition, complications, blood indexes and pulmonary function between different stages of silicosis patients. Results: Among all the participants, 144 cases were diagnosed as silicosis, and the incidence of silicosis was 4.51%, including 138 stage I cases (95.83%) , 4 stage II cases (2.78%) , and 2 stage III cases (1.39%) . Both systolic blood pressure (154.17±14.74 mmHg) and diastolic blood pressure (96.67±10.25 mmHg) of silicosis patients in stage II-III were significantly higher than that of patients in stage I (134.64±15.41 mmHg, 82.20±10.62 mmHg) (P<0.05) . Hypertension (54 cases, 37.50%) were the most commonly complication of silicosis patients, followed by abnormal electrocardiogram (38 cases, 26.39%) , diabetes (22 cases, 15.28%) , malignant tumor (10 cases, 6.94%) , hepatitis B (8 cases, 5.56%) and tuberculosis (8 cases, 5.56%) . The serum levels of red blood cell count (RBC) , hemoglobin (Hb) , albumin (ALB) , magnesium (Mg) and sodium (Na) in stage I silicosis patients were significantly higher than stage II-III patients (P<0.05) . While the serum levels of C-reactive protein (CRP) and carbohydrate antigen 199 (CA199) in stage I silicosis patients were significantly lower than stage II-III patients (P<0.05) . The proportions of normal pulmonary function, mild airflow limitation, moderate airflow limitation and severe airflow limitation in silicosis patients were 44.03% (59/134) , 18.66% (25/134) , 29.85% (40/134) and 7.46% (10/134) , respectively. Conclusion: Our data obviously show that the excavation workers has a high incidence of silicosis. It is necessary to strengthen the occupational health surveillance of silicosis patients, reduce complications and improve their quality of life.

Using the human blood index to investigate host biting plasticity: a systematic review and meta-regression of the three major African malaria vectors.

BACKGROUND: The proportion of mosquito blood-meals that are of human origin, referred to as the 'human blood index' or HBI, is a key determinant of malaria transmission. METHODS: A systematic review was conducted followed by meta-regression of the HBI for the major African malaria vectors. RESULTS: Evidence is presented for higher HBI among Anopheles gambiae (M/S forms and Anopheles coluzzii/An. gambiae sensu stricto are not distinguished for most studies and, therefore, combined) as well as Anopheles funestus when compared with Anopheles arabiensis (prevalence odds ratio adjusted for collection location [i.e. indoor or outdoor]: 1.62; 95% CI 1.09-2.42; 1.84; 95% CI 1.35-2.52, respectively). This finding is in keeping with the entomological literature which describes An. arabiensis to be more zoophagic than the other major African vectors. However, analysis also revealed that HBI was more associated with location of mosquito captures (R2 = 0.29) than with mosquito (sibling) species (R2 = 0.11). CONCLUSIONS: These findings call into question the appropriateness of current methods of assessing host preferences among disease vectors and have important implications for strategizing vector control.

Study on Impact of Iron and Folic Acid on the Plasma Trace Minerals in Pregnant Anemic Women.

Iron deficiency anemia is one of the causes that lead to significant mortality and morbidity among pregnant women and fetus. The present study was undertaken to explore oral iron supplementation can modify the metal contents in pregnant anemic women. Iron and folic acid supplementations was given to 500 anemic women (mild = 200, moderate = 200, and severe = 100) and 100 age matched non-anemic controls daily for 100 days. Blood index values and plasma trace minerals were estimated as per standard protocols. Haemoglobin and ferritin levels were found significantly increased (p < 0.001) in anemic and control subjects after treatment. Moreover, the serum transferring receptor levels and total iron binding capacity were found significantly decreased in all treated groups. Iron (Fe), zinc (Zn) and copper (Cu) levels were found increased (p < 0.01) after oral iron supplementation groups. Moreover, selenium (Se) manganese (Mn) and were found to be decreased in all treated groups. Data provides the conclusion that iron and folic acid supplementation recovered the essential trace minerals, except manganese, which may lead to various complications including peroxidation of vital body molecules resulting in increased risk for pregnant women as well as fetus.

Effects of Replacement of Methionine in Diets with Taurine on Growth Performance and Blood Index in Broilers.

