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Giardia duodenalis is one of the most prevalent enteric parasites of dogs. Point-of-care antigen tests (POC) are rapid and do not require additional equipment, or a specialised diagnostic laboratory. The aim of this study was to compare diagnostic tests available in veterinary practices and in a diagnostic laboratory for the detection of G. duodenalis on a cohort of group-housed dogs from New South Wales, Australia. Two different POC tests were used for the detection of G. duodenalis. Laboratory tests used were the multiplexed-tandem PCR panel (MT-PCR) that includes detection of G. duodenalis DNA, and two reference tests (an in-house TaqMan real-time PCR and a direct immunofluorescence assay, DFA). Canine faecal samples (n = 40) were tested simultaneously for the detection of G. duodenalis. Using either DFA or TaqMan real-time PCR as reference tests, 77.5% (31/40) and 82.5% (33/40) of dogs tested positive, respectively. Agreement (Kappa) between the DFA and TaqMan real-time PCR was 0.84 (95% CI 0.64 to 1.00). There was substantial G. duodenalis test outcome agreement between the two POC tests, Kappa = 0.75. Combining the two POC tests yielded 77% sensitivity and 100% specificity with DFA as reference, and for TaqMan real-time PCR it was 73% sensitivity and 100% specificity. The MT-PCR was in excellent agreement with each reference test, DFA or TaqMan real-time PCR. Due to the high specificity of both POC tests, they can be confidently used as rule-in diagnostics. Confirmatory testing that detects different biological parameters such as DNA, e.g. PCR (inc. MT-PCR), should be implemented before concluding that a dog is negative for the presence of G. duodenalis. [Display omitted] • POC tests for G. duodenalis in dogs are rapid and equipment-free. • Veterinary POC tests show substantial agreement with reference tests. • POC tests provide high specificity for rule-in diagnostics. • Confirmatory tests like PCR (e.g. MT-PCR) are essential for conclusive results. [ABSTRACT FROM AUTHOR]

The present study evaluated, in laboratory and field, the efficacy and safety of formulations of Pelargonium graveolens (geranium – G), Origanum majorana (oregano - O) commercial essential oils (EO) and thymol (T) to control of Rhipicephalus sanguineus sensu lato. In the laboratory, three formulas (A: 2% tween 80%, B: powder and C: nanoemulsion) by a mixture of these components (GOT) were prepared and evaluated, and the best one was used to assess its safety and field application against R. sanguineus s. l. on naturally infested dogs. Besides the major compounds of the EO used were identified. The results of the lab study showed that formula A (2.5 g of each G + O + T + 2% tween 80 to complete 100 mL) was significantly more effective than the other two formulas tested and exhibited highly effective adulticidal, larvicidal, and ovicidal activity against R. sanguineus s.l. Significant LC 50 and LC 90 values of GOT were evaluated (13.4 and 21.5 mg/mL, respectively) for the adulticidal activity, (2.81 and 4.46 mg/mL, respectively) for ovicidal activity and (2.44 and 4.45 mg/mL, respectively) for larvicidal activity. The safety of formula A has been proven by the absence of its cytotoxicity on a cell line of human epidermoid carcinoma. Citronella and carvacrol were the major compounds identified in the commercial essential oils of P. graveolens and O. majorana, respectively. Formula A was used in a field control trial for almost 8 months, during the tick infestation season (April to November, 2022). Fourteen naturally infested dogs were divided into two groups, each with seven dogs. One group received formula A spraying five times during an experiment that continued for 8 months, while the other group received treatment with commercially available malathion acaricide. The animals were sprayed on five occasions throughout the experiment (April, June, July, August, and September). The results showed a substantial percentage of effectiveness after the first application of formula A with a 99.3% reduction in tick count at day 28 post-application (PA). In the case of severe infestation 60 days after the first application of formula A (more than 180 ticks per dog), the second application was done, achieving an efficacy of 54.9% at day 3 PA, so an emergency spray was done at day 5 PA to combat the rest of the tick infestation, achieving efficacy of 99% after 3 days. Consequently, a regular spray (third, fourth, and fifth application) was done every 35 days. This regular spray revealed 100% effectiveness at 14 days PA. Biochemical parameters of treated dogs were evaluated to confirm the safety of formula A. Creatinine, ALT, and albumin of the dogs treated with formula A were within the normal range of dogs, while urea and AST were higher than the normal range. In conclusion, formula A can safely treat R. sanguineus s.l. infestations in dogs with regular application every 5 weeks. [Display omitted] • Three formulations containing geranium-oregano-thymol (GOT) (2.5%) were prepared. • Citronella and carvacrol were the major compounds of these essential oils. • Formula A demonstrated promising acaricide activities against R. sanguineus s.l. • Laboratory and field experiments demonstrated that formula A is safe for dogs. • Regular application of formula A revealed effective control of brown dog tick. [ABSTRACT FROM AUTHOR]

Giardia duodenalis (syn. G. intestinalis or G. lamblia) is a parasitic protozoan that infects the upper intestinal tract of a broad range of hosts, including humans and domestic animals. Thus, it has raised concerns about the public health risk due to companion animals. Recently, with the improvement of living standards and increasing contacts between pets and humans, the zoonotic transmission of Giardia has dramatically increased. From a genetic point of view, G. duodenalis should be viewed as a complex species that includes eight different species-specific genetic assemblages. The laboratory diagnosis is mainly based on the finding of microscopic cysts in stool samples by coprological examination. Other methods include the detection of antigens, immunoassays or PCR protocols, which allow the identification of Giardia assemblages. The study aimed to compare the performance of Direct Fluorescence Antibody test (DFA), zinc sulfate flotation technique (ZnSO4), rapid diagnostic test (RDT), end-point PCR amplification (PCR) for the detection of Giardia and to identify the concerning assemblages in a canine population from Central Italy. Direct fluorescence antibody test is the reference standard for laboratory diagnosis of Giardia in fecal samples from dogs, despite the microscopic examination after flotation remains the most useful method in many veterinary diagnostic centers. The present findings demonstrate the high performance of DFA and ZnSO4 in detecting Giardia, while RDT may be useful as alternative or complementary method to the DFA and ZnSO4. PCR performance was low, but it allowed determining Giardia B zoonotic assemblage in 25% of the PCR-positive specimens (15 out of 60), while the remaining PCR-positive isolates belonged to the dog-specific assemblage C. The 26% prevalence of G. duodenalis detected by DFA in owned dogs and the identification of potentially zoonotic assemblages underline the potential risk for public health and indicate frequent cross-species transmission of the parasite between humans and dogs. • The performance of four laboratory tests for detection of Giardia has been assessed. • High performance of DFA in detecting Giardia. • RDT alternative diagnostic method. • PCR-RFLP protocol allowed the identification of Giardia assemblages. [ABSTRACT FROM AUTHOR]

Otodectes cynotis , commonly known as "the ear mite," is a highly contagious ectoparasite and a significant cause of otitis externa in canines. The objective of the current study was to determine the efficacy of the isoxazoline afoxolaner (Nexgard®), and the combination of afoxolaner with milbemycin oxime (Nexgard Spectra®), in dogs naturally infested with O. cynotis. In total, 32 infested client-owned dogs from two different sites in Greece were included. The animals were randomly divided into four equal groups based on their infestation score. Group 1 served as the negative control, group 2 received one oral administration of Nexgard (Day 0), group 3 received two monthly oral administrations of Nexgard (Days 0, 30), and group 4 received two monthly oral administrations of Nexgard Spectra (Days 0, 30), according to label instructions. Otoscopic examinations for mites and observations on debris/cerumen in the ears were carried out on Days 0, 15, 30, and 45. A quantitative assessment of ear mites by ear duct flushing and live mite counts was performed on Day 45. The results demonstrated that a single oral dose of afoxolaner and two monthly doses of afoxolaner or afoxolaner with milbemycin oxime resulted in a 99.9% reduction in live mite counts compared to the untreated control group by Day 45. Additionally, treated dogs showed improved clinical symptoms, such as ear cerumen/debris decrease, while untreated dogs experienced worsening symptoms over the study duration. No adverse events were reported. Overall, these results support the use of afoxolaner-based products to treat O. cynotis infestation in dogs. • Afoxolaner and afoxolaner with milbemycin reduce Otodectes cynotis mites in dogs by 99.9%. • Treatments with afoxolaner reduce ear cerumen and debris in naturally infested dogs. • Both treatments are safe, well-tolerated by the dogs and with no adverse events. [ABSTRACT FROM AUTHOR]

Dirofilaria repens is a paradigmatic example of an emerging vector-borne pathogen (VBP) in both human and veterinary fields. The spatial expansion and the increasing zoonotic impact of this VBP can be related to several drivers including the genetic structure of parasite populations. Italy is one of the European countries traditionally endemic with the highest incidence of canine and human cases of subcutaneous dirofilariosis. The present study aimed to assess the genetic identity and variability of D. repens isolates of human and canine origin from areas of Central Italy, compared with those isolated from different areas of Europe by sequence analysis of mtDNA genes (i.e., 12 S rDNA and cox1). A total of twenty isolates of D. repens were obtained from biopsies of subcutaneous and ocular cases of dirofilariosis occurring in 10 dogs and 10 humans. The sequence analysis of 12 S rDNA showed that all the sequences obtained clustered as a monophyletic group with a strong nodal support, indicating that all sequence types represented D. repens. The cox1 and the 12 S sequence analysis did not show host-related polymorphisms between human and dog-derived specimens. The sequence analysis of cox1 was performed including 8 additional sequences previously obtained from human and canine isolates in the same areas. Out of the 28 sequences analyzed, 20 were grouped in a haplogroup comprising 15 haplotypes (i.e., DR1, DR2, DR4, DR5, DR7, DR8, DR10-DR18), 2 sequences matched to DR9, reported for the first time in Italy, and 6 showed peculiar polymorphisms that were not previously described. The results obtained have implications for a better understanding of the epidemiology and phylogeography of this emerging vector-borne zoonotic parasite. [Display omitted] • Dirofilaria repens as paradigmatic example of an emerging vector-borne pathogen. • Investigation on mtDNA genes of Dirofilaria repens from Italian isolates. • Expanding knowledge on the genetic diversity of D. repens in Europe. [ABSTRACT FROM AUTHOR]

