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Host genetic variability can modulate infection resistance, although its role in infection clearance remains unclear. Hookworm disease ( Uncinaria sp.) is the leading cause of pup mortality in several otariid species, although the parasite can be cleared through immune-mediated processes. We evaluated the association of host genetic diversity, body condition and immune response with hookworm resistance and/or clearance in the South American fur seal ( Arctocephalus australis ). Uninfected pups had higher heterozygosity than parasitized individuals, indicating a negative relationship between heterozygosity and the chances of infection. Likewise, pups that died of hookworm infection had lower heterozygosity than those that died of non-infectious causes. Interestingly, once infected, pups that survived hookworm infection had heterozygosities similar to pups that died of hookworm disease. However, pups that cleared the infection had a higher body mass and parasite-specific immunoglobulin G levels than those that did not recover or died of hookworm disease. Thus, although heterozygosity predicted resistance to and mortality from hookworm infections, it did not affect parasite clearance, which was facilitated by better body condition and adaptive immune responses. This demonstrates that host genetic variability and host-environment interactions influence disease dynamics, acting at different, well-defined stages of infection.

The present study aims to assess the performance of different molecular targets using various matrices of samples for the detection of Uncinaria stenocephala (US) in hookworm infected dogs. To this end, the DNA extraction was performed on the following matrices of samples: (i) larvae of US obtained from experimentally infected dogs with US with different larvae counts per microliter (µl); (ii) pure US eggs suspension in distilled water with different egg counts per µl; (iii) spiked dog fecal samples with different US eggs per gram (EPG) of feces; (iv) feces from dogs naturally infected with hookworm eggs; (v) fecal suspension with hookworm eggs recovered from the FLOTAC apparatus. All the samples were tested with four different PCR protocols targeting specific regions for the detection of both hookworms US and AC as follows: Protocol A (ITS1, 5.8 S, ITS2) and Protocol B (18 S) for the detection of both species, Protocol C (ITS1) for the detection of AC and Protocol D (ITS1) for the detection of US. The best results were obtained with DNA extracted from US larvae matrix obtained from experimentally infected dogs, showing a detection limit of 3.5 larvae/ml for the protocols A, B and D. A moderate correlation was found between the FLOTAC technique and PCR protocols B and D with respect to fecal samples from dogs naturally infected with hookworms. Indeed, PCR protocols B (18 S) and D (ITS1) gave the best results for feces and fecal suspension from naturally infected dogs. However, all the PCR protocols used showed lower sensitivity than FLOTAC technique. Perhaps, isolating US eggs in advance could help to obtain better quality and quantity of DNA, avoiding some notable factors such as inhibitors present in faecal samples. However, a further study is needed to evaluate and standardise a protocol for the recovery of parasitic elements, that could be applied prior to DNA extraction. Therefore, this could lead to a better amplification of US eggs DNA. In conclusion, our results showed that the type of sample (sample-matrix) used for the DNA extraction samples is crucial, as this affects the diagnostic sensitivity of the technique., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Lavinia Ciuca reports was provided by University of Naples Federico II. Lavinia Ciuca reports a relationship with University of Naples Federico II that includes: employment. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper, (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Helminthiasis due to hookworm infestations in humans and Haemonchus contortus in sheep cause untold levels of disease and economic losses worldwide. Drug resistance is an ever-growing problem with pathogenic helminths. Thus, there is a critical need for new treatment strategies for hookworms and H. contortus that depends on animal models. Because hook- worms and H. contortus are obligate parasites, they can only be maintained using live animals. This review describes use of the Syrian golden hamster to propagate hookworms and Mongolian gerbil to propagate H. contortus .

Recent studies demonstrated that Greyhounds are commonly infected with Ancylostoma caninum and these infections have been shown to be resistant to anthelmintics. This study evaluated samples submitted to a commercial reference laboratory (IDEXX Laboratories) for canine fecal flotation zinc sulfate centrifugation and coproantigen immunoassay between January 1, 2019, and July 30, 2023 for evidence that Greyhounds were more often positive for Ancylostoma spp. (hookworms) compared to other breeds. The purpose of the study was to determine if Greyhounds were more likely to be hookworm-positive compared to other breeds, if Greyhounds on preventives with efficacy against hookworm infections are more likely to test positive than other breeds, if their infections take longer to resolve, to estimate how long this takes and to assess whether the proportion of hookworm positive tests for all breeds is increasing over time. Records of 25,440,055 fecal results were obtained representing 17,671,724 unique dogs. Of these, 49,795 (∼0.3%) were Greyhounds. The overall odds ratio (OR) of 15.3 (p < 0.001) suggests that Greyhounds are at significantly higher risk than other breeds for hookworm positive float findings, and the OR of 14.3 (p < 0.001) suggests significantly higher risk for hookworm antigen positive results. The median time to negative testing event from the Turnbull distribution estimate was in the interval of 1-2 days for other breeds and 71-72 days for Greyhounds. These results provide evidence that anthelmintic resistant A. caninum strains may be having population-level impacts on the frequency and duration of infections in Greyhounds. The findings have broader health implications beyond Greyhounds as MADR A. caninum strains could spread to other breeds and even pet owners., Competing Interests: Declaration of competing interest The study was funded by IDEXX Laboratories, Inc. Westbrook, ME, USA., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Hookworms are blood-feeding nematodes that infect millions of people and animals worldwide. The most common species infecting dogs in Europe are representatives of the family Ancylostomatidae, which differ in invasiology, biology and morphological features. However, the differentiation of invasions of species such as Uncinaria stenocephala and Ancylostoma caninum based on a coproscopic examination is problematic. For this reason, it is recommended to use molecular diagnostics for this purpose. The authors of the article investigated the prevalence of U. stenocephala in dogs kept in various living conditions in Slovakia and developed a two-step morphology-molecular analysis-based strategy to identify the genus and the species of eggs and larvae of the Ancylostomatidae family in dogs., Conclusions: In our opinion, this work is very much needed as it shows how to effectively diagnose hookworm infestations. However, we do not agree with the information in the title of the article because such studies have already been carried out in Central Europe by other authors., (© 2023. The Author(s).)

Hookworms are the most common intestinal nematode parasites of dogs in Australia. The control of these parasites relies mostly on regular deworming with anthelmintics, with pyrantel-based dewormers being a relatively low cost and readily-available option for dog owners. Pyrantel resistance in canine hookworms in Australia was first reported in 2007, however pyrantel-based dewormers are still used against hookworm infection in dogs across Australia. The present study was conducted to evaluate the efficacy of pyrantel against hookworms infecting dogs housed in a shelter facility in Southeast Queensland which receives rescued or surrendered animals from greyhound rescue centres and dog shelters across this region. A total of 10 dogs were examined using the faecal egg count reduction test (FECRT). There was no reduction in FEC in any of the dogs following pyrantel treatment, with drug efficacies ranging from -0.9% to -283.3%. Given that these dogs originated from various sites across Southeast Queensland, the present study suggests that pyrantel resistance is widespread in this region, and hence this anthelmintic may not be a useful option for treatment of hookworm infections in dogs., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:, (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Hookworm infections remain a significant public health concern in tropical and subtropical regions, including Thailand. This study investigated the species and genetic diversity of hookworm infections in domestic dogs from northeastern Thailand. The molecular analysis focused on amplifying and sequencing specific regions of ribosomal RNA genes (ITS1-5.8S-ITS2 region) and the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene in hookworm larvae recovered from 21 domestic dog stool samples. Among 21 larvae (one larva per infected dog) analyzed, 14 had sequences identical to Ancylostoma caninum, and 7 showed sequences almost identical to Ancylostoma ceylanicum. Phylogenetic analysis of cox1 sequences placed A. caninum and A. ceylanicum in separate clades. The median-joining network of A. caninum cox1 sequences from Thailand showed high haplotype diversity and belonged to the same cluster as sequences from Australia while forming separate clusters from those of A. caninum samples from the USA. The available published A. ceylanicum cox1 sequences (n = 33), in combination with seven sequences in the present study, represented 15 haplotypes distributed among three clusters. Interestingly, A. ceylanicum sequences from dogs and humans shared the same haplotypes. These findings are crucial for recognizing the potential for zoonotic transmission, highlighting the necessity for targeted control measures, and increasing awareness among pet owners and healthcare professionals to mitigate the risk of hookworm transmission to humans., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Hookworm infection affects millions globally, leading to chronic conditions like malnutrition and anaemia. Among the hookworm species, Ancylostoma ceylanicum stands out as a generalist, capable of infecting various hosts, including humans, cats, dogs and hamsters. Surprisingly, it cannot establish in mice, despite their close phylogenetic relationship to hamsters. The present study investigated the development of A. ceylanicum in immunodeficient NSG mice to determine the contribution of the immune system to host restriction. The infections became patent on day 19 post-infection (PI) and exhibited elevated egg production which lasted for at least 160 days PI. Infective A. ceylanicum larvae reared from eggs released by infected NSG mice were infectious to hamsters and capable of reproduction, indicating that the adults in the NSG mice were producing viable offspring. In contrast, A. ceylanicum showed limited development in outbred Swiss Webster mice. Furthermore, the closely related canine hookworm Ancylostoma caninum was unable to infect and develop in NSG mice, indicating that different mechanisms may determine host specificity even in closely related species. This is the first report of any hookworm species completing its life cycle in a mouse and implicate the immune system in determining host specificity in A. ceylanicum .

