Pyrantel resistance in canine hookworms in Queensland, Australia.

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.

Animal and Human Dirofilariasis in India and Sri Lanka: A Systematic Review and Meta-Analysis.

Dirofilariasis is an emerging vector-borne tropical disease of public health importance that mainly affects humans and dogs. Dirofilaria immitis and D. repens are the two well-documented dirofilariasis-causing filarioid helminths of both medical and veterinary concerns in India and Sri Lanka. This systematic review and meta-analysis aimed to describe and summarize the current evidence of dirofilariasis prevalence and distribution in India and Sri Lanka. Interestingly, D. repens is reported to circulate in both dogs (prevalence of 35.8% (95% CI: 11.23-60.69)) and humans (97% of published case reports) in India and Sri Lanka, but D. immitis is reported to be present in the dog populations in India (prevalence of 9.7% (95% CI: 8.5-11.0%)), and so far, it has not been reported in Sri Lanka. This peculiar distribution of D. immitis and D. repens in the two neighbouring countries could be due to the interaction between the two parasite species, which could affect the pattern of infection of the two worm species in dogs and thus influence the geographical distribution of these two filarial worms. In medical and veterinary practice, histopathology was the most commonly used diagnostic technique (31.3%; 95% CI 2.5-60.2%). The low specificity of histopathology to speciate the various Dirofilaria spp. may lead to misdiagnosis. It was identified in this study that several regions of India and Sri Lanka have not yet been surveyed for dirofilariasis. This limits our understanding of the geographical distribution and interspecies interactions of the two parasites within these countries. Parasite distribution, disease prevalence, and interspecies interactions between the vectors and the host should be targeted for future research.

Associations between canine hookworm infection and dog owners' awareness, perception, and behaviour: A cross-sectional study in Brisbane, Queensland, 2019-2020.

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.

Prevalence and distribution of Anaplasma phagocytophilum in ticks collected from dogs in the United Kingdom.

BACKGROUND: Anaplasma phagocytophilum is the etiological agent of canine granulocytic anaplasmosis in dogs and causes human granulocytic anaplasmosis (HGA). Tick-borne anaplasmosis has been recognised as an emerging zoonotic health concern worldwide. The aim of the present study was to determine the prevalence of A. phagocytophilum in ticks collected from dogs in the UK and map its distribution. Routine surveillance of tick-borne disease is essential as part of a "One Health" approach to infectious disease management. METHODS: Tick DNA samples collected in 2015 as part of a large-scale tick surveillance programme were analysed using a previously validated diagnostic quantitative PCR for A. phagocytophilum. RESULTS: PCR analysis indicated that 138 out of 2994 tick DNA samples analysed were positive for A. phagocytophilum, a prevalence of 4.6% (95% CI: 3.89-5.42). Among these 138 tick DNA samples, 131 were from Ixodes ricinus, six were from Ixodes hexagonus and one was from Ixodes canisuga. Three of the I. ricinus tick DNA samples positive for A. phagocytophilum DNA were also positive for Borrelia spp. DNA and one was positive for Babesia spp. DNA, indicating co-infection. The ticks positive for the pathogen DNA were found widely distributed throughout the UK. CONCLUSIONS: These data provide important information on the prevalence and wide distribution of A. phagocytophilum in ticks infesting dogs within the UK.

Perceptions of dog owners towards canine gastrointestinal parasitism and associated human health risk in Southeast Queensland.