We studied effects of replacement of methionine with taurine on growth performance and blood index of AA+ broilers. Six hundred 1 day broilers were divided into 5 groups, with 3 replicates of 40 broilers in each. The experiment lasted for 42 days.The control group were fed on formulated diets containing 2% methionine; the other groups were offered feed with equal nitrogen and calories to the control group, but contained 25, 50, 75 and 100% taurine in place of methionine.Compared with the control group, no significant differences were observed in growth performance of 1-21 days broilers, or the serum LDL-C, TC, IgG and SOD of the experimental groups (P> 0.05). ADG and F/G from days 1-42, ADG, ADFI and F/G from days 22-42 were significantly different between the experimental groups and the control group (P < 0.05). ADFI and Mortality in 50, 75 and 100% taurine groups were significantly different compared with the control group (P < 0.05). IgM and GSH-PX of 50 and 75% taurine groups were significantly different compared with the control group (P < 0.05). Serum HDL-C, T-AOC levels in 50, 75 and 100% taurine groups were significantly different compared with the control group (P < 0.05). Based on the quadratic regression analysis, the best replacement ratios were 58%, 61% and 61% on days 1-21, 22-42, and 1-42, respectively. In conclusion, appropriate levels of taurine supplement can improve growth performance, immune system, T-AOC, and lipid metabolism.

Determinants of host feeding success by Anopheles farauti.

BACKGROUND: The proportion of blood meals that mosquitoes take from a host species is a function of the interplay of extrinsic (abundance and location of potential hosts) and intrinsic (innate preference) factors. A mark-release-recapture experiment addressed whether host preference in a population of Anopheles farauti was uniform or if there were anthropophilic and zoophilic subpopulations. The corresponding fitness associated with selecting different hosts for blood meals was compared by measuring fecundity. METHODS: The attractiveness of humans for blood meals by An. farauti in the Solomon Islands was compared to pigs using tent traps. Host fidelity was assessed by mark-release-recapture experiments in which different colour dusts were linked to the host to which the mosquito was first attracted. Outdoor resting An. farauti were captured on barrier screens and the human blood index (HBI) as well as the feeding index were calculated. The fecundity of individual An. farauti after feeding on either humans or pigs was assessed from blood-fed mosquitoes held in individual oviposition chambers. RESULTS: Anopheles farauti were more attracted to humans than pigs at a ratio of 1.31:1.00. The mark-release-recapture experiment found evidence for An. farauti being a single population regarding host preference. The HBI of outdoor resting An. farauti was 0.93 and the feeding index was 1.29. Anopheles farauti that fed on a human host laid more eggs but had a longer oviposition time compared to An. farauti that had blood fed on a pig. CONCLUSIONS: One of the strongest drivers for host species preference was the relative abundance of the different host species. Here, An. farauti have a slight preference for humans over pigs as blood meal sources. However, the limited availability of alternative hosts relative to humans in the Solomon Islands ensures a very high proportion of blood meals are obtained from humans, and thus, the transmission potential of malaria by An. farauti is high.

A New Formula Based on Simple Blood Indices to Differentiate Beta Thalassemia Trait from Iron Deficiency Anemia.

Background: Thalassemia is an inherited blood disorder with a defect in the sufficient production of a protein called hemoglobin. We aimed to investigate the simple blood indices of patients with Beta Thalassemia Trait (BTT) and Iron Deficiency Anemia (IDA) to propose a new formula using logistic regression for differentiate two characteristics from each other. Methods: Among the 702 records of the BTT Counseling Center (Khoy, Iran-2022), 292 cases (219 iron deficiency anemia (IDA) and 73 BTT) were eligible for the study. Blood indices such as RBC, HGB, HbA2 described and used to diagnose two types of participants. Blood indices had high multicollinearity that was modified. Logistic regression for blood indices fitted and goodness of fit indices with Area Under ROC curve (AUC) estimated. Results: The average age of the participants was 24.56 yr. The status of Multicollinearity between independent variables was modified. The HGB, MCV, HbA2, and HbA variables were used in the model and only HbA2 status was significant (P<0.001). According to the output of the model, for each unit increase in HbA2, the chance of having BTT was about 8.5 times higher than IDA. The sensitivity, specificity, AUC curve, and accuracy of the final model were estimated to be 97, 72, 84, and 93%, respectively. A regression formula to differentiate BTT from IDA proposed. Conclusion: In studies related to the differentiation of the BTT from IDA, the presence of the HbA2 index in the model and prediction is very necessary.

