The life cycle of Dermacentor nuttalli from the Qinghai-Tibetan Plateau under laboratory conditions and detection of spotted fever group Rickettsia spp.

Dermacentor nuttalli has been a focus of study because tick-borne pathogens have been widely identified in this tick from northern and southwestern China. The aim of this study was to characterize the life cycle of D. nuttalli under laboratory conditions and to detect spotted fever group (SFG) Rickettsia in the midgut and salivary glands of both field-collected and first laboratory generation adults. D. nuttalli ticks were collected in the field on the Qinghai-Tibetan Plateau from March to April 2021 and their life cycle was studied under laboratory conditions. Tick identify was molecularly confirmed, and SFG Rickettsia were detected in the midgut and salivary glands of males and females by PCR targeting different rickettsial genes. The results showed that the life cycle of D. nuttalli under laboratory conditions was completed in an average of 86.1 days. High positivity of Rickettsia spp. was detected in the midgut and salivary glands of both males (92.0%) and females (93.0%) of field-collected D. nuttalli ticks. However, a relatively lower positivity (4.0–6.0%) was detected in first laboratory generation adults. Furthermore, sequencing analysis showed that the Rickettsia sequences obtained in this study shared 98.6 to 100% nucleotide identity with Rickettsia slovaca and Rickettsia raoultii isolated from Dermacentor spp. in China. Phylogenetic analysis of Rickettsia spp. based on the gltA, ompA, ompB and sca4 genes revealed that the Rickettsia sequences obtained could be classified as belonging to R. slovaca and R. raoultii clades. This study described for the first time the life cycle of D. nuttalli from the Qinghai-Tibetan Plateau under laboratory conditions. Two species of SFG Rickettsia were detected in the midgut and salivary glands of males and females in both field-collected and first laboratory-generation adults of D. nuttalli. Our study provides new insights into pathogen detection in ticks in the Qinghai-Tibet Plateau, and the relationships among hosts, ticks, and pathogens

First description of Blastocystis sp. and Entamoeba sp. infecting zoo animals in the Qinghai-Tibetan plateau area, China

Protozoan parasites are a well-known threat to human health, particularly for people working at or visiting zoos, and potentially cause zoonotic diseases in humans. Captive wildlife may be potential reservoirs for human infection with protozoan parasites. Therefore, focusing on zoonotic protozoan infections in zoo animals is critical. However, there is no report on this topic in the Qinghai-Tibetan Plateau region. In this study, a total of 167 and 103 fecal samples were collected from 12 animal species from Qinghai-Tibet Plateau Wildlife Park in winter and summer, respectively, to detection the prevalence of infections and subtype distribution with Entamoeba sp., Cryptosporidium sp., Giardia duodenalis, Enteromicrosporidia bieneusi sp., Blastocystis sp. by PCR assay. The results showed that a total of 21 fecal samples collected in winter, including from 2 white-lipped deer, 8 Sika deer, 6 blue sheep, 2 wolves and 3 bears, were positive for Entamoeba, with a 12.6% (21/167) positive rate. However, 4.9% (5/103) of animals in summer were positive for Entamoeba, including 1 snow leopard, 1 tiger, 1 Tibetan argali and 2 mouflon. Moreover, 1 white-lipped deer and 1 bear were found to be positive for Blastocystis sp., one zoonotic STs (ST10) was identified and found in white-lipped deer. We found no effect on season on Blastocystis sp. and Entamoeba sp. colonization. To the best of our knowledge, this study is the first description of Blastocystis sp. and Entamoeba sp. infecting zoo animals in the plateau area. The findings provide the latest data on Entamoeba sp. and Blastocystis sp. in zoo animals in China

Epidemiology of atrial fibrillation and risk of CVD mortality among hypertensive population: A prospective cohort study in Northeast China