Current recommendations to diagnose cardiopulmonary dirofilariosis in dogs caused by Dirofilaria immitis involves tandem antigen and circulating microfilariae tests. The modified Knott's test is an important tool in heartworm diagnosis, allowing identification of circulating microfilariae. However, the subjective nature of the modified Knott's test affects its accuracy and diagnostic laboratories usually do not provide a quantitative outcome. Quantitative enumeration of microfilariae enables clinicians to track treatment progress and acts as a proxy for detecting emerging macrocyclic lactone resistance. There is a need for better diagnostic tools suitable for routine use to efficiently and accurately quantify the presence of D. immitis microfilaremia. The aim of this study was to determine whether the quantitative modified Knott's test can be substituted by multiplex quantitative polymerase chain reaction (qPCR) targeting D. immitis and associated Wolbachia endosymbiont DNA in canine blood samples. To do this, genomic DNA samples (n = 161) from Australian dogs, collected as part of a previous 2021 study, were assessed in a TaqMan qPCR targeting DNA of D. immitis , Wolbachia sp. and Canis lupus familiaris. Of the 161 genomic DNA samples, eight were considered positive for D. immitis microfilariae. The qPCR assay demonstrated good efficiency (E = 90 to 110%, R2 > 0.94). Considering the performance and efficient use of bench time, this TaqMan qPCR assay is a suitable alternative to the modified Knott's test for quantitative enumeration of microfilariae (Cohen's kappa coefficient [κ]: κ = 1 using D. immitis qPCR marker, κ = 0.93 using Wolbachia qPCR marker). The qPCR data demonstrated a comparable result to that of the quantitative modified Knott's test in a 2022 survey of D. immitis in Australian dogs (n = 23) before and after macrocyclic lactone (ML) administration. Improving the detection and diagnosis of canine heartworm infections will assist veterinarians in better managing and controlling disease outcomes and will be valuable for tracking the spread of ML resistance in Australia. • Rapid detection of quantitative D. immitis microfilariae levels were achieved by multiplex qPCR. • Comparable outcomes can be achieved between the qPCR assay and quantitative modified Knott's test. • SNP analysis to detect ML resistance in USA matched with the ML susceptible reference genome. [ABSTRACT FROM AUTHOR]

Cystic echinococcosis, which is caused by the Echinococcus granulosus. Carnivores, as final hosts, contain adult tapeworms in the small intestine, and a variety of mammals, including humans, harbor the metacestod. This study was designed to investigate the miRNA-based biomarkers for early and accurate diagnosis of E. granulosus in experimentally infected dogs. A liver with an obvious hydatid cyst was obtained from a slaughterhouse and then protoscoleces were collected. Following, viable protoscoleces were administred to three experimental dogs (ED1, ED2 and ED3) and another uninfected control dog (UCD) was kept as control without infection. Stool samples of all groups were collected during 50 days from the beginning of the experimental infection and stored at − 80 °C till work. Total miRNA was isolated from all individual stool samples. The qRT-PCR method was used to determine the differences in the expression levels of E. granulosus specific miRNAs which were egr-let-7–5p, egr-miR-2b-5p, egr-miR-71–5p and egr-miR-125–5p. All miRNAs were found to be expressed from the first day in all infected dogs. In the stool samples of the UCD, the egr-miR-71–5p was detected, while the other miRNAs (egr-let-7–5p, egr-miR-2b-5p, egr-miR-125–5p) were not expressed. The expression of egr-let-7–5p and egr-miR-125–5p was significantly increased in ED1 compared to UCD on all days. In particular, for the first time, the expression levels of egr-let-7–5p and egr-miR-125–5p increased significantly between days 15 and 19. Similarly, the increase in let-7–5p and miR-125–5p expression was statistically significant in ED2. In ED3, egr-let-7–5p, egr-miR2b-5p and egr-miR-125–5p expressions were significantly increased on all days. In particular, egr-let-7–5p expression levels increased significantly for the first time between days 15 and 19. In addition, egr-mir-125–5p expression levels were found to increase at a high level for the first time on day 16. In conclusion, especially egr-let-7–5p and egr-miR-125–5p can be used as early diagnostic biomarkers in dogs infected with E. granulosus. • Three experimentally infected and a control dog's stools were collected during 50 days. • egr-let-7-5p, egr-miR-2b-5p, egr-miR-71-5p and egr-miR-125-5p were investigated. • All miRNAs were found to be expressed from the first day in all infected dogs. • Especially egr-let-7-5p and egr-miR-125-5p can be used as early diagnostic biomarkers. [ABSTRACT FROM AUTHOR]

Intestinal parasites, including cestodes like Dipylidium caninum , are common in dogs in the United States of America (USA), but fecal flotation consistently, and, at times, dramatically, fails to identify many of these infections. To determine the extent to which including coproantigen testing for D. caninum would improve the identification of dogs infected with this cestode, we evaluated fecal samples from 877 dogs (589 pet and 288 from municipal shelters) from six USA states using zinc sulfate (specific gravity 1.24) fecal flotation with centrifugation along with coproantigen detection for Giardia sp., hookworms, ascarids, and Trichuris vulpis. For D. caninum , PCR of perianal swabs was included. Intestinal parasite infections were identified, using centrifugal fecal flotation or coproantigen, in 265 dogs (13.2 % pet, 64.9 % shelter). Dipylidium caninum infection was detected in 5.6 % of dogs with the combination of coproantigen and centrifugal fecal flotation, and 7.3 % of dogs when perianal swab results were included; prevalence varied by diagnostic method, population, and geographic region. In pet dogs, D. caninum infection was identified by fecal flotation (0), coproantigen (2.2 %), or perianal swabs (1.2 %). The same methods revealed infection in 0.3 %, 12.5 %, and 11.1 % of shelter dogs, respectively. Frequent use of praziquantel in shelter dogs (116/288; 40.3 %) may have reduced prevalence. Positive and negative agreement of D. caninum coproantigen with perianal swab PCR in pet dogs was 85.7 % and 98.8 %, respectively. Multiple logistic regression analysis accounting for region, population, and age found D. caninum infection to be more common in shelter dogs relative to pet (adjusted OR 4.91 [2.48, 10.24]) and in the Southcentral and Southeast regions relative to North (adjusted OR 9.59 [1.92, 174.13] and 17.69 [3.67, 318.09] respectively). Coproantigen testing also enhanced the detection of other intestinal parasites over fecal flotation alone, including Giardia sp. (14.7 % vs 3.3 %), hookworms (13.8 % vs 8.4 %), ascarids (2.9 % vs 2.2 %), and T. vulpis (2.9 % vs 1.4 %). Together, these data indicate that the coproantigen assay employed increases detection of D. caninum infections several fold, supporting the use of this test in clinical practice, and add to a growing body of research documenting enhanced diagnosis through implementation of multiple laboratory-based methods. [Display omitted] • Canine Dipylidium caninum infections readily identified by coproantigen assay. • Dipylidium caninum detected in > 20 % of shelter dogs from southern states. • Intestinal parasites found in > 10 % of pet and > 60 % of shelter dogs tested. [ABSTRACT FROM AUTHOR]

Animal infection models are crucial for studying various aspects of Ehrlichia canis infections. To understand the pathogenesis of the first Chinese isolate of E. canis and simulate the natural progression of canine ehrlichiosis, we developed a model with 18 Beagle dogs that consisted of E. canis initial infection (days 0–17), treatment with doxycycline or rifampicin (days 18–32), recovery (days 33–66), E. canis reinfection (days 67–91), and Babesia vogeli superinfection (days 92–116). We measured body weight and rectal temperature every other day, drew blood every 4 days for routine hematology and biochemistry tests, and for quantification of E. canis and B. vogeli by quantitative PCRs. In this study, the first isolate of E. canis from China was used to experimentally infect dogs, and the infected dogs exhibited clinical signs of acute severe ehrlichiosis, including high fever, loss of appetite, dehydration, and body weight loss, confirming the similar pathogenicity of E. canis in China as compared to isolates from other regions. Infection with E. canis and B. vogeli led to reduced body weight and fever in dogs. Doxycycline treatment led to absence of E. canis DNA in infected dogs, while rifampicin treatment lowered the blood E. canis copy number up to 1.5 folds. E. canis -free infected dogs after doxycycline treatment were successfully re-infected with E. canis , indicating dogs with antibodies are still at risk of re-infection. Super-infection with B. vogeli resulted in higher fever, more severe anemia, and a reduced number of platelets. Splenectomized dogs showed significantly higher E. canis numbers during recovery and re-infection than intact dogs. The histological changes were observed in brain, lung, kidney, liver and spleen of the infected dogs. The findings in this study provide insights into clinical and hematologic responses, as well as effective treatment options, for dogs infected with the first Chinese isolate of E. canis , and may contribute to our understanding of the diagnosis and prevention of tick-borne diseases in dogs, including canine monocytic ehrlichiosis. • Experimentally infected model to study pathogenicity of first E. canis strain from China. • Doxycycline eliminated E. canis while rifampicin lowered E. canis numbers. • E. canis -free dogs after doxycycline treatment were successfully reinfected. • Splenectomized dogs showed higher E. canis numbers than intact dogs. • B. vogeli super-infection led to higher fever, more severe anemia, lower platelet counts. [ABSTRACT FROM AUTHOR]

Parasitic diseases are considered to be a cause of oxidative stress which leads to oxidative damage of various molecules including DNA. This can result in mutations, replication errors, and genome instability. Therefore, aim of this study was to measure DNA damage induced by Dirofilaria immitis in the single cells such as dogs' leukocytes using the comet assay. Also, we monitored the effects of antiparasitic treatment on mitigation of sensitivity to DNA damage in leukocytes treated with H 2 O 2 using the in vivo and ex vivo comet assay. The whole blood samples from 34 dogs from Serbia were used, both males and females, from one to 13 years old, both pure and mixed-breeds. A rapid immunochromatographic test (Antigen Rapid Heartworm Ag 2.0 Test Kit, Bionote, Minnesota, USA) was used for the detection of D. immitis antigens. The modified Knott's test and PCR were used in the aim of detecting D. immitis microfilariae in dogs' blood, and evaluating the number of circulating microfilariae during the treatment. The genotoxicity evaluation showed that D. immitis infection resulted in DNA damage in naturally infected dogs, with the highest DNA damage occurring in the group of dogs with severe clinical signs. Treatment with ivermectin and doxycycline decreased DNA damage in leukocytes of dogs in all groups, as the intensity of infection decreased due to applied therapy. Ex vivo comet assay results showed that leukocytes exhibited decreased sensitivity to H 2 O 2 -induced DNA damage during treatment. The results of the modified Knott's test and PCR in our study showed that treatment with ivermectin and doxycycline was successful in decreasing the average number of microfilariae during the time and at the end eliminating them from the dogs' blood. [Display omitted] • D. immitis infection causes genotoxic effects observed in the Comet assay. • Elimination of the parasites by the applied therapy reduced DNA damage. • Leukocytes of dogs exhibited decreased susceptibility to H2O2 damage after therapy. [ABSTRACT FROM AUTHOR]

