Search

Background: Relapsing fever borreliosis is an infectious disease caused by bacteria of the genus Borrelia, inflicting recurrent episodes of fever and spirochetemia in humans. Borrelia persica, the causative agent of relapsing fever in Israel, is prevalent over a broad geographic area that extends from India to Egypt. It is transmitted by the soft tick Ornithodoros tholozani and causes disease in humans as well as domestic cats and dogs. The goal of this study was to survey domestic dogs and cats in Israel for infection with B. persica., Methods: Blood, sera and demographic and clinical data were collected from dogs and cats brought for veterinary care in central Israel. PCR followed by DNA sequencing was used to detect B. persica DNA in blood samples, and an enzyme-linked immunosorbent assay (ELISA) was used to detect antibodies reactive with B. persica antigens in sera from the same animals. This is the first serological survey of B. persica in dogs and the first survey for antibodies reactive with a relapsing fever Borrelia sp. in cats globally., Results: Four of the 208 dogs (1.9%) and three of 103 cats (2.9%) sampled were positive by PCR for B. persica DNA, and 24 dogs (11.5%) and 18 cats (17.5%) were seropositive for B. persica antigen by ELISA. The ratio between PCR-positivity and seropositivity in both the dog and cat populations was 1:6. All four PCR-positive dogs and two of three PCR-positive cats were seronegative, suggesting a probable recent infection. Thrombocytopenia showed significant association with seropositivity in dogs (P = 0.003). In cats, anemia had a significant association with seropositivity (P = 0.0001), and thrombocytopenia was associated with the combined prevalence of seropositivity or PCR-positivity (P = 0.022)., Conclusions: Borrelia persica infection is more prevalent and widespread in domestic canine and feline populations in Israel than previously thought. Dogs and cats may play a role as reservoirs and sentinels for human infection. Precautions should be taken to prevent transfusion-transmitted infection between blood donor and recipient animals., (© 2022. The Author(s).)

Background: Three species of Leishmania cause disease in humans in Israel and are endemic in the Middle East: Leishmania infantum, Leishmania tropica and Leishmania major. These species infect dogs and cats, but little is known about their prevalence in pet populations and their clinical manifestations. A study on dog and cat Leishmania infection was conducted in a focus of human L. tropica infection in central Israel with the aim of getting insight on leishmaniosis in pets in an area where human infection is highly prevalent., Methods: Blood, demographic and clinical data were collected from dogs and cats brought for veterinary care in a focus of human L. tropica infection during 2018-2020. kDNA PCR and internal transcribed spacer1 high-resolution melt analysis PCR (ITS1 HRM PCR) with DNA sequencing were performed for the detection of Leishmania and species determination., Results: Forty-three of 189 dogs (22.8%) and 44 of 152 cats (28.9%) were positive for Leishmania spp. infection by kDNA PCR. The ITS1 HRM PCR detected six dogs (3.3%) infected with L. infantum and one (0.5%) with L. tropica, whereas six cats (3.9%) were found infected by L. infantum and five (3.3%) by L. tropica. Four of the five L. tropica-positive cats suffered from weight loss, four had azotemia, two with mild and two with severe azotemia and progressive renal disease. Three cats had gingivostomatitis; three had skin lesions with abscess and ulcers in two and scales and hair loss in another cat, which was also FIV +. This is the first report of feline L. tropica infection in Israel. Clinical information on cats with this infection from previous studies elsewhere is scarce., Conclusions: A high rate of Leishmania spp. infection, mostly estimated as sub-clinical, was found in dogs and cats admitted for veterinary care in an L. tropica focus. Among the animals in which infection could be characterized to the species level, more dogs were infected with L. infantum than with L. tropica while 5 of 11 cats were infected with L. tropica and had signs of systemic and skin disease not described before in feline L. tropica infection., (© 2022. The Author(s).)

Canine filarioids are zoonotic vector-borne parasitic nematodes distributed mostly in the tropics and subtropics. The aim of this study was to determine the prevalence, distribution, etiology and genetic variation of canine filarioid infections by three different PCR techniques in four Southern Romanian counties. Blood samples of 300 shelter dogs were screened for infections with canine filarioids by real-time PCR. To determine filarioid species and coinfections, samples positive in the initial screening were further tested by conventional PCR and sequenced. Results indicated that 17% of the tested dogs were positive for at least one filarioid species. The prevalence of D. repens infection was 11.7%, significantly higher than that of D. immitis (4.7%) and A. reconditum (1.3%) (p ≤ 0.003). The high prevalence of canine filarioid infections represents a challenge to animal and human health in the South of Romania, and they should be constantly monitored., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Background: Accurate diagnosis is imperative in dogs with clinical signs of parvovirus infection (CPV-2)., Objectives: To assess quantitative real-time PCR (qRT-PCR) for the diagnosis of CPV-2 infection, and determine the optimal sampling site. Secondarily, to compare qRT-PCR with a point-of-care PCR kit (PCRun), and to assess sensitivity of serology for CPV diagnosis., Animals: Sixty dogs with naturally acquired parvovirus infection, 44 unvaccinated puppies, of which 16 were followed after first and second vaccination, 15 adult dogs, of which 10 were followed also after a booster vaccine, and 9 dogs with distemper virus infection., Methods: Prospective study. Samples from the rectum, blood, and pharynx were obtained for PCR., Results: All dogs with a clinical diagnosis of parvovirus infection were positive by qRT-PCR in at least 1 sampling site (ie, rectum, blood, pharynx), and 50 (83%) of 60 were positive in all sites. qRT-PCR was negative in 67 (99%) of 68 healthy puppies (before-vaccination), puppies with distemper, and healthy adult dogs. Ten days after initial vaccination of puppies, 62% (fecal), 31% (blood), and 12% (pharyngeal) of samples were positive for CPV-2 on qRT-PCR. The proportion of positive pharyngeal samples decreased 20 days after vaccination and all sites were negative 12-28 days after second vaccination. Vaccinated adults were negative before and after booster vaccination., Conclusions and Clinical Importance: Molecular detection of CPV is sensitive, but specificity is hampered temporarily during the vaccination period. Blood, feces, and pharynx are suitable sampling sites. Fecal samples had the lowest sensitivity in sick dogs and highest positivity in puppies after vaccination., (© 2022 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals LLC on behalf of American College of Veterinary Internal Medicine.)

Background: Ticks are important vectors of many pathogens that have contributed to the morbidity and mortality of humans and domestic animals worldwide. Wildlife species have also been implicated as reservoir hosts of a variety of tick-borne pathogens. The objective of this study was to determine which tick-transmitted pathogens were present in the animals harvested from the forest in Trinidad for human consumption., Methods: Thin blood smears from 43 neotropical animals were examined microscopically for tick-borne pathogens. Additionally, DNA extraction and PCR amplification of the 16S rRNA gene were used for amplification of Anaplasma and Ehrlichia while the gltA gene was used for Bartonella, and Rickettsia spp. and the 18S rRNA gene for Babesia, Hepatozoon and Theileria species., Results: Pathogen DNA was amplified from four samples (a deer, collared peccary and two agoutis). Sequencing of the amplified products from the deer and collared peccary revealed 99.8% homology to Anaplasma bovis and 98.8% homology to Ehrlichia canis, respectively. Sequences from two agoutis revealed 90.4% homology to Theileria spp. DNA of Hepatozoon spp., Bartonella spp. Babesia spp. and Rickettsia spp. was not detected in any of the screened samples. An incidental finding in this study was the presence of bacteria in the blood of animals., Conclusions: The results indicate that the DNA of tick-transmitted pathogens is present at a frequency of about 10% in the study population and suggests that neotropical mammals may serve as a source for the potential transmission of tick-borne pathogens to domestic animals and humans. In addition, physicians and hunters should be aware of the symptoms associated with zoonotic tick-borne pathogens so that these infections can be recognised, diagnosed and treated promptly. Bacteria present in carcasses can pose a food safety hazard and hunters should be trained in proper harvesting and handling of carcasses., (© 2022. The Author(s).)