BackgroundDetermining risk factors of cardiovascular disease (CVD)-related mortality and evaluating their influence are important for effectively reducing corresponding mortality. However, few research findings have estimated the relationship between atrial fibrillation (AF) and CVD-related mortality among hypertension individuals.ObjectiveThe objective of this study was to investigate the epidemiology of AF in a hypertension population and determine the relationship between AF and CVD-related mortality.MethodsUsing a multistage, stratified, and cluster random sampling method, the prospective cohort study with a median follow-up of 3.51 years enrolled 10,678 hypertensive participants at baseline. The prevalence, awareness, and anticoagulation data of AF in this focal population were carefully assessed. Stepwise logistic regression and Cox regression analysis were respectively performed to evaluate the determinants of AF and the association between AF and CVD-related mortality.ResultsThe overall prevalence of AF was 1.3% (95% CI, 1.1%−1.6%) in the hypertensive population, and it was higher in men than in women (1.8% vs. 1.0%, respectively; p=0.001). The awareness of AF was 53.1%, and the rate of oral anticoagulant (OAC) therapy was only 4.2%, although all AF participants should have required according to the European Society of Cardiology guidelines. The determinants of AF included elder, male, and history of coronary heart disease in the hypertensive population. Besides, compared with individuals without AF, the risk of CVD-related mortality significantly increased in the hypertensive population with AF (HR 3.37, 95% CI 2.10–5.40).ConclusionOur results indicated a huge burden of AF and underuse of OAC therapy for them in a community-based hypertensive population. Considering that most of the risk factors of AF were unmodifiable in hypertensive individuals, as well as its high risk of mortality, long-term interventions including AF education, timely screening, and widespread use of OACs should be emphasized in the focal populations

Serological Analysis of IgG and IgM Antibodies against Anaplasma spp. in Various Animal Species of the Qinghai-Tibetan Plateau

Anaplasma genus infects the blood cells of humans and animals by biting, causing zoonotic anaplasmosis. However, limited data are available on carrier animals for Anaplasma spp. antibodies in the Qinghai–Tibetan Plateau Area. Therefore, a serological indirect ELISA diagnostic method based on the major surface protein 5 (MSP5), derived from Anaplasma phagocytophilum, was developed in this study to analyze both IgG and IgM antibodies of Anaplasma spp. in a total of 3952 animals from the Qinghai–Tibetan Plateau, including yaks (Bos grunniens), cows (Bos taurus), cattle (Bos taurus domesticus), Tibetan sheep (Ovis aries), horses (Equus ferus caballus), pigs (Sus domesticus), chickens (Gallus gallus domesticus), donkeys (Equus asinus), stray dogs (Canis sp.), and stray cats (Felis sp.). The results showed that recombinant MSP5 protein was expressed and was successfully used to establish the indirect ELISA methods. The overall positivity for Anaplasma IgG and IgM antibodies was 14.6% (578/3952) and 7.9% (312/3952), respectively, and a total of 123 animals (3.1%) were both IgG- and IgM-positive. Moreover, the most prevalent Anaplasma IgG positivity was exhibited by donkeys (82.5%), followed by stray dogs, Tibetan sheep, pigs, chickens, horses, yaks, cows, cattle, and stray cats. The analysis for IgM antibody positivity revealed that IgM positivity was the most prevalent in the stray dogs (30.1%), followed by horses, yaks, Tibetan sheep, cows, stray cats, and cattle. Moreover, the results revealed significant differences (p < 0.05) at different altitudes in Anaplasma-specific IgG in the yaks, Tibetan sheep, and horses, and in IgM in the yaks and Tibetan sheep. In conclusion, this study is the first to demonstrate that yaks, cows, cattle, Tibetan sheep, horses, donkeys, stray dogs, stray cats, pigs, and chickens living in the Qinghai–Tibet Plateau are carrier animals for Anaplasma spp. IgG or IgM antibodies. The current findings provide valuable current data on the seroepidemiology of anaplasmosis in China and for plateau areas of the world

Prevalence of prediabetes and risk of CVD mortality in individuals with prediabetes alone or plus hypertension in Northeast China: insight from a population based cohort study