Background: The zoonotic hookworms Ancylostoma caninum and Uncinaria stenocephala are widespread soil-transmitted helminths in dogs in Europe. Given the veterinary and public health importance of hookworms in dogs and the recent changes in the molecular epidemiology of some species, there is a need to continuously monitor the epidemiological and molecular prevalence of these parasites also at the "local" level. The present study aimed to update the epidemiological scenario of hookworm infections in both owned and stray dogs in southern Italy and to discriminate between different hookworm species (A. caninum and U. stenocephala) through molecular analyses. For this purpose, a retrospective analysis was performed over 10 years (2011-2021), including a total of 7008 owned dogs and 5642 stray dogs referred to our laboratory for copromicroscopic examinations. Moreover, 72 faecal samples, from dogs naturally infected by hookworms, were used to discriminate between A. caninum and U. stenocephala using two PCR protocols. Prior to molecular analyses, a subsample of 40/72 positive faecal samples was used for morphometric investigations on hookworm eggs., Results: The results of the ten-year retrospective analysis (2011-2021) showed an overall prevalence of hookworm infection of 9.16%, specifically 5.1% in owned dogs and 14.2% in stray dogs. Logistic regression showed a significant association between positivity to hookworms and the variable "puppies" both in stray (13.84%; OR = 2.4) and owned (7.07%; OR = 2.2) dogs. The results of molecular analyses showed that positivity was confirmed only in 21/72 samples, specifically, 6 samples using protocol A and 19 with protocol B. Sequencing revealed 15 samples positive to U. stenocephala and 6 to A. caninum., Conclusions: The findings of this study showed a high prevalence of hookworm infections in dogs in southern Italy, updating the epidemiological scenario of the last decade. Moreover, the results of the study revealed the first identification of hookworm species in dogs in Italy by molecular studies, highlighting that U. stenocephala is more prevalent than A. caninum., (© 2023. BioMed Central Ltd., part of Springer Nature.)

Hookworms (Ancylostomatidae) are well-known parasites in dogs due to their health impacts and zoonotic potential. While faecal analysis is the traditional method for detection, improvements in husbandry and deworming have decreased their prevalence in urban owned dogs. Drug resistance in Ancylostoma caninum is becoming a discussion point in small animal practices across the region. This study aimed to identify hookworm species present in Australian and New Zealand dogs using molecular techniques. The ITS-2 and isotype-1 β-tubulin assays were used to identify and quantify hookworm species. Results showed absence of coinfection in Australian samples from Greater Sydney region belonging either to A. caninum or Uncinaria stenocephala, while New Zealand samples were a mixture of A. caninum and U. stenocephala. The amplified isotype-1 β-tubulin sequences exhibited susceptibility to benzimidazole drugs. Rare mutations were identified in A. caninum and U. stenocephala sequences, representing a small percentage of reads. This study highlights the importance of molecular techniques in accurately identifying and quantifying hookworm species in dog populations., (© 2023 The Authors. Australian Veterinary Journal published by John Wiley & Sons Australia, Ltd on behalf of Australian Veterinary Association.)

Canine hookworms are found globally, and infections have been recorded in domestic dogs visiting dog parks in Queensland, Australia. Some canine hookworms also present a risk of zoonotic transmission to humans. Potential transmission of hookworms can occur in the household and at public places because of lack of owner awareness and poor coverage of canine deworming. Between April 2019 and March 2020, faecal samples from owned dogs were collected from 39 dog parks in metropolitan Brisbane, Queensland, and tested for the presence of hookworm eggs using faecal floatation technique. Dog owners who provided samples were requested to complete a survey on their awareness, perceptions and behaviour regarding the risks and consequences of canine parasitic infection. Associations between dog owner demographics and responses to the survey were measured using Goodman and Kruskal's gamma. Statistical associations between canine hookworm infection in dogs and their owner's responses were quantified using a two-sample Wilcoxon rank-sum test. A total of five canine faecal samples from 175 respondents were found positive for hookworm eggs, corresponding to a sample- and park-level prevalence of 2.86% and 12.82%, respectively. Female dog owners were found to be more aware of the importance of regular deworming of their dogs (|γ| = 0.405). Our results indicate that hookworm infection was associated with a lower awareness of the importance of deworming (p = 0.007), less diligence in administration of deworming (p = 0.004), lower awareness of the risk of acquiring parasites from raw meat (p = 0.010), less likelihood of cooking meat before feeding it to their dogs (p = 0.028), and less likelihood to properly dispose their dog's faeces (p = 0.027). This study not only indicates a need for improving owner education towards the importance of deworming but also a need for changing owner's behaviours to reduce potential environmental contamination with infective hookworms. The latter in particular indicates a broader public health risk of disease transmission in public places due to improper disposal of dog faeces, especially in areas with higher canine hookworm infections., (© 2023 The Authors. Zoonoses and Public Health published by Wiley-VCH GmbH.)

Hookworms (genera Ancylostoma and Necator) are amongst the most prevalent and important parasites of humans globally. These intestinal parasites ingest blood, resulting in anemia, growth stunting, malnutrition, and adverse pregnancy outcomes. They are also critical parasites of dogs and other animals. In addition, hookworms and hookworm products are being explored for their use in treatment of autoimmune and inflammatory diseases. There is thus a significant and growing interest in these mammalian host-obligate parasites. Laboratory research is hampered by the lack of good means of cryopreservation and recovery of parasites. Here, we describe a robust method for long-term (≥3 year) cryopreservation and recovery of both Ancylostoma and Necator hookworms that is also applicable to two other intestinal parasites that passage through the infective L3 stage, Strongyloides ratti and Heligmosomoides polygyrus bakeri. The key is a revised recovery method, in which cryopreserved L1s are thawed and raised to the infective L3 stage using activated charcoal mixed with uninfected feces from a permissive host. This technique will greatly facilitate research on and availability of gastrointestinal parasitic nematodes with great importance to global health, companion animal health, and autoimmune/inflammatory disease therapies., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Detection and identification standard of hookworm-Hookworm larvae coproculture techniques ( WS/T 791-2021 ) is the first recommended technical standard for hookworm detection and species identification using the hookworm larvae coproculture technique in China. This standard was issued on November 23, 2021, and had been in effect since May 1, 2022. This article provides a detailed interpretation pertaining to the background, drafting process, main contents, and dos and don'ts for better understanding and application of this standard among professionals working in disease control and prevention institutions and medical institutions.

The canine hookworm Ancylostoma caninum is one of the most prevalent parasitic nematodes in dogs worldwide and has the potential for zoonotic transmission to humans, including the development of cutaneous larva migrans. Recent confirmation of anthelmintic resistance (AR) in A. caninum to several anthelmintic classes, mainly in the USA, indicates the potential for this scenario in Canada. We consider various factors that may lead to resistant isolates in Canada, such as the widespread use of antiparasitic drugs without the assessment of efficacy; increased A. caninum prevalence in various Canadian provinces; and the importation of dogs, mostly from the USA, with a history of persistent infection by A. caninum . Our objective was to review factors that influence A. caninum to develop AR and raise awareness regarding the need for a strategic plan to control this parasitic nematode through the appropriate use of anthelmintics., (Copyright and/or publishing rights held by the Canadian Veterinary Medical Association.)

Background: Anthelmintic resistance to benzimidazole has been detected in the canine hookworm, Ancylostoma caninum. Benzimidazole resistance is believed to have developed originally in greyhounds, but has also been detected in non-greyhound pet dogs. The aim of this study was to validate a probe-based allele-specific real-time PCR tests for the F167Y polymorphism on the β-tubulin isotype-1 gene and to determine the geographic distribution., Methods: Allele-specific real-time PCR tests were established and validated to detect the codon 167 polymorphism in the Ancylostoma caninum β-tubulin isotype-1gene. Additionally, real-time PCR tests were validated for Ancylostoma spp. and Uncinaria stenocephala. Two nucleic acid extraction protocols were validated including mechanical disruption of parasite structures in stool. The frequency of the F167Y single nucleotide polymorphism (SNP) was determined in hookworm confirmed stool samples. Samples with the resistant 167Y genotype were confirmed by β-tubulin gene sequencing and allele frequencies were determined., Results: The Ancylostoma spp. and A. caninum F167Y allele-specific real-time PCR tests were highly sensitive and specific when tested against synthetic DNA, spiked samples, and characterized parasites. Using an optimized total nucleic acid extraction protocol, 54 of 511 (10.6%) were found to contain the benzimidazole resistance allele. All 55 samples containing hookworms with the resistance mutation were confirmed by β-tubulin gene sequencing. The majority of resistant hookworms (44 resistant, 183 tested; 24.4%) originated from Florida, five from California (103 tested, 4.9%), three from Idaho (40 tested, 7.5%), two from Nevada (22 tested, 9.1%), and one sample from Hawaii (13 tested, 7.7%). Resistant genotypes were found in 14 different dog breeds including eight in Greyhounds. Allele-frequency determination revealed resistance allele frequencies between 1 and 100% with 58% above 50%., Conclusions: This data strongly supports recent findings of benzimidazole resistant canine hookworms present throughout the general US pet dog population., (Copyright © 2023. Published by Elsevier Ltd.)