The parasitic filarioid Onchocerca lupi causes ocular disease characterized by conjunctivitis and nodular lesions. This nematode was first described in 1967 in a wolf from Georgia, and since then cases of infection from dogs and cats with ocular onchocercosis and sporadically from humans also with subcutaneous and cervical lesions caused by O. lupi have been reported from the Middle East, Europe, and North America. Due to its zoonotic potential, this parasitic infection has gained attention in the past 20 years. Phylogenetic studies have highlighted the recent divergence of O. lupi from other Onchocerca spp. and the importance of domestication in the evolutionary history of this worm. Moreover, the finding of an O. lupi genotype associated with subclinical and mild infection in the Iberian Peninsula, raises important questions about the pathogenicity of this presently enigmatic parasite., (Copyright ©2021, Yale Journal of Biology and Medicine.)

Background: Canine leishmaniosis (CanL) is a zoonotic disease caused by Leishmania infantum. Although usually transmitted by phlebotomine sand flies, infection by vertical transmission and by blood transfusion have also been reported., Methods: We describe the very early onset of clinical leishmaniosis, starting from 2 months of age, in a litter of pups born to an infected dam and sire. Seven pups from the litter of nine living in different households showed alopecic, exfoliative dermatitis and ulcerative cutaneous lesions. All pups and both parents were tested on at least one occasion both serologically, by enzyme-linked immunosorbent assay (ELISA), and by polymerase chain reaction (PCR) targeting the Leishmania ribosomal operon internal transcribed spacer 1 region and a short fragment of the kinetoplast minicircle; positive amplicons were sequenced., Results: All nine pups were PCR positive for L. infantum verified by DNA sequencing, seven were positive by conjunctival, five by blood, four by lymph node, and one by skin PCR from an ulcerative lesion. Both pups with no clinical signs were seronegative, while five of the seven pups with dermatologic abnormalities were seropositive by ELISA. The sire had typical clinical dermatologic and visceral findings of CanL, was seropositive and PCR positive for L. infantum in the lymph node and fluid from the vas deferens tested after the testes were removed by castration. The dam was sub-clinically infected and seronegative, but positive by blood, lymph node and conjunctival PCR for L. infantum. Allopurinol administered to all clinically affected dogs resulted in clinical recovery., Conclusions: Infection with L. infantum in both parents, the very early age of clinical onset among most of the pups, and the fact that the puppies were born and detected with signs of leishmaniosis in the winter, which is a season without sand fly activity in Israel, strongly suggest vertical transmission. Awareness of the possibility of vertical transmission of L. infantum and infection in littermates should be increased. It is recommended that littermates of young dogs with clinical leishmaniosis should be tested for sub-clinical infection as they may also be infectious to sand flies and thus to other dogs and to humans. Restricting the mating of infected bitches should also be considered to prevent the vertical transmission of the infection.

A questionnaire survey of animal and human health authorities in Europe revealed that leishmaniases are not notifiable in all countries with autochthonous cases. Few countries implement surveillance and control targeting both animal and human infections. Leishmaniases are considered emergent diseases in most countries, and lack of resources is a challenge for control.

Borrelia persica , transmitted by the argasid tick Ornithodoros tholozani , causes human tick-borne relapsing fever in the Middle East and Central Asia. Infection is acquired often when visiting tick-infested caves and reported to be transmitted mainly transovarially between ticks, occasionally infecting humans. To study the epidemiology of this infection, ticks were trapped in 24 caves in 12 geographic zones covering all of Israel and identified morphologically. DNA was extracted from larvae, nymphs, and adult stages from each location and PCR followed by DNA sequencing was performed to identify Borrelia infection, tick species, and tick blood meal sources. We collected 51,472 argasid ticks from 16 of 24 caves surveyed. We analyzed 2,774 O. tholozani ticks, and 72 (2.6%) from nine caves were PCR positive for B. persica Infection rates in male, female, and nymphal ticks (4.4%, 3%, and 3.2%, respectively) were higher than in larva ( P <  0.001), with only 3 (0.04%) positive larvae. Presence of blood meal was associated with B. persica infection in ticks ( P =  0.003), and blood meals of golden jackals, red foxes, and Cairo spiny mouse were associated with infection ( P ≤  0.043). PCR survey of 402 wild mammals revealed B. persica infection with the highest rates in social voles (22%), red foxes (16%), golden jackals (8%), and Cairo spiny mice (3%). In conclusion, although transovarial tick transmission of B. persica occurs at low levels, ticks apparently acquire infection mainly from wildlife canid and rodents and may eventually transmit relapsing fever borreliosis to humans who enter their habitat. IMPORTANCE Borrelia persica is a spirochete that causes tick-borne relapsing fever in humans in an area that spans from India to the Mediterranean. Until now, it was thought that the soft tick vector of this infection, Ornithodoros tholozani , is also its main reservoir and it transmits B. persica mostly transovarially between tick generations. This study showed that tick infection with B. persica is associated with feeding blood from wild jackals, foxes, and rodents and that transovarial transmission is minimal. Since O. tholozani ticks are found in isolated caves and ruins, it is assumed that wild canids who migrate over long distances have a major role in the transmission of B. persica between remote tick populations, and it is then maintained locally also by rodents and eventually transferred to humans during tick bites. Prevention of human infection could be achieved by restricting entrance of canines and humans to habitats with O. tholozani populations., (Copyright © 2021 American Society for Microbiology.)

Ehrlichia canis is the major causative agent of canine monocytic ehrlichiosis (CME). Its morulae might be detected during the acute disease phase, usually within peripheral blood monocytes, but were uncommonly described within peripheral blood lymphocytes. This report describes two unrelated puppies, naturally infected with E. canis. In both, examination of stained peripheral blood smears revealed one to several cytoplasmic inclusions, characteristic of typical E. canis morulae, exclusively within lymphocytes. Ehrlichia canis infection was confirmed in both cases by blood sample real-time PCR. Both dogs were young and had comorbidities. One dog, based on whole blood PCR, was co-infected with Anaplasma platys and Babesia vogeli. The other had no other concurrent tick-borne infection based on PCR, but had bacterial cholangiohepatitis. These comorbidities, and the dogs' young age possibly contributed to the uncommon presence of E. canis morulae within peripheral blood lymphocytes rather than their typical presence in monocytes., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Spirocerca lupi is a parasitic nematode of canids that induces a myriad of clinical manifestations in its host and, in 25% of infections, leads to the formation of sarcomas. The description of the protein composition of the excretory and secretory products ( Sl -ESP) of S. lupi has shed light on its possible interactions with the host environment, including migration within the host and mechanisms of immunomodulation. Despite this, the process by which S. lupi induces cancer in the dog remains poorly understood, and some hypotheses have arisen regarding these possible mechanisms. In this review, we discuss the role of specific ESP from the carcinogenic helminths Clonorchis sinensis, Opisthorchis viverrini and Schistosoma haematobium in inducing chronic inflammation and cancer in their host's tissues. The parasitic worms Taenia solium, Echinococcus granulosus, Heterakis gallinarum, Trichuris muris and Strongyloides stercoralis , which have less-characterized mechanisms of cancer induction, are also analyzed. Based on the pathological findings in spirocercosis and the mechanisms by which other parasitic helminths induce cancer, we propose that the sustained inflammatory response in the dog´s tissues produced in response to the release of Sl -ESP homologous to those of other carcinogenic worms may lead to the malignant process in infected dogs.

