Molecular detection of Rickettsia felis in fleas and ticks collected from dogs and cats of Puebla, Mexico.

Fleas and ticks represent the two main groups of ectoparasites that infest companion animals. In particular, the flea Ctenocephalides felis felis and several members of the Rhipicephalus sanguineus complex are the main vectors of a wide range of pathogens on the American continent. They are competent vectors for several members of the genus Rickettsia, which encompass at least 15 pathogenic obligate intracellular bacteria that colonize the endothelial cells of vertebrates. In Mexico, 10 species of Rickettsia belonging to three groups have been detected in six species of ectoparasites from dogs in 9 of the 32 states of the country. However, in some larger regions of the country, active epidemiological surveillance has not been carried out. For this reason, the aim of this study was to identify the presence of members of the genus Rickettsia in fleas and ticks of dogs and cats in the state of Puebla, Mexico. A cross-sectional study was carried out to collect ectoparasites of dogs and cats during August to November 2019. Samples were fixed in 70% ethanol and examined to identify the presence of Rickettsia DNA by the amplification and sequencing of specific fragments of the gltA and ompB genes using conventional PCR. The recovered sequences were compared with those deposited in GenBank, and phylogenetic analyses were carried out to identify the position of the pathogens detected with respect to the valid species previously reported worldwide. Additionally, ecological parameters of the ectoparasite infestations were also calculated. We recovered 196 ectoparasites belonging to two species, 33 C. felis felis and 163 R. sanguineus s.l. (Rhipicephalus linnaei), parasitizing 46 hosts (42 dogs and 4 cats) in 11 localities of the state of Puebla. We detected the presence of Rickettsia felis in three pools of C. felis felis, and five from R. sanguineus s.l. Our work provides the first record of R. felis in hard ticks of Mexico and Central America, with new collection localities for this pathogen in central Mexico.

Factors associated with endoparasites and ectoparasites in domiciled dogs in the metropolitan area of Toluca, México / Factores asociados con la presencia de endoparásitos y ectoparásitos en perros domiciliados de la zona metropolitana de Toluca, México.

Introduction: Endoparasites and ectoparasites in dogs are of global distribution. The close relationship between dogs and man implies a risk for the transmission of zoonotic parasites. Therefore, it is necessary to determine the parasites hosted by dogs in specific areas and the factors associated with their presence. Objectives: To identify and to estimate the prevalence of endoparasites and ectoparasites in domiciled dogs in the Metropolitan area of Toluca, México, and the prevalence of D. caninum in fleas of the genus Ctenocephalides spp. Materials and methods: We collected samples from 402 domiciled dogs in four reference hospitals in the area in Toluca. We diagnosed endoparasites using direct smear, flotation, and sedimentation techniques and we performed the taxonomic identification of ectoparasites. Finally, the molecular diagnosis of D. caninum in fleas was made using the polymerase chain reaction technique (PCR). Results: A total of 37.2% of dogs were positive for endoparasites; the genera or species identified were Toxocara spp., Giardia spp., Ancylostoma spp., Cystoisospora spp., D. caninum, Taenia spp., and Trichuris vulpis; the prevalence of ectoparasites was 13.13%. We identified fleas of the species Ctenocephalides felis, Ctenocephalides canis; only one animal was parasitized with Rhipicephalus sanguineus and another one with Trichodectes canis; the prevalence of D. caninum in fleas was 9.5%. Conclusion: The prevalence of endoparasites was 37.2% while that of ectoparasites was 13.1%; this is the first analysis of endoparasites and ectoparasites conducted in the same population of dogs in México together with the molecular diagnosis of D. caninum in fleas.

