Waterborne diseases are a major global problem, resulting in high morbidity and mortality, and massive economic costs. The ability to rapidly and reliably detect and monitor the spread of waterborne ...diseases is vital for early intervention and preventing more widespread disease outbreaks. Pathogens are, however, difficult to detect in water and are not practicably detectable at acceptable concentrations that need to be achieved in treated drinking water (which are of the order one per million litre). Furthermore, current clinical-based surveillance methods have many limitations such as the invasive nature of the testing and the challenges in testing large numbers of people. Wastewater-based epidemiology (WBE), which is based on the analysis of wastewater to monitor the emergence and spread of infectious disease at a population level, has received renewed attention in light of the current coronavirus disease 2019 (COVID-19) pandemic. The present review will focus on the application of WBE for the detection and surveillance of pathogens with a focus on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and the waterborne protozoan parasites
Cryptosporidium
and
Giardia.
The review highlights the benefits and challenges of WBE and the future of this tool for community-wide infectious disease surveillance.
The enteric parasite, Cryptosporidium is a major cause of diarrhoeal illness in humans and animals worldwide. No effective therapeutics or vaccines are available and therefore control is dependent on ...understanding transmission dynamics. The development of molecular detection and typing tools has resulted in the identification of a large number of cryptic species and genotypes and facilitated our understanding of their potential for zoonotic transmission. Of the 44 recognised Cryptosporidium species and >120 genotypes, 19 species, and four genotypes have been reported in humans with C. hominis, C. parvum, C. meleagridis, C. canis and C. felis being the most prevalent. The development of typing tools that are still lacking some zoonotic species and genotypes and more extensive molecular epidemiological studies in countries where the potential for transmission is highest are required to further our understanding of this important zoonotic pathogen. Similarly, whole-genome sequencing (WGS) and amplicon next-generation sequencing (NGS) are important for more accurately tracking transmission and understanding the mechanisms behind host specificity.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, UILJ, UKNU, UL, UM, UPUK
is an enteric parasite that is transmitted via the faecal-oral route, water and food. Humans, wildlife and domestic livestock all potentially contribute
to surface waters. Human encroachment into ...natural ecosystems has led to an increase in interactions between humans, domestic animals and wildlife populations. Increasing numbers of zoonotic diseases and spill over/back of zoonotic pathogens is a consequence of this anthropogenic disturbance. Drinking water catchments and water reservoir areas have been at the front line of this conflict as they can be easily contaminated by zoonotic waterborne pathogens. Therefore, the epidemiology of zoonotic species of
in free-ranging and captive wildlife is of increasing importance. This review focuses on zoonotic
species reported in global wildlife populations to date, and highlights their significance for public health and the water industry.
Eimeria
is an important coccidian enteric parasite that infects a wide range of hosts and can cause substantial economic losses in the poultry and livestock industries. It is common for multiple
...Eimeria
species to infect individual hosts, and this can make species identification difficult due to morphological similarities between species and mixed chromatograms when using Sanger sequencing. Relatively few studies have applied next-generation amplicon sequencing (NGS) to determining the genetic diversity of
Eimeria
species in different hosts. The present study screened 408 faecal samples from a range of hosts including livestock and wildlife using a previously developed quantitative polymerase chain reaction (qPCR) at the 18S locus and conducted amplicon NGS on the positives using a ~ 455-bp fragment of the 18S locus. A total of 41 positives (10.1%) were identified by qPCR from various hosts and NGS was successful for 38 of these positives. Fifteen
Eimeria
species and three genotypes were detected by NGS:
E. ferrisi
,
E. kanyana
,
E. potoroi
,
E. quokka
,
E. setonicis
,
E. trichosuri
,
E. reichenowi
,
E. angustus
,
E. ahsata
,
E. auburnensis
,
E. bovis
,
E. brasiliensis
,
E. christenseni
,
E. crandallis
,
E. ovinoidalis
,
Eimeria
sp. (JF419345),
Eimeria
sp. (JF419349) and
Eimeria
sp. (JF419351). Mixed infections were detected in 55.3% (21/38) of positive samples. The most striking finding was the identification of the same species in different hosts. This could be due to contamination and/or mechanical transmission or may provide support for previous studies suggesting that
Eimeria
species can infect not just closely related hosts but different genera and further research is required. This is also the first study to audit
Eimeria
populations in livestock (sheep and cattle) by NGS and could be applied in the future to determine the extent of pathogenic species and outcomes of
Eimeria
control strategies.