Abstract Background To evaluate the current prevalence of prediabetes in northeast China, and further determine the association between prediabetes alone or coexistent with hypertension and cardiovascular disease (CVD) mortality. Methods In the prospective study, 15,557 participants without diabetes among aged ≥40 years in northeast China, were followed for a median of 5.5 years. Following the American Diabetes Association, prediabetes was defined as fasting plasma glucose (FPG) range of 5.6-6.9 mmol/L or glycated hemoglobin (HbA1c) range of 5.7-6.4% in people without diabetes. Results The prevalence of prediabetes was 44.3% among population aged ≥40 years in northeast China. Prediabetes alone did not promote risk of CVD mortality. However, when the subgroups were stratified by hypertension, the CVD mortality risk in prediabetes plus hypertension subjects increased significantly compared with population without prediabetes and hypertension. Multivariate-adjusted hazard ratios for CVD mortality in prediabetes subgroups plus hypertension were 2.28 (95% CI: 1.50, 3.47) for those diagnosed by FPG < 5.6 mmol/L & HbA1c 5.7-6.4%, 2.18 (95% CI: 1.53, 3.10) for those diagnosed by FPG 5.6-6.0 mmol/L & HbA1c < 6.5% and 2.35 (95% CI: 1.65, 3.35) for those diagnosed by FPG 6.1-6.9 & HbA1c < 6.5% compared with the reference group. Moreover, the percentage of hypertension in prediabetes subjects was high (60.4%), but the awareness, treatment and control rates were far from satisfactory (45.3, 35.1 and 4.8%, respectively). Conclusions The prevalence of prediabetes remains high in northeast China, and the CVD mortality was elevated significantly in prediabetes coexistent with hypertension. Considering the high percentage and low control rate of hypertension in prediabetes, strategies focused on HbA1c screening, FPG lowering and blood pressure management should be emphasized in northeast China

Prevalence of atrial fibrillation and the risk of cardiovascular mortality among hypertensive elderly population in northeast China

Abstract Little is known about the epidemiology and impact of atrial fibrillation (AF) on cardiovascular diseases (CVD) mortality among hypertensive elderly population in northeast China. The community‐based study included 4497 hypertensive elderly residents aged ≥65 years who lived in northeast China from September 2017 to March 2019. Information on CVD deaths was obtained from baseline until July 31, 2021. Cox proportional hazard regression models were performed in the evaluation of CVD mortality. We identified 101 persons with AF. The prevalence of AF was 2.2% among elderly hypertensive population, which increased significantly with age. The prevalence of AF was higher in men than in women. The awareness rate was 51.5%, higher in urban areas than in rural areas (68.8% vs 43.5%, P = .018). Only 4.0% patients received oral anticoagulant (OAC) therapy among AF patients. Moreover, diabetes (26.7%) and dyslipidemia (37.6%) were highly prevalent in AF patients. Furthermore, 212 persons died due to CVD (14.7/1000 person‐years) during a median follow‐up of 3.2 years. AF patients had a 3.42 (95% CI: 2.07‐5.63) times higher risk of CVD mortality than the patients without AF in the fully adjusted model. Therefore, the burden of AF among hypertensive elderly population in northeast China was considerable. Long‐term screening and management strategies for AF and related risk factors are required among hypertensive elderly in northeast China

Recurrence Status and Combined Risk Factors of Surviving Stroke Patients Aged Over 40 in Urban Areas