Introducción. Los endoparásitos y ectoparásitos en perros son de distribución mundial. La estrecha relación entre los perros y el hombre implica un riesgo de transmisión de parasitosis zoonóticas, por lo cual es necesario conocer las especies que parasitan a los perros de esta zona y determinar los factores asociados. Objetivos. Estimar la prevalencia de endoparásitos y ectoparásitos, identificarlos en perros domiciliados de la zona metropolitana de Toluca, México, y determinar la prevalencia de Dipyilidium caninum en pulgas del género Ctenocephalides spp. Materiales y métodos. Se recolectaron muestras de 402 perros que fueron llevados a consulta en cuatro hospitales de referencia de Toluca. En el diagnóstico de endoparásitos, se utilizaron las técnicas coproparasitoscópicas de frotis directo, flotación y sedimentación; además, se recolectaron ectoparásitos para su identificación taxonómica. Por último, la detección de D. caninum en pulgas se hizo mediante la reacción en cadena de la polimerasa (PCR). Resultados. El 37,2 % de los perros resultó positivo para endoparásitos. Los géneros o especies identificados fueron Toxocara spp., Giardia spp., Ancylostoma spp., Cystoisospora spp., D. caninum, Taenia spp. y Trichuris vulpis. Se determinó una prevalencia de ectoparásitos de 13,13 %. Se identificaron pulgas de las especies Ctenocephalides felis y C. canis, en tanto que solo un animal presentó parasitosis por Rhipicephalus sanguineus y otro por Trichodectes canis. La prevalencia de D. caninum en pulgas fue del 9,5 %. Conclusión. La prevalencia de endoparásitos fue de 37,2 % y, la de ectoparásitos, de 13,1 %. Por primera vez en México se hizo un análisis de endoparásitos y ectoparásitos en una misma población de perros, así como el diagnóstico molecular de D. caninum.

Factores asociados con la presencia de endoparásitos y ectoparásitos en perros domiciliados de la zona metropolitana de Toluca, México / Factors associated with endoparasites and ectoparasites in domiciled dogs in the metropolitan area of Toluca, México

Resumen | Introducción. Los endoparásitos y ectoparásitos en perros son de distribución mundial. La estrecha relación entre los perros y el hombre implica un riesgo de transmisión de parasitosis zoonóticas, por lo cual es necesario conocer las especies que parasitan a los perros de esta zona y determinar los factores asociados. Objetivos. Estimar la prevalencia de endoparásitos y ectoparásitos, identificarlos en perros domiciliados de la zona metropolitana de Toluca, México, y determinar la prevalencia de Dipyilidium caninum en pulgas del género Ctenocephalides spp. Materiales y métodos. Se recolectaron muestras de 402 perros que fueron llevados a consulta en cuatro hospitales de referencia de Toluca. En el diagnóstico de endoparásitos, se utilizaron las técnicas coproparasitoscópicas de frotis directo, flotación y sedimentación; además, se recolectaron ectoparásitos para su identificación taxonómica. Por último, la detección de D. caninum en pulgas se hizo mediante la reacción en cadena de la polimerasa (PCR). Resultados. El 37,2 % de los perros resultó positivo para endoparásitos. Los géneros o especies identificados fueron Toxocara spp., Giardia spp., Ancylostoma spp., Cystoisospora spp., D. caninum, Taenia spp. y Trichuris vulpis. Se determinó una prevalencia de ectoparásitos de 13,13 %. Se identificaron pulgas de las especies Ctenocephalides felis y C. canis, en tanto que solo un animal presentó parasitosis por Rhipicephalus sanguineus y otro por Trichodectes canis. La prevalencia de D. caninum en pulgas fue del 9,5 %. Conclusión. La prevalencia de endoparásitos fue de 37,2 % y, la de ectoparásitos, de 13,1 %. Por primera vez en México se hizo un análisis de endoparásitos y ectoparásitos en una misma población de perros, así como el diagnóstico molecular de D. caninum.