Canine companion animals can carry a number of zoonotic parasites which can adversely impact both human and animal health. Previous studies in Australia indicated that while parasitic infections in dogs are still common and there is variability in the awareness and perception of zoonotic risks among pet owners, the likely contribution of sociodemographic factors to the variation in awareness and perception needs to be further explored. The primary objective of this study is to quantify the relationship between dog owners' knowledge and beliefs about dog parasites and their sociodemographic characteristics. In this study, we surveyed a total of 281 dog owners in SE Queensland between April 2019 to March 2020 and the relationship between dog owners' perception of gastrointestinal parasite infection was assessed using an adaptation of the Health Belief Model, social cognitive framework for health protection. The model looked into the role of dog owners' demography on their perceived severity and susceptibility to zoonotic canine parasites and their likelihood of performing actions associated with worm control of their pets. Our results indicate that owners perceptions about parasitic disease severity in their pets was 26% higher in female dog owners compared to males, in respondents owning dogs over 10 years (27% higher than those owning a dog <3 years) and those owners that regularly deworm their pets and report faeces disposal. Our study indicates that the perceptions of pet owners towards zoonotic canine parasites varies demographically and owner education is important to prevent infection among dogs and control the zoonotic transmission to owners and the community. Finally, there was evidence that increased frequency of visits to veterinary clinics can increase the likelihood of owners performing worm treatment, proper faecal disposal, and cooking meat before feeding it to dogs.

The prevalence of Rickettsia felis DNA in fleas collected from cats and dogs in the UK.

In a large-scale survey in the UK, recruited veterinary practices were asked to inspect client-ownedcats and dogs, selected at random between April and June 2018, following a standardised flea inspection protocol. A total of 326 veterinary practices participated and 812 cats and 662 dogs were examined during the 3-month period. Fleas were collected, identified to species level and fleas of the same species collected from a single animal were pooled together and treated as a single sample. A total of 470 pooled flea samples were screened by PCR and DNA sequence analysis for a subset of Rickettsia species including R. felis and R. typhi. On analysis, 27 (5.7%) of the pooled flea samples were positive for R. felis DNA; these were predominantly in the cat flea, Ctenocephalides felis, but one dog flea, Ctenocephalides canis was also positive for this pathogen.

Pathogens in fleas collected from cats and dogs: distribution and prevalence in the UK.

BACKGROUND: Fleas (Siphonaptera) are the most clinically important ectoparasites of dogs and cats worldwide. Rising levels of pet ownership, climate change and globalisation are increasing the importance of a detailed understanding of the endemicity and prevalence of flea-borne pathogens. This requires continued surveillance to detect change. This study reports a large-scale survey of pathogens in fleas collected from client-owned cats and dogs in the UK. METHODS: Recruited veterinary practices were asked to follow a standardised flea inspection protocol on a randomised selection of cats and dogs brought into the practice in April and June 2018. A total of 326 practices participated and 812 cats and 662 dogs were examined. Fleas were collected, identified to species and pooled flea samples from each host were analysed for the presence of pathogens using PCR and sequence analysis. RESULTS: Overall, 28.1% of cats and 14.4% of dogs were flea infested. More than 90% of the fleas on both cats and dogs were cat fleas, Ctenocephalides felis felis. Fleas of the same species from each infested host were pooled. DNA was amplified from 470 of the pooled flea samples using conventional PCR, 66 of which (14% ± 95% CI 3.14%) were positive for at least one pathogen. Fifty-three (11.3% ± 95% CI 2.85%) of the pooled flea DNA samples were positive for Bartonella spp., 35 were from cats and 4 from dogs, the remainder had no host record. Seventeen of the Bartonella spp. samples were found to be Bartonella henselae, 27 were Bartonella clarridgeiae (of two different strains), 4 samples were Bartonella alsatica and one was Bartonella grahamii; 4 samples could not be identified. Fourteen (3% ± 95% CI 1.53%) of the flea DNA samples were found to be positive for Dipylidium caninum, 10 of the D. caninum-infected samples were collected from cats and one from a dog, the other 3 positive flea samples had no host species record. Only 3 flea samples were positive for Mycoplasma haemofelis or Mycoplasma haemocanis; 2 were collected from cats and one had no host species record. Three fleas were positive for both D. caninum and Bartonella spp. One flea was positive for both Bartonella spp. and M. haemofelis or M. haemocanis. CONCLUSIONS: This study highlights the need for ongoing flea control, particularly given the relatively high prevalence of Bartonella spp., which is of concern for both animal welfare and human health. The study demonstrates the ongoing need to educate pet owners about the effects of both flea infestation and also the pathogen risks these fleas present.