Spirocerca lupi is the etiologic agent of spirocercosis in canids from the tropical and subtropical regions of the world. A 1-year-old dog was presented with weight loss and diarrhea. Upon examination, thickened limbs, a mass in the lung's right middle lobe and an esophageal mass were found. The pulmonary mass aspirate revealed S. lupi eggs surrounded by a purulent infiltrate. Limb bone biopsies showed irregular mineralization and cartilaginous metaplasia. Treatment with doramectin resolved the initial clinical signs. However, regurgitation was reported 5 months later. Therefore, the dog received additional doramectin and later vomited a worm morphologically consistent with a S. lupi adult. Herein we report the esophageal migration of S. lupi with egg deposition to the lung parenchyma, accompanied by secondary hypertrophic osteopathy. This report highlights the presence of S. lupi in Costa Rica and the importance of including this parasite as a differential diagnosis when compatible signs are identified., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Three hundred and two stool samples were collected from municipal shelters and owned dogs in different geographical locations in Israel from December 2016 to September 2017 and examined for Giardia and assemblage type by PCR targeting the 18S rRNA and β-giardin genes. Overall Giardia prevalence was 24.5 % (74/30). Giardia prevalence was 1.9-fold higher in dogs ≤ 6 months old compared to > 6 ≤ 12 months old and older dogs [25/61 (41 %), 18/73 (24.6 %) and 31/166 (18.7 %), respectively, (p = 0.001)], 2.3-fold higher in winter [32/90 (35.5 %)] compared to its prevalence during autumn [15/60 (25 %)], spring [10/62 (16.1 %)] and summer [17/89 (19.1 %), p = 0.003)], and 2.7-fold more frequent among diarrheic dogs [23/43 (53.4 %)] compared to those with formed stools [51/253 (20.1 %)], (p = 0.001)]. The Giardia sp. assemblages detected were C and D. Higher infection rates in young, diarrheic dogs, sampled during winter, and housed in municipal shelters, indicates the need for targeted preventive measures., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

In horses, Neospora caninum and Neospora hughesi have been associated with fetal loss, and neurological disease, respectively. This study investigated the role of Neospora spp. infection in equine abortion in Israel. The presence of anti- Neospora spp. antibodies was evaluated in 31 aborting mares by indirect fluorescent antibody test (IFAT) and the presence of parasite DNA in their aborted fetuses was evaluated by polymerase chain reaction (PCR), using two target loci (ITS1 and Nc5). The seroprevalence found in aborting mares was 70.9% and the prevalence by DNA detection in the aborted fetuses was 41.9%. Transplacental transmission from positive mares to their fetuses was 45.4% (10/22), while 33.3% (3/9) of fetuses of seronegative mares also tested positive for Neospora . The use of two PCR targets improved the sensitivity of parasite detection, and positive samples were identified by sequence analyses as N. caninum . These finding suggest that N. caninum could be a significant cause of abortion in horses, and that transplacental transmission in horses is an important way of transmission of N. caninum . The results presented here demonstrated the necessity to use several tests concurrently, including serological and molecular assays in order to confirm the involvement of Neospora in mare abortions.

The current postexposure prophylaxis regimen for tick-borne relapsing fever (TBRF) consists of 5 days' doxycycline. In this observational study of 77 spelunkers at high risk for TBRF, a single dose of 100 mg doxycycline taken up to 72 hours after exposure to ticks was 100% effective in preventing the disease., (© The Author(s) 2020. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.)

Feline lungworms infect the respiratory tract of wild and domestic cats, causing infection often associated with clinical disease. Until recently, Aelurostrongylus abstrusus has been considered the most relevant species of lungworm, while Troglostrongylus brevior was considered of less significance. Fecal samples of feral cats from Jerusalem, Israel, collected over a year, were examined for first stage lungworm larvae (L1) using the Baermann method. Positive samples were morphologically identified, and their species identity was molecularly confirmed. Forty of 400 (10.0%) cats were lungworm-positive, of which 38/40 (95.0%) shed Troglostrongylus brevior and 6/40 (15.0%) shed Aelurostrongylus abstrusus. Four cats (10.0%) had mixed infections with both lungworm species. L1 shedding was associated with clinical respiratory signs in 11 (19.0%) T. brevior shedding cats of a total of 58 cats manifesting respiratory signs, while 23/342 (6.7%) cats without respiratory signs were L1-positive (p = 0.006). Non-respiratory clinical signs were also found to be more prevalent in L1 shedders (p = 0.012). A young kitten ≤ 4 weeks of age shed T. brevior L1 larvae. DNA sequences of both lungworm species using the ribosomal internal transcribed spacer 2 (ITS2) locus were > 99% similar to other sequences deposited in GenBank, suggesting that T. brevior and A. abstrusus ITS2 sequences are both highly conserved. In conclusion, L1 shedding in feral cats from Jerusalem were mostly caused by T. brevior with only a small proportion involving A. abstrusus, different from many studies from other geographical regions.

Protists embrace many species, some of which may be either occasional or permanent parasites of vertebrate animals. Between the parasite species, several of medical and veterinary importance are vector-transmitted. The ecology and epidemiology of vector-borne parasitoses, including babesiosis, leishmaniasis and malaria, are particularly complex, as they are influenced by many factors, such as vector reproductive efficiency and geographical spread, vectorial capacity, host immunity, travel and human behaviour and climatic factors. Transmission dynamics are determined by the interactions between pathogen, vector, host and environmental factors and, given their complexity, many different types of mathematical models have been developed to understand them. A good basic knowledge of vector-pathogen relationships and transmission dynamics is thus essential for disease surveillance and control interventions and may help in understanding the spread of epidemics and be useful for public health planning., (Copyright © 2020. Published by Elsevier GmbH.)

Spirocerca vulpis is a parasitic nematode of red foxes associated with gastric nodule formation. Since its description in 2018, this species has been reported in red foxes from Spain, Bosnia and Herzegovina, Italy, and Portugal. We present here the analysis of uncharacterized nematodes obtained from gastric nodules of a red fox from Switzerland in 1999. The specimens were identified as S. vulpis based on the observation of teeth-like structures in the buccal capsule and a 99.4% sequence identity to S. vulpis DNA from Spain. Phylogenetic analysis demonstrated the clustering of the Swiss sequences in a different group from specimens of other geographical locations. Altogether, this study constitutes the first report of S. vulpis in Switzerland and a report of the oldest specimen of this species in the world. Our findings highlight the widespread distribution of S. vulpis in Europe which may be facilitated by the free-roaming nature of red foxes.

Recent studies have described Spirocerca lupi-like nematodes in the stomach of red foxes (Vulpes vulpes) in Europe. A phylogenetic analysis of those specimens using mitochondrial DNA and their morphological reexamination allowed their characterization as a different species, Spirocerca vulpis. Between the years of 2010 and 2017, roundworms were collected from seven red foxes of northeastern Portugal found at necropsy with nodular lesions on their stomach wall. Histopathological analysis of four foxes revealed granulomatous lesions of the gastric nodules. On morphological assessment, by light microscopy, nematodes revealed the presence of six triangular teeth-like buccal capsule structures, which are absent in S. lupi. Polymerase chain reaction was run to amplify a 551 bp partial fragment of the cytochrome c oxidase subunit 1 gene. Sequences were 99% similar to S. vulpis (85% coverage) of red foxes from Spain and Bosnia and Herzegovina, 99% similar (99% coverage) to sequences of Spirocerca sp. of red foxes from Denmark and 93% similar (99% coverage) to S. lupi from South Africa. This is the first report of S. vulpis in foxes or any other host from Portugal.