Soil-transmitted helminths (STH) infect 1.5 billion people and countless animals worldwide. In Australian Indigenous communities, STH infections have largely remained endemic despite control efforts, suggesting reservoirs of infection may exist. Dogs fulfil various important cultural, social and occupational roles in Australian Indigenous communities and are populous in these settings. Dogs may also harbour zoonotic STHs capable of producing morbidity and mortality in dogs and humans. This review provides an overview of human and zoonotic STH infections, identifies the Australian Indigenous locations affected and the parasite species and hosts involved. The meta-analysis provides estimates of individual study and pooled true prevalence of STH infections in Australian Indigenous communities and identifies knowledge gaps for further research on zoonotic or anthroponotic potential. A systematic literature search identified 45 eligible studies documenting the presence of Strongyloides stercoralis, Trichuris trichiura, Ancylostoma caninum, Ancylostoma duodenale, Ancylostoma ceylanicum, undifferentiated hookworm, and Ascaris lumbricoides. Of these studies, 26 were also eligible for inclusion in meta-analysis to establish true prevalence in the light of imperfect diagnostic test sensitivity and specificity by Rogan-Gladen and Bayesian methods. These studies revealed pooled true prevalence estimates of 18.9% (95% CI 15.8-22.1) for human and canine S. stercoralis infections and 77.3% (95% CI 63.7-91.0) for canine A. caninum infections indicating continued endemicity, but considerably more heterogenous pooled estimates for canine A. ceylanicum infections, and A. duodenale, undifferentiated hookworm and T. trichiura in humans. This review suggests that the prevalence of STHs in Australian Indigenous communities has likely been underestimated, principally based on imperfect diagnostic tests. Potential misclassification of hookworm species in humans and dogs due to outdated methodology, also obscures this picture. High-quality contemporary studies are required to establish current true prevalence of parasite species in all relevant hosts to guide future policy development and control decisions under a culturally sound One Health framework., Competing Interests: The authors have declared that no competing interests exist.

Purpose: Canine hookworm disease is a global zoonotic parasitic disease caused by a variety of nematodes in families Ancylostomatidae, including Ancylostoma spp., Necator spp., and Uncinaria spp., in the small intestine (mainly the duodenum) of dogs. The disease is widely distributed in China. The purpose of this study is to systematically diagnose and treat canine hookworm disease through the case of miniaturization Schnauzer dog feed infected with A. ceylanicum, so as to provide experimental basis for subsequent prevention and control of canine hookworm disease., Methods: In the current study, we isolated hookworm eggs from a diseased miniature schnauzer, then the polymerase chain reaction (PCR) was used to amplify the ITS1-5.8S-ITS2 gene sequence from genomic DNA extracted from hookworms. Phylogenetic analysis based on ITS1-5.8S-ITS2 gene sequence sequences was inferred using MEGA-X. After phylogenetic analysis, etiologic and symptomatic therapies were used to treat the canine hookworm disease., Results: The sequencing results showed that the length of the ITS1-5.8S-ITS2 gene sequence was approximately 960 bp, and ITS1 and ITS2 were extracted to analyze similarity with other hookworms to build a phylogenetic tree. After phylogenetic analysis, the results showed that the diseased miniature schnauzer was infected by A. ceylanicum. Using etiologic and symptomatic therapies, the sick dog with an A. ceylanicum infection was also treated for 5 days., Conclusions: To our knowledge, this is the first report of diagnosis and treatment for canine hookworm disease in Guangzhou city. In addition, with the improvement of economic level, the scale of pet dog breeding is also increasing. The diagnostic methods and treatment schemes adopted in this report will help to standardize the prevention and control of canine hookworm disease., (© 2022. The Author(s) under exclusive licence to Witold Stefański Institute of Parasitology, Polish Academy of Sciences.)

Background: A large number of studies have assessed risk factors for infection with soil-transmitted helminths (STH), but few have investigated the interactions between the different parasites or compared these between host species across hosts. Here, we assessed the associations between Ascaris, Trichuris, hookworm, strongyle and Toxocara infections in the Philippines in human and animal hosts., Methods: Faecal samples were collected from humans and animals (dogs, cats and pigs) in 252 households from four villages in southern Philippines and intestinal helminth infections were assessed by microscopy. Associations between worm species were assessed using multiple logistic regression., Results: Ascaris infections showed a similar prevalence in humans (13.9%) and pigs (13.7%). Hookworm was the most prevalent infection in dogs (48%); the most prevalent infection in pigs was strongyles (42%). The prevalences of hookworm and Toxocara in cats were similar (41%). Statistically significant associations were observed between Ascaris and Trichuris and between Ascaris and hookworm infections in humans, and also between Ascaris and Trichuris infections in pigs. Dual and triple infections were observed, which were more common in dogs, cats and pigs than in humans., Conclusions: Associations are likely to exist between STH species in humans and animals, possibly due to shared exposures and transmission routes. Individual factors and behaviours will play a key role in the occurrence of co-infections, which will have effects on disease severity. Moreover, the implications of co-infection for the emergence of zoonoses need to be explored further., (© The Author(s) 2022. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene.)

Purpose: Hookworms are hematophagous parasitic nematodes that occur in the intestinal tract of various mammals, including humans. The objective of this work was to develop a two-step morphology-molecular analysis-based strategy to identify the genus and the species of eggs and larvae of the Ancylostomatidae family in dogs, which were kept in various living conditions in Slovakia., Methods: Faecal samples were collected from 270 dogs kept in two different shelters (160 samples) and in a marginalised Roma community (110 samples). Faecal samples were processed using the flotation method. Microscopically positive faecal samples with hookworm eggs were subjected to a coproculture and the hatched larvae were identified morphometrically, prior to molecular testing. The faecal samples with hookworm´s eggs and individual larvae were identified by a molecular assay based on the amplification of the 18S ribosomal RNA gene fragment. Further, species-specific primer sets were designed for the internal transcribed spacer (ITS 1 region) and the mitochondrial cytochrome c oxidase subunit I (COX1) gene section., Results: Hookworm eggs were microscopically detected in 9.6% (26/270) of the total number of faecal samples. The prevalence in the Roma settlement was higher, 14.5% (16/110), than in shelters, 6.3% (10/160). Using PCR and subsequent Sanger sequencing, we identified the canine hookworm species Uncinaria stenocephala in all positive samples., Conclusion: Our results have provided new data on the molecular identification of the neglected species U. stenocephala affecting dogs in Slovakia and supplemented the missing information on the prevalence and incidence of hookworms in dogs in Europe., (© 2021. The Author(s) under exclusive licence to Witold Stefański Institute of Parasitology, Polish Academy of Sciences.)

Hookworms are the most common and voracious blood-sucking parasites of the small intestines of mammalian hosts such as dogs, cats, ruminants and humans. Canine hookworms are endemic in the Southeast Asian countries including Bangladesh. There is scarcity of information on the prevalence of hookworms of stray dogs in Bangladesh. The present study determined the prevalence of canine hookworms using fecal examination followed by morphometric and molecular identification. Fecal samples were collected from 320 stray dogs living in rural areas of Mymensingh district (Gauripur upazila, Mymensingh sadar upazila and Tarakanda upazila) and hookworm eggs were identified using the flotation techniques. The overall prevalence of hookworm was 79.1% through microscopic examination. Estimated fecal prevalence was higher in Gauripur upazila (89.7%) followed by Mymensingh sadar upazila (84.8%) and Tarakanda upazila (53.2%). Five hookworm species were identified based on the morphometric examination, namely, Ancylostoma caninum, Ancylostoma ceylanicum, Ancylostoma tubaeforme, Ancylostoma braziliense and Ancylostoma duodenale, respectively. Polymerase Chain Reaction (PCR) was performed with the genomic DNA by targeting the 5.8S rRNA (~ 404 bp) and Cytochrome oxidase-1 (Cox 1, ~ 450 bp) and confirmed the identification for the first time in Bangladesh. This study reveals that stray dogs may act as reservoir hosts of human hookworm infection. Further detail molecular study is warranted to explore the genetic diversity of hookworms that infect both dogs and human in Bangladesh., (Copyright © 2022. Published by Elsevier B.V.)

