Seasonality and other risk factors for fleas infestations in domestic dogs and cats.

Fleas in the genus Ctenocephalides are the most clinically important parasitic arthropods of dogs and cats worldwide yet risk factors that might increase the risk of infestation in small animals remains unclear. Here we developed a supervised text mining approach analysing key aspects of flea epidemiology using electronic health records from domestic cats and dogs seen at a sentinel network of 191 voluntary veterinary practices across Great Britain between March 2014 and July 2020. Our methods identified fleas as likely to have been present during 22,276 of 1,902,016 cat consultations (1.17%) and 12,168 of 4,844,850 dog consultations (0.25%). Multivariable logistic regression modelling found that animals originating from areas of least deprivation were associated with 50% reductions in odds of veterinary-recorded flea infestation compared to the most deprived regions in England. Age of the animal was significantly associated with flea presentation in both cats and dogs, with cases peaking before animals reached 12 months. Cases were recorded through each study years, peaking between July and October, with fluctuations between each year. Our findings can be used towards healthcare messaging for veterinary practitioners and owners.

Onchocerca lupi in imported dogs in the UK: implications for animal and public health.

BACKGROUND: Onchocerca lupi is a filarial nematode affecting dogs, and occasionally cats and humans, in continental Europe, North Africa, the Middle East, and the USA. Adult worms are usually found in periocular nodules and enucleation is sometimes required if the infection fails to respond to other treatment options. CASE PRESENTATION: Here, we report the presence of O. lupi in the UK for the first time. Of two dogs re-homed from continental Europe, one developed an ocular nodule seven years after arrival from Portugal. The conjunctival perilimbal mass in its left eye was surgically removed but despite anthelminthic treatment, a further nodule developed in the same eye six months later. In the second case - a dog imported from Romania 12 months earlier - a perilimbal mass was excised from the left eye and prior anthelminthic treatment was supplemented with oral prednisolone and doxycycline. However, nodules recurred, and the left globe was subsequently enucleated. Conjunctival hyperaemia then appeared in the right eye and neither additional anthelminthic treatment nor removal of worm masses failed to prevent the further development of lesions. Excised adult worms were identified in both cases as O. lupi based on morphological characteristics, as well as PCR and sequencing of cytochrome c oxidase subunit I and 12S rRNA gene fragments. CONCLUSION: O. lupi parasitosis can apparently remain cryptic in dogs for several years before any clinical signs manifest. Moreover, the progression of infection can be highly aggressive and recalcitrant to both surgical intervention and anthelminthic treatment. Increasingly, former stray dogs of unknown infection status are entering the UK, raising both veterinary and public health concerns.

Symbiosis in an overlooked microcosm: a systematic review of the bacterial flora of mites.

A dataset of bacterial diversity found in mites was compiled from 193 publications (from 1964 to January 2015). A total of 143 mite species belonging to the 3 orders (Mesostigmata, Sarcoptiformes and Trombidiformes) were recorded and found to be associated with approximately 150 bacteria species (in 85 genera, 51 families, 25 orders and 7 phyla). From the literature, the intracellular symbiont Cardinium, the scrub typhus agent Orientia, and Wolbachia (the most prevalent symbiont of arthropods) were the dominant mite-associated bacteria, with approximately 30 mite species infected each. Moreover, a number of bacteria of medical and veterinary importance were also reported from mites, including species from the genera Rickettsia, Anaplasma, Bartonella, Francisella, Coxiella, Borrelia, Salmonella, Erysipelothrix and Serratia. Significant differences in bacterial infection patterns among mite taxa were identified. These data will not only be useful for raising awareness of the potential for mites to transmit disease, but also enable a deeper understanding of the relationship of symbionts with their arthropod hosts, and may facilitate the development of intervention tools for disease vector control. This review provides a comprehensive overview of mite-associated bacteria and is a valuable reference database for future research on mites of agricultural, veterinary and/or medical importance.

Proteomics and in silico approaches to extend understanding of the glutathione transferase superfamily of the tropical liver fluke Fasciola gigantica.