• Combination treatment with moxidectin and doxycycline is adulticidal in dogs naturally infected with D. immitis. • Efficacy is 100% at 9 months from the beginning of treatment. • Efficacy for elimination of microfilariae is 99.9% after 1 month from the beginning of treatment. • Treatment is well tolerated and non-inferior to melarsomine at 12, 18 and 24 month from the beginning of treatment. Heartworm infection (also known as dirofilariosis due to Dirofilaria immitis) in dogs causes chronic pulmonary disease that, if left untreated, can lead to right-side congestive heart failure. Currently, the only registered drug for adulticide therapy in dogs with heartworm disease (HWD) is melarsomine dihydrochloride. The recent targeting of the bacterial endosymbiont Wolbachia , through antibiotic therapy of the infected host, has offered an interesting alternative for the treatment of HWD. Recent reports of the adulticide activity of an ivermectin/doxycycline combination protocol has lead the American Heartworm Society (AHS) to include in its guidelines that, in cases where arsenical therapy is not possible or is contraindicated, a monthly heartworm preventive along with doxycycline for a 4-week period might be considered. In the present study, 20 dogs with confirmed natural D. immitis infection were included following owner consent. Fourteen dogs were treated with a topical formulation containing 10% w/v imidacloprid and 2.5% w/v moxidectin (Advocate®, Advantage Multi®, Bayer), monthly for nine months, associated to doxycycline (10 mg/kg/BID) for the first 30 days. Six dogs were treated with melarsomine (Immiticide®, Merial) (2.5 mg/kg) at enrollment, followed one month later by two injections 24 h apart. The presence of circulating antigens and the number of microfilariae (mf) were evaluated at the moment of enrollment and then at 1, 2, 3, 4, 5, 6, 7, 8, 12, 18, 24 months post enrollment. Echocardiogram and radiographs were performed at month 0, 6, 12, 18, 24. Monthly moxidectin combined with 30 days of doxycycline eliminated circulating microfilariae within one month, thus breaking the transmission cycle very quickly. Furthermore, dogs treated with the combination protocol started to become negative for circulating antigens at 4 months from the beginning of treatment and all except one were antigen negative at 9 months. All dogs treated with melarsomine were antigen negative by 5 months from the beginning of the treatment. No dogs showed worsening of pulmonary patterns or criteria indicative of pulmonary hypertension 12 to 24 months after. For the criteria mf concentration, antigen concentration, radiography and echocardiography at 12, 18 and 24 months the non-inferiority for the moxidectin group could be proven for a non-inferiority margin of 15% for the rate difference. Dogs treated with moxidectin and doxycycline became negative for microfilariae and antigens sooner when compared to melarsomine in the present study and to dogs treated with doxycycline combined with ivermectin in studies previously published. [ABSTRACT FROM AUTHOR]

• Saliva proteome showed 90 differentially expressed proteins in dogs with CanL. • Saliva could be a potential source of diagnosis biomarkers of CanL. • Proteins related to carbohydrates metabolism could play key roles in CanL. In the present study, a quantitative proteomic approach to study changes in saliva proteins associated with canine leishmaniosis (CanL) was performed. For this, canine salivary proteins were analysed and compared between dogs before (T0) and after (T1) experimental infection with Leishmania infantum by high-throughput label-based quantitative LC–MS/MS proteomic approach and bioinformatic analysis of the in silico inferred interactome protein network was created from the initial list of differential proteins. More than 2000 proteins were identified, and of the 90 differentially expressed proteins between T0 and T1, 12 were down-regulated with log2 fold change lower than −0.5849, and 19 were up-regulated with log2 fold change greater than 0.5849. This study provides evidence of changes in salivary proteome that can occur in canine leishmaniosis and revealed biological pathways in saliva modulated in canine leishmaniosis with potential for further targeted research. [ABSTRACT FROM AUTHOR]

• This is the largest cohort of PCR confirmed, natural Babesia rossi infections published. • Clinical, hematological, biochemical and pathology is described. • Odds ratios confirmed which clinical and biochemical measures associat.e with poor outcome. • ROC curves identified TT4, bilirubin, urea and cortisol are most predictive of death. • Cut-offs for these measures allow clinicians and researchers to objectively classify cases. Babesia rossi causes the most severe clinical disease in dogs of all the babesia parasites. We included 320 naturally-infected dogs that presented for care at the Onderstepoort Veterinary Academic Hospital between 2006 and 2016. All dogs had mono-infections confirmed by multiplex PCR. The data allowed more accurate clinical classification of the disease and identified parameters that were associated with disease severity and death. Odds ratios for dying were significant (P < 0.05) for increased band neutrophil count, collapse at presentation; presence of cerebral signs; hypoglycaemia; hyperlactatemia; high urea, high creatinine; hyperbilirubinaemia; hypercortisolaemia; and hypothyroxinaemia. Joint component analysis confirmed that the variables with significant odds ratios grouped together with death. Yet, multivariate logistic regression was unable to identify a group of significant independent predictors of death. Receiver Operator Characteristic curves indicated that low total thyroid hormone, high bilirubin, high serum urea and high cortisol concentrations were the variables with the highest sensitivity and specificity for death. These data provide both the clinician and researcher with a set of easily-measured laboratory and clinical assessments to classify cases into those that are uncomplicated and those that are complicated. The disease is complex and multisystemic and probably involves mechanisms more proximal in the pathogenesis than those that have been evaluated. [ABSTRACT FROM AUTHOR]

• Immunogenetics might explain outcome of Leishmania infantum infection in dogs. • One SNP in CIITA gene associated with infection in case control study. • Polymorphisms in multiple genes associated with susceptibility in infected Beagles. • SNPs were not significant after correction for multiple testing. The outcome of infection with Leishmania infantum in dogs is variable, which is thought to be due to the nature of the immune response mounted by the host. As a consequence, the clinical signs and severity of canine leishmaniosis vary between individual dogs. Host immunogenetic factors might play an important role in determining the outcome of infection. The aim of this study was to examine polymorphisms in innate and adaptive immune response genes, to determine whether any of these were associated with susceptibility or resistance to L. infantum infection. Genomic DNA was obtained from two groups: pet dogs in endemic regions of Europe and a group of Beagles exposed to sand fly infection as part of a vaccine study. Genotyping was performed using a SNP (single nucleotide polymorphism) array for selected immune response genes. The first part of the study compared 62 clinical cases with 101 clinically unaffected dogs that were seronegative for Leishmania antibodies. One SNP in the CIITA gene demonstrated a significantly higher minor allele frequency in the case group, compared with the control group at the individual SNP level after permutation, but was not significant after correction for multiple testing. The second part of the study examined 48 Beagle dogs exposed to L. infantum over two transmission seasons. Twenty-seven dogs with a resistant phenotype (no evidence of clinical disease, seronegative at the end of the study period, negative on lymph node culture and only transiently PCR positive in bone marrow) were compared with 21 dogs demonstrating a susceptible phenotype (clinical disease, seropositive, positive lymph node culture and consistently PCR positive in bone marrow). Three SNPs in TLR3 , two SNPs in PTPN22 and one SNP in TLR4 and IL1A were associated with the susceptible phenotype in the Beagle group at the individual SNP level after permutation analysis, but were not significant after correction for multiple testing. Further validation of these SNPs is required in a larger cohort of dogs, ideally with extreme phenotypes to confirm an association with the outcome of L. infantum infection. [ABSTRACT FROM AUTHOR]

• A randomized trial assessed the effects of Fluralaner in dogs on Triatoma infestans. • Fluralaner exerted no significant effects on bug feeding success or engorgement level. • Insect mortality was ≥99% up to 60 days posttreatment (DPT). • Mortality was high over 90–120 DPT depending on bug stage and susceptibility status. • Fluralaner and xenointoxication are eligible for Phase III efficacy trials. New vector control tools that can fit into a broader integrated vector management strategy are notably lacking. We conducted a seven-month randomized trial to assess the efficacy of a single oral dose of Fluralaner (Bravecto®) administered to dogs on the blood-feeding success, engorgement levels and mortality of pyrethroid-resistant and -susceptible Triatoma infestans third- and fifth-instar nymphs. The trial included 10 Fluralaner-treated and 10 placebo-treated (control) outbred healthy dogs residing in rural houses of the Argentine Chaco. Most (92.7%) of the 3017 triatomines exposed were able to blood-feed. Generalized linear models showed that blood-feeding success was not significantly modified by Fluralaner treatment, time posttreatment and their interaction. However, pyrethroid-susceptible fifth instars blood-fed significantly more frequently than susceptible third instars, and no significant differences were observed between the latter and resistant fifth instars. Engorgement levels were not significantly modified by Fluralaner treatment, time posttreatment and their interaction. Nearly all the triatomines that blood-fed on treated dogs up to 60 days posttreatment (DPT) died within 24 h regardless of pyrethroid susceptibility status combined with bug stage. Cumulative bug mortality over 4 days postexposure remained high over 90–120 DPT (70–81% in susceptible third and fifth instars, and 47–49% in resistant fifth instars), and was virtually nil at 210 DPT. Triatomines that fed on control dogs suffered marginal mortality (0–4%) except at 4 and 30 DPT. Fluralaner and xenointoxication are eligible for Phase III efficacy trials alone or combined with other methods in the frame of an integrated vector management strategy in areas with or without pyrethroid resistance. [ABSTRACT FROM AUTHOR]

Highlights • The questionnaire was sent to 2795 veterinary facilities. The response rate was 25%. • At least one case of Dirofilaria spp. in dogs has been diagnosed in 47% facilities and in 6% for cats. • To diagnose heartworm infection 83.8% of the facilities used serology. • Prevention of D. immitis was begun mainly in April-May. • Prevention of D. immitis in cats was not recommended in 51% of the facilities. Abstract Dirofilaria immitis and D. repens are vector-borne parasites of dogs and cats, with zoonotic potential, endemic in many parts of Europe, including Italy. Control and prevention of dirofilariosis are essential points to reduce their diffusion in animals and humans and veterinarians are the main subjects involved in this "battle". With the aim to better understand current practice by veterinarians, an online questionnaire on different aspects of D. immitis and D. repens was sent to companion animal veterinary facilities registered with the Italian Veterinary chamber. The overall response rate was 25%. Approximately 47% had diagnosed at least one case of Dirofilaria spp. in dogs in the last year and approximately 6% had diagnosed at least one case of Dirofilaria spp. in cats. Of the 662 facilities surveyed, 83.8% used serology to diagnose heartworm infection. For the diagnosis of D. repens infection in dogs and cats, a high percentage of facilities relied on an external laboratory. Most recommended beginning prevention of canine heartworm disease in April-May. Topical and injectable moxidectin and oral ivermectin were the two most commonly used preventives for D. immitis in dogs. The drug most commonly used for heartworm adulticide treatment in dogs was melarsomine. This study show that veterinary facilities ians working in an endemic area for D. immitis and D. repens are aware of recent developments in adulticide therapy. Results would suggest however that diagnosis is not always carried out according to what is currently recommended by international guidelines and that the timing for starting prevention may not be optimal. [ABSTRACT FROM AUTHOR]

Highlights • Foxes may act as wildlife reservoirs of Angiostrongylus vasorum for dogs in Europe. • Considerable genetic diversity was found among A. vasorum from foxes in Greater London. • First report highlighting genetic diversity of A. vasorum within individual foxes. • Foxes may act as reservoirs of genetic diversity of A. vasorum in the UK. Abstract Angiostrongylus vasorum is a nematode parasite of the pulmonary arteries and heart that infects domestic and wild canids. Dogs (Canis familiaris) and red foxes (Vulpes vulpes) are the most commonly affected definitive hosts. Recent studies suggest that angiostrongylosis is an emerging disease, and that red foxes may play an important role in the epidemiology of the parasite. Genetic analyses of parasites collected from dogs and foxes throughout Europe have shown that the same parasite haplotypes are commonly shared between different host species. However, the extent of genetic diversity within local A. vasorum populations and individual hosts is unknown. The objective of the present study was to assess the occurrence of genetic diversity among A. vasorum (a) recovered from different foxes within the Greater London area (a localised population, single worm per fox dataset); and (b) hosted within single foxes (multiple worms per fox dataset). During 2016, A. vasorum worms were collected from foxes culled for other purposes in London. DNA was extracted from each parasite and a partial fragment of the mitochondrial cytochrome oxidase subunit 1 (mtCOI) gene was amplified and sequenced. Sequences from the single worm dataset were compared with those published elsewhere. Combined, 19 haplotypes were described of which 15 were identified from foxes found in London, indicating that considerable genetic diversity can be detected within a local geographic area. Analysis of the multiple worm dataset identified 22 haplotypes defining worms recovered from just six foxes, emphasising the relevance of wild canines as reservoirs of genetic diversity. This is the first study to explore the genetic complexity of individual fox-hosted A. vasorum populations. [ABSTRACT FROM AUTHOR]

