Recently concerns are increasing that dirofilarial nematodes may spread from endemic areas in sou... more Recently concerns are increasing that dirofilarial nematodes may spread from endemic areas in southern, eastern and central Europe to countries in northern regions of Europe. The increasing number of autochthonous cases of canine Dirofilaria repens infections in Germany indicates that worms of this genus may invade new areas, and climate change may be a key factor in this scenario. Thus analysis of long term development of regional temperature is a pivotal factor in risk analysis related to transmission of these worms. Such information is important for suggestions of counteracting strategies, such as definition of periods of increased transmission risk and, consequently, time slots most suited for preventative measures. In this study, mean daily temperature data from 34 geographical clustered weather stations representing all parts of Germany were analyzed. It is concluded that the increasing trend for average daily temperatures observed in the period from 1984 to 2013 has led to cl...
Parasites are a common threat to human and animal health. One option to combat parasites that pro... more Parasites are a common threat to human and animal health. One option to combat parasites that produce infective environmental stages is to inactivate them by chemical disinfection. Standardised laboratory assays that enable proper evaluation of products suspected to be efficient are highly desirable to allow prudent selection and use of such potentially hazardous agents. Here, we present a newly developed in vitro germ carrier assay to evaluate inactivation of oocysts of the model organism Cryptosporidium parvum by chemical disinfectants. Stainless steel discs were used as carrier to mimic surface contamination by C. parvum oocysts. The germ carriers were incubated with approved chemical disinfectant for the specified time (2 h) and rinsed thereafter to remove the disinfectant and recover the exposed oocysts. Recovered oocysts were transferred to HCT-8 monolayers, and 48 h later, genomic DNA was extracted and quantified by real-time PCR targeting the hsp70 gene to estimate parasite reproduction. A panel of commercially available and approved disinfectants were examined and data compared with those of suspension assays and historical data obtained from efficacy assays based on infection of chicken with oocysts of Eimeria tenella. Altogether, data achieved by these divergent assays allowed similar conclusions although the sensitivity of the in vitro assay was higher. Consequently, a threshold of 99.5% inactivation is proposed to evaluate disinfectants in vitro using C. parvum as model organism as compared to the E. tenella animal infection assay (95%).
To analyze the potential of RNA interference (RNAi) in the intestinal nematode Heligmosomoides po... more To analyze the potential of RNA interference (RNAi) in the intestinal nematode Heligmosomoides polygyrus, we delivered double-stranded RNA (dsRNA) of tropomyosin to various life stages of the parasite. Three different methods were examined for their potential use. First, feeding of recombinant bacteria that expressed dsRNA did neither result in phenotypical changes of H. polygyrus nor in a significant reduction of tropomyosin mRNA levels. In contrast, feeding of such bacteria to Caenorhabditis elegans elicited the expected phenotypes. Quantification of bacteria ingested by C. elegans and H. polygyrus larvae (L1) revealed that the parasitic worms took up only a fraction of the bacteria ingested by C. elegans. Second, electroporation of L1 failed to transport siRNA through the cuticle and was lethal to the larvae. However, the cuticle of adult worms was penetrated by dye-labeled RNA, but no systemic spreading was observed. Third, soaking of adult H. polygyrus in tropomyosin dsRNA led to a higher proportion of worms showing symptoms of ageing, such as a disintegrated gut and ovaries, but did not induce reduction of tropomyosin mRNA levels. Database analysis revealed that orthologous proteins involved in dsRNA-uptake and -systemic spread in C. elegans are missing in the parasitic nematodes Brugia malayi and Haemonchus contortus, whereas proteins responsible for dsRNA-processing, -amplification and mRNA-regulation are present. Thus, our data indicate that the study of gene function by RNAi in H. polygyrus is limited, possibly due to deficiencies of genes involved in RNA-uptake and spread.
Developmental and comparative immunology, Jan 27, 2015
Cryptosporidium parvum causes a zoonotic infection with worldwide distribution. Besides humans, c... more Cryptosporidium parvum causes a zoonotic infection with worldwide distribution. Besides humans, cryptosporidiosis affects a wide range of animals leading to significant economic losses due to severe enteritis in neonatal livestock. Neutrophil extracellular trap (NET) formation has been demonstrated as an important host effector mechanism of PMN acting against several invading pathogens. In the present study, C. parvum-mediated NET formation was investigated in human and bovine PMN in vitro. We here demonstrate that C. parvum sporozoites indeed trigger NET formation in a time-dependent manner. Thereby, the classical characteristics of NETs were demonstrated by co-localization of extracellular DNA with histones, neutrophil elastase (NE) and myeloperoxidase (MPO). A significant reduction of NET formation was measured following treatments of PMN with NADPH oxidase-, NE- and MPO-inhibitors, confirming the key role of these enzymes in C. parvum-induced NETs. Additionally, sporozoite-trigg...