Abstract | Introduction: Endoparasites and ectoparasites in dogs are of global distribution. The close relationship between dogs and man implies a risk for the transmission of zoonotic parasites. Therefore, it is necessary to determine the parasites hosted by dogs in specific areas and the factors associated with their presence. Objectives: To identify and to estimate the prevalence of endoparasites and ectoparasites in domiciled dogs in the Metropolitan area of Toluca, México, and the prevalence of D. caninum in fleas of the genus Ctenocephalides spp. Materials and methods: We collected samples from 402 domiciled dogs in four reference hospitals in the area in Toluca. We diagnosed endoparasites using direct smear, flotation, and sedimentation techniques and we performed the taxonomic identification of ectoparasites. Finally, the molecular diagnosis of D. caninum in fleas was made using the polymerase chain reaction technique (PCR). Results: A total of 37.2% of dogs were positive for endoparasites; the genera or species identified were Toxocara spp., Giardia spp., Ancylostoma spp., Cystoisospora spp., D. caninum, Taenia spp., and Trichuris vulpis; the prevalence of ectoparasites was 13.13%. We identified fleas of the species Ctenocephalides felis, Ctenocephalides canis; only one animal was parasitized with Rhipicephalus sanguineus and another one with Trichodectes canis; the prevalence of D. caninum in fleas was 9.5%. Conclusion: The prevalence of endoparasites was 37.2% while that of ectoparasites was 13.1%; this is the first analysis of endoparasites and ectoparasites conducted in the same population of dogs in México together with the molecular diagnosis of D. caninum in fleas.

Molecular Survey and Genetic Diversity of Bartonella spp. in Small Indian Mongooses (Urva auropunctata) and Their Fleas on Saint Kitts, West Indies.

This study aimed to molecularly survey and evaluate the genetic diversity of Bartonella spp. in mongooses and their fleas from St. Kitts. Spleen (n = 54), blood (n = 71), and pooled flea samples, all identified as Ctenocephalides felis (n = 53), were submitted to TaqMan real-time quantitative PCR (qPCR) targeting Bartonella-nuoG fragment (84 bp). Positive samples underwent further conventional PCR assays targeting five loci (gltA, rpoB, fstZ, nuoG, and ITS), subsequent sequencing, and phylogenetic and haplotype analyses. The overall occurrence of Bartonella spp. in mongooses and fleas was 51.2% (64/125 [95% CI (42.1-60.2%)]) and 62.3% (33/53) [95% CI (47.9-75.2%)]), respectively. From samples sequenced across the five loci, 50.8% (33/65) were identified as Bartonella henselae, 26.2% (17/65) were 96.74-99.01% similar by BLAST analysis to an unidentified Bartonella sp. previously reported in Japanese badgers (Meles anakuma), and 23.1% (15/65) were co-infected with both species. Nucleotide polymorphism analysis showed low diversity amongst haplotypes but did concur with phylogenetic analysis, placing the unidentified species in a separate clade from B. henselae by multiple mutational events. Our data confirms that mongooses and Ctenocephalides felis fleas collected from them are not only potential reservoirs for B. henselae but also a novel Bartonella sp. which we propose be called 'Candidatus Bartonella kittensis'.

Urban foci of murine typhus involving cat fleas (Ctenocephalides felis felis) collected from opossums in Mexico City.

Murine typhus, a neglected rickettsiosis caused by Rickettsia typhi, is a common disease in several Latin-American countries. The sylvatic life cycle of R. typhi encompasses the presence of several wild mammals, particularly opossums of the genus Didelphis and their associated fleas. Due to the colonization of wild environments by human populations, the increase in contact with opossum fleas has generated the presence of urban outbreaks of typhus. For this reason, the aim of our study was to identify the presence and diversity of Rickettsia sp. in fleas collected from opossums of an urban reserve in Mexico City. Opossums were captured from February to September 2017. For the detection of Rickettsia DNA, fragments of 800 bp of the citrate synthase (gltA) and the outer membrane protein B (ompB) were amplified. A total of 141 fleas (111 ♀, 30 ♂) of a single species (Ctenocephalides felis felis) were recovered from 31 Didelphis virginiana. Rickettsia DNA was detected in 17.7% (25/141) of the analysed fleas, recovered from seven infested opossums. The Maximum likelihood of sequences exhibited an identity of 99%-100% with sequences of R. typhi from southern United States. This work represents the first record of R. typhi in fleas from opossums in Mexico.

