Evaluation of suitable reference genes for expression profile analyses of target genes in the coffee berry borer, Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae).

The coffee berry borer, Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae), is a major destructive insect pest of coffee, which impacts the coffee crops negatively. As a draft genome has been completed for this insect, most molecular studies on gene transcriptional levels under different experimental conditions will be conducted using real-time reverse-transcription quantitative polymerase chain reactions (RT-qPCR). However, the lack of suitable internal reference genes will affect the accuracy of RT-qPCR results. In this study, the expression stability of nine candidate reference genes was evaluated under different developmental stages, temperature stress, and Beauveria bassiana infection. Data analyses were completed by four commonly used programs, BestKeeper, NormFinder, geNorm, and RefFinder. The result showed that RPL3 and EF1α combination were recommended as the most stable reference genes for developmental stages. EF1α and RPS3a combination were the top two stable reference genes for B. bassiana infection. RPS3a and RPL3 combination performed as the optimal reference genes both in temperature stress and all samples. Our results should provide a good foundation for the expression profile analyses of target genes in the future, especially for molecular studies on insect genetic development, temperature adaptability, and immune mechanism to entomogenous fungi in H. hampei.

Economic burden of dengue fever in China: A retrospective research study.

BACKGROUND: Dengue fever has been a significant public health challenge in China. This will be particularly important in the context of global warming, frequent international travels, and urbanization with increasing city size and population movement. In order to design relevant prevention and control strategies and allocate health resources reasonably, this study evaluated the economic burden of dengue fever in China in 2019. METHODS: The economic burden of dengue fever patients was calculated from both family and the organisation perspectives. A survey was conducted among 1,027 dengue fever patients in Zhejiang, Chongqing, and Yunnan Provinces. Treatment expenses, lost working days, and insurance reimbursement expenses information were collected to estimate the total economic burden of dengue fever patients in 2019. The expenditures related to dengue fever prevention and control from government, Center for Disease Control and Prevention (CDC), communities and subdistrict offices of 30 counties (or districts) in Zhejiang Province and Chongqing City were also collected. RESULTS: The direct, indirect and total economic burden for dengue fever patients in 2019 in the three Provinces were about 36,927,380.00 Chinese Yuan (CNY), 10,579,572.00 CNY and 46,805,064.00 CNY, respectively. The costs for prevention and control of dengue fever for the counties (or districts) without cases, counties (or districts) with imported cases, and counties (or districts) with local cases are 205,800.00 CNY, 731,180.00 CNY and 6,934,378.00 CNY, respectively. The total investment of dengue fever prevention and control in the 30 counties in China in 2019 was approximately 3,166,660,240.00 CNY. CONCLUSION: The economic burden of dengue fever patients is relatively high, and medical insurance coverage should be increased to lighten patients' direct medical economic burden. At the same time, the results suggests that China should increase funding for primary health service institutions to prevent dengue fever transmission.

Association between meteorological factors and the prevalence dynamics of Japanese encephalitis.

Japanese encephalitis (JE) is an acute infectious disease caused by the Japanese encephalitis virus (JEV) and is transmitted by mosquitoes. Meteorological conditions are known to play a pivotal role in the spread of JEV. In this study, a zero-inflated generalised additive model and a long short-term memory model were used to assess the relationship between the meteorological factors and population density of Culex tritaeniorhynchus as well as the incidence of JE and to predict the prevalence dynamics of JE, respectively. The incidence of JE in the previous month, the mean air temperature and the average of relative humidity had positive effects on the outbreak risk and intensity. Meanwhile, the density of all mosquito species in livestock sheds (DMSL) only affected the outbreak risk. Moreover, the region-specific prediction model of JE was developed in Chongqing by used the Long Short-Term Memory Neural Network. Our study contributes to a better understanding of the JE dynamics and helps the local government establish precise prevention and control measures.

The MaCreA Gene Regulates Normal Conidiation and Microcycle Conidiation in Metarhizium acridum.

As a C2H2 type zinc finger transcription factor, CreA is the key in Carbon Catabolism Repression (CCR) pathway, which negatively regulates the genes in carbon sources utilization. As conidiation in filamentous fungi is affected by nutritional conditions, CreA may contribute to fungal conidiation, which has been well studied in filamentous fungi, especially Aspergillus spp., but researches on entomopathogenic fungi are not enough. In this study, we found a homologous gene MaCreA in Metarhizium acridum, and the MaCreA deletion strain showed delayed conidiation, significant decrease in conidial yield, and 96.88% lower conidial production, when compared with the wild-type strain, and the normal conidiation and microcycle conidiation pattern shift was blocked. RT-qPCR showed that the transcription levels of the genes FlbD and LaeA (related to asexual development) were significantly altered, and those of most of the conidiation-related genes were higher in ΔMaCreA strain. The results of RNA-Seq revealed that MaCreA regulated the two conidiation patterns by mediating genes related to cell cycle, cell division, cell wall, and cell polarity. In conclusion, CreA, as a core regulatory gene in conidiation, provides new insight into the mechanism of conidiation in entomopathogenic fungi.

[Surveillance on the dynamics of leptospirosis epidemics in the Chongqing Section of the Three Gorges dam area].

OBJECTIVE: To analyze the changes of leptospirosis epidemic characteristics before and after the Phase 2 'reservoir store water project' in Chongqing section of the Three Gorges dam area and to provide prevention, control and intervention measures to prevent the spread of leptospirosis from infectious focus to the Three Gorges dam area and downstream region of Changjiang River. METHODS: Changshou district and Fengdu county were selected as surveillance sites. We monitored the source of infection through examining the serum antibody of patients, healthy groups together with farm cattle measured by micro agglutination test (MAT). RESULT: Sporadic cases were reported before and after the storage of water in the reservoir. There was no significant difference found between mouse density before and after the Phase 2 reservoir project (chi2 = 1.00, P > 0.05). The main species of rat were Sewer rat before and Insectivorea after the storage of water. The germ-carrying rate of rats was 1.72% (10/583) and positive carrying rate of rats was 16.51% (18/109) when using PCR. Results showed a significant difference when comparing it to culture method (chi2 = 51.80, P < 0.01). Positive rate of leoptopirosis appeared in the serum of patients was 73.33% (33/45) with the major serum group as the Australia group. The rate of infection among the healthy group was 26.84% (233/868). There was significant difference seen between the serum antibody positive rate of epidemic prophase (23.85%) and epidemic anaphase (29.86%) of the healthy group (chi2 = 3.99, P < 0.05). The GMRT of ox serum antibody of leoptopirosis was 29.97 with Bailen group as the predominant microbial population. CONCLUSION: There was no epidemics of leptopirosis occurred in the Three Gorges dam area. There was no significant difference between mouse density before and after the storage of water in the reservoir. However, the major species of rats had a change. The natural infection level of people living in the dam area was low, but there existed potential of leoptopirosis outbreak.