Feline lungworms such as Aerulostrongylus abstrusus and Troglostrongylus brevior are snail-borne pathogens causing respiratory disease in domestic cats. Paratenic hosts such as rodents and reptiles have also been implicated in the epidemiology of these parasites. Although A. abstrusus has been recognized for a long time as the most prevalent lungworm among cats worldwide, T. brevior is of major concern in kittens. Bearing in mind that disease due to T. brevior occurs mainly in pediatric patients younger than 6 months of age, the diagnosis of this parasite in two kittens presenting severe respiratory disease from the garden of one of the authors inspired us to investigate the potential routes of transmission for T. brevior in domestic cats. Of the three queens (A, B and C) that delivered kittens (n = 8), only cat A was positive for T. brevior, presenting her two kittens severe respiratory clinical signs, which lead to the exitus in one of them, 18 days of age. In addition, three kittens, the offspring of queen B, turned to be positive at the coprological examination after suckling from queen A, whereas those from queen C (that suckled only on their own mother) remained negative. A series of coprological, histological and molecular tests were conducted to confirm the presence of T. brevior in the patients as well as in the other cats cohabiting the same garden. Adult nematodes were retrieved from the trachea and bronchi of the dead kitten (kitten 1A), and larvae at the histology of the lung and liver parenchyma associated with bronco pneumonitis and lymphocytic pericholangitis, respectively. Cornu aspersum (n = 60), Eobania vermiculata (n = 30) snails (intermediate hosts) as well as lizards and rats (potential paratenic hosts) were collected from the same garden and processed through tissue digestion and molecular detection. Troglostrongylus brevior larvae were recovered through tissue digestion from two C. aspersum (3.33 %) and it was confirmed by PCR-sequencing approach, which also detected T. brevior DNA in the liver and lungs of one rat and in the coelomatic cavity of one gecko lizard. During the COVID-19 lockdown, when scientists spent more time at home, we grasp the opportunity to decipher T. brevior biology and ecology starting in a small ecological niche, such as the garden of our house. Data herein presented led us to suggest: i) the transmammary transmission of T. brevior in domestic cats; ii) the role of intermediate and paratenic hosts (including reptiles) in the epidemiology of the infection which they transmit; as well as iii) the importance of observational parasitology in studying any event that certainly occurs in small ecological niches, as it could be in our home gardens., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Purpose: The aim of this study was to evaluate the prevalence and distribution of canine babesiosis and hepatozoonosis in the south of Romania as well as to assess if there are differences in the prevalence between regions in that area., Methods: Blood samples from 300 dogs from the south of Romania were screened for the presence of piroplasmids by PCR. Positive samples were further tested for the presence of Babesia spp. and Hepatozoon canis. Pearson's chi-square with Yates correction was used to determine if there is statistical difference between infected populations., Results: Fifty-four percent (163/300) of the dogs were positive for Hepatozoon spp. or piroplasmids. Forty-eight percent (143/300) were infected with H. canis and 9.6% (29/300) were positive to B. canis. Three percent (9/300) of the dogs were co-infected with Babesia spp. and H. canis. The prevalence of H. canis in the south-west of Romania was significantly higher (p < 0.05) than that of three other regions while the prevalence of Babesia spp. infection in the south-east of the country was significantly higher (p < 0.05) compared to its south-west., Conclusions: A high rate of H. canis infection was found in the stray dog population surveyed with the highest infection rate detected in south-western Romania. Babesia canis was found to pose the highest threat in south-eastern Romania, whereas B. vogeli infection was focused in the same region of high H. canis infection in south-western Romania. These infections should be of important concern to veterinarians and dog owners in Romania.

The Tropical Council for Companion Animal Parasites Ltd. (TroCCAP) is a not-for-profit organisation whose mission is to independently inform, guide and make best-practice recommendations for the diagnosis, treatment and control of companion animal parasites in the tropics and sub-tropics, with the aim of protecting animal and human health. In line with this primary mission, TroCCAP recently developed guidelines for the diagnosis, treatment and control of feline and canine parasites in the tropics. The development of these guidelines required unique and complex considerations to be addressed, often inapplicable to developed nations. Much of the tropics encompass middle-to-low income countries in which poor standards of environmental hygiene and large populations of stray dogs and cats coexist. In these regions, a range of parasites pose a high risk to companion animals, which ultimately may place their owners at risk of acquiring parasitic zoonoses. These considerations led to the development of unique recommendations with regard, for example, to deworming and endoparasite testing intervals for the control of both global and 'region-specific' parasites in the tropics. Moreover, the 'off-' or 'extra'-label use of drugs for the treatment and control of parasitic infections is common practice in many tropical countries and many generic products lack manufacturers' information on efficacy, safety, and quality control. Recommendations and advice concerning the use of such drugs and protocols are also addressed in these guidelines. The formation of these guidelines is an important first step towards improving the education of veterinarians specifically regarding best-practice for the diagnosis, treatment and control of canine and feline parasites in the tropics., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Venezuela is a country where human and canine leishmaniosis due to Leishmania infantum, Leishmania braziliensis and other Leishmania spp. is endemic. However, only limited data is available on canine Leishmania infection in Venezuela. The aim of this cross-sectional study was to evaluate the prevalence of Leishmania infection in dogs (n = 152) from the states of Lara (n = 91) and Yaracuy (n = 61) in Venezuela by means of serological and molecular methods. Physical examination was performed and blood samples were collected from all dogs. Serology for antibodies reactive with L. infantum and L. braziliensis antigens was assessed by the enzyme-linked immunosorbent assay (ELISA) and detection of Leishmania DNA from blood samples was evaluated by kinetoplast Leishmania real-time polymerase chain reaction (RT-PCR). In addition, Leishmania internal transcribed spacer (ITS-1) RT-PCR was performed on the samples positive by kinetoplast RT-PCR. The prevalence of Leishmania infection based on serological and/or molecular techniques was 11.8%. The seroprevalence for L. infantum and L. braziliensis antigens were 2.1% (3/144) and 8.3% (12/144), respectively. All dogs from the state of Yaracuy were serologically negative to L. infantum while 4.6% (4/86) of the dogs were reactive to L. braziliensis antigen. Fourteen percent (8/58) of the dogs from the state of Lara were positive to L. infantum and 5.2% (3/58) to L. braziliensis antigen. Three dogs were positive to both Leishmania spp. antigens. By RT-PCR, 6.5% (4/61) and 4.4% (4/91) of the dogs were positive for infection in the states of Lara and Yaracuy, respectively. The RT-PCR product of one dog from the state of Yaracuy was sequenced revealing a 100% identity with L. infantum. However, all RT-PCR positive dogs were seronegative to both Leishmania spp. antigens. In conclusion, the positivity for Leishmania spp. infections observed indicates that dogs are frequently infected by L. infantum, L. braziliensis or related Leishmania spp. in Venezuela., Competing Interests: Declaration of Competing Interest The authors declare that they have no competing interests., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Introduction: Babesiosis is a protozoan tick-borne infection associated with anemia and life-threatening disease in humans, domestic and wildlife animals. Dogs are infected by at least six well-characterized Babesia spp. that cause clinical disease. Infection with a piroplasmid species was detected by light microscopy of stained blood smears from five sick dogs from Israel and prompted an investigation on the parasite's identity., Methods: Genetic characterization of the piroplasmid was performed by PCR amplification of the 18S rRNA and the cytochrome c oxidase subunit 1 (cox1) genes, DNA sequencing and phylogenetic analysis. Four of the dogs were co-infected with Borrelia persica (Dschunkowsky, 1913), a relapsing fever spirochete transmitted by the argasid tick Ornithodoros tholozani Laboulbène & Mégnin. Co-infection of dogs with B. persica raised the possibility of transmission by O. tholozani and therefore, a piroplasmid PCR survey of ticks from this species was performed., Results: The infected dogs presented with fever (4/5), anemia, thrombocytopenia (4/5) and icterus (3/5). Comparison of the 18S rRNA and cox1 piroplasmid gene sequences revealed 99-100% identity between sequences amplified from different dogs and ticks. Phylogenetic trees demonstrated a previously undescribed species of Babesia belonging to the western group of Babesia (sensu lato) and closely related to the human pathogen Babesia duncani Conrad, Kjemtrup, Carreno, Thomford, Wainwright, Eberhard, Quick, Telfrom & Herwalt, 2006 while more moderately related to Babesia conradae Kjemtrup, Wainwright, Miller, Penzhorn & Carreno, 2006 which infects dogs. The piroplasm forms detected included tetrads (Maltese cross), merozoite and trophozoite stages whose average size was larger than stages of other canine Babesia spp. belonging to the Babesia (s.l.) and B. gibsoni Patton, 1910, and smaller than other canine Babesia (sensu stricto) spp. Of 212 O. tholozani ticks surveyed, 11 (5.2%) harbored DNA of the new species of Babesia., Conclusions: Babesia negevi n. sp. is described based on morphological and genetic characterization and phylogenetic analyses. The species is named after the Negev desert of southern Israel, where the first infected dog originated from. Despite co-infection in four dogs, the fifth dog had fatal disease attesting that B. negevi n. sp. infection requires clinical attention. Incriminating O. tholozani or another tick species as the vector of Babesia negevi n. sp., would require additional studies.