The early-to-mid 1900s was an era that marked the first published reports of clinical cases and experimental human infections that provided valuable evidence to inform our current knowledge on the zoonotic nature of the canid and felid hookworms. To this day, descriptions of the pathognomonic syndromes produced in humans by these early pioneers provide vital information on the putative geographical distribution of these hookworm species, even before their discovery in dogs and cats. Cases of hookworm-related cutaneous larvae migrans and Ancylostoma caninum-induced eosinophilic enteritis continue to be reported in the medical literature, most commonly in recent migrants or returned travellers who have spent time in regions in which these canine and feline hookworms remain highly endemic. Ancylostoma ceylanicum, now recognised as the second most common hookworm infecting humans in the Asia Pacific region, is also being reported in regions of Africa and South America previously assumed free of this parasite. Despite the substantial technological progress made in the field of parasite diagnostics, our knowledge on the epidemiology and population-level morbidity impacts of these zoonotic hookworms in humans has remained relatively stagnant over the last few decades, with 'covert' infections continuing to remain undetected by currently available forms of routine diagnostics. Improved diagnostic tools that enable accurate and rapid species-specific diagnosis of zoonotic hookworm infections in humans are required to provide the necessary evidence to advocate for future investments in One Health-based intervention strategies., (Copyright © 2021 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

The prevalence of enteric parasites in cats in metropolitan Bangkok has not been updated in over 13 years. The main objectives of this study include updating the prevalence of endoparasitism in client-owned cats, status of retroviral infections and determining the association between feline hookworm infection and possible risk factors. A total of 509 fecal samples were collected from client-owned cats in 2014-2015 and examined by a wet fecal mount technique. If additional sample remained, a PBS-ethyl acetate sedimentation was done (n = 229), and ZnSO 4 centrifugal flotation was also performed if there was sufficient remaining sample (n = 105). At least one parasite was observed in 32.0% (163/509) of cats, with Ancylostoma being the most common intestinal parasite detected in 21.6% (110/509) of cats. Other parasitic infections detected by fecal examinations included Toxocara (6.9%; 35/509), Platynosomum (3.7%; 19/509), Cystoisospora (3.5%; 18/509), Taenia (2.9%; 15/509), Spirometra (1.6%; 8/509), Dipylidium (0.4%; 2/509), and Opisthorchis-like trematode (0.2%; 1/509). Examination for Giardia infection was conducted with the SNAP® Giardia Test, a coproantigen test, on a subset of the fecal samples (233/509) and revealed a positive result on 3.9% (9/233) of samples. Plasma samples were analyzed using the SNAP® Triple Test detecting antigens of Feline Leukemia Virus (FeLV) and Dirofilaria immitis while also detecting antibodies to Feline Immunodeficiency Virus (FIV). Antigens of FeLV and antibodies to FIV were found in 7.1% (19/269) and 5.2% (14/269) of cats, respectively. None of the cats were found to have circulating antigen of Dirofilaria immitis using this test. No association between retroviral and endoparasitic infections was found. From multivariable logistic regression examining associated factors, the ability of cats to access the outdoors (adjusted OR = 3.22, 95% CI; 1.42-7.87) and having tapeworm segments or adult helminths in feces (adjusted OR = 3.31, 95% CI; 1.34-8.21) were significantly associated with the finding of hookworm eggs in feces. This work presents the most up-to-date data on enteric feline parasite prevalence in the metropolitan Bangkok area from which fecal samples were directly collected from cats. Consequently, this study emphasizes that diagnosis of parasitic infections and the routine use of antiparasitic medications should be encouraged by veterinarians and to owners in order to reduce the reservoir of potentially zoonotic parasites., (Copyright © 2021 Elsevier B.V. All rights reserved.)

A number of gastrointestinal parasites have been reported to infect dogs in Nigeria, some of which have zoonotic potential. Of these, hookworms are the most prevalent, with both Ancylostoma caninum and Uncinaria stenocephala reported in the country. In this study, we subjected 203 hookworm microscopy-positive samples of the 885 individual faecal samples collected from dogs in Nigeria to a recently developed multiplex qPCR for the detection and characterisation of canine hookworm species. The qPCR demonstrated a diagnostic sensitivity of 98% (95% CI 95-99.4) allowing the detection of A. caninum and A. braziliense in 81.3% (165/203, 95% CI 75.3-86.1) and 51.2% (104/203, 95% CI 44.4-58) of the microscopy-positive faecal samples of dogs from Nigeria, respectively and 34.5% (70/203, 95% CI 28.3-41.3) of mixed infections with both hookworm species. The finding of A. braziliense is particularly worrisome given this is a well-known agent of persistent cutaneous larva migrans, commonly referred to as "creeping eruptions" in humans. Although this parasite has been diagnosed in locals and in people travelling in Nigeria suffering from dermatological illnesses, this represents the first molecular identification of A. braziliense in its canine reservoir in the country. These results update the occurrence and distribution of hookworm species affecting dogs in Nigeria highlighting the suitability of the newly developed multiplex qPCR assay as a high-throughput tool for the surveillance of zoonotic hookworms, globally., (Copyright © 2020. Published by Elsevier B.V.)

Background: Animal rescue and animal welfare organizations are relocating thousands of dogs per year following natural disasters and in attempts to provide greater adoption opportunities. Many dogs are sourced from the southeastern USA, which historically has a high prevalence rate for many parasites and parasitic diseases. The Colorado Department of Agriculture Pet Animal Care Facilities Act (PACFA) requires animal shelters and animal welfare organizations to report annually a variety of statistics including the numbers of dogs imported into Colorado from out of state. The Companion Animal Parasite Council (CAPC) provides data nationally, down to the state and county level, on a variety of common parasitic and vector borne diseases. These data make it possible to track changes in parasite prevalence over several years., Methods: Test results for canine roundworm, hookworm and whipworm were collected from the CAPC maps for 2013-2017. Dog importation data for 2014-2017 was collected from PACFA reports. For evaluation of the statistical significance of prevalence changes when comparing 2013 to 2017, 2 × 2 contingency tables were constructed with both positive and negative test results for each year and the data assessed using Chi-square tests to determine if the 2017 prevalence was significantly different than the 2013 prevalence for roundworm, hookworm and whipworm., Results: Significant increases in intestinal nematode prevalence occurred in Colorado from 2013 to 2017. The prevalence of canine roundworm rose 35.60%, the prevalence of canine hookworm rose 137.33% and the prevalence of canine whipworm rose 63.68%. From 2014 to 2017, over 114,000 dogs were transported into Colorado from out-of-state, by more than 130 animal shelters and rescue organizations. Three of the larger organizations reported that the majority of their dogs were obtained from New Mexico, Texas and Oklahoma. Texas and Oklahoma have historically much higher parasite prevalence than Colorado., Conclusions: Veterinarians in areas with historically low parasite prevalence where dogs from high parasite prevalence areas are arriving for adoption may need to reevaluate their recommendations regarding fecal examination and deworming frequencies as historic levels of intestinal parasite infection may no longer be accurate assessments of future infection risks.

Ancylostoma caninum is the most prevalent intestinal nematode of dogs, and has a zoonotic potential. Multiple-drug resistance (MDR) has been confirmed in a number of A. caninum isolates, including isolate Worthy 4.1F3P, against all anthelmintic drug classes approved for hookworm treatment in dogs in the United States (US). The cyclooctadepsipeptide emodepside is not registered to use in dogs in the US, but in a number of other countries/regions. The objective of this study was to evaluate the efficacy of emodepside + praziquantel, as well as three commercial products that are commonly used in the US for treatment of hookworms, against a suspected (subsequently confirmed) MDR A. caninum isolate Worthy 4.1F3P. 40 dogs infected on study day (SD) 0 with 300 third-stage larvae, were randomly allocated to one of five treatment groups with eight dogs each: pyrantel pamoate (Nemex®-2), fenbendazole (Panacur® C), milbemycin oxime (Interceptor®), emodepside + praziquantel tablets and non-treated control. Fecal egg counts (FEC) were performed on SDs 19, 20, 22, 27, 31 and 34. All treatments were administered as per label requirements on SD 24 to dogs in Groups 1 through 4. Two additional treatments were administered on SDs 25 and 26 to dogs in Group 2 as per label requirements. Dogs were necropsied on SD 34 and the digestive tract was removed/processed for worm recovery and enumeration. The geometric mean (GM) worm counts for the control group was 97.4, and for the pyrantel pamoate, fenbendazole, milbemycin oxime, and emodepside + praziquantel groups were 74.8, 72.0, 88.9, and 0.4, respectively. These yielded efficacies of 23.2%, 26.1%, and 8.8%, and 99.6%, respectively. These data support previous findings of the MDR status of Worthy 4.1F3P as treatments with pyrantel pamoate, fenbendazole and milbemycin oxime lacked efficacy. In sharp contrast, Worthy 4.1F3P was highly susceptible to treatment with emodepside + praziquantel., Competing Interests: Declaration of competing interest The study reported here was funded by Bayer US LLC, Animal Health, Research and Development, Shawnee, KS, USA and was performed at TRS Labs Inc., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The long-term fidelity of pinniped hosts to their natal rookery site suggests the genetic architecture of their Uncinaria spp. hookworms should be strongly structured by host breeding biology. However, historical events affecting host populations may also shape parasite genetic structure. Sequences of the mitochondrial cytochrome c oxidase 1 (COI) gene of 86 Uncinaria lucasi individuals were obtained to assess genetic variation and structure of nematodes from 2 host species (68 hookworms from northern fur seals; 18 hookworms from Steller sea lions) and rookeries from 3 widely separated geographic regions: the western Bering Sea and Sea of Okhotsk, eastern Bering Sea, and the eastern Pacific Ocean. High COI haplotype (h = 0.96-0.98) and nucleotide (π = 0.014) diversity was found. The haplotype network showed a star-shaped pattern with a large number of haplotypes separated by few substitutions. The network did not show separation of U. lucasi by geographic region or host species. Fst values between U. lucasi individuals representing geographic regions showed no differentiation, consistent with the absence of genetic structure. At face value, this lack of genetic structure in U. lucasi suggests high gene flow but could also be explained by recent (post-glacial) population expansions of northern fur seals and their hookworms., (© American Society of Parasitologists 2020.)