Effects of Dietary Protein Level and Rumen-Protected Methionine and Lysine on Growth Performance, Rumen Fermentation and Serum Indexes for Yaks.

This study investigated the effects of the dietary protein level and rumen-protected methionine and lysine (RPML) on the growth performance, rumen fermentation, and serum indexes of yaks. Thirty-six male yaks were randomly assigned to a two by three factorial experiment with two protein levels, 15.05% and 16.51%, and three RPML levels: 0% RPML; 0.05% RPMet and 0.15% RPLys; and 0.1% RPMet and 0.3% RPLys. The trial lasted for sixty days. The results showed that the low-protein diet increased the DMI and feed conversion ratio of yaks. The diet supplemented with RPML increased the activities of IGF1 and INS and nutrient digestibility. The high-protein diet decreased the rumen butyrate concentration and increased the rumen isovalerate concentration. The low-protein diet supplemented with RPML increased the rumen pH and the concentrations of total volatile fatty acids, butyrate and NH3-N; the high-protein diet supplemented with a high level of RPML decreased the rumen pH and the concentrations of isobutyrate, isovalerate, propionate and NH3-N. The low-protein diet supplemented with RPML increased the total antioxidant capacity and glutathione peroxidase activity, along with the concentrations of malondialdehyde and amino acids such as aspartic acid, lysine, cysteine, etc. In conclusion, a low-protein diet supplemented with RPML is beneficial for rumen and body health, physiological response, and metabolic status in yaks.

Optimizing malaria vector control in the Greater Mekong Subregion: a systematic review and mathematical modelling study to identify desirable intervention characteristics.

BACKGROUND: In the Greater Mekong Subregion (GMS), new vector-control tools are needed to target mosquitoes that bite outside during the daytime and night-time to advance malaria elimination. METHODS: We conducted systematic literature searches to generate a bionomic dataset of the main malaria vectors in the GMS, including human blood index (HBI), parity proportion, sac proportion (proportion with uncontracted ovary sacs, indicating the amount of time until they returned to host seeking after oviposition) and the resting period duration. We then performed global sensitivity analyses to assess the influence of bionomics and intervention characteristics on vectorial capacity. RESULTS: Our review showed that Anopheles minimus, An. sinensis, An. maculatus and An. sundaicus display opportunistic blood-feeding behaviour, while An. dirus is more anthropophilic. Multivariate regression analysis indicated that environmental, climatic and sampling factors influence the proportion of parous mosquitoes, and resting duration varies seasonally. Sensitivity analysis highlighted HBI and parity proportion as the most influential bionomic parameters, followed by resting duration. Killing before feeding is always a desirable characteristic across all settings in the GMS. Disarming is also a desirable characteristic in settings with a low HBI. Repelling is only an effective strategy in settings with a low HBI and low parity proportion. Killing after feeding is only a desirable characteristic if the HBI and parity proportions in the setting are high. CONCLUSIONS: Although in general adopting tools that kill before feeding would have the largest community-level effect on reducing outdoor transmission, other modes of action can be effective. Current tools in development which target outdoor biting mosquitoes should be implemented in different settings dependent on their characteristics.

Laboratory scoring system to predict hepatic indocyanine green clearance ability during fluorescence imaging-guided laparoscopic hepatectomy.