Highlights • Renal azotemia was diagnosed at presentation in 14% of all dogs. • Based on urine specific gravity, renal azotemia was an underdiagnosed event. • Based on urine osmolality, most azotemic dogs had renal azotemia. • Old age and hyperphosphatemia could help to identify concomitant renal azotemia. Abstract The occurrence of acute kidney injury in canine babesiosis is not well documented. Furthermore, interpretation of urine specific gravity (USG) to assess renal concentrating ability is hampered by the frequent presence of hemoglobinuria in this disease. This cross-sectional study aimed to test the hypothesis that renal azotemia (RA) is underdiagnosed according to current canine babesiosis literature by determining its occurrence at presentation, using urine osmolality instead of USG to measure urinary concentration. The second objective was to examine potential associations between the presence of RA and selected clinical and laboratory variables at presentation. Medical records available from 3 previously performed prospective data collections were reviewed retrospectively. Client-owned dogs that were diagnosed with babesiosis caused by Babesia rossi, were included if a complete blood count, biochemistry profile, and urinalysis was performed at admission. Urine osmolality was measured to identify dogs with RA. Differences between dogs with RA and dogs without RA were assessed by nonparametric statistics. One hundred and fifty-two dogs were included, of which 26 (17%) were azotemic at admission. The occurrence of RA was 14% (21/152), hence 81% (21/26) of all azotemic dogs were diagnosed with RA. In contrast, when diagnosis of RA was based on an admission USG < 1.030, only 23% (6/26) of the azotemic dogs would have been considered to have RA. Several signalment and clinicopathological findings were found to be associated with the presence of RA, including older age, and the presence of collapse, hypoglycemia, hyperphosphatemia, cerebral babesiosis, and acute respiratory distress syndrome. Lastly, survival at discharge was significantly lower in dogs diagnosed with RA at presentation. Our results clarified that RA is more common than previously reported in B. rossi. This study also demonstrated that USG determination is not a reliable method to evaluate renal concentrating ability in azotemic dogs with babesiosis. Thus, if available, urine osmolality should be part of the diagnostic work-up of dogs infected with B. rossi to avoid misclassification of dogs with RA as having prerenal azotemia. If urine osmolality cannot be measured, clinicians should realize that most azotemic dogs with B. rossi infection have RA. [ABSTRACT FROM AUTHOR]

Canine leishmaniasis (CanL) is a disease caused by Leishmania infantum that can vary from a subclinical infection to a severe disease. Dogs affected with CanL present varying degrees of renal dysfunction. Unfortunately, traditional biomarkers such as urea and creatinine detect renal damage in advanced stages of the disease, so more accurate biomarkers are needed. Hence, we aimed to study how urinary cystatin C (CysC) and N-acetyl-beta-D-glucosaminidase (NAG), behave in dogs with CanL at different stages of the disease. Eighty-six CanL infected dogs were classified according to LeishVet stages: LI (16 dogs), LIIa (12 dogs), LIIb (12 dogs), LIII (16 dogs) and LIV (30 dogs); as a control, 17 healthy dogs were studied. Blood samples were collected for complete haematological and biochemistry analysis including plasma cystatin C. Urine analysis included urine specific gravity (USG), urine protein to creatinine ratio (UPC), CysC and NAG expressed as a ratio with creatinine uCysCc (μg/g) and uNAGc (IU/g). The haematological, biochemical and urinary analysis coincided with the LeishVet guidelines. The statistical study of the uCysCc ratio and the uNAGc, showed significant increase when compared against control starting from group LI (p < 0.05). Interestingly, when the cut-off values were calculated using the ROC curve, uCysCc (258.85 µg/g) and uNAGc (2.25 IU/g) 75 % of the dogs included in LI groups surpassed the threshold. Hence our study indicates that uCysCc and uNAGc, could help to detect early renal damage in CanL affected dogs. • Dogs infected with Leishmania spp. present varying degrees of renal dysfunction. • Urinary Cystatin C and plasma Neutrophil Gelatinase-Associated Lipocalin were studied. • Cut-off in urine was 258,8 µg/g for CysC and 2,25 UI/g for NAG in non-azotemic dogs. • Plasma CysC > 0,22 mg/l detected renal damage in azotemic dogs. [ABSTRACT FROM AUTHOR]

Cystic echinococcosis is endemic both in livestock and humans in many parts of Kenya. However, very little data exists on Echinococcus infections in dogs, and therefore their role in maintaining the transmission cycles and environmental contamination with eggs of Echinococcus species is unknown. The study aimed to establish the prevalence and distribution of Echinococcus granulosus sensu lato causing infection in dogs in Kenya. A total of 1621 dog faecal samples were collected from the environment in four different regions and examined microscopically for the presence of taeniid eggs. Up to 20 individual taeniid eggs per faecal sample were picked, lysed and genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing of the NADH dehydrogenase subunit 1 ( nad1 ) gene. Eleven percent (178/1621) of faecal samples had taeniid eggs, while 4.4% (71/1621) contained Echinococcus spp. eggs. Area-wise, the faecal prevalence of Echinococcus spp. was 9.2% (48/524) in Turkana, 4.0% (20/500) in Maasai Mara, 0.7% (2/294) in Isiolo and 0.3% (1/303) in Meru. E. granulosus sensu stricto (s. s.) was the dominant Echinococcus taxon, followed by E. canadensis (G6/7) that was detected in 51 and 23 faecal samples, respectively. E. ortleppi was detected in only 5 faecal samples. We report for the first time the presence of E. felidis eggs in two dog faecal samples (from Maasai Mara region). Mixed infections of these taxa were also found in faecal samples, including: E. granulosus s. s. and E. canadensis (G6/7) (n = 7), E. granulosus s. s. and E. ortleppi (n = 1) and all three species (n = 1). The dog data presented here confirm the differences in diversity and abundance of Echinococcus spp. between regions of Kenya, correspond well with previously published data from livestock, and tentatively suggest a role of domestic dogs as a link between domestic and sylvatic cycles of Echinococcus spp. [ABSTRACT FROM AUTHOR]

In Australia, tick paralysis in dogs (caused by a toxin in the saliva of Ixodes species during feeding) is a serious, distressing condition, and untreated it is often fatal. The aim of this study was to quantify the association between parkland (recreational or natural) in an urban area and the occurrence of canine tick paralysis. Brisbane, as a large urban centre located within the zone of paralysis tick habitat along the east coast of Australia, was selected as the study area. Postcodes selected for inclusion were those defined as being of an urban character (Australian Bureau of Statistics). The number of natural and recreational parkland polygons and total land area per postcode were derived. Tick paralysis case data for the selected postcodes were extracted from a national companion animal disease surveillance database. Between October 2010 and January 2017, 1650 cases of tick paralysis in dogs were reported and included in this study. Significant correlations were found between the number of reported cases per postcode and parklands: natural counts, 0.584 ( P  < 0.0001); natural area, 0.293 ( P =  0.0075); recreational counts, 0.297 ( P =  0.0151); and recreational area, 0.241 ( P =  0.0286). Four disease clusters were also detected within the study area. All of these were located on the edges of the study area – either coastal or on the urban fringe; no clusters were identified within the core urban zone of the study area. Of the disease cases included in this study, strong seasonality was evidence: 68% of all cases were identified in spring. Within urban environments, areas of natural vegetation in particular appear to pose a risk for tick paralysis in dogs. This evidence can be used by veterinarians and dog owners to reduce the impact of tick paralysis by raising awareness of risk areas so as to enhance prevention via chemoprophylaxis and targeted searches of pet dogs for attached ticks. [ABSTRACT FROM AUTHOR]

This guideline is intended as an aid in the design, implementation and interpretation of studies for the assessment of drug efficacy against Eimeria in cattle and sheep, Cystoisospora in pigs and dogs, and Cryptosporidium in cattle. It deals with the most important aspects of how to conduct both experimental and field studies for dose determination, dose confirmation and assessment of field effectiveness. Also, guidance on the selection of animals, diagnostic techniques, statistical evaluation and methods for the preparation, maintenance and use of parasites is provided. The specific management conditions that may influence the course of natural infections and consequently determine treatment schemes are mentioned and suggestions for best practice in sampling and evaluation of data prior to conducting of efficacy studies are given. The guideline is also intended to assist investigators in carrying out specific studies, provide relevant information for registration authorities involved in the decision-making process, assist in the approval of anticoccidial drugs in the target species, and facilitate the world-wide adoption of standard procedures. Although currently not implemented, issues of drug resistance testing and alternative methods for drug testing are also discussed as future issues in drug testing against mammalian coccidia. [ABSTRACT FROM AUTHOR]

We report Taenia ovis infection in Danish sheep for the first time. In spring 2016, the metacestode stage of T. ovis was at slaughter observed in heart muscles, diaphragm and skeletal muscles from approx. a third of all sheep from one specific farm localised in South Jutland. The diagnosis was confirmed by molecular typing of the mitochondrial cytochrome c oxidase I (cox1) gene. Three newly imported dogs were suspected but the definitive host was unidentifiable. The finding is not regulated in the meat control procedures. However, infected meat is usually condemned due to aesthetic reasons causing economic losses. Thus, finding of T. ovis is of concern to sheep meat producers in the area, as the infection could have spread further on to other farms. [ABSTRACT FROM AUTHOR]

Gastrointestinal parasites, protozoa and helminths, remain a common important finding in dogs presented to veterinary medical practices. Moreover, dogs may play an active role in the transmission of enteric parasites to humans, given their cohabitation, making canine gastrointestinal parasitism an issue of major concern for public health. To get an overview of the current state of canine gastrointestinal parasites’ prevalence in Greece, a total of 1036 faecal samples were collected from dogs either presented in veterinary clinics or in animal shelters. Samples were examined by a combined sedimentation–flotation technique. Possible risk factors (age, ownership status, co-existence with other animals, coinfection with other parasites) were assessed using binary regression models for each one of the most prevalent parasites. The overall gastrointestinal parasite prevalence in dogs’ faecal samples was 39%. A total of 11 intestinal parasitic species were detected and up to five different parasites were isolated in the same faecal sample. Toxocara canis was the most prevalent parasite in the studied population, followed by Isospora spp., Giardia spp., Uncinaria spp., Trichuris vulpis , Ancylostoma spp., Toxascaris leonina , and Dipylidium caninum . Additionally, other genera ( Taenia , Capillaria and Angiostrongylus ) were also detected in very low percentages. Young, stray, living without other dogs, coinfected with T. vulpis or T. canis and free of Isospora spp. dogs were more likely to be infected by Giardia spp. Likewise, young, stray, coinfected with T. vulpis or T. canis but free of Giardia spp. dogs were more likely to be infected by Isospora spp. T. canis infections were more common in dogs coinfected with Isospora spp., Giardia spp., D. caninum , Τaenia spp., T. vulpis and T. leonina Finally, T. vulpis more often infected older dogs and dogs coinfected with Isospora spp., Giardia spp. and T. canis Consequently, the prevalence of parasitized dogs was high and the diversity of parasites found was notable, which calls for a greater awareness among veterinarians and pet owners. The proposed prediction models should be taken under consideration in diagnostic approach of clinical cases, as well as in planning sustainable antiparasitic strategies. [ABSTRACT FROM AUTHOR]