Does bathing affect tick and flea burdens and ectoparasiticide effectiveness of a spot-on formulation (fipronil + (S)-methoprene) for dogs?

This work evaluated tick and flea burdens and the efficacy of a single topical application of 10% fipronil + 9% (S)-methoprene spot-on against experimental infestations of Rhipicephalus sanguineus sensu lato and Ctenocephalides felis felis on dogs that were submitted to regular baths for 56-days post-treatment. Four treatments (n = 24) were evaluated: no chemical treatment and no bathing (T01); no chemical treatment + bathing (T02); chemical treatment and no bathing (T03) and chemical treatment + bathing (T04). Dogs were infested with adult ticks and fleas seven days pre-treatment (-7). The ectoparasites were then counted and removed on day -5, followed by a new infestation on day -2 and treatment on day 0. Ticks and fleas were then counted without removal on day 1 and counted with removal on day 2. The dogs were then repeatedly infested with both ectoparasites on days 5, 12, 19, 26, 33, 40, 47 and 54 post-treatment. Baths and ectoparasite counts with removal were performed every 48 h before and after experimental infestation, respectively. Animals of T04 had lower (P ≤ 0.05) tick burden than animals from T01 and T02 from day 2 to day 28. For fleas, the counts varied among days of the study. Dogs from T03 presented lower (P ≤ 0.05) flea burden than dogs from T01 from day 1 until day 56. The same was true for T04 when compared to T01 from day 1 to day 42. Animals of T02 greater (P ≤ 0.05) flea burden than animals of T04 in all dates post treatment. In addition, dogs from T02 presented higher (P ≤ 0.05) flea burden than T01 from day 7 to day 28 and from day 49 to day 56. In sum, the efficacy of the spot-on formulation used declined over the time against both ectoparasites. The efficacy for ticks ranged from 0.0 to 96.5% and 0.0 to 98.52% for T03 and T04, respectively. While for fleas, the efficacy of the formulation ranged from 67.41 to 100% for T03 and 40.18 to 100% for T04. So, bathing dogs that were not treated increased C. f. felis burden but not R. sanguineus s. l. burden. The residual efficacy of the product had a shorter duration against these ectoparasites for dogs that received subsequent bathing.

Bartonella in dogs and fleas from Tulancingo, Hidalgo, Mexico.

Bartonella sp. infection is quite common in free-roaming dogs in many tropical countries. However, limited information is available of the presence of these pathogens in Mexico. The present study looked at prevalence of Bartonella exposure and/or infection in dogs and their fleas in Central Mexico. Blood samples were collected from 31 stray dogs in August 2014 at the municipal pound, Tulancingo, Mexico, as well as fleas on 26 of them. Bartonella seropositivity was 46.9%, including 35.5% for Bartonella henselae, 45% for Bartonella clarridgeiae and 32.2% for Bartonella vinsonii subsp. berkhoffii. Three (9.7%) dogs were polymerase chain reaction (PCR) positive for the Bartonella gltA gene. Partial sequencing of that gene revealed that these three dogs were infected with B. henselae. In total, 86 fleas were collected from 26 dogs (range 1-9 fleas per dog), including 52 Ctenocephalides felis and 34 Ctenocephalides canis. Of 40 pools of fleas (20 pools of C. canis and 20 pools of C. felis), five (12.5%) were PCR positive for the Bartonella sp. gltA gene, including three C. canis pools (five fleas) and two C. felis pools (three fleas). All sequences showed 99.25% to 100% homology with B. henselae Houston I.