Recycled wastewater can carry human-infectious microbial pathogens and therefore wastewater treatment strategies must effectively eliminate pathogens before recycled wastewater is used to supplement ...drinking and agricultural water supplies. This study characterised the bacterial composition of four wastewater treatment plants (WWTPs) (three waste stabilisation ponds and one oxidation ditch WWTP using activated sludge treatment) in Western Australia. The hypervariable region 4 (V4) of the bacterial 16S rRNA (16S) gene was sequenced using next-generation sequencing (NGS) on the Illumina MiSeq platform. Sequences were pre-processed in USEARCH v10.0 and denoised into zero-radius taxonomic units (ZOTUs) with UNOISE3. Taxonomy was assigned to the ZOTUs using QIIME 2 and the Greengenes database and cross-checked with the NCBI nr/nt database. Bacterial composition of all WWTPs and treatment stages (influent, intermediate and effluent) were dominated by Proteobacteria (29.0–87.4%), particularly Betaproteobacteria (9.0–53.5%) and Gammaproteobacteria (8.6–34.6%). Nitrifying bacteria (Nitrospira spp.) were found only in the intermediate and effluent of the oxidation ditch WWTP, and denitrifying and floc-forming bacteria were detected in all WWTPs, particularly from the families Comamonadaceae and Rhodocyclales. Twelve pathogens were assigned taxonomy by the Greengenes database, but comparison of sequences from genera and families known to contain pathogens to the NCBI nr/nt database showed that only three pathogens (Arcobacter venerupis, Laribacter hongkongensis and Neisseria canis) could be identified in the dataset at the V4 region. Importantly, Enterobacteriaceae genera could not be differentiated. Family level taxa assigned by Greengenes database agreed with NCBI nr/nt in most cases, however, BLAST analyses revealed erroneous taxa in Greengenes database. This study highlights the importance of validating taxonomy of NGS sequences with databases such as NCBI nr/nt, and recommends including the V3 region of 16S in future short amplicon NGS studies that aim to identify bacterial enteric pathogens, as this will improve taxonomic resolution of most, but not all, Enterobacteriaceae species.
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•Waste water samples were screened with bacterial 16S next-generation sequencing.•The V4 region of 16S could not differentiate Enterobacteriaceae.•Only three pathogens could be identified to the species level.•Erroneous taxa in the 16S Greengenes database were identified.•NCBI nr/nt database comparisons provided more accurate taxonomic assignments.
Cryptosporidiosis, caused by parasite species of the genus Cryptosporidium, is a major diarrhoeal disease in both people and animals globally, with C. hominis and C. parvum the main species infecting ...humans. Environmentally robust oocysts which are shed in high numbers in the faeces of infected individuals are resistant to disinfectants, including levels of chlorine normally used in drinking water. As a result, Cryptosporidium is a major cause of waterborne and foodborne outbreaks. Interestingly, C. hominis is responsible for the majority of waterborne outbreaks typed to date with C. parvum responsible for the majority of foodborne outbreaks. No vaccine and few treatments are currently available, which has greatly limited control of this disease to date. Livestock are both an important reservoir and source of human infections and improved husbandry and management practices as well as a One Health integrated molecular typing approach across both veterinary and public health systems are essential to improve our ability to control this disease.
•Update on waterborne and foodborne outbreaks caused by Cryptosporidium over the last 10 years.•Impact of cryptosporidiosis on livestock.•Evidence for livestock as a source of human infections.•Importance of a One Health integrated molecular typing approach to limit transmission.
Repairing and replacing of tumor tissues have been grown due to the genetic changes and environmental factors. Conventional cancer therapies apply the systemic approaches for delivery of anticancer ...agents in which normal and cancer cells are not discriminated. Therefore, the risk of cancer recurrence in the resection site increases. In this paper, we studied a graphene reinforced gelatin hydrogel for tissue engineering and local controlled release of chemotherapeutic agents. Protein-integrated graphene (PIG) synthesized by ultrasonic-supported technique was incorporated into gelatin matrix, where the swelling ratio of hydrogels decreased with increasing the PIG concentration. In hydrogels, doxorubicin (DOX) was released in a controlled and pH-sensitive manner and the release rate was controlled by PIG concentration. The effect of PIG on the controlled release system was detected in MCF-7 cell viability. Moreover, 3D tumor spheroid test studied the extracellular matrix (ECM) and cell aggregation as an in vivo model. Generally, we introduced a novel hydrogel nanocomposite, which is a suitable candidate for post-surgery treatment and localized therapy.
The Bellinger River snapping turtle (
Myuchelys georgesi
) is endemic to Australia and is confined to a highly restricted distribution in the Bellinger River in New South Wales. Routine veterinary ...health examinations of 17 healthy turtles were undertaken, along with the collection and analysis of blood samples, during conservation efforts to save the species following a catastrophic population decline. Microscopy analysis of blood films detected Haemoproteidae parasites that morphologically resembled
Haemocystidium chelodinae
inside turtle erythrocytes. Of the 17 turtles examined, 16 were positive for infection with
H. chelodinae
by both light microscopy and PCR. DNA sequencing of a partial fragment of the mitochondrial cytochrome b (
cytb
) gene and phylogenetic analysis identified two different
H. chelodinae-
like genotypes. The phylogenetic relationship of
H. chelodinae
-like to other Haemoproteidae species based on
cytb
sequences grouped
H. chelodinae
-like into the reptile clade, but revealed the
Haemocystidium
genus to be paraphyletic as the clade also contained
Haemoproteus
, thus supporting a re-naming of
Haemoproteus
species from reptiles to
Haemocystidium
species
.