The aim of this study was to evaluate the combined use of different chemical (albendazole, ivermectin, glycerine and Vaseline) and biological (Monacrosporium thaumasium) compounds in the control of Ancylostoma caninum. Infective larvae of A. caninum were obtained from coprocultures of positive faeces from naturally infected dogs. We used 1% ivermectin, 1% albendazole, 100% glycerine, 100% Vaseline and an isolate of the nematophagous fungus M. thaumasium (NF34), alone or in combinations. Next, an experimental test was set up with 16 groups in microtubes, with a 24-h interaction. The groups (G1 to G15) that contained any chemical or biological compound (NF34) and/or their combined use (chemical + biological) showed a difference in relation to the control group, except G5 - Vaseline 100% without combinations. It was concluded that, even on an experimental basis, the combined use of anthelmintic drugs with biological control was efficient; however, more studies must be carried out in order to elucidate the synergistic action between chemical and biological compounds to be used in the effective control of hookworms in the future.

Definitive diagnosis of hookworm infection is usually based on the microscopic detection of eggs in a stool sample; however, several cases display a low or irregular egg output. Serodiagnosis can be a useful tool to identify these cases, but conventional tests do not differentiate past from active infections. The aim of this study was to obtain and apply egg yolk polyclonal immunoglobulin (IgY) antibodies to detect immune complexes (ICs) in serum samples from patients infected with hookworm. Hens were immunized with Ancylostoma ceylanicum saline extract, their eggs were collected and then IgY antibodies were extracted and purified. Antibody purity was tested by 12% sodium dodecyl sulphate polyacrylamide gel electrophoresis and specificity was assessed by immunoblotting and immunofluorescence. IgY production was evaluated by kinetics enzyme-linked immunosorbent assay (ELISA). Sandwich ELISA tested the ability of IgY to detect ICs in serum samples, from which diagnostic parameters were calculated. Antibody responses increased steadily from day 7 to 42. In the immunoblotting assay, IgY recognized two protein complexes. The immunofluorescence assay showed no staining in control samples. The sandwich ELISA presented a very high diagnostic value, with a sensitivity of 90% and a specificity of 86.7%. Our pioneer strategy highlights the potential use of egg yolk IgY as a diagnostic test to detect active hookworm infection.

Background: Ancylostomatids ('hookworms') are among the most important zoonotic nematode parasites infecting dogs worldwide. Ancylostoma caninum and Uncinaria stenocephala are two of the most common hookworm species that infect dogs. Both immature and adult stages of hookworms are voracious blood feeders and can cause death in young dogs before infection can be detected by routine fecal examination. Hence, treatment of both immature and adult stages of hookworms will decrease the risk of important clinical disease in the dog as well as the environmental contamination caused by egg-laying adults, which should reduce the risk of infection for both dogs and humans. The studies presented here were conducted to evaluate the efficacy of a novel, oral chewable tablet containing sarolaner, moxidectin and pyrantel (Simparica Trio™), against induced larval (L 4 ), immature adult (L 5 ) and adult A. caninum, and adult U. stenocephala infections in dogs., Methods: Eight negative-controlled, masked, randomized laboratory studies were conducted. Two separate studies were conducted against each of the target parasites and stages. Sixteen or 18 purpose bred dogs, 8 or 9 in each of the two treatment groups, were included in each study. Dogs experimentally infected with the target parasite were dosed once on Day 0 with either placebo tablets or Simparica Trio™ tablets to provide minimum dosages of 1.2 mg/kg sarolaner, 24 µg/kg moxidectin and 5.0 mg/kg pyrantel (as pamoate salt). Timing of dosing relative to parasite inoculation allowed for efficacy to be evaluated primarily against the target parasite stage. Worm counts were conducted 7 or 8 days after treatments during necropsy. Efficacy was based on the number of worms recovered at necropsy compared to placebo control., Results: Based on geometric mean worm counts, efficacy of Simparica Trio™ was ≥ 98.4% against L 4 larval stage of A. caninum, ≥ 99.8% against immature adult (L 5 ) A. caninum, and 100% against adult A. caninum and adult U. stenocephala., Conclusions: These studies confirm the efficacy of a single oral dose of a novel, chewable tablet containing sarolaner, moxidectin and pyrantel (Simparica Trio™) against L 4 larval and immature adult (L 5 ) A. caninum, and adult A. caninum and U. stenocephala infections in dogs.

Introduction: Hookworm infections are neglected tropical diseases of humans and animals worldwide. A meta-analysis and systematic review was designed to evaluate the prevalence of hookworm infection in animal and human hosts in Asia until July 2018., Methods: The available online articles of five English databases (PubMed, Scopus, Science Direct, Web of Science and Google Scholar) were explored., Results: The most parasitized carnivores were jackal (48%, CI: 4 to 91%), followed by dog (41%, CI: 29 to 53%), cat (26%, CI: 14 to 38%) and the red fox (19%, CI: 13 to 24%). The weighted prevalence of Ancylostoma braziliensis, A. caninum, A. ceylanicum, A. tubaeforme and Uncinaria stenocephala isolated from different canids were found to be 27% (CI: 21 to 33%), 23% (CI: 7.0 to 53%), 24% (CI: 12 to 35%), 44% (CI: 37 to 51%) and 37% (CI: 18 to 55%), respectively. In total, 98 records were obtained for human hookworms from 3209 760 examined individuals and the calculated weighted prevalence in this population was 19% (CI: 17 to 20%)., Conclusions: These findings highlight a desirable ecological milieu for parasite survival and transmission in such territories, which implicates revisiting control programs and public health infrastructures in those areas., (© The Author(s) 2020. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Hookworm infection is globally prevalent among dogs and cats representing a major public health risk. Although previous studies have surveyed canine and feline hookworms in Guangzhou city, the status of these infection needs to be further explored in other regions of South China. To investigate the prevalence and zoonotic risk of canine and feline hookworms in eight cities (Guangzhou, Foshan, Shenzhen, Huizhou, Zhongshan, Shaoguan, Shantou and Chaozhou) of Guangdong province, China, we developed specific PCR methods based on ITS sequence for identifying three common hookworm species. The results showed that the prevalence of hookworms from stray dogs and cats was 20.23% (142/702) and 15.26% (47/308), respectively. The established PCR methods could identify Ancylostoma ceylanicum, A. caninum and A. tubaeforme. The mixed infections of A. caninum and A. ceylanicum were detected in stray dogs of Guangzhou and Shaoguan, with the rate of 8.3% and 21.2%, respectively. Among the stray dogs in Foshan, the infection rate of A. ceylanicum was higher than that of A. caninum. The stray cats in four of five investigated cities were infected with A. ceylanicum. The different region, age and rearing environments had an impact on the hookworm infection rates of stray dogs and cats. In conclusion, the reported higher infection rate of A. ceylanicum than other hookworm species in stray dogs and cats poses a potential risk to public health., (Copyright © 2019. Published by Elsevier B.V.)