BACKGROUND: Indocyanine green (ICG) fluorescence played an important role in tumor localization and margin delineation in hepatobiliary surgery. However, the preoperative regimen of ICG administration was still controversial. Factors associated with tumor fluorescence staining effect were unclear. AIM: To investigate the preoperative laboratory indexes corelated with ICG fluorescence staining effect and establish a novel laboratory scoring system to screen specifical patients who need ICG dose adjustment. METHODS: To investigate the predictive indicators of ICG fluorescence characteristics in patients undergoing laparoscopic hepatectomy from January 2018 to January 2021 were included. Blood laboratory tests were completed within 1 wk before surgery. All patients received 5 mg ICG injection 24 h before surgery for preliminary tumor imaging. ImageJ software was used to measure the fluorescence intensity values of regions of interest. Correlation analysis was used to identify risk factors. A laboratory risk model was established to identify individuals at high risk for high liver background fluorescence. RESULTS: There were 110 patients who were enrolled in this study from January 2019 to January 2021. The mean values of fluorescence intensity of liver background (FI-LB), fluorescence intensity of gallbladder, and fluorescence intensity of target area were 18.87 ± 17.06, 54.84 ± 33.29, and 68.56 ± 36.11, respectively. The receiver operating characteristic (ROC) curve showed that FI-LB was a good indicator for liver clearance ability [area under the ROC curve (AUC) = 0.984]. Correlation analysis found pre-operative aspartate aminotransferase, alanine aminotransferase, gamma-glutamyl transpeptidase, adenosine deaminase, and lactate dehydrogenase were positively associated with FI-LB and red blood cell, cholinesterase, and were negatively associated with FI-LB. Total laboratory risk score (TLRS) was calculated according to ROC curve (AUC = 0.848, sensitivity = 0.773, specificity = 0.885). When TLRS was greater than 6.5, the liver clearance ability of ICG was considered as poor. CONCLUSION: Preoperative laboratory blood indicators can predict hepatic ICG clearance ability. Surgeons can adjust the dose and timing of ICG preoperatively to achieve better liver fluorescent staining.

Effects of Diets Containing Different Levels of Copper, Manganese, and Iodine on Rumen Fermentation, Blood Parameters, and Growth Performance of Yaks.

Copper, manganese, and iodine are part of a yak's required trace elements. However, knowledge about their dietary requirements is scarce. Therefore, an experiment was conducted to evaluate rumen fermentation, blood parameters, and growth performance and screen out the optimum levels of trace elements in yaks' diet. Here, 18 three-year-old castrated yaks were randomly divided into four groups, which fed with diets containing basal (CON: 4.40, 33.82, and 0 mg/kg) and low-level (LL: 10.00, 40.00, and 0.30 mg/kg), middle-level (ML: 15.00, 50.00, and 0.50 mg/kg), and high-level (HL: 20.00, 60.00, and 0.70 mg/kg) copper, manganese, and iodine for 30 days. With the increase in trace elements, yaks' daily weight gain (DWG), rumen pH, ammonia nitrogen, microbial protein (MCP), and volatile fatty acids levels and serum triglycerides and urea nitrogen levels showed firstly increasing and then decreasing trends and reached the highest values in ML, and serum ceruloplasmin and total superoxide dismutase (T-SOD) activities showed continuously increasing trends. Yaks' DWG, rumen MCP, butyrate, and valerate levels and serum triglycerides, urea nitrogen, ceruloplasmin, and T-SOD levels in ML were significantly higher than CON. Therefore, the recommended levels of copper, manganese, and iodine in growing yaks' diet are 15.00, 50.00, and 0.50 mg/kg (ML), respectively.

Dietary protein to starch metabolizable energy ratios alter growth performance and gastrointestinal microbiota of calves.