Background Stroke has become the 3rd leading cause of death among urban residents in China, and the combined rate ofrisk factorssuch as hypertension, diabetes, and dyslipidemia is high among urban high-risk groups. Liaoning Province is a high incidence area of stroke in China, however, there are a few studies on the recurrence of stroke and its combined risk factors in the northern urban areas. Objective To explore the recurrence status of surviving stroke patients aged over 40 and its combined risk factors in urban areas of Liaoning Province, in orderto provide a theoretical basis for formulating effective secondary prevention strategies. Methods From September 2018 to December 2020, 6 349 permanent urban residents aged over 40 in Liaoyang, Dandong, Benxi, and Chaoyang Cities of Liaoning Province were selected according to the economic and regional distribution using stratified cluster random samplingand investigated. A total of 488 surviving stroke patients were included and their recurrence rate and associated risk factors were analyzed in this study. Results There were 244 males and 244 females among the 488 surviving stroke patients, the average age was (65.8±7.5) years old and the proportion of patients aged 60-69 was the largest (43.7%) ; There were 352 (72.1%) first-onset stroke patients, 136 (27.9%) recurrent stroke patients, and a recurrence rate of 27.9% among surviving stroke patients; The recurrence rates of male and female surviving stroke patients were 29.5% (72 cases) and 26.2% (64 cases) , respectively, and the difference was not statistically significant (&#x03C7;2=0.652, P=0.419) . The recurrence rate of stroke among people aged 40-49, 50-59, 60-69, 70-79 and ≥80 years old was 4/11, 23.4% (22/94) , 25.4% (54/213) , 35.9% (47/131) and 23.1% (9/39) , respectively, there was no significant difference in recurrence rate between male and female surviving stroke patients in different age groups (P&gt;0.05) .The current drinking rate of the relapsed patients was lower than that of the first-onset stroke patients, and the awareness rate of hypertension, dyslipidemia, the treatment rate of dyslipidemia, and the control rate of dyslipidemia were higher than those of the first-onset stroke patients, and the difference was statistically significant (P&lt;0.05) . Conclusion The recurrence rate of surviving stroke patients aged over 40 in urban areas of Liaoning Province is high, and patients with recurrence have higher rates of smoking, physical inactivity, overweight or obesity, and poor control of combined risk factors such as hypertension and diabetes mellitus, which require comprehensive and effective secondary prevention measures to reduce the risk of recurrence, disability and death in stroke patients

Image_5_Application of Toxoplasma gondii-specific SAG1, GRA7 and BAG1 proteins in serodiagnosis of animal toxoplasmosis.tif

Toxoplasmosis is a zoonotic disease caused by the obligate intracellular protozoan parasite T. gondii which is widely prevalent in humans and animals worldwide. The diagnosis of toxoplasmosis and distinguishing acute or chronic T. gondii infections have utmost importance for humans and animals. The TgSAG1, TgGRA7, and TgBAG1 proteins were used in the present study to develop the serological rSAG1-ELISA, rGRA7-ELISA and rBAG1-ELISA methods for the testing of T. gondii specific IgG and IgM antibodies and differentiating acute or chronic toxoplasmosis in 3733 animals, including Tibetan sheep, yaks, pigs, cows, cattle, horses, chickens, camels and donkeys from the Qinghai-Tibetan Plateau. The ELISA tests showed that the overall positivity of IgG antibody was 21.1% (786/3733), 15.3% (570/3733) and 18.2% (680/3733) for rSAG1-, rGRA7- and rBAG1-ELISA, respectively, and the positivity of IgM antibody was 11.8% (439/3733), 13.0% (486/3733) and 11.8% (442/3733) for rSAG1-, rGRA7- and rBAG1-ELISA, respectively. A total of 241 animals (6.5%) positive for all rSAG1-, rGRA7- and rBAG1-IgG were found in this study, and the 141 animals (3.8%) tested were anti-T. gondii IgM positive in all three ELISAs. Moreover, the 338, 284 and 377 animals were IgG positive in rSAG1 + rGRA7-, rBAG1 + rGRA7- and rSAG1 + rBAG1- ELISAs respectively, and the 346, 178 and 166 animals in rSAG1 + rGRA7-, rBAG1 + rGRA7- and rSAG1 + rBAG1-ELISAs were IgM positive respectively. The results confirmed that the application of SAG1, GRA7, and BAG1 recombinant antigens could successfully be used in the detection of specific IgG and IgM antibodies for distinguishing between acute or chronic T. gondii infections. It is inferred that the forms in which current animal species in the plateau area were infected with T. gondii, and the period of infection or the clinical manifestations of the current infections may be different. The present study provides substantial clinical evidence for the differential diagnosis of toxoplasmosis, and the classification of acute and chronic T. gondii infections.</p

Image_4_Application of Toxoplasma gondii-specific SAG1, GRA7 and BAG1 proteins in serodiagnosis of animal toxoplasmosis.tif