Spectrophotometric analysis of lipid used to examine the phenology of the tick Ixodes ricinus.

BACKGROUND: Ticks store lipid as an energy souce, which depletes progressively between blood meals. The amount of lipid and rate of lipid depletion can be used as a good indicator of the feeding history and assist in explaining the phenology of tick populations. However, existing gravimetric approaches to lipid measurement are relatively imprecise. To improve our ability to accurately measure lipid accumulation and metabolism in individual ticks, a microquantity colorimetric sulfophosphovanillan method of lipid estimation was standardised and used to explore the seasonal variations in the lipid content of I. ricinus nymphs. RESULTS: Lipid values for field-derived questing ticks, collected by blanket dragging, varied between 5-45 µg and clear patterns of lipid depletion were demonstrated under controlled laboratory conditions. For field populations collected monthly over two years, the results indicate that two different cohorts of nymphs enter the questing tick population in autumn and spring, with very few nymphs joining the population in summer. CONCLUSIONS: The data illustrate the seasonal change in lipid content of nymphal ticks, reflecting their feeding history and highlight the utility of the spectrophotometric technique for analysis of lipid in ticks in helping to improve our understanding of seasonal activity patterns.

Anaplasma phagocytophilum, Bartonella spp., haemoplasma species and Hepatozoon spp. in ticks infesting cats: a large-scale survey.

BACKGROUND: Ticks derived from cats have rarely been evaluated for the presence of pathogens. The aim of this study was to determine the prevalence of Anaplasma phagocytophilum, Bartonella spp., haemoplasma species and Hepatozoon spp. in ticks collected from cats in the UK. METHODS: Five hundred and forty DNA samples extracted from 540 ticks collected from cats presenting to veterinarians in UK practices were used. Samples underwent a conventional generic PCR assay for detection of Hepatozoon spp. and real-time quantitative PCR assays for detection of Anaplasma phagocytophilum and three feline haemoplasma species and a generic qPCR for detection of Bartonella spp. Feline 28S rDNA served as an endogenous internal PCR control and was assessed within the haemoplasma qPCR assays. Samples positive on the conventional and quantitative generic PCRs were submitted for DNA sequencing for species identification. RESULTS: Feline 28S rDNA was amplified from 475 of the 540 (88.0%) ticks. No evidence of PCR inhibition was found using an internal amplification control. Of 540 ticks, 19 (3.5%) contained DNA from one of the tick-borne pathogens evaluated. Pathogens detected were: A. phagocytophilum (n = 5; 0.9%), Bartonella spp. (n = 7; 1.3%) [including Bartonella henselae (n = 3; 0.6%) and Bartonella clarridgeiae (n = 1; 0.2%)], haemoplasma species (n = 5; 0.9%), "Candidatus Mycoplasma haemominutum" (n = 3; 0.6%), Mycoplasma haemofelis (n = 1; 0.2%), "Candidatus Mycoplasma turicensis" (n = 1; 0.2%), Hepatozoon spp. (n = 2; 0.4%), Hepatozoon felis (n = 1; 0.2%) and Hepatozoon silvestris (n = 1; 0.2%). CONCLUSION: These data provide important information on the prevalence of tick-borne pathogens in ticks infesting cats, with the identification of haemoplasma species, A. phagocytophilum, H. felis and Bartonella spp. (including B. henselae and B. clarridgeiae). This study also documents the first report of H. silvestris in ticks collected from domestic cats.

Contributions to the phylogeny of Ixodes (Pholeoixodes) canisuga, I. (Ph.) kaiseri, I. (Ph.) hexagonus and a simple pictorial key for the identification of their females.