The diet structure is very important for the growth and development of calves. This study aimed to investigate the effects of dietary protein-to-starch metabolizable energy ratios (DPSRs) on growth performance, blood index, and gastrointestinal microbiota of calves. Forty-eight Holstein bull calves were fed six dietary DPSRs including A20-35 (20% CP and 35% starch), B20-30, C20-25, D22-35, E22-30, and F22-25 at d 4 to d 60, and then changed to another six dietary DPSRs at d 61 to d 180 (A18-30, B18-27, C18-24, D20-30, E20-27, and F20-24). Twelve calves (d 60) from groups A20-35, C20-25, D22-35, and F22-25 (n = 3) and another twelve calves (d 180) from groups A18-30, C18-24, D20-30, and F20-24 (n = 3) were euthanized. The growth performance parameters were measured. Blood, ruminal fluid, and cecum digesta were collected for further analysis. Results showed heart girth gain of B18-27 was significantly higher than A18-30, C18-24, and heart girth gain (d 180) was significantly affected by protein × starch (DPSRs; p < 0.05). Blood urea nitrogen (BUN; d 60) in C20-25 was significantly higher than A20-35 and B20-30 (p < 0.05). The BUN (d 180) in D20-30 was significantly higher than A18-30 (p < 0.05). The BUN was significantly affected by protein × starch (p < 0.05) on d 60. The albumin (ALB) levels in C20-25 and C18-24 were significantly higher than that in A20-35 on d 60 and A18-30 on d 180, respectively (p < 0.05). The ALB level in D22-35 on d 60 and E20-27 on d 180 was significantly higher than that in other groups (p < 0.05). The ALB level was significantly affected by protein and starch, respectively, on d 60 (p < 0.05). In the rumen, the genera Roseburia (C20-25) and Dialister (D22-35), Prevotellaceae UCG-001 (C18-24), Erysipelotrichaceae UCG-002, and Anaerovorax (F20-24) were found in significant higher relative abundances than those in other groups (p < 0.05). In the cecum, the genera Bacteroides and Eisenbergiella (F22-25), Ruminiclostridium_1 and Candidatus Stoquefichus (A18-30), Erysipelotrichaceae UCG-004 and Tyzzerella 4 (D20-30), and Prevotellaceae UCG-003 and Klebsiella (F20-24) were found in significant higher abundances than those in other groups (p < 0.05). Collectively, these results indicated that the heart girth, BUN, ALB, and gastrointestinal microbiota responded distinctly to differing DPSRs.

Spatiotemporal distribution and insecticide resistance status of Aedes aegypti in Ghana.

BACKGROUND: Vector control is the main intervention used to control arboviral diseases transmitted by Aedes mosquitoes because there are no effective vaccines or treatments for most of them. Control of Aedes mosquitoes relies heavily on the use of insecticides, the effectiveness of which may be impacted by resistance. In addition, rational insecticide application requires detailed knowledge of vector distribution, dynamics, resting, and feeding behaviours, which are poorly understood for Aedes mosquitoes in Africa. This study investigated the spatiotemporal distribution and insecticide resistance status of Aedes aegypti across ecological extremes of Ghana. METHODS: Immature mosquitoes were sampled from containers in and around human dwellings at seven study sites in urban, suburban, and rural areas of Ghana. Adult Aedes mosquitoes were sampled indoors and outdoors using Biogents BG-Sentinel 2 mosquito traps, human landing catches, and Prokopack aspiration. Distributions of immature and adult Aedes mosquitoes were determined indoors and outdoors during dry and rainy seasons at all sites. The phenotypic resistance status of Aedes mosquitoes to insecticides was determined using World Health Organization susceptibility bioassays. The host blood meal source was determined by polymerase chain reaction. RESULTS: A total of 16,711 immature Aedes were sampled, with over 70% found in car tyres. Significantly more breeding containers had Aedes immatures during the rainy season (11,856; 70.95%) compared to the dry season (4855; 29.05%). A total of 1895 adult Aedes mosquitos were collected, including Aedes aegypti (97.8%), Aedes africanus (2.1%) and Aedes luteocephalus (0.1%). Indoor sampling of adult Aedes yielded a total of 381 (20.1%) and outdoor sampling a total of 1514 (79.9%) mosquitoes (z = - 5.427, P = 0.0000) over the entire sampling period. Aedes aegypti populations were resistant to dichlorodiphenyltrichloroethane at all study sites. Vectors showed suspected resistance to bendiocarb (96-97%), permethrin (90-96%) and deltamethrin (91-96%), and were susceptible to the organophosphate for all study sites. Blood meal analysis showed that the Aedes mosquitoes were mostly anthropophilic, with a human blood index of 0.9 (i.e. humans, 90%; human and dog, 5%; dog and cow, 5%). CONCLUSIONS: Aedes mosquitoes were found at high densities in all ecological zones of Ghana. Resistance of Aedes spp. to pyrethroids and carbamates may limit the efficacy of vector control programmes and thus requires careful monitoring.

Growth Performance and Hematological Changes in Growing Sika Deers Fed with Spent Mushroom Substrate of Pleurotus ostreatus.