The prevalence data of Leishmania infantum infection in cats are characterized by a large variability mainly attributed to the differences in diagnostic techniques. In the absence of consensus about the method of choice for diagnosing feline leishmaniosis, the performance of a new immunofluorescence antibody test (IFAT) was herein analytically described by the comparison with IFAT commonly used for the diagnosis of canine leishmaniosis (i.e., IFAT-OIE) and a laboratory enzyme-linked immunosorbent assay (ELISA). Sera of cats living in visceral leishmaniosis-endemic (n = 105) and visceral leishmaniosis-non-endemic (n = 50) areas were tested by the above methodologies and real-time PCR (qPCR). The most frequent result was represented by triple negativity to the three tests (IFAT-OIE, ELISA, and qPCR) in 42.9% and 80% cats from endemic and non-endemic areas, respectively. Bayes latent class analysis gave an output probability of 34.1% (posterior standard deviation, psd = 5.4%) of true L. infantum cases (TCL) which represent the true estimated prevalence of infection. The sensitivity of each variable contributing to define the TCL was 24% (psd = 6.3%) for qPCR, 78.8% (psd = 8.7%) for ELISA and 91.8% (psd = 5.2%) for IFAT-OIE. The probability to be a TCL was 94.5% for the sample from an endemic area. The cross-validation of the new IFAT by a logistic model correctly identified as positive 80.7% of subjects defined as TCL and negative 89.9% as not TCL, respectively, by the Bayesian model. The study results estimate a good accuracy of the IFAT in predicting cats exposed to L. infantum. Therefore, this procedure may be beneficial for screening cat populations for a better understanding of the epidemiology of feline leishmaniosis.

Infectious diseases by pathogens, including those of zoonotic concern, may act as a primary or contributory cause of threat to wildlife conservation and may represent a risk for human health, mainly for people working at, or visiting the zoological parks. Given the paucity of data on pathogens infecting wild tigers, we investigated the occurrence of infectious agents in this animal species, with a special focus on those of zoonotic concern. Blood and serum samples from tigers (n = 20) living in a wildlife safari park of southern Italy were screened by serological and molecular tests. All animals scored positive for antibodies against Toxoplasma gondii (100%), whereas they displayed different prevalence of seropositivity for Rickettsia conorii (30%), Bartonella henselae (15%) and Leptospira interrogans sv Icterohaemorrhagiae and/or Leptospira kirschneri sv Grippotyphosa (15%). No antibodies against Coxiella burnetii were detected. In addition, 8 tigers (40%) tested molecularly positive to " Candidatus Mycoplasma haemominutum", and 3 (15%) to Hepatozoon canis . No DNA of R. conorii , Bartonella spp., Ehrlichia / Anaplasma in given environments may represent a relevant health issue considering the close association among animals and humans visiting, or working at, the wildlife safari park. Preventative measures are advocated in order to control ectoparasites and other sources of infection (e.g., small rodents), thus for minimizing the risk of infection for animals as well as for humans.T. gondii, R. conorii, L. interrogans sv Icterohaemorrhagiae, " Candidatus Mycoplasma haemominutum" and B. henselae in given environments may represent a relevant health issue considering the close association among animals and humans visiting, or working at, the wildlife safari park. Preventative measures are advocated in order to control ectoparasites and other sources of infection (e.g., small rodents), thus for minimizing the risk of infection for animals as well as for humans., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2020 The Authors.)

Background: An outbreak of leishmaniosis was studied in cats and dogs housed together with no separation in an animal shelter in Israel., Methods: The study included recording of clinical signs, serology for Leishmania infection by ELISA, PCR of blood for Leishmania DNA by ITS1 HRM and kDNA PCR, parasite quantification, and trapping of sand flies around the shelter., Results: Thirty-seven % (22/60) of the dogs and 75% (50/67) of the cats were seropositive to L. infantum with a significantly higher seropositivity rate in the cat population (χ 2 = 42.160, P < 0.0001). Twenty-five percent (15/60) of the dogs were positive for Leishmania by blood PCR, 12% by the Leishmania ITS1 HRM PCR and 22% by kDNA PCR. Of the cats, 16% (11/67) were positive by kDNA PCR and none by ITS1 HRM PCR. All the PCR-positive animals were infected by L. infantum verified by DNA sequencing and there was no significant difference between the PCR-positivity in the dog and cat populations. Altogether, 43% (26/60) of the dogs and 79% (53/67) of the cats were positive by serology or PCR for L. infantum. The average Leishmania parasite load in the blood of PCR-positive dogs (42,967 parasites/ml) was significantly higher than in PCR-positive cats (1259 parasites/ml) (t (12) = 2.33, P = 0.037). Dogs that were positive by the Leishmania ITS1 HRM PCR and kDNA PCR had significantly higher parasite loads than dogs positive only by the kDNA PCR (t (11) = - 3.186580, P < 0.009). No significant effect was found for FIV seropositivity on Leishmania infection in the cats (χ 2 = 0.506, P = 0.777). A higher percentage of Leishmania-positive dogs showed clinical signs compatible with leishmaniosis compared to Leishmania-positive cats (100 vs 52.8%, χ 2 =15.242, P < 0.0001). Phlebotomus perfiliewi, a proven vector of L. infantum, comprised 92% of trapped sand flies., Conclusions: Comparisons of populations of cats and dogs exposed to sand flies and L. infantum under the same conditions indicated that although a high rate of exposure was detected in cats as manifested by a significantly greater degree of seropositivity, dogs had significantly higher blood parasite loads, and were likely to be more infectious to sand flies than cats.

This report presents a novel canine condition in 32 dogs in which aberrant migration of Spirocerca lupi larvae through mesenteric arteries, instead of gastric arteries, led to small or large intestinal infarction. This form of spirocercosis was first recognized in Israel in 2013 and is currently ongoing. Typical clinical signs were anorexia and weakness of 3 to 4 days and, less frequently, vomiting and diarrhea, followed by collapse, bloody diarrhea, and severe vomiting. Exploratory laparotomy showed 1 or more infarcted and often perforated intestinal segments in all cases. Microscopically, there was intestinal mucosal to transmural coagulative necrosis and mesenteric multifocal necrotizing eosinophilic arteritis, thrombosis, hemorrhage, and early fibroplasia. Third-stage S. lupi larvae were identified by morphologic features in 9 of 32 (28%) cases, and the species was confirmed by polymerase chain reaction in 4 cases. Nearly 50% of the dogs had been receiving prophylactic therapy, which did not prevent this form of spirocercosis.