Tick paralysis has a major impact on pet dog and cat populations in southeastern Australia. It results from envenomation by Ixodes holocyclus and Ixodes cornuatus ticks, the role of Ixodes cornuatus in the epidemiology of this disease in Australia being unclear. The aim of this study was to describe the geographical distribution of tick paralysis cases in southeastern Australia using data from a national disease surveillance system and to compare characteristics of “endemic” cases with those reported outside this endemic zone (“sporadic” cases). Data were collated and a proportional symbol map of all cases by postcode was created. A 15-case isopleth was developed based on descriptive spatial statistics (directional ellipses) and then kernel smoothing to distinguish endemic from sporadic cases. During the study period (January 2010–December 2015) 12,421 cases were reported, and 10,839 of these reported by clinics located in 434 postcodes were included in the study. Endemic cases were predominantly reported from postcodes in coastal southeastern Australia, from southern Queensland to eastern Victoria. Of those cases meeting selection criteria, within the endemic zone 10,767 cases were reported from 351 (88%) postcodes and outside this zone 72 cases were reported from 48 (12%) postcodes. Of these latter 48 postcodes, 18 were in Victoria (26 cases), 16 in New South Wales (28 cases), 7 in Tasmania (9 cases), 5 in South Australia (7 cases) and 2 in Queensland (2 cases). Seasonal distribution in reporting was found: 62% of endemic and 52% of sporadic cases were reported in spring. The number of both endemic and sporadic cases reported peaked in October and November, but importantly a secondary peak in reporting of sporadic cases in April was found. In non-endemic areas, summer was the lowest risk season whilst in endemic areas, autumn was the lowest risk season. Two clusters of sporadic cases were identified, one in South Australia (P = 0.022) during the period 22 May to 2 June 2012 and another in New South Wales (P = 0.059) during the period 9 October to 29 November 2012. Endemic and sporadic cases did not differ with respect to neuter status (P = 0.188), sex (P = 0.205), case outcome (P = 0.367) or method of diagnosis (P = 0.413). However, sporadic cases were 4.2-times more likely to be dogs than cats (P < 0.001). The endemic tick paralysis zone described is consistent with previous anecdotal reports. Sporadic cases reported outside this zone might be due to a history of pet travel to endemic areas, small foci of I. holocyclus outside of the endemic zone, or in the case of southern areas, tick paralysis caused by I. cornuatus . [ABSTRACT FROM AUTHOR]

Interferon-Gamma (IFN-γ) Release Assays (IGRAs) are easy tests that allow rapid screening of primed memory T-cells immunity in response to antigen. The aim of this study was to use IGRA to assess IFN-γ release in response to Soluble Leishmania infantum antigen (SLA) in whole blood of dogs living in endemic area of visceral leishmaniasis and to interpret IGRA results according to clinical examination, specific anti- Leishmania humoral response and presence of L. infantum DNA in blood. The study was carried out on 56 dogs living in greater Tunis area. Physical examination, quantitative serology and PCR on blood were used to characterize dogs’ status in relation to Leishmania infection and disease. IGRA consisted on testing by ELISA for IFN-γ-secretion in whole blood after a 20-h challenge with SLA. PBS and Phytohemagglutinin (PHA) stimulations were used as controls. Four groups of dogs were characterized: 31 were negative by both serology and PCR, two had doubtful serology, 10 presented no to mild clinical signs but low antibodies levels and 13 were affected by Canine Leishmaniasis (CanL). In seronegative dogs, IGRA was little contributory in 4 puppies (age <6 months) and 5 old dogs (median age = 72 months, IQR: 45–84 months) that didn’t respond to PHA stimulation, IGRA was negative in 19 and positive in three animals with lymph node enlargement. In dogs with doubtful serology, IGRA was positive in one dog and negative in the other. In infected dogs with no to mild clinical signs, one dog exhibited high level of IFN-γ in absence of antigenic stimulation and all the other were positive by IGRA. CanL dogs showed variable IGRA results. Negative IGRAs (n = 4) were shown in animals with the highest parasitic burden whereas positive IGRAs (n = 5) were shown in dogs with negative PCR or low parasitic load. The 4 remaining dogs either didn’t respond to PHA (n = 2) or showed non-specific secretion in PBS tube (n = 2). The results of this study showed that IGRA is a useful new tool that can assess exposure to Leishmania in dogs with no to mild clinical signs in endemic area. Further comparative investigations using assays exploring cellular immunity are needed to determine its accuracy. [ABSTRACT FROM AUTHOR]

Several endoparasites of dogs cannot only be detrimental to their primary host but might also represent a threat to human health because of their zoonotic potential. Due to their high dog population densities, metropolitan areas can be highly endemic for such parasites. We aimed to estimate the prevalence of endoparasites in dogs in the Austrian capital of Vienna by examining a representative number of canine faecal samples and to compare the prevalences with two neighbouring peri-urban and rural regions. In addition we analysed whether the density of dog populations and cleanliness of dog zones correlated with parasite occurrence. We collected 1001 anonymous faecal samples from 55 dog zones from all 23 districts of the federal state of Vienna, as well as 480 faecal samples from the Mödling district and Wolkersdorf with a peri-urban and rural character, respectively. Faeces were examined by flotation and by Baermann technique. Additionally we evaluated 292 Viennese, 102 peri-urban and 50 rural samples for Giardia and Cryptosporidium by Giardia FAST est ® and Crypto FAST est ® . Samples from “clean” dog zones were compared to samples from “dirty” zones. The infection rate of Toxocara was surprisingly low, ranging from 0.6% to 1.9%. Trichuris was the most frequent helminth (1.8–7.5%) and Giardia the most frequent protozoan (4.0–10.8%). Ancylostomatidae , Crenosoma, Capillaria, Taeniidae , Cystoisospora and Sarcocystis were found in 1.8–2.2%, 0–0.9%, 0–0.9%, 0–0.6%, 0.3–3.1% and 0–0.6% of the samples, respectively. Samples from “dirty” dog zones in Vienna showed a significantly higher rate of parasites overall (p = 0.003) and of Trichuris (p = 0.048) compared to samples from “clean” dog zones. There were no statistically significant differences in densely vs. less densely populated areas of Vienna. Samples from the rural region of Wolkersdorf had significantly higher overall parasite, Trichuris and Cystoisospora prevalences than the peri-urban Mödling district and Vienna (p = 0.000–0.039), while samples from the Mödling district had a significantly higher Giardia, Crenosoma and Capillaria prevalence than those from Vienna (p = 0.002–0.047). Parasite excretion is dynamic and representative sampling and monitoring are necessary for parasite surveillance. Dog owners should be informed about the zoonotic risk and encouraged to remove dog faeces and dispose of them properly to reduce the infection risk for both other dogs and humans. [ABSTRACT FROM AUTHOR]

Five diagnostic tests were compared for the diagnosis of Giardia duodenalis in fecal samples of young dogs. Fecal samples were collected from 136 healthy dogs <1 year old and examined using immunofluorescence antibody microscopy (IFA) after sucrose gradient centrifugation, zinc sulfate centrifugal flotation technique (ZSCT), SNAP ® Giardia test, and ProSpecT ® Giardia EZ Microplate assay. In addition, polymerase chain reaction (PCR) of the 16S rRNA gene was performed. Kappa (κ) statistic was calculated to assess diagnostic agreement between the IFA and each test. Using the IFA as the gold standard, the relative sensitivity and specificity of each test were determined. Subsequently, a Bayesian approach was used to estimate the sensitivity and specificity of each test in comparison to the IFA results. Giardia duodenalis was detected in 41% of the samples examined by IFA. The ZSCT resulted in 37% of positive samples, with a relative sensitivity and specificity of 86 and 98%, respectively. The SNAP ® Giardia test was positive in 40% of the samples, with a relative sensitivity and specificity of 91 and 96%, respectively. The ProSpecT ® test was positive in 51% of the samples, with a relative sensitivity and specificity of 100 and 83%, respectively. The relative sensitivity and specificity for PCR were 58 and 56%, respectively, with 55% of samples being PCR-positive. While the sensitivity and specificity estimates of each test in comparison to the IFA changed when using a Bayesian approach, the conclusions remained the same. While the ProSpecT ® test was the most sensitive test in this study, it is not designed for dogs and more costly than the other tests. The SNAP ® Giardia test performed similar to the ZSCT but may be more favorable because it is fast and easy to perform. Performance of the PCR was poor and the benefit of PCR may be in determining genotypes for evaluating zoonotic transfer between dogs and humans. [ABSTRACT FROM AUTHOR]

The rapid speed of kill of a spot-on, combination of fipronil-permethrin (Effitix ® , Virbac) was shown against infestations of Rhipicephalus sanguineus and Ctenocephalides felis on dogs. Efficacy was determined against new infestations at weekly intervals for one month after treatment. Dogs were allocated randomly to either an untreated control or to a single administration, given on Day 0, of either topical fipronil-permethrin (6.7–13.4 mg/kg and 60–120 mg/kg, respectively) or oral afoxolaner (2.72–6.8 mg/kg), based on pre-treatment, host-suitability flea counts. Dogs were infested with 50, unfed, adult R. sanguineus on Days 7, 14, 21 and 28, and with 100 C. felis on Days 8, 15, 22 and 29. Tick counts were performed 0.5, 2, 6, 12 and 24 h, and flea counts were performed 0.5 and 24 h after each infestation. No treatment-related adverse reactions occurred. Dogs in the untreated group maintained viable infestations throughout the study. Following infestation, live tick and flea counts for dogs treated with fipronil-permethrin compared with untreated dogs were rapidly and significantly reduced with efficacy apparent at 0.5 h after infestation. Flea efficacies (arithmetic mean counts) at 0.5 h after infestation on Day 7 (Day 28) were significantly greater for fipronil-permethrin, 70% (34%) compared with 8% (18%) for afoxolaner (P ≤ 0.05). Tick efficacies at 2 h on Day 7 (Day 28) were 74% (63%) for fipronil-permethrin compared with 10% (0%) for afoxolaner (P ≤ 0.05). Efficacies for tick repellency as indicated by counts of ticks off the dogs at 2 h on Day 7 (Day 28) were greater for fipronil-permethrin, 32% (22%) compared with afoxolaner, 0% (0%) (P ≤ 0.05). Anti-attachment efficacies at 12 h were greater for fipronil-permethrin compared with afoxolaner. Tick efficacies at 24 h, based on arithmetic (geometric) means, were significantly greater on Day 28 for fipronil-permethrin compared with afoxolaner (P ≤ 0.05), 74% (87%) and 45% (60%), respectively, and were similar (P > 0.05) on Days 7, 14 and 21. Flea efficacies, 24 h after infestation were >98% and similar for both treated groups on all infestation days (P > 0.05). The topically applied fipronil-permethrin containing ectoparasiticide Effitix ® offers rapid efficacy against R. sanguineus and C. felis which persists for one month after a single administration in dogs. Afoxolaner is also effective although speed of kill is slower. The rapid and sustained speed of kill of both parasites by fipronil-permethrin should contribute to effective management not only of these parasites and their direct adverse effects including irritancy and allergy, but also to reducing the risk of transmitting infections. [ABSTRACT FROM AUTHOR]