Candidatus Rickettsia senegalensis in Cat Fleas (Siphonaptera: Pulicidae) Collected from Dogs and Cats in Cauca, Colombia.

Rickettsia typhi and Rickettsia felis (Rickettsiales: Rickettsiaceae) are flea-transmitted pathogens. They are important causes of acute febrile illness throughout the world. We, therefore, sought to identify the rickettsial species present in the fleas of dogs and cats in the department of Cauca, Colombia. In this study, we collected 1,242 fleas from 132 dogs and 43 fleas from 11 cats. All fleas were morphologically identified as Ctenocephalides felis (Bouché) adults and organized in pools for DNA extraction (234 pools from dogs and 11 from cats). The gltA gene from rickettsiae was targeted for screening amplification using conventional PCR. In total, 144 of the 245 pools (58.7%) were positive. The positive samples were then processed for the amplification of the 17kDa antigen gene (144/144; 100% positive) and sca5 gene (140/144; 97.2% positive). In addition, restriction enzyme length polymorphism analysis using NlaIV on the amplified product of the sca5 gene demonstrated several organisms: 21/140 (15%) were R. felis, 118/140 (84.3%) were Rickettsia asemboensis, and 1/140 (0.7%) were Candidatus Rickettsia senegalensis. Subsequent sequencing confirmed Candidatus Rickettsia senegalensis in C. felis collected from dogs the first reported from Colombia.

Efficacy of afoxolaner in the flea control in experimentally infested cats / Eficácia do afoxolaner no controle de pulgas em gatos experimentalmente infestados

Abstract The aim of this study was to evaluate the efficacy of a single dose of oral afoxolaner in controlling fleas in cats. Fourteen cats were used. The cats were given identification numbers, housed individually, artificially infested with Ctenocephalides felis felis, and treated (or not) with afoxolaner. Were divided into a treatment group and a control group (n = 7/group), on the basis of the fleas count hours after an infestation applied on Day (one-by-one allocation after ordering by count). At the start of the experimental protocol (designated day 0), the treated group received afoxolaner in a single dose of 2.5 mg/kg and the control group animals received a placebo. All animals were infested with 100 C. felis felis fleas two days before day 0, as well as on days 5, 12, 19, 26, 33, 40, 47, 54, and 63, parasite loads being evaluated at 48 h after each infestation. The efficacy of afoxolaner was 100% on day 2 and remained above 98% until day 42, decreasing to 95.3% by day 63. The findings confirm that a single dose of oral afoxolaner was effective in controlling C. felis felis in cats, and there were no observed adverse events.

Resumo O objetivo do estudo foi avaliar a eficácia de uma dose única de afoxolaner oral no controle de pulgas em gatos. Foram utilizados 14 gatos. Os animais foram identificados, alojados individualmente, infestados artificialmente com C. felis felis e tratados (ou não) com afoxolaner. Foram divididos em um grupo de tratamento e um grupo controle (n = 7/ grupo), com base na contagem de pulgas, horas após a infestação aplicada no dia (alocação de um por um após o período por contagem). No início do protocolo experimental (dia 0), o grupo tratado recebeu afoxolaner em dose inicial de 2,5 mg / kg e os animais do grupo controle receberam um placebo. Todos os animais foram infestados com 100 pulgas C. felis felis dois dias antes do dia 0, assim como nos dias 5, 12, 19, 26, 33, 40, 47, 54 e 63, sendo avaliadas as cargas parasitárias às 48 h após cada infestação. A eficácia do afoxolaner foi de 100% no dia 2 e permaneceu acima de 98% até o dia 42, diminuindo para 95,3% no dia 63. Os resultados confirmam que uma dose única de afoxolaner oral foi eficaz no controle de C. felis felis em gatos, e não houve eventos adversos observados.