Fasciolosis is an important foodborne, zoonotic disease of livestock and humans, with global annual health and economic losses estimated at several billion US$. Fasciola hepatica is the major species in temperate regions, while F. gigantica dominates in the tropics. In the absence of commercially available vaccines to control fasciolosis, increasing reports of resistance to current chemotherapeutic strategies and the spread of fasciolosis into new areas, new functional genomics approaches are being used to identify potential new drug targets and vaccine candidates. The glutathione transferase (GST) superfamily is both a candidate drug and vaccine target. This study reports the identification of a putatively novel Sigma class GST, present in a water-soluble cytosol extract from the tropical liver fluke F. gigantica. The GST was cloned and expressed as an enzymically active recombinant protein. This GST shares a greater identity with the human schistosomiasis GST vaccine currently at Phase II clinical trials than previously discovered F. gigantica GSTs, stimulating interest in its immuno-protective properties. In addition, in silico analysis of the GST superfamily of both F. gigantica and F. hepatica has revealed an additional Mu class GST, Omega class GSTs, and for the first time, a Zeta class member.

Diffusive uptake rates for passive air sampling: Application to volatile organic compound exposure during FIREX-AQ campaign.

Passive (diffusive) sampling using sorbents is an economical and versatile method of measuring pollutants in air, including volatile organic compounds (VOCs). Diffusive uptake rates (UTRs) are needed for each analyte to obtain average concentrations during a specific passive sampling time duration. Here, a simultaneous active/diffusive ambient air sampling technique on Tenax®TA was employed to measure 24-hours, 7, 14 and 28-days UTRs of up to 27 VOCs, including benzene, toluene, ethylbenzene, xylenes (BTEX), C6-C12 hydrocarbons, benzenes derivatives, tetrachloroethylene, pinenes and limonene. Samples were analyzed via thermal desorption-gas chromatography-mass spectrometry (TD-GC-MS) for desired analytes. Seven-day UTR values ranged from 0.17 to 0.59 mL/min and many compounds exhibited a linear relationship with UTR and time duration up to 14 or 28 days. This may be the most comprehensive UTR tabulation of VOCs on Tenax®TA for time periods of 24 hours -28 days available. These rates were applied to VOC data measured during the 2019 NASA/NOAA Fire Influence on Regional to Global Environments and Air Quality (FIREX-AQ) campaign, with goals to determine the chemical composition of western US wildfire smoke and to assess human exposure to air toxics. Summer 2019 exposure levels of BTEX at five Northwestern cities were low and the cancer risk due to benzene was assessed during FIREX-AQ to be background or 1 × 10-6. The UTRs derived here can be useful in applications of diffusive sampling, including estimation of sub-chronic to chronic human exposure risk of air toxics and wildfire smoke.

Health Risk Implications of Volatile Organic Compounds in Wildfire Smoke During the 2019 FIREX-AQ Campaign and Beyond.

Fire Influence on Regional to Global Environments and Air Quality was a NOAA/NASA collaborative campaign conducted during the summer of 2019. The objectives included identifying and quantifying wildfire composition, smoke evolution, and climate and health impacts of wildfires and agricultural fires in the United States. Ground based mobile sampling via sorbent tubes occurred at the Nethker and Williams Flats fires (2019) and Chief Timothy and Whitetail Loop fires (2020) in Idaho and Washington. Air samples were analyzed through thermal desorption-gas chromatography-mass spectrometry for a variety of volatile organic compounds to elucidate both composition and health impacts. Benzene, toluene, ethylbenzene, xylenes, butenes, phenol, isoprene and pinenes were observed in the wildfire smoke, with benzene ranging from 0.04 to 25 ppbv. Health risk was assessed for each fire by determining sub-chronic (wildfire event) and projected chronic inhalation risk exposure from benzene, a carcinogen, as well as other non-carcinogenic compounds including toluene, ethylbenzene, xylenes, and hexane. The cancer risk of benzene from sub-chronic exposure was 1 extra cancer per million people and ranged from 1 to 19 extra cancers per million people for the projected chronic scenarios, compared to a background level of 1 extra cancer per million people. The hazard index of non-carcinogenic compounds was less than one for all scenarios and wildfires sampled, which was considered low risk for non-cancer health events.