This survey was conducted in four Brazilian cities, from three federative units (Pernambuco, Minas Gerais, and Federal District), representing different socioeconomic settings, to determine the presence of antibodies to or antigens and DNA of selected pathogens in privately owned dogs. From January to April 2015, blood and serum samples were collected and assayed using different tests. In particular, antibodies to Anaplasma spp., Borrelia burgdorferi sensu lato , and Ehrlichia spp., and antigens of Dirofilaria immitis were detected using a rapid enzyme-linked immunosorbent assay, whereas antibodies to Babesia spp. were detected by an immunofluorescence antibody assay. Moreover, the presence of Leishmania DNA in blood samples was assessed by real-time polymerase chain reaction. Overall, 208 (69.3%) out of 300 dogs were positive for at least one tested pathogen (intended here as antibodies, antigen or DNA, as abovementioned), with 139 (66.8%) of them being positive to two or more pathogens. No dog presented antibodies to B. burgdorferi s.l., and D. immitis antigens were detected exclusively in dogs from Pernambuco. Among positive dogs, the most common clinical signs were lymphadenomegaly (45.2%), onychogryphosis (41.3%), dermatitis (34.1%), pale mucous membranes (19.7%), weight loss (14.9%), fever (12.5%), alopecia (11.1%), and lethargy (4.8%). Tick and flea infestations were recorded in 41.7% and 29.3% of the dogs, respectively, with 49 (16.3%) dogs being co-infested by both ticks and fleas. Most of the tick- and flea-infested dogs presented high level of infestation (>10 ticks and >20 fleas). The level of tick infestation varied significantly among federative units, being highest in Minas Gerais (68.0%), followed by Pernambuco (36.0%) and Federal District (21.0%). On the other hand, the level of flea infestation was higher in Pernambuco (50.0%), followed by Minas Gerais (29.0%) and Federal District (9.0%). The number of dog owners reporting the use of ectoparasiticides (on dogs and/or in the environment) varied significantly, ranging from 6.0% in Pernambuco to 60.0% in Federal District. This study reveals disparate results in terms of dog exposure to fleas and ticks in the studied communities, which may be related to factors such as the owners’ capability/willingness to afford the use of ectoparasiticides on their dogs and the dog’s level of restriction. Further research is needed to establish the relationship between dog owners’ socioeconomic situation and the level of exposure to ectoparasites and their transmitted pathogens. [ABSTRACT FROM AUTHOR]

During six years (April 2010–April 2016) we examined individual feces samples collected from 1314 dogs located in Center and Northwest Romania (Transylvania). Stool samples were analyzed by saturated salt flotation, sedimentation technique and modified Ziehl-Nielsen staining method. The overall prevalence of endoparasitic infections was 66.6% (n = 875). Sixteen species/genera of endoparasites were identified. The most prevalent species were Ancylostoma caninum/Uncinaria stenocephala (33.0%) ( p = 0.0001) followed by Trichocephalus vulpis (25.0%). Mixed infections, were significantly more frequent ( p = 0.0001) than single species infections. The age and the living condition/service of dogs were identified as the main risk factors for infection with endoparasites. [ABSTRACT FROM AUTHOR]

Echinococcosis and taeniasis are important helminth diseases that carry considerable impact on human and animal health. Domestic dogs and other canids are definitive hosts for several parasites of this group, including Echinococcus granulosus , Taenia multiceps , T. ovis , T. hydatigena and E. multilocularis. Detection of infection in dog populations is imperative for estimating the risk to susceptible humans and animals, and for its mitigation through prevention measures in dogs, other animals and humans. To date, identification of taeniid eggs, antigens or DNA in fecal samples are the most practical diagnostic modalities available for canine definitive hosts. Although widely used for this purpose, there is limited information comparing copro PCR and combined coproscopy-PCR protocols for the detection of taeniids. In the current study, a widely used multiplex PCR was performed on a large number of dog fecal samples using DNA extracted directly from feces. The samples were also tested by fecal flotation and coproscopy, eggs were isolated from microscopically-positive samples and extracted DNA was tested using the same multiplex PCR. The total number of taeniid positive samples detected using both methods was 46/317 (14.5%), including 10/317 (3.2%) E. granulosus positive samples. Both copro PCR and coproscopy have identified an equal number of samples as taeniid positive (n = 32). However, for the purpose of identification to species level, the copro PCR was significantly more sensitive than coproscopy followed by PCR on isolated eggs (sensitivity 0.7 vs. 0.41, p = 0.012), with 32/317 (10.1%) and 19/317 (6%) positive samples identified, respectively. The difference in identification of E. granulosus was highly apparent, as the majority of the E. granulosus positive samples (8/10) were detected by the copro PCR only. Coproscopy and egg PCR have identified 5/317 (1.6%) positive samples not detected by the copro PCR, including only a single sample (0.3%) positive for E. granulosus. Adding these positive samples to those identified by the copro PCR did not significantly improve the overall sensitivity (p = 0.074). Therefore, using both copro PCR and coproscopy in parallel may not be advantageous for taeniid detection and identification, at least until the egg PCR is further optimized and performs better. These results should be weighed against the different advantages that coproscopy based approach may offer. [Display omitted] • Copro PCR and coproscopy followed by PCR for the detection of Echinococcus and Taenia spp. in dog fecal samples were compared. • Both Copro PCR and coproscopy demonstrated sensitivity of 0.7 for initial screening of taeniid-positive samples. • The sensitivity of coproscopy followed by taeniid egg isolation and PCR was significantly lower at 0.41. • The copro PCR detected nine Echinococcus positive samples while the egg PCR detected only two. [ABSTRACT FROM AUTHOR]

In Morocco no data has been published on canine exposure to Anaplasma spp., Borrrelia burgdorferi, and Ehrlichia spp., and only one report is available on the occurrence of Dirofilaria immitis in dogs. Therefore, the aim of this study was to collect current data on the canine exposure to these vector-borne pathogens (VBPs) in Morocco. A total of 217 urban (n = 57), rural (n = 110) and military (n = 50) dogs from seven Moroccan locations were screened for Anaplasma spp., B. burgdorferi and Ehrlichia spp. antibodies and for D. immitis antigens using a commercial in-clinic ELISA test. Of these dogs, 182 (83.9%) tested positive for at least one pathogen and positivity to two or three pathogens was found in 14.3% and 2.3% of the dogs, respectively. Ehrlichia spp. antibodies (34.6%) were the most frequently detected followed by Anaplasma spp. antibodies (16.6%) and D. immitis antigens (16.1%). None of the dogs was tested seropositive to B. burgdorferi . Statistically significant differences in seropositivity rates were found for Ehrlichia spp. and D. immitis in rural dogs especially those from the north central region (p < 0.001) but not for Anaplasma spp. No significant difference was found according to the health status of the dog. This study demonstrates that Moroccan dogs are at high risk of acquiring a vector-borne infection. [ABSTRACT FROM AUTHOR]

Praziquantel (PZQ) is an anthelmintic drug used both in humans and animals that can be administered through various routes. There are transdermal formulations for cats, but only oral or subcutaneous dosage forms for dogs. Given the fact that the cat's skin and the dog's skin have different characteristics, which in turn affect bioavailability, we developed a PZQ spot-on formulation for dogs. This study was aimed at determining the plasmatic behavior of topically administered PZQ (Labyes ® ) in adult dogs. Dogs were administered PZQ (14.5 mg/kg PZQ, from a solution of 100 mg/ml). Blood samples were drawn before treatment onset and at the following time points after PZQ administration: 1, 2, 4, 6, 12, 24 and 48 h. PZQ plasma concentration was determined by ultra-high performance liquid chromatography (UPLC) coupled to tandem mass spectrometry (MS/MS). Observed maximum concentration (C max ), area under the concentration–time curve from the time of drug administration to infinity (AUC inf ) and time to maximum concentration (T max ) were calculated for each animal, and mean ± SD for each parameter was obtained. Results were as follows: C max = 56.0 ± 15 ng/ml; AUC inf = 910.2 ± 220 ng* h/ml, T max = 5.0 ± 1.1 h. This is the first study to provide pharmacokinetic data of a praziquantel spot-on formulation for dogs. [ABSTRACT FROM AUTHOR]

In the present study, Leishmania braziliensis enolase was cloned and the recombinant protein (rEnolase) was evaluated for the serodiagnosis of canine and human visceral leishmaniosis (VL). For the canine VL diagnosis, this study examined serum samples of Leishmania infantum -infected dogs, from non-infected animals living in endemic or non-endemic areas of leishmaniosis, as well as those from Leish-Tec ® -vaccinated dogs and Trypanosoma cruzi or Ehrlichia canis experimentally infected animals. For the human VL diagnosis, this study analyzed serum samples from VL patients, from non-infected subjects living in endemic or non-endemic areas of leishmaniosis, as well as those from T. cruzi -infected patients. In the results, an indirect ELISA method using rEnolase showed diagnostic sensitivity and specificity values of 100% and 98.57%, respectively, for canine VL serodiagnosis, and of 100% and 97.87%, respectively, for human VL diagnosis. These results showed rEnolase with an improved diagnostic performance when compared to the recombinant A2 protein, the crude soluble Leishmania antigenic preparation, and the recombinant K39-based immunochromatographic test. In conclusion, preliminary results suggest that the detection of antibodies against rEnolase improves the serodiagnosis of human and canine visceral leishmaniosis. [ABSTRACT FROM AUTHOR]

Hepatozoon parasites comprise intracellular apicomplexan parasites transmitted to vertebrate animals by ingestion of arthropods definitive hosts. The present work aimed to investigate the occurrence of Hepatozoon spp. in wild animals, domestic dogs and their respective ectoparasites, in southern Pantanal region, central-western Brazil, by molecular techniques. Between August 2013 and March 2015, 31 coatis ( Nasua nasua ), 78 crab-eating foxes ( Cerdocyon thous ), seven ocelots ( Leopardus pardalis ), 42 dogs ( Canis lupus familiaris ), 110 wild rodents (77 Thichomys fosteri, 25 Oecomys mamorae, and 8 Clyomys laticeps ), 30 marsupials (14 Thylamys macrurus, 11 Gracilinanus agilis, 4 Monodelphis domestica and 1 Didelphis albiventris ), and 1582 ticks and 80 fleas collected from the sampled animals were investigated. DNA samples were submitted to PCR assays for Hepatozoon spp. targeting 18S rRNA gene. Purified amplicons were directly sequenced and submitted to phylogenetic analysis. A high prevalence of Hepatozoon among carnivores ( C. thous [91.02%], dogs [45.23%], N. nasua [41.9%] and L. pardalis [71.4%]) was found. However, ticks and fleas were negative to Hepatozoon PCR assays. By phylogenetic analysis based on 18S rRNA sequences, Hepatozoon sequences amplified from crab-eating foxes, dogs, coatis and ocelots clustered with sequences of H. canis, H. americanum and H. felis. The closely related positioning of Hepatozoon sequences amplified from wild rodents and T. macrurus marsupial to Hepatozoon from reptiles and amphibians suggest a possible transmission of those Hepatozoon species between hosts by ectoparasites or by predation. Hepatozoon haplotypes found circulating in wild rodents seem to present a higher degree of polymorphism when compared to those found in other groups of animals. Although rodents seem not to participate as source of Hepatozoon infection to wild carnivores and domestic dogs, they may play an important role in the transmission of Hepatozoon to reptiles and amphibians in Pantanal biome. [ABSTRACT FROM AUTHOR]