Controle de pulgas em cervo-do-pantanal (Blastocerus dichotomus) de cativeiro com uso de coleira com imidacloprida e flumetrina / Control of fleas in captive marsh deer (Blastocerus dichotomus) with imidacloprid and flumetrin collar / Control de pulgas en ciervos de los pantanos (Blastocerus dichotomus) cautivo con uso de collar con imidacloprida e flumetrina

O cervo-do-pantanal (Blastocerus dichotomus) é o maior cervídeo do continente sul-americano e figura na lista de animais ameaçados de extinção. Dentre os principais problemas dessa espécie em ambiente artificial está a puliciose. Essa infestação pode causar anemia expressiva, inapetência e óbito. O controle desses parasitos é realizado de diversas formas, com combate das pulgas nos animais e no ambiente, sendo que o controle conjunto é o mais eficaz. No entanto, o controle das pulgas em cervídeos é bastante restrito, tanto pelo temperamento dos animais, quanto pela inexistência de um produto eficaz e de longa ação. Esse trabalho testou uma coleira à base de imidacloprida e flumetrina, preconizada para uso em cães domésticos, por ser de aplicação única e de longa duração. A coleira se mostrou eficiente por seis meses na eliminação das pulgas, associada à aspersão de deltametrina no recinto; sem ocorrência de efeitos adversos.(AU)

The marsh deer (Blastocerus dichotomus) is the largest deer in the South American continent and is listed in the red list of endangered animals. Among the main problems of this species “ex situ” we have puliciosis. This infestation can cause significant anemia, weariness and even death. The control of these parasites is carried out in several ways, with flea combat in the hosts and in the environment, and joint control is the most effective method. However, the control of fleas in animals is quite restricted, both by the behavior of the animal, and by the nonexistence of an effective and long-acting anti parasitic product. This work tested a collar impregnated with imidacloprid and flumethrin recommended for use in domestic dogs, because it is of single application and long duration. The collar showed to be efficient for six months, in the elimination of fleas, associated with spraying of deltamethrin in the enclosure; without occurrence of adverse effects.(AU)

El ciervo de los pantanos (Blastocerus dichotomus), es el ciervo más grande del continente sudamericano. Entre los principales problemas dela especie "ex situ" está la puliciosis. Esta infestación puede causar anemia significativa, debilidad, pérdida de apetito y incluso la muerte. El control de los parásitos se lleva de varias maneras, con el combate de las pulgas en el huésped y el medio ambiente, en el que el conjunto de control es más eficaz. Sin embargo, el control de pulgas en animales salvajes es muy restringido, por el comportamiento agresivo del animal, como la ausencia de un producto eficaz y de acción prolongada. Este estudio probó un collar impregnado con imidacloprida y flumetrina, com uso recomiendado en perros domésticos, en una sola aplicación y con largo plazo. El producto ha demostrado eficaz durante seis meses en la eliminacion de pulgas, asociado con la pulverizacion de deltametrina en la habitación; sin efectos adversos en los animales salvajes.(AU)

Infection by Rickettsia felis in Ctenocephalides felis felis Fleas from North of Colombia.

BACKGROUND: Rickettsia felis is an emergent Rickettsial agent whose main vector is Ctenocephalides felis, but ticks, mites and lice are also infected. We aimed to search for molecular evidence of Rickettsia spp. in fleas collected from dogs and wild rodents (Heteromys anomalous) from three villages of Córdoba and Antioquia provinces (Northern of Colombia), where outbreaks of rickettsioses have occurred, and discuss the possible role of fleas on endemic/enzootic regions for rickettsia. METHODS: During 2010 and 2012, 649 Ctenocephalides felis felis and 24 Pulex irritans fleas were removed from dogs and wild rodents (Heteromys anomalous), respectively, in 3 locations from Córdoba and Antioquia provinces (Colombia). These fleas were tested into pools for Rickettsial infection by PCR, targeting gltA, ompB, and ompA Rickettsial genes. RESULTS: Almost 20% (30/153) of C. felis felis pools contained Rickettsial DNA. The fragments of ompB gene showed high identity values between sequences from Necocli and Los Cordobas with R. felis strain from Senegal (100% and 99.7% respectively) and all were highly related by phylogenetic analyses. Rickettsial DNA in pools of P. irritans was not detected. CONCLUSION: Our findings highlighted the endemicity of the infection by R. felis in fleas from northern of Colombia and showed the likely importance of dogs as hosts of C. felis felis fleas and their potential role as reservoirs of R. felis.