House dust and storage mite contamination of dry dog food stored in open bags and sealed boxes in 10 domestic households.

Dry pet food is a potential source of exposure to house dust and storage mite allergens in canine atopic dermatitis. This study evaluated contamination of house dust and dry dog food stored in paper bags, sealable plastic bags and sealable plastic boxes in 10 households for 90 days using Acarex(®) tests for guanine, a Der p 1 ELISA and mite flotation. Acarex(®) tests were negative in all the food samples but positive in all the house dust samples. The Der p 1 levels and mite numbers significantly increased in food from paper bags (P = 0.0073 and P = 0.02, respectively), but not plastic bags or boxes. Mite numbers and Der p 1 levels were 10-1000 times higher in house dust than the corresponding food samples (P < 0.0001). There were significant correlations between Der p 1 in house dust and food from the paper (P < 0.0001) and plastic bags (P = 0.003), and mite numbers in house dust and food from the paper bags (P = 0.0007). Bedding and carpets were significantly associated with Der p 1 levels in house dust (P = 0.015 and P = 0.01, respectively), and food from the paper (both P = 0.02) and plastic bags (P = 0.03 and P = 0.04, respectively). Mites were identified in six of 10 paper bag, three of 10 plastic bag, one of 10 plastic box and nine of 10 house dust samples. These comprised Dermatophagoides (54%), Tyrophagus (10%; all from food) and unidentified mites (36%). Storage of food in sealable plastic boxes largely prevented contamination for 3 months. Exposure to mites and mite proteins in all the stored food, however, appeared to be trivial compared with house dust.

Automatic barcode gap discovery reveals diverse clades of Rhipicephalus spp. and Haemaphysalis spp. ticks from small mammals in 'Asir, Saudi Arabia.

BACKGROUND: The ixodid tick genera Rhipicephalus and Haemaphysalis contain several species of medical and/or veterinary importance, but their diversity in some regions of the world remains under-explored. For instance, very few modern studies have been performed on the taxonomy of these genera on the Arabian Peninsula. METHODS: In this study, we trapped small mammals in the 'Asir Mountains of south-western Saudi Arabia and collected tick specimens for morphological examination and molecular barcoding, targeting three mitochondrial loci: cox1, 16S rRNA and 12S rRNA. RESULTS: We obtained a total of 733 ticks (608 Haemaphysalis spp. and 125 Rhipicephalus spp.) from 75 small mammal hosts belonging to six species. All tick specimens were immature except for nine adults recovered from a hedgehog (Paraechinus aethiopicus). Morphologically, the Rhipicephalus ticks resembled R. camicasi, but the Haemaphysalis ticks showed differences in palp morphology compared with species previously described from Saudi Arabia. Phylogenetic analysis and automatic barcode gap discovery identified a novel clade of Rhipicephalus sp. representing most of the nymphs. This was most closely related to R. leporis, R. guilhoni and R. linnaei. The adult ticks and a small proportion of nymphs clustered with R. camicasi sequences from a previous study. Finally, the Haemaphysalis nymphs formed two distinct clades that were clearly separated from all reference sequences but closest to some African species. CONCLUSIONS: This apparent high level of tick diversity observed in a single study site of only ~ 170 km2, on a relatively small number of hosts, highlights the potential for the discovery of new tick species on the Arabian Peninsula.

Bacterial Pathogens and Symbionts Harboured by Ixodes ricinus Ticks Parasitising Red Squirrels in the United Kingdom.

Red squirrels (Sciurus vulgaris) are native to most of Eurasia; in much of the United Kingdom, they have been supplanted by the non-native grey squirrel, and are considered an endangered species. Very little is known about the range of tick-borne pathogens to which UK red squirrels are exposed. As part of trap-and-release surveys examining prevalence of Mycobacterium spp. in red squirrel populations on two UK islands, Ixodes ricinus ticks were removed from squirrels and PCR screened for Borrelia spp., intracellular arthropod-borne bacteria and the parasitic wasp Ixodiphagus hookeri. At both sites, the most commonly encountered tick-transmitted bacterium was Borrelia burgdorferi sensu lato (overall minimum prevalence 12.7%), followed by Anaplasma phagocytophilum (overall minimum prevalence 1.6%). Single ticks infected with Spiroplasma were found at both sites, and single ticks infected with Borrelia miyamotoi or an Ehrlichia sp. at one site. Ticks harbouring Wolbachia (overall minimum prevalence 15.2%) were all positive for I. hookeri. Our study shows that UK red squirrels are potentially exposed to a variety of bacterial pathogens via feeding ticks. The effects on the health and survival of this already vulnerable wildlife species are unknown, and further studies are needed to evaluate the threat posed to red squirrels by Borrelia and other tick-borne pathogens.