The present study was aimed to determine the effect on liver, associated oxidative stress, trace element and vitamin alteration in dogs with sarcoptic mange. A total of 24 dogs with clinically established diagnosis of sarcoptic mange, divided into two groups, severely infested group (n = 9) and mild/moderately infested group (n = 15), according to the extent of skin lesions caused by sarcoptic mange and 6 dogs as control group were included in the present study. In comparison to healthy control hemoglobin, PCV, and TEC were significantly (P < 0.05) decreased in dogs with sarcoptic mange however, significant increase in TLC along with neutrophilia and lymphopenia was observed only in severely infested dogs. The albumin, glucose and cholesterol were significantly (P < 0.05) decreased and globulin, ALT, AST and bilirubin were significantly (P < 0.05) increased in severely infested dogs when compared to other two groups. Malondialdehyde (MDA) levels were significantly (P < 0.01) higher in dogs with sarcoptic mange, with levels highest in severely infested groups. Activity of superoxide dismutase (SOD) (P < 0.05) and catalase were significantly (P < 0.01) lower in sarcoptic infested dogs when compared with the healthy control group. Zinc and copper levels in dogs with sarcoptic mange were significantly (P < 0.05) lower when compared with healthy control group with the levels lowest in severely infested group. Vitamin A and vitamin C levels were significantly (P < 0.05) lower in sarcoptic infested dogs when compared to healthy control. From the present study, it was concluded that sarcoptic mange in dogs affects the liver and the infestation is associated with oxidant/anti-oxidant imbalance, significant alteration in trace elements and vitamins [ABSTRACT FROM AUTHOR]

Thelazia callipaeda (Spirurida, Thelaziidae) has been documented as agent of ocular infection in domestic animals (dogs and cats), wildlife (e.g., foxes, hares, rabbits), and humans. In the last two decades, this parasitosis has been increasingly reported in several European countries. Both adult and larval stages of the eyeworm are responsible for symptoms ranging from mild (e.g., lacrimation, ocular discharge, epiphora) to severe (e.g., conjunctivitis, keratitis, and corneal opacity or ulcers). The present study evaluated the clinical efficacy and safety of imidacloprid 10% and moxidectin 2.5% spot on (Advocate ® , Bayer Animal Health) in comparison to milbemycin oxime/praziquantel tablets (Milbemax ® , Novartis-Animal Health), as positive control, in the treatment of canine thelaziosis in naturally infected dogs and, a third group was used as an untreated control. Forty-seven dogs (27 females and 20 males) harbouring at least one live adult worm of T. callipaeda in one eye were enrolled from an endemic area of southern Italy. Each dog was then weighed and assigned in accordance with a random treatment allocation plan to one of the treatment groups (G1: imidacloprid 10% and moxidectin 2.5% spot on, G2: Untreated control and G3: milbemycin oxime/praziquantel tablets). On Day (D) 7, 14, 28 and 35 dogs were physically examined and the infection level was assessed by examination of both eyes, including conjunctival pouch and third eyelid for live adult T. callipaeda count and clinical scores. Dogs in G1 were treated on D0 and D28, whereas those in G3 on D0 and D7. Efficacy in G1 was 100% at each day post treatment (p < 0.01). For the G3 group efficacy was 57.39% on D7 (p < 0.05), 92.79% on D14 and 100% on D28 and D35 (p < 0.01). The application of the spot on formulation moxidectin 2.5% and imidacloprid 10% was highly effective in the treatment of canine thelaziosis caused by T. callipaeda . Advocate ® spot on can be recommended for the control of T. callipaeda infection, considering that this formulation is currently licensed in Europe for the treatment of a wide range of parasites affecting dogs. [ABSTRACT FROM AUTHOR]

The objective of the present study was to analyze the skin (nasal surface and ear regions), lymph nodes (popliteal and pre-scapular), spleen and liver of dogs with visceral leishmaniasis (VL), in order to investigate the relationship between the parasite load measured as DNA copy number of Alpha gene of DNA polymerase of Leishmania infantum by quantitative PCR and the number of M2 macrophages by immunohistochemistry. A set of 29 naturally infected dogs from an endemic area for VL were sampled and another set of six dogs negative for VL and from a non-endemic area were analyzed as the control group (C). The spleen presented the highest number of Leishmania DNA copies, with significant differences between the groups G1 and G2 (with and without skin lesions, respectively). The M2 phenotype immunostaining predominated among the macrophages in granulomas and inflammatory infiltrates of samples from the skin, lymph nodes and spleens examined. The presence of M2 macrophages in dogs from infected group differed significantly from the control group, in all organs analyzed, excepted liver. The highest proportion of M2 macrophages coincided with the highest parasitism loads found in more susceptible organs of VL dogs, even in the skin, considered a more resistant organ, while the liver showed low parasitism load and low immunostaining for M2 macrophages with no significant differences between infected and negative groups. It was concluded that the predominance of M2 phenotype in VL dogs favored the multiplication of Leishmania infantum in organs of dogs that are more susceptible to Leishmania infection, as skin, lymph nodes and spleen. [ABSTRACT FROM AUTHOR]

Main objective of the present nationwide study was to assess the impact of the ESCCAP guideline for the control of worm infections in dogs and cats 8–10 years after its first publication in Germany. A secondary aim was to determine the prevalence of canine and feline cardiopulmonary nematodes and intestinal protozoa. Faecal samples of 53,693 dogs and 26,491 cats in 2004–2006 as well as of 129,578 dogs and 45,709 cats in 2015–2017 routinely submitted by veterinarians to a private veterinary laboratory were examined using appropriate parasitological methods. In dogs, the prevalence of Toxocara and taeniid egg shedding was significantly lower in 2015–2017 (3.8 % and 0.16 %, respectively) than in 2004–2006 (4.6 % and 0.27 %, respectively). The prevalence of hookworm and Capillaria eggs was higher in the second study period (2.3 % and 0.77 %, respectively) than in the first (1.3 % and 0.6 %, respectively). For Toxascaris leonina (0.55–0.6 %) and Trichuris (0.8–0.9 %), the difference was not significant between the study periods. Dogs shed more often Angiostrongylus vasorum larvae in the second study (3.1 %) than in the first (1.0 %), whereas the prevalence of Crenosoma vulpis did not change significantly (2.2–2.6 %). Cystoisospora canis and C. ohioensis -like infections were less detected in the second study period (1.0 % and 2.1 %, respectively) than in the first (1.8 % and 2.7 %, respectively). Neospora -like oocysts and Sarcocystis sporocysts were more prevalent in the second study period (0.19 % and 0.13 %, respectively) than in the first (0.13 % and 0.06 %, respectively). The percentage of Giardia or Cryptosporidium coproantigen-positive samples was lower in the second study period (18.9 % and 6.7 %, respectively) than in the first (22.8 % and 10.0 %, respectively). In cats, the prevalence of egg shedding of T. cati , Capillaria and taeniids was significantly lower in 2015–2017 (3.5 %, 0.25 % and 0.1 %, respectively) than in 2004–2006 (4.8 %, 0.54 % and 0.22 %, respectively). No difference was recorded for hookworms (0.12–0.13 %) and Ts. leonina (0.04–0.05 %). Aelurostrongylus -like larvae were detected more often in the second study period (6.5 %) than in the first (2.6 %). Infections with Cystoisospora felis , C. rivolta , Toxoplasma -like coccids and Sarcocystis were less prevalent in the second study period (1.9 %, 0.7 %, 0.24 % and 0.02 %, respectively) than in the first (2.7 %, 1.1 %, 0.36 % and 0.1 %, respectively). The percentage of Giardia or Cryptosporidium coproantigen-positive samples was significantly lower in the second study period (10.6 % and 4.8 %, respectively) than in the first (15.4 % and 8.3 %, respectively). Although these results indicate a decline of the occurrence of most canine and feline intestinal parasites in Germany over the years, a transmission risk of zoonotic parasites remains. Therefore, the control of helminth infections in domestic dogs and cats continues to be a challenge for veterinarians and pet owners. • Toxocara eggs were the most common intestinal helminth stages found. • The prevalence of Toxocara and taeniid egg shedding was significantly higher in in the first study period. • The prevalence of Hookworms, Capillaria, Sarcocysti and Angiostrongylus was significantly higher in the second study period. [ABSTRACT FROM AUTHOR]

These revised guidelines have been developed to assist in the design, execution, and interpretation of studies to assess the efficacy of anthelmintic drugs against internal parasites in dogs and cats. The design and execution of studies are outlined and discussed. Considerations for specific targeted parasites are included. Information is provided on the principles of selection of animals, procedures for randomization, housing, feeding, necropsy procedures, and record keeping for dose determination dose confirmation studies and field studies. Complementary to the WAAVP general anthelmintic guidelines, these species-specific guidelines should assist investigators in the evaluation of anthelmintic drugs in dogs and cats by using comparable and standardized procedures in studies with appropriate numbers of animals. • Scientific guidance for evaluation of anthelmintic efficacy in cats and dogs. • Includes updated recommendations for specific parasites. • Efficacy evaluation mainly based on worm and faecal counts. • Revised guideline to ensure consistency over the different host animal species. [ABSTRACT FROM AUTHOR]

Angiostrongylus vasorum is a widely distributed cardiopulmonary parasite of canids in Europe. Clinical signs in dogs can be highly variable and diagnostically challenging. A correct and early diagnosis is hence indispensable to adequately manage affected patients. First-stage larvae (L1) are excreted in the faeces of definitive hosts and conventionally identified using the Baermann technique. Moreover, ELISAs for the detection of circulating antigen and specific antibodies have been presented. The current study aimed at i) quantitatively assessing larval migration in the Baermann funnel after 12 h and 24 h; ii) investigating the influence of sample storage at 4 °C over the course of three days on the number of detected L1; iii) evaluating potential associations of adult worm burdens with larval shedding in dogs and ELISA optical density (OD) values for circulating parasite antigen and specific antibodies. Faecal samples were obtained from naturally infected dogs (n = 21) and Baermann funnels were set up in duplicate over the course of four consecutive days (days 0–3) starting with the day of sample collection. Funnels were harvested on days 1–4 after 12 and 24 h, respectively, and the number of L1 per gram faeces (LPG) was determined. The LPG did not differ between larval harvest after 12 h from harvest after 24 h. Storage of faecal samples at 4 °C for two and three days entailed a considerable decrease in LPG. Adult worm burdens and larval excretion data from previous experiments demonstrated a correlation between worm burden and LPG. In contrast, no correlations between worm burden and the level of parasite antigen and specific antibody OD values, respectively, were identified. Thus, OD values of both antigen and antibody ELISA did not allow for conclusions on infection intensity reflected by the number of adult parasites. For the detection of L1 in faeces, 12 or 24 h of larval migration time was not discriminating for A. vasorum positivity. Thus, early processing of faecal samples is essential, since larval detection and hence sensitivity of the approach considerably decreased over the course of three days of storage. Therefore, the common recommendation to collect faecal samples for three consecutive days and to subsequently analyse them needs to be reconsidered. The results of this study can be readily translated into precise recommendations for daily practice to adequately assess A. vasorum infected dogs. [Display omitted] • Baermann technique: 12 vs. 24 of larval migration is not diagnostically relevant. • Early examination of faecal samples is crucial for the sensitivity of the method. • Larval excretion reflects adult worm burden in infected dogs. • Serological ELISA OD values do not reflect adult worm burden in infected dogs. [ABSTRACT FROM AUTHOR]