Babesia is tick-transmitted protozoan parasites that infect mammalian hosts and have a major impact on farm and pet health-associated costs worldwide. This study aimed to test the prevalence of Babesia spp. infection in a small cohort of dogs at a veterinary hospital and to perform molecular characterization of the Babesia species causing the infection. For the PCR assay, 5 mL of blood was collected by venipuncture of the cephalic or radial veins in 300 dogs of different ages, sex, and breeds, which were presented to the veterinary hospital of the Federal University of Uberlândia between March 2015 and April 2016. In addition, a drop of blood was collected from the marginal blood vessels of the ear of dogs included in this study. Ninety-two (30.67%) were positive for Babesia spp., as determined by microscopic observation of the blood smear, revealing the presence of intra-erythrocyte merozoites. For molecular characterization by PCR, 17 samples were chosen from dogs who were tested positive for Babesia spp. by blood smears. Among them, B. vogeli was found to infect all 17 dogs, as determined by 99-100% sequence identity (closest GenBank match KT246307) using primers PIRO A/PIRO B. Our results indicate that the species observed in these dogs was B. vogeli.

Prevention of canine Leishmania infantum infection is critical to management of visceral leishmaniasis in people living in endemic areas of Brazil. A bill (PL 1738/11), currently under consideration, proposes to establish a national vaccination policy against canine leishmaniasis in Brazil. However, there is no solid scientific evidence supporting the idea that this could reduce transmission from infected vaccinated dogs to sand flies to a level that would significantly reduce the risk of L. infantum infection or visceral leishmaniasis in humans. Thus, we advocate that insecticide-impregnated collars should the first line protective measure for public health purposes and that vaccines are applied on a case-by-case, optional basis for individual dog protection., (Copyright © 2020 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

Cardiopulmonary dirofilariosis in dogs and other carnivores is caused by Dirofilaria immitis, while Dirofilaria repens usually causes a subcutaneous infection. The importance of red foxes and golden jackals in the epidemiology of dirofilariosis remains unknown. Thus, the aim of this study was to conduct a cross-sectional molecular survey of Dirofilaria species in stray dogs, red foxes and golden jackals from the endemic region of Vojvodina, Serbia, in order to determine and update data on their prevalence and provide insight into the epidemiological importance of wild canids. A total of 59 blood samples from stray dogs, 94 from red foxes and 32 from golden jackals were collected and screened by real-time PCR targeting a 115-bp fragment of the mitochondrial 12S gene of filarioids and by conventional PCR assay targeting a 484-524-bp fragment of 5.8S-ITS2-28S locus of filarioids. The cross-sectional molecular survey detected the filarioid mitochondrial 12S gene fragment in stray dogs (27.1 %), red foxes (8.5 %) and golden jackals (6.3 %) in the same endemic region of Vojvodina, Serbia. Only D. immitis was detected in stray dogs, while both D. immitis and D. repens were detected in populations of red foxes and golden jackals. These results outline a possible interaction of D. immitis infection between the dog population and the wild canid populations, while D. repens was found to circulate mostly in golden jackals and red foxes populations., Competing Interests: Declaration of Competing Interest The authors declare that they have no competing interest., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Giardia duodenalis causes gastro-intestinal (GI) disease and carries a zoonotic risk. The risk for infection depends on local prevalence and individual immunity, but clinical signs are inconsistent and the role of G. duodenalis in other GI diseases is unknown. The current study aims were to evaluate the prevalence of G. duodenalis in dogs presented to a university veterinary teaching hospital in Israel, investigate risk factors for infection and its associations with other diseases. Fecal samples from 163 dogs were tested for G. duodenalis by an antigen assay (FASTest® Giardia strip). Prevalence of G. duodenalis infection was 11.9 %. It was significantly associated with young age (≤1year) and canine parvoviral enteritis. Housing conditions, signs of GI disease or other systemic diseases were unassociated with infection. In conclusion, G. duodenalis infection prevalence is relatively low in the study population. Young age and diagnosis of co-infections are common, warranting screening for G. duodenalis., Competing Interests: Declaration of Competing Interest None., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The main objective of the present study was to evaluate the acute phase protein (APP) concentrations that dogs naturally infected with Leishmania infantum show in different clinical stages of disease staged according to the Leishvet and CLWG classifications. In addition, these classifications are compared with the groups based on APP recommendations published recently. Medical records of 458 dogs with leishmaniosis whose samples were submitted to our laboratory for clinical pathology evaluation were reviewed and 77 cases met the inclusion criteria. All dogs were classified according to the CLWG system and the majority of the dogs (33.8%) were classified in stage D. Although some dogs (41.6%) could not be classified by the Leishvet system since it includes only dogs with clinical disease, most of the classified dogs (27.3%) were at Leishvet stage II. According to the APP classification, the majority of dogs (32.5%) were classified in stage 3a. Dogs in the more advanced stages of Leishvet and CWLG classifications had significant increases in serum ferritin and C-reactive protein (CRP) and decrease in Paraoxonase 1 (PON1). These findings indicate that APPs show more significant changes in the more advanced stages of Leishvet and CWLG classifications corresponding with more severe cases of canine leishmaniosis., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Spirocerca lupi is a nematode transmitted by dung beetles that infects domestic and wild canids in tropical and subtropical regions and is associated with neoplasia. It produces a distinctive pathology with the formation of esophageal nodules classified as inflammatory, preneoplastic, or neoplastic with metastasis to distant organs. Aberrant central nervous system migration of this nematode is also responsible for severe neurological manifestations. Reports of spirocercosis have increased over the last two decades showing spread of this canine helminth in five continents. S. lupi from different geographical locations is genetically distinct with two genotypes, genotype I from Africa, Asia, and Australia, and genotype II from Europe, and recently separated from Spirocerca vulpis, a new species described in red foxes from Europe., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Dogs are the main reservoir of Leishmania infantum and in some countries have been regularly culled as part of government policy to control visceral leishmaniasis. At the 13th Symposium of the Companion Vector-Borne Diseases World Forum in Windsor, UK, March 19-22, 2018, we consolidated a consensus statement regarding the usefulness of dog culling as a means of controlling visceral leishmaniasis. The statement highlighted the futility of culling infected dogs, whether healthy or sick, as a measure to control the domestic reservoir of L. infantum and reduce the risk for visceral leishmaniasis.