Threat of exotic worms in dogs imported from Romania.

More and more dogs are legally entering the UK from overseas, bringing with them the risk of importing unusual parasites, some of which are zoonotic. Here, Ian Wright and colleagues describe three cases of parasitic worms found in Romanian dogs.

Ecological and microbiological diversity of chigger mites, including vectors of scrub typhus, on small mammals across stratified habitats in Thailand.

BACKGROUND: Scrub typhus, caused by a bacterial pathogen (Orientia spp.), is a potentially life-threatening febrile illness widely distributed in the Asia-Pacific region and is emerging elsewhere. The infection is transmitted by the larval stage of trombiculid mites ("chiggers") that often exhibit low host specificity. Here, we present an analysis of chigger ecology for 38 species sampled from 11 provinces of Thailand and microbiomes for eight widespread species. RESULTS: In total, > 16,000 individual chiggers were collected from 1574 small mammal specimens belonging to 18 species across four horizontally-stratified habitat types. Chigger species richness was positively associated with higher latitudes, dry seasonal conditions, and host maturity; but negatively associated with increased human land use. Human scrub typhus incidence was found to be positively correlated with chigger species richness. The bacterial microbiome of chiggers was highly diverse, with Sphingobium, Mycobacterium, Neisseriaceae and various Bacillales representing the most abundant taxa. Only Leptotrombidium deliense was found to be infected with Orientia and another potential pathogen, Borrelia spp., was frequently detected in pools of this species. ß-diversity, but not α-diversity, was significantly different between chigger species and geographic regions, although not between habitat types. CONCLUSION: Our study identified several key environmental and host-derived correlates of chigger species richness across Thailand, which in turn impacted on human scrub typhus incidence. Moreover, this first extensive field survey of the chigger microbiome revealed species- and province-level variation in microbial ß-diversity across the country, providing a framework for future studies on interactions between pathogens and other symbionts in these understudied vectors.

Multi-locus sequence typing of Ixodes ricinus and its symbiont Candidatus Midichloria mitochondrii across Europe reveals evidence of local co-cladogenesis in Scotland.

Ticks have relatively complex microbiomes, but only a small proportion of the bacterial symbionts recorded from ticks are vertically transmitted. Moreover, co-cladogenesis between ticks and their symbionts, indicating an intimate relationship over evolutionary history driven by a mutualistic association, is the exception rather than the rule. One of the most widespread tick symbionts is Candidatus Midichloria, which has been detected in all of the major tick genera of medical and veterinary importance. In some species of Ixodes, such as the sheep tick Ixodes ricinus (infected with Candidatus Midichloria mitochondrii), the symbiont is fixed in wild adult female ticks, suggesting an obligate mutualism. However, almost no information is available on genetic variation in Candidatus M. mitochondrii or possible co-cladogenesis with its host across its geographic range. Here, we report the first survey of Candidatus M. mitochondrii in I. ricinus in Great Britain and a multi-locus sequence typing (MLST) analysis of tick and symbiont between British ticks and those collected in continental Europe. We show that while the prevalence of the symbiont in nymphs collected in England is similar to that reported from the continent, a higher prevalence in nymphs and adult males is apparent in Wales. In general, Candidatus M. mitochondrii exhibits very low levels of sequence diversity, although a consistent signal of host-symbiont coevolution was apparent in Scotland. Moreover, the tick MLST scheme revealed that Scottish specimens form a clade that is partially separated from other British ticks, with almost no contribution of continental sequence types in this north-westerly border of the tick's natural range. The low diversity of Candidatus M. mitochondrii, in contrast with previously reported high rates of polymorphism in I. ricinus mitogenomes, suggests that the symbiont may have swept across Europe recently via a horizontal, rather than vertical, transmission route.