Toxoplasmosis is a zoonotic disease caused by the obligate intracellular protozoan parasite T. gondii which is widely prevalent in humans and animals worldwide. The diagnosis of toxoplasmosis and distinguishing acute or chronic T. gondii infections have utmost importance for humans and animals. The TgSAG1, TgGRA7, and TgBAG1 proteins were used in the present study to develop the serological rSAG1-ELISA, rGRA7-ELISA and rBAG1-ELISA methods for the testing of T. gondii specific IgG and IgM antibodies and differentiating acute or chronic toxoplasmosis in 3733 animals, including Tibetan sheep, yaks, pigs, cows, cattle, horses, chickens, camels and donkeys from the Qinghai-Tibetan Plateau. The ELISA tests showed that the overall positivity of IgG antibody was 21.1% (786/3733), 15.3% (570/3733) and 18.2% (680/3733) for rSAG1-, rGRA7- and rBAG1-ELISA, respectively, and the positivity of IgM antibody was 11.8% (439/3733), 13.0% (486/3733) and 11.8% (442/3733) for rSAG1-, rGRA7- and rBAG1-ELISA, respectively. A total of 241 animals (6.5%) positive for all rSAG1-, rGRA7- and rBAG1-IgG were found in this study, and the 141 animals (3.8%) tested were anti-T. gondii IgM positive in all three ELISAs. Moreover, the 338, 284 and 377 animals were IgG positive in rSAG1 + rGRA7-, rBAG1 + rGRA7- and rSAG1 + rBAG1- ELISAs respectively, and the 346, 178 and 166 animals in rSAG1 + rGRA7-, rBAG1 + rGRA7- and rSAG1 + rBAG1-ELISAs were IgM positive respectively. The results confirmed that the application of SAG1, GRA7, and BAG1 recombinant antigens could successfully be used in the detection of specific IgG and IgM antibodies for distinguishing between acute or chronic T. gondii infections. It is inferred that the forms in which current animal species in the plateau area were infected with T. gondii, and the period of infection or the clinical manifestations of the current infections may be different. The present study provides substantial clinical evidence for the differential diagnosis of toxoplasmosis, and the classification of acute and chronic T. gondii infections.</p

Image_1_Application of Toxoplasma gondii-specific SAG1, GRA7 and BAG1 proteins in serodiagnosis of animal toxoplasmosis.tif

Toxoplasmosis is a zoonotic disease caused by the obligate intracellular protozoan parasite T. gondii which is widely prevalent in humans and animals worldwide. The diagnosis of toxoplasmosis and distinguishing acute or chronic T. gondii infections have utmost importance for humans and animals. The TgSAG1, TgGRA7, and TgBAG1 proteins were used in the present study to develop the serological rSAG1-ELISA, rGRA7-ELISA and rBAG1-ELISA methods for the testing of T. gondii specific IgG and IgM antibodies and differentiating acute or chronic toxoplasmosis in 3733 animals, including Tibetan sheep, yaks, pigs, cows, cattle, horses, chickens, camels and donkeys from the Qinghai-Tibetan Plateau. The ELISA tests showed that the overall positivity of IgG antibody was 21.1% (786/3733), 15.3% (570/3733) and 18.2% (680/3733) for rSAG1-, rGRA7- and rBAG1-ELISA, respectively, and the positivity of IgM antibody was 11.8% (439/3733), 13.0% (486/3733) and 11.8% (442/3733) for rSAG1-, rGRA7- and rBAG1-ELISA, respectively. A total of 241 animals (6.5%) positive for all rSAG1-, rGRA7- and rBAG1-IgG were found in this study, and the 141 animals (3.8%) tested were anti-T. gondii IgM positive in all three ELISAs. Moreover, the 338, 284 and 377 animals were IgG positive in rSAG1 + rGRA7-, rBAG1 + rGRA7- and rSAG1 + rBAG1- ELISAs respectively, and the 346, 178 and 166 animals in rSAG1 + rGRA7-, rBAG1 + rGRA7- and rSAG1 + rBAG1-ELISAs were IgM positive respectively. The results confirmed that the application of SAG1, GRA7, and BAG1 recombinant antigens could successfully be used in the detection of specific IgG and IgM antibodies for distinguishing between acute or chronic T. gondii infections. It is inferred that the forms in which current animal species in the plateau area were infected with T. gondii, and the period of infection or the clinical manifestations of the current infections may be different. The present study provides substantial clinical evidence for the differential diagnosis of toxoplasmosis, and the classification of acute and chronic T. gondii infections.</p