Parasites can cause chronic stress in some animal species, and this type of stress response has been associated with adverse consequences for the host. In order to know whether parasitism elicited a stress response associated with decreased host fitness, hookworm (Uncinaria sp.) infection was studied in a colony of South American fur seals (Arctocephalus australis) in which hookworms infect nearly all pups born in a reproductive season. A parasite-free group was generated by treating a subset of pups with an antiparasitic drug before they developed patent hookworm infection. Stress and metabolic hormones, energy balance, and humoral and cellular immune parameters were measured in this group and hookworm-infected pups. Hookworms elicited a marked increase in plasma cortisol levels in fur seal pups. These hookworm-infected pups were able to maintain constant glucose levels, despite losing body mass over the course of infection potentially because of increased protein catabolism. Infected pups were able to mount an effective immune response against the parasite and eliminated hookworms from the intestine, recovering partial body mass lost as a result of hookworm infection at the end of the study period. As shown in previous studies, adequate glucose levels are critical for proper T lymphocyte reactivity, and it is possible that, through activation of a stress response, energy can be readily available for immune response against the parasite contributing to early recovery from infection. Although there are potential fitness costs to mounting a sustained stress response, these could also be adaptive and promote survival during critical life-history stages.

The presence and distribution of various species of canine hookworms in Africa are poorly known. The main objective of this study, therefore, was to identify the hookworm species present in canine faecal samples from Morogoro, Tanzania, using molecular techniques. Faecal samples from 160 local dogs were collected and hookworm positive samples processed to recover larvae for further molecular characterization. DNA was extracted from pools of larvae from individual samples (n = 66), which were analysed subsequently using two different molecular approaches, polymerase chain reaction-linked restriction fragment length polymorphism (PCR-RFLP) and species-specific PCR coupled with Sanger sequencing. The PCR-RFLP technique detected only the presence of the ubiquitous Ancylostoma caninum in the 66 samples. However, by species-specific PCR coupled with Sanger sequencing we identified ten samples with A. braziliense, two with Uncinaria stenocephala and five with A. ceylanicum. Thus, all four known species of canine hookworms were identified in Morogoro, Tanzania. To our knowledge this is the first report of the detection of the presence of U. stenocephala and A. ceylanicum in Africa using molecular techniques. In addition to their veterinary importance, canine hookworms have zoonotic potential and are of public health concern.

Soil-transmitted nematodes infect over a billion people and place several billion more at risk of infection. Hookworm disease is the most significant of these soil-transmitted nematodes, with over 500 million people infected. Hookworm infection can result in debilitating and sometimes fatal iron-deficiency anemia, which is particularly devastating in children and pregnant women. Currently, hookworms and other soil-transmitted nematodes are controlled by administration of a single dose of a benzimidazole to targeted populations in endemic areas. While effective, people are quickly re-infected, necessitating frequent treatment. Widespread exposure to anthelmintic drugs can place significant selective pressure on parasitic nematodes to generate resistance, which has severely compromised benzimidazole anthelmintics for control of livestock nematodes in many areas of the world. Here we report, to our knowledge, the first naturally occurring multidrug-resistant strain of the canine hookworm Ancylostoma caninum. We reveal that this isolate is resistant to fenbendazole at the clinical dosage of 50 mg/kg for 3 days. Our data shows that this strain harbors a fixed, single base pair mutation at amino acid 167 of the β-tubulin isotype 1 gene, and by using CRISPR/Cas9 we demonstrate that introduction of this mutation into the corresponding amino acid in the orthologous β-tubulin gene of Caenorhabditis elegans confers a similar level of resistance to thiabendazole. We also show that the isolate is resistant to the macrocyclic lactone anthelmintic ivermectin. Understanding the mechanism of anthelmintic resistance is important for rational design of control strategies to maintain the usefulness of current drugs, and to monitor the emergence of resistance. The isolate we describe represents the first multidrug-resistant strain of A. caninum reported, and our data reveal a resistance marker that can emerge naturally in response to heavy anthelminthic treatment., (Copyright © 2019 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

To investigate the prevalence of canine and feline hookworms in South China, and to assess the risk of zoonotic hookworms to humans, one pair of primers (HRM-F/HRM-R) was designed to establish a high-resolution melting (HRM) method based on internal transcribed spacer 1 (ITS-1) rDNA for the detection of Ancylostoma ceylanicum, A. caninum and A. tubaeforme infection. The results showed that the HRM for the three hookworms produced different melting-curve profiles, where melting temperature (Tm) values were 84.50°C for A. ceylanicum, 82.25°C for A. caninum and 81.73°C for A. tubaeforme, respectively. The reproducibility of intra- and inter-assay melting curves was almost perfect. The lowest concentration detected was about 5.69 ×10-4 g/μl. The HRM detection results from 18 canine and feline hookworm samples were in complete accordance with their sequencing results. The HRM method was more sensitive than the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique in the detection of 98 clinical samples. It is concluded that the HRM method can differentiate between A. ceylanicum, A. caninum, A. tubaeforme and their mixed infections, which may provide important technical support for the zoonotic risk assessment and molecular epidemiological survey of canine and feline hookworms.

All canine hookworms are known to be zoonotic, causing infections ranging from transient skin irritations to prolonged 'creeping eruptions', eosinophilic enteritis and even patent intestinal infections. There is little information on canine hookworm species and their public health significance in sub-Saharan Africa. This study determined the prevalence and species of hookworms in dogs from different climatic zones of Kenya. Dog faecal samples were collected from the environment, and hookworm eggs were isolated by zinc chloride flotation and subjected to DNA extraction. Polymerase chain reaction (PCR) assays targeting the internal transcribed spacer (ITS) 1 and 2, 5.8S and 28S ribosomal RNA of Ancylostoma spp. and Uncinaria stenocephala were performed, and hookworm species were identified by PCR-restriction fragment length polymorphism (RFLP) or DNA sequencing. Hookworm eggs were detected by microscopy in 490/1621 (30.23%, 95% CI 28.01-32.54) faecal samples. Estimates of faecal prevalence were high in counties receiving higher rainfall (Narok 46.80%, Meru 44.88%) and low in those with a more arid climate (Isiolo 19.73%, Turkana 11.83%). In a subset of 70 faecal samples, Ancylostoma caninum (n = 59) was the most common species, followed by A. braziliense (n = 10) and A. cf. duodenale (n = 1). This study reports for the first time the detection of A. cf. duodenale in dog faeces and zoonotic hookworm species in Kenyan dogs. These findings emphasize the need for control measures such as enforcing laws for restraining stray dogs, regular deworming of dogs, and public health awareness programmes aimed at informing communities on outdoor use of footwear.

There is a paucity of information on hookworm species in humans, domestic animals and wildlife in southern Africa. Our study aimed to identify hookworm species from stray dogs, humans, and selected wildlife from South Africa. A total of 356 faecal samples were screened for the presence of hookworm-like eggs and subsequently coproculture from the positive samples was carried out to obtain larvae. Hookworm-like eggs were detected in 23.03% (82/356) of samples. Of these samples, 78/296 were from dogs, 3/50 from humans and 1/10 from wildlife. DNA was then isolated from the larvae of 55 positive samples, which were subjected to polymerase chain reaction (PCR), polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing of the nuclear ribosomal internal transcribed spacer (ITS1) and 5.8S rRNA region. Presence of Ancylostoma caninum, A. braziliense and A. ceylanicum-like species was recorded in stray dogs and A. caninum was recorded in wildlife and humans, using PCR-RFLP. Although PCR-RFLP results pointed to the presence of A. ceylanicum, we did not get a sequence that matched with A. ceylanicum from GenBank. This may have been due to the low proportion of A. ceylanicum larvae in our samples. Twenty-two of the 27 positive amplicons from stray dogs matched with A. caninum, three with A. braziliense and two had mixed infections of A. braziliense and A. caninum. Sequences from a lion and three humans matched with A. caninum. This is the first confirmation of a patent A. caninum infection in humans as evidenced by the presence of eggs in faeces, with the subsequent larvae from coproculture being identified as A. caninum.

Hookworms of the genus Uncinaria parasitize pinniped pups in various locations worldwide. Four species have been described, two of which parasitize pinniped pups in the southern hemisphere: Uncinaria hamiltoni parasitizes Otaria flavescens and Arctocephalus australis from the South American coast, and Uncinaria sanguinis parasitizes Neophoca cinerea from the Australian coast. However, their geographical ranges and host specificity are unknown. Uncinaria spp. are morphologically similar, but molecular analyses have allowed the recognition of new species in the genus Uncinaria. We used nuclear genetic markers (internal transcribed spacer (ITS) and large subunit (LSU) rDNA) and a mitochondrial genetic marker (cytochrome c oxidase subunit I (COI)) to evaluate the phylogenetic relationships of Uncinaria spp. parasitizing A. australis and O. flavescens from South American coasts (Atlantic and Pacific coasts). We compared our sequences with published Uncinaria sequences. A Generalized Mixed Yule Coalescent (GMYC) analysis was also used to delimit species, and principal component analysis was used to compare morphometry among Uncinaria specimens. Parasites were sampled from A. australis from Peru (12°S), southern Chile (42°S), and the Uruguayan coast, and from O. flavescens from northern Chile (24°S) and the Uruguayan coast. Morphometric differences were observed between Uncinaria specimens from both South American coasts and between Uncinaria specimens from A. australis in Peru and southern Chile. Phylogenetic and GMYC analyses suggest that south-eastern Pacific otariid species harbour U. hamiltoni and an undescribed putative species of Uncinaria. However, more samples from A. australis and O. flavescens are necessary to understand the phylogenetic patterns of Uncinaria spp. across the South Pacific.