Interactions among different species of parasites co-infecting the same host could be synergistic or antagonistic. These interactions may modify both the frequency of infected hosts and their infectiousness, and therefore impact on transmission dynamics. This study determined the infectiousness of Trypanosoma cruzi -seropositive dogs (using xenodiagnosis) and their parasite load (quantified by qPCR), and tested the association between both variables and the presence of concomitant endoparasites. A cross-sectional serosurvey conducted in eight rural villages from Pampa del Indio and neighboring municipalities (northeastern Argentina) detected 32 T. cruzi -seropositive dogs out of 217 individuals examined for infection. Both the infectiousness to the vector Triatoma infestans and parasite load of T. cruzi -seropositive dogs examined were heterogeneous. A statistically significant, nine-fold higher mean infectiousness was registered in T. cruzi -seropositive dogs co-infected with Ancylostoma caninum and a trematode than in T. cruzi -seropositive dogs without these infections. The median parasite load of T. cruzi was also significantly higher in dogs co-infected with these helminths. An opposite trend was observed in T. cruzi -seropositive dogs that were serologically positive to Toxoplasma gondii or Neospora caninum relative to dogs seronegative for these parasites. Using multiple logistic regression analysis with random effects, we found a positive and significant association between the infectiousness of T. cruzi -seropositive dogs and co-infections with A. caninum and a trematode. Our results suggest that co-infections may be a modifier of host infectiousness in dogs naturally infected with T. cruzi . [ABSTRACT FROM AUTHOR]

Mycoplasma haemocanis (Mhc) and ‘ Candidatus Mycoplasma haematoparvum’ (CMhp) are canine haemoplasma species that can induce anaemia in immunocompromised and/or splenectomised dogs. This study aimed to determine the prevalence and phylogeny of canine haemoplasma species in dogs from Nigeria and describe any risk factors for infection. Canine haemoplasma species-specific and generic haemoplasma qPCR assays were used. The species-specific qPCR assays found Mhc infection in 18 of 245 dogs (7.3%), and CMhp infection in only one dog (0.4%). The generic haemoplasma qPCR assays were positive in 44 of 245 (17.9%) dogs. Twenty-five dogs had discordant qPCR results in that they were generic haemoplasma qPCR positive but species-specific qPCR negative. Further evaluation of these dogs by 16S rDNA sequencing gave limited results but 5 were confirmed to be infected with non-haemoplasma species: 2 Anaplasma phagocytophilum , 1 Anaplasma ovis , 1 Serratia marcescens and 1 Aerococcus spp. The 16S rRNA gene sequences from Mhc species showed > 99.8% identity with each other and > 99.6% identity with GenBank sequences, and resided in a single clade with other global Mhc and Mycoplasma haemofelis sequences, indicating low 16S rRNA genetic variability amongst this canine haemoplasma species. [ABSTRACT FROM AUTHOR]

Several parasites including the protozoa Blastocystis sp. and Cryptosporidium spp. may be causative agents of gastrointestinal symptoms in domestic dogs, and there may be a potential risk of transmission to owners. While France is one of the largest European countries in terms of its canine population, little data is available about the molecular epidemiology of these two parasites. The purpose of this study was to determine the prevalence of intestinal parasites in household dogs in France, and to evaluate the zoonotic risk of Blastocystis sp. and Cryptosporidium spp. by genotyping the corresponding isolates. To this end, 116 faecal samples were collected from household dogs regardless of breed, age or gender, living in the Lyons area, France. Various intestinal protozoa and helminths were identified by light microscopy. Screening for Blastocystis sp. and Cryptosporidium spp. were subsequently performed by PCR targeting the small subunit (SSU) rDNA coding region, followed by direct sequencing of the PCR products and analysis of the sequences obtained for genotyping. The overall prevalence of dogs infected with at least one gastrointestinal parasite was 42.2% (49/116). After light microscopy examination of faecal samples, the most common parasites found were the protozoa Giardia sp. (25.0%) and Cystoisospora sp. (19.8%). Using molecular methods, four dogs (3.4%) were shown to be infected by Blastocystis sp. and carried either subtype (ST) 2, commonly identified in various animal groups, or ST10, frequently found in bovids. Three dogs (2.6%) were positive for C. canis , infecting humans episodically. The low prevalence of both parasites, combined with the identification of C. canis and Blastocystis sp. ST2 and ST10 in the canine population, strongly suggests that dogs play a negligible role as zoonotic reservoirs for both parasites and do not seem to be natural hosts of Blastocystis sp. [ABSTRACT FROM AUTHOR]

The collection of 729 tick specimens ( Ixodes ricinus , 88.6%; Ixodes hexagonus , 9.2%; Dermacentor reticulatus, 2.2%) removed from 373 dogs and 78 cats, along with 201 ticks from vegetation ( I. ricinus , 75.6%; D. reticulatus, 24.4%), allows one to say that pets play an important role in maintaining tick life cycles in different urban area. It shows the lack of statistical differences between tick intensity in high-impact anthropogenic areas (HIAA), low-impact anthropogenic areas (LIAA) and mixed areas designed, in an objective way, by GIS techniques. The comparable (statistically insignificant) level of infection with Borrelia spp. of I. ricinus from pets (22.5%) and vegetation (24.8%), shows that dogs and cats do not have zooprophylactic competence for Borrelia spp. in different urban areas. Moreover, Borrelia spp. was detected in I. hexagonues (1.5%) collected from pets, and in D. reticulatus (2%) obtained from vegetation. The presence of D. reticulatus in the Wrocław Agglomeration confirms its expansion and the distribution range in Poland. [ABSTRACT FROM AUTHOR]

Diagnosis and detection of Echinococcus granulosus (sensu lato) infection in animals is a prerequisite for epidemiological studies and surveillance of echinococcosis in endemic, re-emergent or emergent transmission zones. Advances in diagnostic approaches for definitive hosts and livestock, however, have not progressed equally over the last 20 years. Development of laboratory based diagnostics for canids using coproantigen ELISA and also coproPCR, have had a huge impact on epidemiological studies and more recently on surveillance during hydatid control programmes. In contrast, diagnosis of cystic echinococcosis (CE) in livestock still relies largely on conventional post-mortem inspection, despite a relatively low diagnostic sensitivity especially in early infections, as current serodiagnostics do not provide a sufficiently specific and sensitive practical pre-mortem alternative. As a result, testing of dog faecal samples by coproantigen ELISA, often combined with mass ultrasound screening programmes for human CE, has been the preferred approach for monitoring and surveillance in resource-poor endemic areas and during control schemes. In this article we review the current options and approaches for diagnosis of E. granulosus infection in definitive and animal intermediate hosts (including applications in non-domesticated species) and make conclusions and recommendations for further improvements in diagnosis for use in epidemiological studies and surveillance schemes. [ABSTRACT FROM AUTHOR]

Over the last few decades, the world has witnessed radical changes in climate, landscape, and ecosystems. These events, together with other factors such as increasing illegal wildlife trade and changing human behaviour towards wildlife, are resulting into thinning boundaries between wild canids and felids and their domestic counterparts. As a consequence, the epidemiology of diseases caused by a number of infectious agents is undergoing profound readjustements, as pathogens adapt to new hosts and environments. Therefore, there is a risk for diseases of wildlife to spread to domestic carnivores and vice versa , and for zoonotic agents to emerge or re-emerge in human populations. Hence, the identification of the hazards arising from the co-habitation of these species is critical in order to plan and develop adequate control strategies against these pathogens. In the first of this two-part article, we review the role that wild canids and felids may play in the transmission of protozoa and arthropod-borne agents to dogs and cats in Europe, and provide an account of how current and future progress in our understanding of the ecology and epidemiology of parasites, as well as of host-parasite interactions, can assist efforts aimed at controlling parasite transmission. [ABSTRACT FROM AUTHOR]

An available enzyme-linked immunosorbent assay (ELISA) was studied for the detection of anti- Dioctophyma renale antibodies in the sera of dogs using, detection of parasite eggs in urine sediment as a reference test. ELISA uses a soluble antigenic preparation of esophagus of D. renale and the optimal dilutions of the antigen, serum and conjugate were determined by means of checker board titration, using positive ( n = 13) and negative ( n = 27) reference serum. The specificity and sensitivity of the ELISA were 93.8% and 92.3% respectively and the kappa index was good (0.76). These results suggest that ELISA described may prove to be an effective serological test for detecting dogs infected and exposed to this parasite mainly dogs that are not eliminating parasite eggs through their urine. [ABSTRACT FROM AUTHOR]

Two controlled clinical trials were carried out to assess the anti-feeding and adulticidal effects of a spot-on combining fipronil and permethrin (Effitix ® , Virbac, Carros, France) against Phlebotomus perniciosus in dogs. The first study (Exp. A) was a dose-determination study in which 3 doses of permethrin (30 mg/kg, 60 mg/kg and 120 mg/kg) were compared. The second study (Exp. B) was an efficacy study using commercial dose of permethrin contained in Effitix ® (the minimum dose of permethrin applied to dogs was 60 mg/kg). Twenty four and twelve Beagle dogs with equal sensitivity to sandflies were included in Exp. A and in Exp. B, respectively. Dogs were challenged with female sandflies (50 per dogs in Exp. A and 80 in Exp. B) for 60 ± 5 min on Days – 7, 1, 7, 14, 21 and 28 (Day 0 being treatment day). Counts and engorgement determination of dead and alive sandflies were performed after each exposure to treated and untreated dogs. Dead sandflies were also counted 24 h after exposure. In Exp. A, the repellency induced by an administration of 30 mg/kg of permethrin to dogs was above 91% for the first two weeks and then dropped to 82.2, 83.1 and 81.1% on Days 14, 21 and 28, respectively. For dogs receiving 60 mg/kg of permethrin, the repellency was a bit higher with 95.8, 97.6, 92.1, 91.4, and 86.8%, for Days 1, 7, 14, 21 and 28, respectively. The repellency induced by 120 mg/kg of permethrin was significantly higher than that induced by 60 mg/kg of permethrin on Day 14 only. In Exp. B the anti-feeding effect of the spot-on formulation was 94.1, 97.8, 96.3, 90.8 and 87% on Days 1, 7, 14, 21 and 28, respectively. The mortality effect was 98.9, 99.1, 99.8, 97.0 and 89.7% on Days 1, 7, 14, 21 and 28, respectively. At each challenge point, the mortality and anti-feeding effects on sandflies were significantly different between control and treatment group ( p < 0.05). The results indicate that a monthly administration of this new combination of permethrin and fipronil could be used as an effective sandfly control strategy in dogs and therefore recommended for use in an integrated leishmaniosis prevention program. [ABSTRACT FROM AUTHOR]