This study reports for the first time the genetic characterisation of
H. chelodinae
-like organisms isolated from a new Testudine host species, the Bellinger River snapping turtle. As evidence grows, further research will be necessary to understand the mode of transmission and to investigate whether these parasites are pathogenic to their hosts.
Cryptosporidium is one of the most common zoonotic waterborne parasitic diseases worldwide and represents a major public health concern of water utilities in developed nations. As animals in ...catchments can shed human-infectious Cryptosporidium oocysts, determining the potential role of animals in dissemination of zoonotic Cryptosporidium to drinking water sources is crucial. In the present study, a total of 952 animal faecal samples from four dominant species (kangaroos, rabbits, cattle and sheep) inhabiting Sydney's drinking water catchments were screened for the presence of Cryptosporidium using a quantitative PCR (qPCR) and positives sequenced at multiple loci. Cryptosporidium species were detected in 3.6% (21/576) of kangaroos, 7.0% (10/142) of cattle, 2.3% (3/128) of sheep and 13.2% (14/106) of rabbit samples screened. Sequence analysis of a region of the 18S rRNA locus identified C. macropodum and C. hominis in 4 and 17 isolates from kangaroos respectively, C. hominis and C. parvum in 6 and 4 isolates respectively each from cattle, C. ubiquitum in 3 isolates from sheep and C. cuniculus in 14 isolates from rabbits. All the Cryptosporidium species identified were zoonotic species with the exception of C. macropodum. Subtyping using the 5' half of gp60 identified C. hominis IbA10G2 (n = 12) and IdA15G1 (n = 2) in kangaroo faecal samples; C. hominis IbA10G2 (n = 4) and C. parvum IIaA18G3R1 (n = 4) in cattle faecal samples, C. ubiquitum subtype XIIa (n = 1) in sheep and C. cuniculus VbA23 (n = 9) in rabbits. Additional analysis of a subset of samples using primers targeting conserved regions of the MIC1 gene and the 3' end of gp60 suggests that the C. hominis detected in these animals represent substantial variants that failed to amplify as expected. The significance of this finding requires further investigation but might be reflective of the ability of this C. hominis variant to infect animals. The finding of zoonotic Cryptosporidium species in these animals may have important implications for the management of drinking water catchments to minimize risk to public health.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Apicomplexan tick-borne pathogens that cause disease in companion animals include species of Babesia Starcovici, 1893, Cytauxzoon Neitz & Thomas, 1948, Hepatozoon Miller, 1908 and Theileria ...Bettencourt, Franca & Borges, 1907. The only apicomplexan tick-borne disease of companion animals that is known to occur in Australia is babesiosis, caused by Babesia canis vogeli Reichenow, 1937 and Babesia gibsoni Patton, 1910. However, no molecular investigations have widely investigated members of Apicomplexa Levine, 1980 in Australian ticks that parasitise dogs, cats or horses, until this present investigation.
Ticks (n = 711) removed from dogs (n = 498), cats (n = 139) and horses (n = 74) throughout Australia were screened for piroplasms and Hepatozoon spp. using conventional PCR and Sanger sequencing. The tick-borne pathogen B. vogeli was identified in two Rhipicephalus sanguineus Latreille ticks from dogs residing in the Northern Territory and Queensland (QLD). Theileria orientalis Yakimov & Sudachenkov, 1931 genotype Ikeda was detected in three Haemaphysalis longicornis Neumann ticks from dogs in New South Wales. Unexpectedly, the exotic tick-borne pathogen Hepatozoon canis James, 1905 was identified in an Ixodes holocyclus Neumann tick from a dog in QLD. Eight novel piroplasm and Hepatozoon species were identified and described in native ticks and named as follows: Babesia lohae n. sp., Babesia mackerrasorum n. sp., Hepatozoon banethi n. sp., Hepatozoon ewingi n. sp., Theileria apogeana n. sp., Theileria palmeri n. sp., Theileria paparinii n. sp. and Theileria worthingtonorum n. sp. Additionally, a novel cf. Sarcocystidae sp. sequence was obtained from Ixodes tasmani Neumann but could not be confidently identified at the genus level.
Novel species of parasites in ticks represent an unknown threat to the health of companion animals that are bitten by these native tick species. The vector potential of Australian ticks for the newly discovered apicomplexans needs to be assessed, and further clinical and molecular investigations of these parasites, particularly in blood samples from dogs, cats and horses, is required to determine their potential for pathogenicity.
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