Increases in ocean temperature are associated with changes in the distribution of fish stocks, and the foraging regimes and maternal attendance patterns of marine mammals. However, it is not well understood how these changes affect offspring health and survival. The maternal attendance patterns and immunity of South American fur seals were assessed in a rookery where hookworm disease is the main cause of pup mortality. Pups receiving higher levels of maternal attendance had a positive energy balance and a more reactive immune system. These pups were able to expel hookworms through a specific immune mediated mechanism and survived the infection. Maternal attendance was higher in years with low sea surface temperature, therefore, the mean hookworm burden and mortality increased with sea surface temperature over a 10-year period. We provide a mechanistic explanation regarding how changes in ocean temperature and maternal care affect infectious diseases dynamics in a marine mammal., Competing Interests: MS, FM, DP, JG, HP, AM, CV, EH, VM, NG No competing interests declared, (© 2018, Seguel et al.)

Tissues of South American fur seal pups naturally infected with hookworms ( Uncinaria sp) were examined. Hookworm infection was found in nearly all pups examined (132/140, 94%), and hookworm enteritis with secondary bacteremia was considered the cause of death in 46 (35%) pups. Common findings in these pups included severe hemorrhagic enteritis and numerous (mean intensity = 761.8) hookworms in the jejunum. Hookworms were recovered from the abdominal cavity in 12 of 55 pups (22%) examined through peritoneal wash; these pups had an average of 1343.3 intestinal hookworms and marked fibrinohemorrhagic peritonitis. In all pups that died as a consequence of hookworm infection, the intestinal villi were short, blunt, and fused, and there were variable numbers of free and intrahistiocytic gram-negative bacteria in submucosal hookworm feeding tracks, mesenteric lymph nodes, spleen, blood vessels, and liver sinusoids. Pups that died of causes unrelated to the hookworm infection (trauma) had hookworm feeding tracks confined to the apical portions of the mucosa, and moderate to marked catarrhal eosinophilic enteritis. The number of hookworms was negatively correlated with intestinal villous length and number of leukocytes in the intestine. Pups with hookworm peritoneal penetration had nematodes with little or no blood in the hookworm intestine, suggesting that lack of food for the nematode could be associated with peritoneal penetration. Findings suggest that the initial burden of larval infection, the level of the host tissue response, or a combination determine the number of nematodes in the intestine, the severity of hookworm tissue damage, and pup mortality.

Background: Humans can get infected through direct or indirect contact with infective stages of zoonotic parasites shed to the environment through dog faeces., Objectives: This study was designed to investigate the presence of gastrointestinal parasites present in dog faeces shed on the street of Ibadan metropolis, one of the largest cities in Africa., Methods: Twenty-three locations were randomly selected using grid-sampling method. A total of 203 faecal samples collected from the streets of selected areas were processed for detection of helminth eggs and protozoan oocysts using flotation technique. Eggs/oocysts per gram of faeces was counted using modified McMaster technique., Results: The prevalence of gastrointestinal parasites was 43.3% (88/203). Single and multiple infections were 69 (78.4%) and 19 (21.6%) respectively. The parasites detected were Ancylostoma sp. 24.6% (50/88) Isospora sp. 14.2% (29/88), Toxocara sp. 9.8% (20/88), Uncinaria sp. 2.5% (5/88) and Strongyloides sp, 3.9% (8/88). Ancylostoma sp. (320 × 10 2 epg) and Uncinaria sp. (5 × 10 2 epg) had the highest and least intensity respectively. Streets within residential areas having markets had the highest number of positive samples. All the genera of parasites detected in this study have zoonotic potential., Conclusion: The high prevalence of zoonotic parasites detected in dog faeces from Ibadan metropolis showed that infected stray dogs roam the streets and constitute potential risk to human health. This study suggests the need for enforcement of laws restraining roaming or straying dogs and proper veterinary care of dogs., Funding: None declared., Competing Interests: Conflict of interest: None declared

Hookworm and threadworm infections are major public health problems in developing countries. A cross sectional study comprising 843 participants (346 males and 497 females) was conducted in three populations: i) Thai residents (TR) of Ubon Ratchathani Province, Thailand; ii) Laotian immigrant workers (LI) in the same province; and iii) Laotian residents (LR) in Champasak Province, Lao PDR. Participants were interviewed based on a structured questionnaire regarding their health status. Stool samples from participants and 300 samples from domestic animals (277 dogs and 23 cats) living in the participants households were collected and examined for parasitic infection using a formalin-ether concentration and a Harada-Mori filter paper culture techniques. Approximately one-third of TR and LI populations and domestic animals in Thailand were positive for parasitic infections, while almost half of LR population and domestic animals were positive. We confirmed by PCR and DNA sequencing a case of Ancylostoma ceylanicum infection in a Thai man. We also observed infections of other parasites, such as Taenia spp and Opisthorchis viverrini. Multivariate analysis indicated that risk factors for hookworm infection were population group and walking barefoot. Factors associated with threadworm infection were population group, adult male, lack of previous antiparasitic treatment and of knowledge of parasitic infection, and failure to wash hands after contact with domestic animals. Our results highlight the high prevalence of both hookworm and threadworm infections especially among LI population and domestic animals in both countries. Our findings emphasize the need for public health intervention to control the spread of parasitic infections in Thailand and Lao PDR.

Background: Hookworms (Necator americanus and Ancylostoma duodenale) remain a major public health problem worldwide. Infections with hookworms (e.g., A. caninum, A. ceylanicum and A. braziliense) are also prevalent in dogs, but the role of dogs as a reservoir for zoonotic hookworm infections in humans needs to be further explored., Methodology/principal Findings: As part of an open-label community based cluster-randomized trial in a tribal area in Tamil Nadu (India; 2013-2015), a total of 143 isolates of hookworm eggs from human stool were speciated based on a previously described PCR-RFLP methodology. The presence of hookworm DNA was confirmed in 119 of 143 human samples. N. americanus (100%) was the most prevalent species, followed by A. caninum (16.8%) and A. duodenale (8.4%). Because of the high prevalence of A. caninum in humans, dog samples were also collected to assess the prevalence of A. caninum in dogs. In 68 out of 77 canine stool samples the presence of hookworms was confirmed using PCR-RFLP. In dogs, both A. caninum (76.4%) and A. ceylanicum (27.9%) were identified. Additionally, to determine the contamination of soil with zoonotic hookworm larvae, topsoil was collected from defecating areas. Hookworm DNA was detected in 72 out of 78 soil samples that revealed presence of hookworm-like nematode larvae. In soil, different hookworm species were identified, with animal hookworms being more prevalent (A. ceylanicum: 60.2%, A. caninum: 29.4%, A. duodenale: 16.6%, N. americanus: 1.4%, A. braziliense: 1.4%)., Conclusions/significance: In our study we regularly detected the presence of A. caninum DNA in the stool of humans. Whether this is the result of infection is currently unknown but it does warrant a closer look at dogs as a potential reservoir.

The tetraprimer ARMS-PCR technique is efficient for SNP detection and can be used to search for polymorphisms associated with drug resistance. However, the establishment of this methodology is not always straightforward because of the constraints on primer design due to the restrictions of the polymorphic regions. Here, we describe the standardization of the tetraprimer ARMS-PCR methodology for the detection of a SNP at codon 198 of the Ancylostoma caninum β-tubulin gene. This SNP is associated with resistance to albendazole in various nematodes. The methodology was used to screen 327 individuals from 6 different locations. No mutation was found in any of the samples. This methodology will be useful for screening for the E198A SNP in the β-tubulin gene of canine hookworms in a broader population to determine whether this SNP is associated with benzimidazole resistance in this species. The method could also be adapted for the analysis of other SNPs in other nematode species., (Copyright © 2016 Elsevier B.V. All rights reserved.)

There are many known benefits of social grooming among primates, including maintenance of social relationships, removal of ectoparasites, and improved physiological condition. Recently, however, researchers have noted that social grooming and social contact may also present a significant cost by facilitating transmission of some parasites and pathogens. We investigated whether the number of social grooming partners varied based on infection status for gastrointestinal parasites. We used focal animal sampling and continuous recording to collect data on the number of grooming partners for known individual vervet monkeys (Chlorocebus aethiops). We collected non-invasive faecal samples and examined them using faecal flotation, faecal sedimentation, and immunofluorescence microscopy. We detected 6 parasites: Trichuris sp. (92%), hookworm (71%), spirurids (68%), Oesophagostomum sp. (84%), Strongyloides sp. (24%), and Entamoeba coli (92%). The number of grooming partners varied significantly based on infection with hookworm and sex. No significant relationships were detected for other parasites. Associations between host behavioural variation and some parasite taxa (specifically Trichuris, Oesophagostomum, and Entamoeba spp.) were impossible to explore due to an extremely high prevalence among hosts. This is the first report that we are aware of that has detected an association between social grooming behaviours and infection with hookworm., (© 2016 S. Karger AG, Basel.)