Spirocerca lupi is a parasitic and carcinogenic nematode of canids distributed in tropical and subtropical regions around the world. The excretion-secretion proteins (PES) of S. lupi have been suggested to play a role in the pathogenesis of its infection. We aimed to identify the PES of different stages of S. lupi and search for proteins that would be useful for diagnostic, therapeutic and vaccination purposes as well as understand their functions. A nano-UPLC mass spectrometry de novo analysis was performed on proteins collected from cultures of S. lupi L3 larvae, L4 females, adult females and adult males from naturally infected hosts. A total of 211 proteins were identified in all cultures. Accordingly, 117, 130, 99 and 116 proteins were detected in L3 larva, L4 females, adult females and adult males, respectively, with a strong correlation in the biological replicates (Pearson coefficients > 0.73). Fourty-four proteins were detected in all developmental stages, 64 were stage-specific and 49 were exclusively identified in L4 females. Cell compartment enrichment analysis revealed that proteins common to all stages were cytoplasmatic (p < 9.x10 -6 ), whereas L4 unique proteins were in collagen trimers, and macromolecular complexes (p < 0.00001). Functional enrichment analysis of proteins showed significant enrichment in lipid metabolism in L3-unique proteins (p<0.00005), in mannose metabolism and protein de-glycosylation for L4-unique proteins (p < 0.00004), and in phosphorus metabolism in proteins shared by all stages (p <  2.1 x10 -9 ). Interestingly, annexin 6, associated with cancer in humans, was detected in all life stages, but in a larger abundance in L4 females and adults. These findings indicate that S. lupi establishes complex interactions with its hosts by an arsenal of proteins expressed in different patterns in each life stage which influence the pathogenesis and oncogenesis of S. lupi and may be used as potential targets for diagnostic assays, drug targets or vaccine candidates., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Vector-borne diseases (VBDs) of domestic and wild carnivores are of major public health concern both in industrialized and developing countries, especially in poor socioeconomic settings. War-torn areas specifically suffer from absence of veterinary surveillance of VBDs, resulting in lack of scientific knowledge on this topic. To investigate occurence and prevalence of several vector-borne pathogens (VBPs) in some carnivore species from Iraq, blood samples (n = 397) were obtained from 190 canids [97 stray dogs (Canis familiaris), 55 jackals (Canis aureus) and 38 red foxes (Vulpes vulpes)] and 207 stray cats (Felis catus) collected during a feral animal control and zoonotic disease surveillance program in several United States military bases in Iraq. The presence of Babesia spp., Hepatozoon spp., Ehrlichia spp., Anaplasma spp., Dirofilaria spp. and Leishmania spp. DNA was molecularly investigated. Out of 397 animals tested, 176 (44.3%; 95% CI: 39.5-49.2%) were positive for at least one pathogen with the highest prevalence in foxes (73.7%; 95% CI: 58-85%), followed by jackals (54.5%; 95% CI: 41.5-67%), dogs (38.1%; 29.1-48.1%) and cats (39.1%; 95% CI: 32.7-45.9%). Up to five pathogens were diagnosed in dogs. Hepatozoon canis was the most prevalent VBP in jackals (49.1%; 95% CI: 36.4-61.9%), foxes (47.3%; 95% CI: 32.5-62.7%) and dogs (33%; 95% CI: 24.4-42.8%), whereas Hepatozoon felis was the only species detected in cats (39.1%; 95% CI: 32.7-45.9%). A species of Babesia related to but different from Babesia lengau and designated as Babesia sp. MML was detected in six foxes (15.8%; 95% CI: 7.4-30.4%) and in one jackal (1.8%; 95% CI: 0.3-9.6%). This finding suggested the existence of a new species in the genus Babesia as inferred by molecular and phylogenetical analysis. Further, Babesia vulpes was identified only in two foxes (5.3%; 95% CI: 1.5-17.3%). All samples were negative for Leishmania spp. and Ehrlichia spp. Co-infection with H. canis and Babesia spp. was the most prevalent (5/176, 2.8%, i.e., 4 foxes and 1 jackal), followed by H. canis and Dirofilaria immitis (1/176, 1.3%, i.e., in 1 jackal), H. canis and Dirofilaria repens or Acanthocheilonema reconditum (1/176, 1.3%, i.e., in one dog, each). Data presented fill gaps into knowledge of VBPs in dogs, cats and wild canids in Iraq, indicating that different pathogens circulate amongst animal populations living in the same areas, possibly sharing the same tick vectors. Large-scale surveys are urgently needed to further assess VBPs distribution in Iraq and establish preventative strategies in domestic animals to minimize the risk of infection for animals and humans., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Feline lung worm infection is increasingly reported in recent years, and recognized as a cause for respiratory disease in cats. Aelurostrongylus abstrusus is regarded as the most prevalent cause of such cases. Infective L3 larvae carried in gastropods and paratenic hosts infect felines, developing to adult worms that reside in the lungs' parenchyma and may cause verminous pneumonia. The L1 larvae hatch from eggs deposited in the lung, and are released to the environment by either feces or sputum. While the majority of epidemiological information regarding A. abstrusus originates in European countries, recent studies have shown that it is also found around the Mediterranean basin, as far east as Turkey and Cyprus. A local domestic cat from Israel showing signs of respiratory illness was diagnosed with aelurostrongylosis, confirmed by both morphological and molecular tools. Presence in Israel of this nematode was previously reported in 1949, with no further mentions since. ITS-2 sequence of the isolated larvae was highly similar to that of A. abstrusus from domestic cats from Italy. These findings show that distribution of A. abstrusus stretch to the eastern shores of the Mediterranean, and that this nematode should be considered as a cause for respiratory disease in cats in Israel and the surrounding countries., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Leishmaniosis due to Leishmania infantum is a complex infection that can affect both humans and dogs, and present a wide range of clinical signs and clinicopathological abnormalities. The conventional treatment of this disease is challenging due to the fact that complete parasitological cure commonly does not occur. Furthermore, treatment of the disease with the conventionally used drugs has several shortcomings. These include the need for long-term treatment, side effects and the formation of drug resistance. Moreover, it is important to highlight that the host immune responses play a crucial role in the outcome of this infection. For this reason, the use of immunotherapy in clinical leishmaniosis to improve the result of treatment with the conventional anti-leishmanial drugs by enhancing the immune response is imperative. The aim of this review is to provide a comparative overview of the wide range of immunotherapeutical approaches and strategies for the treatment of L. infantum infection in animals focusing on dogs., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Infectious haemolytic anaemia (IHA) in dogs share similar clinical signs including fever, lethargy, icterus, paleness of mucous membranes and splenomegaly. Postmortal findings are similar and, without additional diagnostic methods, an accurate aetiological diagnosis is difficult to achieve. In order to investigate causes of lethal IHA in Croatian dogs, we performed a retrospective study on archived formalin-fixed, paraffin-embedded tissue blocks (FFPEB) from dogs that died due to haemolytic crisis, using microscopic and molecular diagnostic tools to determine the aetiological cause of disease. Molecular analysis was performed on kidney, lung, myocardium and spleen on FFPEB from all dogs. The originally stated aetiological diagnosis of B. canis or leptospirosis was confirmed in only 53% of the dogs. PCR and sequencing revealed that, in addition to the expected pathogens, B. canis and Leptospira interrogans, the presence of previously undiagnosed "new" pathogens causing anaemia including Candidatus Neoehrlichia mikurensis and Anaplasma phagocytophilum. Furthermore, Theileria capreoli was detected for the first time in a dog with postmortal descriptions of lesions. Intensive extravascular hemolysis was noticeable as jaundice of the mucosa, subcutis and fat tissue, green or yellow discoloration of renal parenchyma caused by bilirubin excretion in the renal tubules and bile accumulation within the liver in 90% of the dogs. This work highlights the value of molecular diagnostics to complement traditional ante-mortem and post-mortem diagnostic protocols for the aetiological diagnosis of pathogens associated with IHA., (Copyright © 2019. Published by Elsevier Ltd.)

Aberrant migration of Spirocerca lupi into the spinal cord is an important cause of severe progressive neurological dysfunction in dogs. Although early diagnosis is essential to prevent deterioration, ante-mortem diagnosis of this condition remains challenging. The aim of this study was to evaluate the detection of the 18S ribosomal DNA (rDNA) S. lupi gene in the cerebrospinal fluid (CSF) of presumptively-affected dogs using polymerase chain reaction (PCR). Dogs with a non-compressive spinal cord lesion, pleocytosis with presence of eosinophils in the CSF and a characteristic clinical presentation were included. CSF samples from eight dogs were available for the study, of which seven were definitively diagnosed with intraspinal spirocercosis by PCR of either the CSF samples (6/7) or tissue samples obtained at necropsy examination (3/7), or both (2/7). Of these seven positive cases, only one dog had a negative CSF PCR, indicating a sensitivity of 86% for detecting nematode DNA in the CSF of infected dogs using this PCR protocol. The nematode DNA sequences obtained from the CSF of six dogs and the spinal cord tissue of three dogs were 98-100% identical to the publicly available sequences of S. lupi, confirming the diagnosis. These findings indicate that PCR targeting the 18S rDNA of S. lupi in CSF is useful for the ante-mortem diagnosis of canine intraspinal spirocercosis., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Extrachromosomal genomes of the adeleorinid parasite Hepatozoon canis infecting an Israeli dog were investigated using next-generation and standard sequencing technologies. A complete apicoplast genome and several mitochondrion-associated sequences were generated. The apicoplast genome (31,869 bp) possessed two copies of both large subunit (23S) and small subunit (16S) ribosomal RNA genes (rDNA) within an inverted repeat region, as well as 22 protein-coding sequences, 25 transfer RNA genes (tDNA) and seven open reading frames of unknown function. Although circular-mapping, the apicoplast genome was physically linear according to next-generation data. Unlike other apicoplast genomes, genes encoding ribosomal protein S19 and tDNAs for alanine, aspartic acid, histidine, threonine and valine were not identified. No complete mitochondrial genome was recovered using next-generation data or directed PCR amplifications. Eight mitochondrion-associated (215-3523 bp) contigs assembled from next-generation data encoded a complete cytochrome c oxidase subunit I coding sequence, a complete cytochrome c oxidase subunit III coding sequence, two complete cytochrome B coding sequences, a non-coding, pseudogene for cytochrome B and multiple fragmented mitochondrial rDNA genes (SSUA, SSUB, SSUD, LSUC, LSUG, RNA6, RNA10, RNA14, RNA18). The paucity of NGS reads generating each of the mitochondrion-like sequences suggested that a complete mitochondrial genome at typically high copy number was absent in H. canis. In contrast, the complete nuclear rDNA unit sequence of H. canis (18S rDNA to 28S rDNA, 6977 bp) had >1000-fold next-generation coverage. Multiple divergent (from 93.6% to 99.9% pairwise identities) nuclear 18S rDNA contigs were generated (three types with 10 subtypes total). To our knowledge this is the first apicoplast genome sequenced from any adeleorinid coccidium and the first mitochondrion-associated sequences from this serious pathogen of wild and domestic canids. These newly generated sequences may provide useful genetic loci for high-resolution species-level genotyping that is currently impossible using existing nuclear rDNA targets., (Copyright © 2019 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