BACKGROUND: In Europe, hard ticks of the subgenus Pholeoixodes (Ixodidae: Ixodes) are usually associated with burrow-dwelling mammals and terrestrial birds. Reports of Pholeoixodes spp. from carnivores are frequently contradictory, and their identification is not based on key diagnostic characters. Therefore, the aims of the present study were to identify ticks collected from dogs, foxes and badgers in several European countries, and to reassess their systematic status with molecular analyses using two mitochondrial markers. RESULTS: Between 2003 and 2017, 144 Pholeoixodes spp. ticks were collected in nine European countries. From accurate descriptions and comparison with type-materials, a simple illustrated identification key was compiled for adult females, by focusing on the shape of the anterior surface of basis capituli. Based on this key, 71 female ticks were identified as I. canisuga, 21 as I. kaiseri and 21 as I. hexagonus. DNA was extracted from these 113 female ticks, and from further 31 specimens. Fragments of two mitochondrial genes, cox1 (cytochrome c oxidase subunit 1) and 16S rRNA, were amplified and sequenced. Ixodes kaiseri had nine unique cox1 haplotypes, which showed 99.2-100% sequence identity, whereas I. canisuga and I. hexagonus had eleven and five cox1 haplotypes, respectively, with 99.5-100% sequence identity. The distribution of cox1 haplotypes reflected a geographical pattern. Pholeoixodes spp. ticks had fewer 16S rRNA haplotypes, with a lower degree of intraspecific divergence (99.5-100% sequence identity) and no geographical clustering. Phylogenetic analyses were in agreement with morphology: I. kaiseri and I. hexagonus (with the similar shape of the anterior surface of basis capituli) were genetically more closely related to each other than to I. canisuga. Phylogenetic analyses also showed that the subgenus Eschatocephalus (bat ticks) clustered within the subgenus Pholeoixodes. CONCLUSIONS: A simple, illustrated identification key is provided for female Pholeoixodes ticks of carnivores (including I. hexagonus and I. rugicollis) to prevent future misidentification of these species. It is also shown that I. kaiseri is more widespread in Europe than previously thought. Phylogenetic analyses suggest that the subgenus Pholeoixodes is not monophyletic: either the subgenus Eschatocephalus should be included in Pholeoixodes, or the latter subgenus should be divided, which is a task for future studies.

Prevalence of ticks and tick-borne pathogens: Babesia and Borrelia species in ticks infesting cats of Great Britain.

In a study of tick and tick-borne pathogen prevalence, between May and October 2016, 278 veterinary practices in Great Britain examined 1855 cats. Six-hundred and one cats were found to have attached ticks. The most frequently recorded tick species was Ixodes ricinus (57.1%), followed by Ixodes hexagonus (41.4%) and Ixodes trianguliceps (1.5%). Male cats, 4-6 years of age living in rural areas were most likely to be carrying a tick; hair length and tick treatment history had no significant association with attachment. For cats that were parasitized by ticks in large urban areas, I. hexagonus was the most frequent species recorded. Molecular analysis was possible for 541 individual tick samples, others were too damaged for analysis; Babesia spp., and Borrelia burgdorferi sensu lato were identified in 1.1% (n=6) and 1.8% (n=10) of these, respectively. Babesia spp. included Babesia vulpes sp. nov./Babesia microti-like (n=4) in I. hexagonus and Babesia venatorum (n=2) in I. ricinus. Borrelia burgdorferi s.l. species included Borrelia garinii (n=6) and Borrelia afzelii (n=4). The majority of B. burgorferi s.l. cases were found in I. ricinus, with B. afzelii in one I. hexagonus nymph. No Borrelia or Babesia spp. were present in I. trianguliceps. To determine a true prevalence for ticks on cats, practices that only submitted questionnaires from cats with ticks and practices that submitted fewer than 5 returns per week were removed; amongst those considered to have adhered strictly to the collection protocol, feline tick prevalence amongst cats that had access to the outdoors was 6.6%. These results show that ticks can be found on cats throughout Great Britain, which harbour a range of species of Babesia and B. burgdorferi s.l. and that cats, particularly in green spaces within urban areas, may form an important host for I. hexagonus, a known vector of pathogens.

Ticks infesting domestic dogs in the UK: a large-scale surveillance programme.