The purpose of this experiment is to expand the feed of growing sika deer and to explore the effects on growing sika deer of the spent mushroom substrate of Pleurotus ostreatus (SMS-MP). Twelve immature female growing sika deer were randomly assigned to four groups. The ratios of SMS-MP to replace concentration supplements were 0%, 10%, 20%, and 30%, respectively, and the growth performance, feed intake and apparent digestibility, serum biochemical indexes, blood physiological indexes, serum immune globulin and plasma amino acid of growing sika deer were measured. The results of the current study confirmed the applicability of SMS-MP as a feed ingredient in growing sika deer diets. There was no significant change in growth performance and hematology of growing sika deer when the concentrate supplement was replaced with 10-20% SMS-MP. However, replacing 30% of concentrate supplements with SMS-MP in the growing sika deer diet resulted in significantly decreased Hb and HCT levels. It can be concluded that, as a waste resource, adding a small amount of SMS-MP has no significant effect on the growth of sika deer, and at the same time can reduce the consumption of concentrate supplements, thereby improving the economic benefits of sika deer breeding.

The Association between Blood Indexes and Immune Cell Concentrations in the Primary Tumor Microenvironment Predicting Survival of Immunotherapy in Gastric Cancer.

The tumor microenvironment plays a vital role in tumor progression and treatment response. However, the association between immune cell concentrations in primary tumor and blood indexes remains unknown. Thus, we enrolled patients with gastric cancer (GC) in two cohorts. We used multiplexed immunohistochemistry to quantify in situ proteins covering rare cell types at sub-cellular resolution in 80 patients with GC in the first cohort. A high correlation between the LMR (lymphocyte-to-monocyte ratio)/NLR (neutrophil-to-lymphocyte ratio) and tumor immune microenvironment was found. The density of exhausted CD8 T cells including CD8+PD1−TIM3+, CD8+LAG3+PD1+, CD8+LAG3+PD1−, CD8+LAG3+PD1+TIM3− was negatively associated with LMR and positively associated with NLR (p < 0.05). Additionally, the higher density of macrophages in tumor core was associated with a higher platelet-to-lymphocyte ratio and systemic immune-inflammation index. Furthermore, we validated the prognostic value of LMR and NLR in an independent cohort of 357 gastric cancer patients receiving immunotherapy. Higher LMR at baseline was significantly associated with superior immune-related PFS (irPFS) and a trend of superior immune-related OS (irOS). Higher NLR was associated with inferior irOS. In conclusion, blood indexes were associated with immune cells infiltrating in primary tumors of GC. NLR and LMR are associated with the density of exhausted CD8+ T immune cells, which leads to prognostic values of immunotherapy.

The resting behavior of malaria vectors in different ecological zones of Ghana and its implications for vector control.

BACKGROUND: In sub-Saharan Africa there is widespread use of long-lasting insecticidal nets and indoor residual spraying to help control the densities of malaria vectors and decrease the incidence of malaria. This study was carried out to investigate the resting behavior, host preference and infection with Plasmodium falciparum of malaria vectors in Ghana in the context of the increasing insecticide resistance of malaria vectors in sub-Saharan Africa. METHODS: Indoor and outdoor resting anopheline mosquitoes were sampled during the dry and rainy seasons in five sites in three ecological zones [Sahel savannah (Kpalsogo, Pagaza, Libga); coastal savannah (Anyakpor); and forest (Konongo)]. Polymerase chain reaction-based molecular diagnostics were used to determine speciation, genotypes for knockdown resistance mutations (L1014S and L1014F) and the G119S ace1 mutation, specific host blood meal origins and sporozoite infection in the field-collected mosquitoes. RESULTS: Anopheles gambiae sensu lato (s.l.) predominated (89.95%, n = 1718), followed by Anopheles rufipes (8.48%, n = 162) and Anopheles funestus s.l. (1.57%, n = 30). Sibling species of the Anopheles gambiae s.l. revealed Anopheles coluzzii accounted for 63% (95% confidence interval = 57.10-68.91) and 27% (95% confidence interval = 21.66-32.55) was Anopheles gambiae s. s.. The mean resting density of An. gambiae s.l. was higher outdoors (79.63%; 1368/1718) than indoors (20.37%; 350/1718) (Wilcoxon rank sum test, Z = - 4.815, P < 0.0001). The kdr west L1014F and the ace1 mutation frequencies were higher in indoor resting An. coluzzii and An. gambiae in the Sahel savannah sites than in the forest and coastal savannah sites. Overall, the blood meal analyses revealed that a larger proportion of the malaria vectors preferred feeding on humans (70.2%) than on animals (29.8%) in all of the sites. Sporozoites were only detected in indoor resting An. coluzzii from the Sahel savannah (5.0%) and forest (2.5%) zones. CONCLUSIONS: This study reports high outdoor resting densities of An. gambiae and An. coluzzii with high kdr west mutation frequencies, and the presence of malaria vectors indoors despite the use of long-lasting insecticidal nets and indoor residual spraying. Continuous monitoring of changes in the resting behavior of mosquitoes and the implementation of complementary malaria control interventions that target outdoor resting Anopheles mosquitoes are necessary in Ghana.