Background: Information on the status of vector-borne pathogens among canines in Northeast India is lacking, particularly for the states of Mizoram and Tripura close to the Myanmar border. Blood samples collected from 130 dogs, 80 from Mizoram and 50 from Tripura, were examined in this study., Methods: Polymerase chain reaction (PCR) was performed for filariid worms, Babesia, Hepatozoon, Ehrlichia and Anaplasma spp. and DNA sequencing was then carried out to identify pathogens at the species level., Results: Vector-borne pathogens were detected in 52% (68/130) of the sampled dogs. The most prevalent pathogen was Babesia gibsoni detected in 56/130 (43%) dogs, followed by Hepatozoon canis in 50/130 (38%), Anaplasma platys in 4/130 (3%), B. vogeli in 4/130 (3%), Acanthochelionema reconditum in 3/130 (2%) and Dirofilaria immitis in 2/130 (2%). Forty-four dogs (34%) were co-infected with two or more pathogens. The most common co-infection observed was with B. gibsoni + H. canis (34%) followed by triple-infection with B. gibsoni + H. canis + A. platys (3%), and B. gibsoni + B. vogeli + H. canis (3%). The infection rate was higher in Mizoram (58%) than in Tripura (44%)., Conclusions: The high prevalence of infection in the studied dog population, especially with B. gibsoni and H. canis, indicates that vector-borne diseases pose a serious threat to the health of dogs in this area of Northeast India. Prevention of vector-borne diseases by using topical acaricides and heartworm preventative treatment would be of great benefit for reducing the threat of vector-borne diseases in the study area.

Background: Canine babesiosis is a severe disease caused by several Babesia spp. A number of names have been proposed for the canine-infecting piroplasmid pathogen initially named Theileria annae Zahler, Rinder, Schein & Gothe, 2000. It was shown to be a member of the Babesia (sensu lato) group infecting carnivores and is also closely related to the Babesia microti group. Subsequently, the same parasite species was reclassified as a member of the genus Babesia and the name Babesia vulpes Baneth, Florin-Christensen, Cardoso & Schnittger, 2015 was proposed for it. However, both names do not meet the requirements of the International Code of Zoological Nomenclature (no accompanying descriptions, no deposition of type-specimens) and cannot be recognized as available names from the nomenclatural point of view. The purpose of this study was to further characterize this parasite in order to confirm its validity, to provide its description and to introduce zoological nomenclature for it with the name Babesia vulpes n. sp., Results: Morphological description of the parasite in canine erythrocytes demonstrated that it takes the shape of small (1.33 × 0.98 µm), round to oval forms reminiscent of the pyriform and ring shapes of other small canine Babesia spp., such as Babesia gibsoni Patton, 1910 and Babesia conradae Kjemtrup, Wainwright, Miller, Penzhorn & Carreno, 2006. However, these parasite forms were overall smaller than those measured for the latter two species and no tetrad (Maltese cross) form was reported. Furthermore, phylogenetic analysis using the cytochrome c oxidase subunit 1 (COX1) amino acid sequences substantiates the species identity of this parasite as previously demonstrated based on phylogenetic analysis of the 18S rRNA and β-tubulin genes. The holotype of the parasite species was designated and deposited in an accessible public collection., Conclusions: This study ratifies the name Babesia vulpes n. sp. proposed for the parasite previously referred to as Theileria annae Zahler, Rinder, Schein & Gothe, 2000, Babesia annae (Zahler, Rinder, Schein & Gothe, 2000) or Babesia vulpes Baneth, Florin-Christensen, Cardoso & Schnittger, 2015, or mentioned as "Babesia microti-like piroplasm", "Babesia Spanish dog isolate" and Babesia cf. microti.

Peritoneal larval cestodiasis caused by Mesocestoides spp. is a rare infection in dogs. A 6-year-old female dog was presented for veterinary care with urinary incontinence which started 1 year earlier. After performing hematology, ultrasound, and computerized tomography, an exploratory laparotomy revealed canine peritoneal larval cestodiasis (CPLC) with the presence of Mesocestoides vogae (syn. Mesocestoides corti) tetrathyridia confirmed by morphological identification and PCR and DNA sequencing. Parasitic cysts were found around the urinary bladder and appeared to inhibit its normal function. An initial treatment with 5 mg/kg praziquantel subcutaneously every 2 weeks for four treatments failed to alleviate the clinical signs, and only treatment with fenbendazole at 100 mg/kg P.O. twice daily for 28 days was associated with the disappearance of ascites and regaining of urinary control. This is the first report of CPLC associated with urinary incontinence in dogs and the first description of this cyclophyllidean cestode in dogs in Israel.

Background: Hosts repeatedly bitten by sand flies develop antibodies against sand fly saliva and screening of these immunoglobulins can be employed to estimate the risk of Leishmania transmission, to indicate the feeding preferences of sand flies, or to evaluate the effectiveness of vector control campaigns. Previously, antibodies to sand fly saliva were detected using whole salivary gland homogenate (SGH) or recombinant proteins, both of which also have their disadvantages. This is the first study on sand flies where short peptides designed based on salivary antigens were successfully utilized for antibody screening., Methodology/principal Findings: Specific IgG was studied in hosts naturally exposed to Phlebotomus orientalis, the main vector of Leishmania donovani in East Africa. Four peptides were designed by the commercial program EpiQuest-B, based on the sequences of the two most promising salivary antigens, yellow-related protein and ParSP25-like protein. Short amino acid peptides were synthesised and modified for ELISA experiments. Specific anti-P. orientalis IgG was detected in sera of dogs, goats, and sheep from Ethiopia. The peptide OR24 P2 was shown to be suitable for antibody screening; it correlated positively with SGH and its specificity and sensitivity were comparable or even better than that of previously published recombinant proteins., Conclusions/significance: OR24 P2, the peptide based on salivary antigen of P. orientalis, was shown to be a valuable tool for antibody screening of domestic animals naturally exposed to P. orientalis. We suggest the application of this promising methodology using species-specific short peptides to other sand fly-host combinations., Competing Interests: Sergey V. Litvinov is an employee of Aptum Biologics Ltd. Aptum Biologics Ltd. is a participant in the Consortium that produced this work, and provided the software and technology for the sequence analysis, a unique one. Aptum Biologics Ltd. has not involved any patents, consultancy, employment or any other forms of financial affiliation or benefits from this work.