BACKGROUND: Recent changes in the distribution of tick vectors and the incidence of tick-borne disease, driven variously by factors such as climate change, habitat modification, increasing host abundance and the increased movement of people and animals, highlight the importance of ongoing, active surveillance. This paper documents the results of a large-scale survey of tick abundance on dogs presented to veterinary practices in the UK, using a participatory approach that allows relatively cost- and time-effective extensive data collection. METHODS: Over a period of 16 weeks (April-July 2015), 1094 veterinary practices were recruited to monitor tick attachment to dogs and provided with a tick collection and submission protocol. Recruitment was encouraged through a national publicity and communication initiative. Participating practices were asked to select five dogs at random each week and undertake a thorough, standardized examination of each dog for ticks. The clinical history and any ticks were then sent to the investigators for identification. RESULTS: A total of 12,000 and 96 dogs were examined and 6555 tick samples from infested dogs were received. Ixodes ricinus (Linnaeus) was identified on 5265 dogs (89 %), Ixodes hexagonus Leach on 577 (9.8 %) and Ixodes canisuga Johnston on 46 (0.8 %). Ten dogs had Dermacentor reticulatus (Fabricius), one had Dermacentor variabilis (Say), three had Haemaphysalis punctata Canesteini & Fanzago and 13 had Rhipicephalus sanguineus Latreille. 640 ticks were too damaged for identification. All the R. sanguineus and the single D. variabilis were on dogs with a recent history of travel outside the UK. The overall prevalence of tick attachment was 30 % (range 28-32 %). The relatively high prevalence recorded is likely to have been inflated by the method of participant recruitment. CONCLUSION: The data presented provide a comprehensive spatial understanding of tick distribution and species abundance in the UK against which future changes can be compared. Relative prevalence maps show the highest rates in Scotland and south west England providing a valuable guide to tick-bite risk in the UK.

Bacillus cereus food poisoning: international and Indian perspective.

Food borne illnesses result from eating food or drinking beverages that are contaminated with chemical matter, heavy metals, parasites, fungi, viruses and Bacteria. Bacillus cereus is one of the food-borne disease causing Bacteria. Species of Bacillus and related genera have long been troublesome to food producers on account of their resistant endospores. Their spores may be present on various types of raw and cooked foods, and their ability to survive high cooking temperatures requires that cooked foods be served hot or cooled rapidly to prevent the growth of this bacteria. Bacillus cereus is well known as a cause of food poisoning, and much more is now known about the toxins produced by various strains of this species, so that its significance in such episodes are clearer. However, it is still unclear why such cases are so rarely reported worldwide.

Esterase profile of Rhipicephalus (Boophilus) microplus populations collected from Northern India exhibiting varied susceptibility to deltamethrin.

Rhipicephalus (Boophilus) microplus is an economically important ectoparasite of cattle. Chemical acaricides remain the most practical method for control of these pests. During past two decades there have been increasing reports of resistance development against synthetic pyrethroids in tick populations of this species throughout the world. A study was conducted to determine the level of susceptibility of R. (B.) microplus to deltamethrin collected from different geographical locations of northern India. LPT bioassay results revealed LC(50) values of deltamethrin ranging from 0.035 to 0.00037 % A.I. Esterase profile of the tick larval extracts using native PAGE, revealed 5 bands of esterase activity designated EST-5 to EST-1A. Inhibitory tests recognized EST-1, EST-2 and EST-3 as Acetylcholinesterases (AchEs), EST-4 and EST-5 as Carboxylesterases (CaEs). The band intensity varied between tick populations of various locations, being more intense in case of the resistant populations. An extra band of esterase activity (EST-1A) was obtained in larval extracts of ticks from 3 locations. This increased esterase activity may be involved in the resistance development in these tick populations. Acaricide resistance is a multi-factorial phenomenon, thus other causes of increased resistance like sodium channel mutation and reduced drug penetration (e.g. cuticle thickening) and behavioural changes (e.g. avoiding the pesticides) are to be tested in future in order to confirm the basic cause of the resistance development in these acaricide resistant tick populations.