Lack of serologic evidence of Neospora caninum in humans, England.

Retrospective testing of 3,232 serum samples from the general population and 518 serum samples from a high-risk group showed no evidence of human exposure to Neospora caninum in England. Results were obtained by using immunofluorescence antibody testing and ELISA to analyze frequency distribution.

Elmslie-Trillat procedure for the treatment of recurrent patellar instability.

PURPOSE: The purpose of this study was to evaluate the mid-term results of the Elmslie-Trillat procedure for the treatment of patella instability. Patella instability can be addressed by proximal, distal, and combined approaches. The Elmslie-Trillat procedure is a combined bony distal realignment with medial soft tissue technique offering a more rapid recovery than more extensive distal realignments. METHODS: A consecutive series of patients with recurrent patellar instability treated with a modified Elmslie-Trillat procedure were assessed subjectively by visual analog scale, Tegner scale, and both pre-and postoperatively by physical examination, International Knee Documentation Committee activity, Lysholm, and Fulkerson knee scores. Inclusion criteria were patients with recurrent lateral dislocations or increasingly frequent subluxations who failed nonoperative therapy. Exclusion criteria were open growth plates, patellofemoral arthritis, and concurrent cruciate ligament or meniscal injuries. The primary endpoint was evidence of recurrent instability. The secondary endpoint was the functional scoring status. RESULTS: Thirty-five knees were evaluated for an average of 98 months (range, 25 to 209) postsurgery. The average age was 27.7 years. Twelve had failed a previous surgery to correct the patellar instability. The mean Lysholm and Fulkerson scores improved from 44.5 and 42.4 to 83.4 and 84.3, respectively. Follow-up Tegner score was 3.8, and the average visual analog scale score was 7.9 out of 10. Two patients had recurrent dislocations and 1 had a single subluxation 6 weeks after surgery. CONCLUSIONS: The Elmslie-Trillat procedure successfully eliminated recurrent patellar instability in 91.4%. Functionally, all patients were improved at an average of 98 months after surgery. LEVEL OF EVIDENCE: Level IV, therapeutic case series.

Insertion force of articular cartilage transplantation systems.

This study compared the insertion force, plug harvest consistency, and recipient site creation consistency of 4 different articular cartilage transplantation systems (COR, OATS, Mosaicplasty, and New COR2) during plug insertion using a single-impaction technique. Maximum insertion forces fell into 3 statistically different groups: group 1, OATS 8-mm (238 N) and 10-mm (215 N) systems; group 2, COR 6-mm (133 N) and 8-mm (176 N), Mosaicplasty, 6.5-mm (147 N) and 8.5-mm (134 N), and OATS 6-mm (137 N) systems; and group 3, New COR2 6-mm (68 N), 8-mm (55 N), and 10-mm (54 N) systems (P < or = .05). OATS compaction pressures were 172 N (6 mm), 353 N (8 mm), and 550 N (10 mm). COR, New COR2, and Mosaicplasty donor plugs were created consistently, but the Mosaicplasty system required toggling. OATS plugs had inconsistent lengths but required no toggling. Insertion forces with the New COR2 system were statistically the lowest. Compaction significantly increased surface forces.

A biomechanical study of Achilles tendon repair augmentation using GraftJacket matrix.