Bloodmeal sources and feeding behavior of anopheline mosquitoes in Bure district, northwestern Ethiopia.

BACKGROUND: Mosquito bloodmeal sources determine the feeding rates, adult survival, fecundity, hatching rates, and developmental times. Only the female Anopheles mosquito takes bloodmeals from humans, birds, mammals, and other vertebrates for egg development. Studies of the host preference patterns in blood-feeding anopheline mosquitoes are crucial to determine malaria vectors. However, the human blood index, foraging ratio, and host preference index of anopheline mosquitoes are not known so far in Bure district, Ethiopia. METHODS: The origins of bloodmeals from all freshly fed and a few half-gravid exophagic and endophagic females collected using Centers for Disease Control and Prevention light traps were identified as human and bovine using enzyme-linked immunosorbent assay. The human blood index, forage ratio, and host feeding index were calculated. RESULTS: A total of 617 specimens belonging to An. arabiensis (n = 209), An. funestus (n = 217), An. coustani (n = 123), An. squamosus (n = 54), and An. cinereus (n = 14) were only analyzed using blood ELISA. Five hundred seventy-five of the specimens were positive for blood antigens of the host bloods. All anopheline mosquitoes assayed for a bloodmeal source had mixed- rather than single-source bloodmeals. The FR for humans was slightly > 1.0 compared to bovines for all Anopheles species. HFI for each pair of vertebrate hosts revealed that humans were the slightly preferred bloodmeal source compared to bovines for all species (except An. squamosus), but there was no marked host selection. CONCLUSIONS: All anopheline mosquitoes assayed for bloodmeal ELISA had mixed feeds, which tends to diminish the density of gametocytes in the mosquito stomach, thereby reducing the chance of fertilization of the female gamete and reducing the chances of a malaria vector becoming infected. Moreover, An. coustani was the only species that had only human bloodmeals, meaning that this species has the potential to transmit the disease. Therefore, combination zooprophylaxis should be reinforced as a means of vector control because the study sites are mixed dwellings.

Host feeding patterns of Nyssorhynchus darlingi (Diptera: Culicidae) in the Brazilian Amazon.

Nyssorhynchus darlingi (Root) is the dominant malaria vector in the Brazilian Amazon River basin, with additional Anophelinae Grassi species involved in local and regional transmission. Mosquito blood-feeding behavior is an essential component to define the mosquito-human contact rate and shape the transmission cycle of vector-borne diseases. However, there is little information on the host preferences and blood-feeding behavior of Anophelinae vectors in rural Amazonian landscapes. The barrier screen sampling (BSS) method was employed to sample females from 34 peridomestic habitats in 27 rural communities from 11 municipalities in the Brazilian Amazon states of Acre, Amazonas, Pará and Rondônia, from August 2015 to November 2017. Nyssorhynchus darlingi comprised 97.94% of the females collected resting on barrier screens, and DNA sequence comparison detected 9 vertebrate hosts species. The HBI index ranged from 0.03-1.00. Results revealed the plasticity of Ny. darlingi in blood-feeding on a wide range of mainly mammalian hosts. In addition, the identification of blood meal sources using silica-dried females is appropriate for studies of human malaria vectors in remote locations.