Recently concerns are increasing that dirofilarial nematodes may spread from endemic areas in sou... more Recently concerns are increasing that dirofilarial nematodes may spread from endemic areas in southern, eastern and central Europe to countries in northern regions of Europe. The increasing number of autochthonous cases of canine Dirofilaria repens infections in Germany indicates that worms of this genus may invade new areas, and climate change may be a key factor in this scenario. Thus analysis of long term development of regional temperature is a pivotal factor in risk analysis related to transmission of these worms. Such information is important for suggestions of counteracting strategies, such as definition of periods of increased transmission risk and, consequently, time slots most suited for preventative measures. In this study, mean daily temperature data from 34 geographical clustered weather stations representing all parts of Germany were analyzed. It is concluded that the increasing trend for average daily temperatures observed in the period from 1984 to 2013 has led to cl...
Parasites are a common threat to human and animal health. One option to combat parasites that pro... more Parasites are a common threat to human and animal health. One option to combat parasites that produce infective environmental stages is to inactivate them by chemical disinfection. Standardised laboratory assays that enable proper evaluation of products suspected to be efficient are highly desirable to allow prudent selection and use of such potentially hazardous agents. Here, we present a newly developed in vitro germ carrier assay to evaluate inactivation of oocysts of the model organism Cryptosporidium parvum by chemical disinfectants. Stainless steel discs were used as carrier to mimic surface contamination by C. parvum oocysts. The germ carriers were incubated with approved chemical disinfectant for the specified time (2 h) and rinsed thereafter to remove the disinfectant and recover the exposed oocysts. Recovered oocysts were transferred to HCT-8 monolayers, and 48 h later, genomic DNA was extracted and quantified by real-time PCR targeting the hsp70 gene to estimate parasite reproduction. A panel of commercially available and approved disinfectants were examined and data compared with those of suspension assays and historical data obtained from efficacy assays based on infection of chicken with oocysts of Eimeria tenella. Altogether, data achieved by these divergent assays allowed similar conclusions although the sensitivity of the in vitro assay was higher. Consequently, a threshold of 99.5% inactivation is proposed to evaluate disinfectants in vitro using C. parvum as model organism as compared to the E. tenella animal infection assay (95%).
To analyze the potential of RNA interference (RNAi) in the intestinal nematode Heligmosomoides po... more To analyze the potential of RNA interference (RNAi) in the intestinal nematode Heligmosomoides polygyrus, we delivered double-stranded RNA (dsRNA) of tropomyosin to various life stages of the parasite. Three different methods were examined for their potential use. First, feeding of recombinant bacteria that expressed dsRNA did neither result in phenotypical changes of H. polygyrus nor in a significant reduction of tropomyosin mRNA levels. In contrast, feeding of such bacteria to Caenorhabditis elegans elicited the expected phenotypes. Quantification of bacteria ingested by C. elegans and H. polygyrus larvae (L1) revealed that the parasitic worms took up only a fraction of the bacteria ingested by C. elegans. Second, electroporation of L1 failed to transport siRNA through the cuticle and was lethal to the larvae. However, the cuticle of adult worms was penetrated by dye-labeled RNA, but no systemic spreading was observed. Third, soaking of adult H. polygyrus in tropomyosin dsRNA led to a higher proportion of worms showing symptoms of ageing, such as a disintegrated gut and ovaries, but did not induce reduction of tropomyosin mRNA levels. Database analysis revealed that orthologous proteins involved in dsRNA-uptake and -systemic spread in C. elegans are missing in the parasitic nematodes Brugia malayi and Haemonchus contortus, whereas proteins responsible for dsRNA-processing, -amplification and mRNA-regulation are present. Thus, our data indicate that the study of gene function by RNAi in H. polygyrus is limited, possibly due to deficiencies of genes involved in RNA-uptake and spread.
Developmental and comparative immunology, Jan 27, 2015
Cryptosporidium parvum causes a zoonotic infection with worldwide distribution. Besides humans, c... more Cryptosporidium parvum causes a zoonotic infection with worldwide distribution. Besides humans, cryptosporidiosis affects a wide range of animals leading to significant economic losses due to severe enteritis in neonatal livestock. Neutrophil extracellular trap (NET) formation has been demonstrated as an important host effector mechanism of PMN acting against several invading pathogens. In the present study, C. parvum-mediated NET formation was investigated in human and bovine PMN in vitro. We here demonstrate that C. parvum sporozoites indeed trigger NET formation in a time-dependent manner. Thereby, the classical characteristics of NETs were demonstrated by co-localization of extracellular DNA with histones, neutrophil elastase (NE) and myeloperoxidase (MPO). A significant reduction of NET formation was measured following treatments of PMN with NADPH oxidase-, NE- and MPO-inhibitors, confirming the key role of these enzymes in C. parvum-induced NETs. Additionally, sporozoite-trigg...
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Papers by Matthias Lendner