BACKGROUND: Ruptured Achilles tendons benefit from primary repair by decreasing re-rupture rates and allowing earlier range of motion. A stronger repair might allow for more aggressive rehabilitation decreasing postoperative stiffness, calf atrophy, and repair site gapping. The hypothesis of this study was that human dermal allograft augmentation of an Achilles repair would significantly increase repair strength and stiffness. This study evaluated strength and stiffness of an Achilles tendon repair augmented with a human dermal allograft (GraftJacket). MATERIALS AND METHODS: Eight matched pairs of human cadaver legs were used. Simulated Achilles tendon ruptures were created 4 cm proximal to the calcaneal insertion. All tendons were repaired with a Krackow locking loop stitch. One of each matched pair was augmented with GraftJacket. Each construct was pre-loaded at 10 N and cyclically loaded (20 cycles) from 2 N to 30 N at a rate of 5 N/sec on an Instron machine. This was followed by testing to failure at a displacement rate of 6 mm/sec. RESULTS: The ultimate failure load in the control group was 217 N +/- 31 compared to 455 N +/- 76.5 in the GraftJacket group (p < 0.001). The mean stiffness in the control group was 4.3 +/- 0.83 N/mm which was significantly less than the 12.99 +/- 5.34; N/mm in the GraftJacket group (p = 0.002). CONCLUSION: The augmentation of an Achilles tendon repair with GraftJacket significantly increased repair strength and stiffness. CLINICAL SIGNIFICANCE: These findings suggest that a GraftJacket augmented Achilles tendon repair could acutely withstand a more aggressive rehabilitation program, potentially decreasing ankle stiffness and allowing earlier return to full activities.

Genomes of trombidid mites reveal novel predicted allergens and laterally transferred genes associated with secondary metabolism.

Background: Trombidid mites have a unique life cycle in which only the larval stage is ectoparasitic. In the superfamily Trombiculoidea ("chiggers"), the larvae feed preferentially on vertebrates, including humans. Species in the genus Leptotrombidium are vectors of a potentially fatal bacterial infection, scrub typhus, that affects 1 million people annually. Moreover, chiggers can cause pruritic dermatitis (trombiculiasis) in humans and domesticated animals. In the Trombidioidea (velvet mites), the larvae feed on other arthropods and are potential biological control agents for agricultural pests. Here, we present the first trombidid mites genomes, obtained both for a chigger, Leptotrombidium deliense, and for a velvet mite, Dinothrombium tinctorium. Results: Sequencing was performed using Illumina technology. A 180 Mb draft assembly for D. tinctorium was generated from two paired-end and one mate-pair library using a single adult specimen. For L. deliense, a lower-coverage draft assembly (117 Mb) was obtained using pooled, engorged larvae with a single paired-end library. Remarkably, both genomes exhibited evidence of ancient lateral gene transfer from soil-derived bacteria or fungi. The transferred genes confer functions that are rare in animals, including terpene and carotenoid synthesis. Thirty-seven allergenic protein families were predicted in the L. deliense genome, of which nine were unique. Preliminary proteomic analyses identified several of these putative allergens in larvae. Conclusions: Trombidid mite genomes appear to be more dynamic than those of other acariform mites. A priority for future research is to determine the biological function of terpene synthesis in this taxon and its potential for exploitation in disease control.

Introducing forensic entomology in cases of suspected animal neglect.

Cases of arthropod-infested, abandoned or abused animals are sometimes brought to the attention of veterinarians by animal welfare authorities, with the requirement for a full postmortem examination towards criminal or civil proceedings. In these situations, entomology is an important support tool for the pathologists' investigation since the presence of arthropod life cycle stages serve as reliable forensic markers, especially for blowflies which form the first waves of activity following death. In the present study, 70 cadavers from a total of 544 referred to the Institute of Veterinary Science, University of Liverpool, between 2009 and 2014 displayed evidence of infestation. Here, the authors introduce principles of applied entomology and simplified approaches for estimating the minimum time since death, relevant in the context of routine submissions and the broad remit of individual cases. Despite often limited availability of scene of the crime and local thermal data, the interpretation of the minimum postmortem interval has nonetheless proved valuable as an adjunct to the expert pathology report. However, future developments and enhanced accuracy in this area of animal welfare require resource and training in expertise, and agreed standardisation of both laboratory and field procedures.

Three cases of imported eyeworm infection in dogs: a new threat for the United Kingdom.

In July 2016 we described the first known case of canine ocular thelaziosis in the UK in a dog recently imported from Romania. Here we confirm our initial diagnosis using PCR followed by sequence analysis, and we report a further two clinical cases in dogs with recent history of travel to Italy and France. In view of the presence in the UK of the vector for Thelazia callipaeda, namely Phortica spp, we discuss the significance of these three cases in the context of the UK government's pet travel scheme, disease control and both animal and public health in the UK.