This study describes the first record of Globocephalus samoensis (Nematoda: Ancylostomatidae) recovered in wild boars from southwestern regions of Korea. Gastrointestinal tracts of 111 Korean wild boars (Sus scrofa coreanus) hunted from mountains in Suncheon-si, Gwangyang-si, and Boseong-gun between 2009 and 2012 were examined for their visceral helminths. G. samoensis, as identified by morphological characteristics of the head and tail, were recovered from the small intestine of 51 (45.9%) wild boars. Worms were found from 7 of 28 wild boars (25.0%) from Suncheon-si, 40 of 79 (50.6%) from Gwangyang-si, and all 4 (100%) from Boseong-gun. The length of adult females was 7.2±0.5 mm, and the thickest part of the body measured the average 0.47±0.03 mm, while those of males were 6.52±0.19 and 0.37±0.02 mm, respectively. The buccal cavity was equipped with a pair of large and bicuspid subventral lancets near the base of the capsule. The average length of spicules of males was 0.45±0.02 mm. By the present study, G. samoensis is recorded for the first time in southwestern regions of Korea. Additionally, morphological characteristics and identification keys provided in the present study will be helpful in the faunistic and taxonomic studies for strongylid nematodes in both domestic and wild pigs. The infection of G. samoensis apparently did not elicit pathologic lesions, as revealed by macroscopic observation during the autopsy of all wild boars in this study.

The protozoan parasite Giardia duodenalis causes a waterborne diarrhoeal disease in animals and humans, yet many Giardia-infected hosts remain asymptomatic. Mixed parasite infections are common in both animals and humans with unknown consequences for Giardia or other parasites. We compared the composition and diversity of bacterial communities from 40 dogs, including free-roaming dogs, and 21 surrendered cats from Australia. The dog cohort included 17 (42.5%) dogs positive for Giardia and 13 (32.5%) dogs positive for dog hookworm (Ancylostoma caninum). The cat samples included eight positive for Giardia and eight positive for Cystoisospora. The V4 region of 16S rRNA was sequenced at an average of 36,383 high quality sequences (>200 bp) per sample using the Ion Torrent PGM platform. In dogs we found significant (P<0.05, AnoSim) difference between the Giardia-positive and -negative groups when evaluating bacterial genera. No such difference was demonstrated between Ancylostoma-positive and -negative dogs. However, there was a modest but not significant separation of the Giardia-negative and -positive dogs (P=0.09, UniFrac) using principal coordinate analysis. Removal of dogs with hookworms further separated Giardia-positive and -negative groupings (P=0.06, UniFrac). In cats, the presence of Giardia was not associated with a significant difference based on bacterial genera (P>0.05, AnoSim). Cystoisospora-positive cats, however, exhibited significantly different profiles from Cystoisospora-negative cats (P=0.02, AnoSim) and UniFrac showed significant separation of Cystoisospora-positive and -negative samples (P<0.01). The results suggest that in clinically heathy dogs and cats, helminths and protozoa are associated with different microbiomes and possibly variable gut microbiota functions. Understanding the association of parasites and microbiomes has important consequences for the administration of antiparasitic drugs in animals and humans., (Copyright © 2015 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.)

Background: Differentiation of canine hookworm species is crucial from both a veterinary and public health standpoint. In Vietnam, three hookworm species, namely Ancylostoma caninum, Ancylostoma braziliense and Uncinaria stenocephala are reported to infect dogs. In light of the emerging distribution of A. ceylanicum in Asia, this study aims to re-evaluate the status of Ancylostoma in dogs in Vietnam., Methods: Faecal samples collected from 200 community dogs in Dak Lak province were subjected to faecal floatation for the detection of hookworm eggs. Hookworm-positive samples were subjected to a PCR-Restriction Fragment Length Polymorphism (PCR-RFLP) assay targeting the internal transcribed spacer (ITS) region of rDNA for hookworm species identification. A subset of hookworm-positive samples was also subject to haplotype characterisation at the cytochrome oxidase-1 (COX-1) gene. Detailed morphological criteria were utilised in addition to molecular markers, to identify adult hookworms recovered from necropsied dogs., Results: Of 200 canine faecal samples, 111 (55.5 %) were positive for hookworm eggs on faecal flotation. Of these, 94/111 (84.7 %) were successfully amplified and assigned species status by PCR-RFLP targeting the ITS region. In total, 54.3 % (51/94) dogs harboured single infections with A. ceylanicum, 33.0 % (31/94) with A. caninum, and 12.7 % (12/94) harboured mixed infections with both A. ceylanicum and A. caninum. Adult worms recovered from necropsied dogs matched morphological description provided for A. ceylanicum, Looss (1911) for which the mediolateral and posteriolateral rays are parallel. Characterisation of the COX-1 gene placed all Vietnamese canine isolates of A. ceylanicum within the 'zoonotic' haplotype., Conclusion: Based on this information, it is apparent that the hookworms present in dogs in Vietnam are those of A. ceylanicum and not A. braziliense. Owing to the endemic nature of this significant zoonosis in dogs, the study strongly advocates for specific identification of this hookworm in human hookworm surveys.

A placebo-controlled study was used to investigate the effectiveness of ivermectin to treat hookworm (Uncinaria sanguinis) and lice (Antarctophthirus microchir) infections in free-ranging Australian sea lion (Neophoca cinerea) pups and to test the hypotheses that these parasitic infections cause anaemia, systemic inflammatory responses, and reduced growth, and contribute towards decreased pup survival. Ivermectin was identified as an effective and safe anthelmintic in this species. Pups administered ivermectin had significantly higher erythrocyte counts and significantly lower eosinophil counts compared to controls at 1-2 months post-treatment, confirming that U. sanguinis and/or A. microchir are causatively associated with disease and demonstrating the positive effect of ivermectin treatment on clinical health parameters. Higher growth rates were not seen in ivermectin-treated pups and, unexpectedly, relatively older pups treated with ivermectin demonstrated significantly reduced growth rates when compared to matched saline-control pups. Differences in survival were not identified between treatment groups; however, this was attributed to the unexpectedly low mortality rate of recruited pups, likely due to the unintended recruitment bias towards pups >1-2 months of age for which mortality due to hookworm infection is less likely. This finding highlights the logistical and practical challenges associated with treating pups of this species shortly after birth at a remote colony. This study informs the assessment of the use of anthelmintics as a tool for the conservation management of free-ranging wildlife and outlines essential steps to further the development of strategies to ensure the effective conservation of the Australian sea lion and its parasitic fauna.

Background: A major constraint for the effective control and management of helminth parasites is the lack of rapid, high-throughput, routine diagnostic tests to assess the health status of individual animals and herds and to identify the parasite species responsible for these helminthoses. The capability of a multiplex platform for the simultaneous detection of three pasture associated parasite species was evaluated and compared to existing ELISAs., Methods: The recombinant antigens 14.2 kDa ES protein for Cooperia oncophora, major sperm protein for Dictyocaulus viviparus and Cathepsin L1 for Fasciola hepatica were recombinantly expressed either in Escherichia coli or Pichia pastoris. Antigens were covalently coupled onto magnetic beads. Optimal concentrations for coupling were determined following the examination of serum samples collected from experimentally mono-infected animals, before and after their infection with the target species. Absence of cross-reactivity was further determined with sera from calves mono-infected with Haemonchus contortus, Ostertagia ostertagi and Trichostrongylus colubriformis. Examination of negative serum samples was characterised by low median fluorescence intensity (MFI)., Results: Establishment of the optimal serum dilution of 1:200 was achieved for all three bead sets. Receiver Operating Characteristic analyses were performed to obtain cut-off MFI values for each parasite separately. Sensitivity and specificity at the chosen cut-off values were close to, or 100% for all bead sets. Examination of serum samples collected on different days post infection from different animals showed a high reproducibility of the assays. Serum samples were additionally examined with two already established ELISAs, an in-house ELISA using the recombinant MSP as an antigen and a DRG ELISA using Cathepsin L1 for liver fluke. The results between the assays were compared and kappa tests revealed an overall good agreement., Conclusions: A versatile bead-based assay using fluorescence detection (xMAP technology) was developed to simultaneously detect antibodies against C. oncophora, D. viviparus and F. hepatica in cattle serum samples. This platform provides rapid, high-throughput results and is highly sensitive and specific in comparison to existing serological as well as coproscopical diagnostic techniques.