Detection of Rickettsia spp. and Bartonella spp. in Ctenocephalides felis fleas from free-ranging crab-eating foxes (Cerdocyon thous).

Fleas are insects with a worldwide distribution that have been implicated in the transmission of several pathogens. The present study aimed to investigate the presence of Rickettsia spp. (Rickettsiales: Rickettsiaceae) and Bartonella spp. (Rhizobiales: Bartonellaceae) in fleas from free-ranging crab-eating foxes Cerdocyon thous (Linnaeus, 1766) (Carnivora: Canidae) from Rio Grande do Sul, southern Brazil. Fleas were collected manually from animals and used for the molecular detection of Rickettsia spp. and Bartonella spp. Twenty-nine C. thous were sampled in six municipalities. Four foxes were parasitized by 10 fleas, all of which were identified as Ctenocephalides felis (Bouché, 1935) (Siphonaptera: Pulicidae). DNA from Rickettsia felis Bouyer et al., 2001 and Rickettsia asembonensis Maina et al., 2016 were found in three and eight fleas, respectively. In four fleas, DNA of Bartonella sp. was identified. Phylogenetic analysis grouped Bartonella sp. together with other genotypes previously reported in C. felis worldwide. The scenario described in the present study highlights a Neotropical canid parasitized by the invasive cosmopolitan cat flea, which in turn, is carrying potentially invasive vector-borne microorganisms. These findings suggest that C. felis is adapted to wild hosts in wilderness areas in southern Brazil, hypothetically exposing the Neotropical fauna to unknown ecological and health disturbances.

Cellular and Humoral Responses to Cte f 2, a Cat Flea Allergen, in Children with Papular Urticaria.

BACKGROUND: Papular urticaria (PU) is a common insect bite skin hypersensitivity in tropical countries. In order to gain insight into its causal allergens, we aimed to evaluate cellular and humoral immune responses to the recombinant salivary antigen Cte f 2 from the cat flea Ctenocephalides felis. METHOD: Sixty patients with PU and 27 healthy controls were included in this study. Specific IgE, IgG, IgG1, and IgG4 against Cte f 2 and C. felis extract were determined by ELISA. The T-cell response was analyzed using a carboxyfluorescein succinimidyl ester (CFSE)-based dilution assay and Th1/Th2/Th17 cytokine measurements. In addition, a proteomic analysis of IgG and IgE reactive spots of C. felis extract was performed. RESULTS: The frequency of IgE sensitization to Cte f 2 was similar between patients (36.7%) and controls (40.7%). The specific IgE, IgG1, and IgG4 responses to Cte f 2 and C. felis extract were not significantly different between patients and controls. Among the 3 conditions (i.e., Cte f 2, C. felis extract, and only medium) Cte f 2 was the strongest inducer of CD3+CD4+ proliferation in the patients; however, the mean response was not significantly different from those in controls (Cte f 2: 4.5 vs. 2.5%; p = 0.46). No salivary proteins were identified in C. felis, and most of the spots were identified as muscle-skeletal components (tropomyosin, actin, myosin, and ankirin). CONCLUSIONS: Cte f 2 induces IgE and IgG production as well as T-cell proliferation in children living in a geographical area where PU induced by a flea bite is common. The use of C. felis extract is not recommended for the study of bite-induced hypersensitivity disease since salivary antigens are not well represented.

Controle de pulgas em cervo-do-pantanal (Blastocerus dichotomus) de cativeiro com uso de coleira com imidacloprida e flumetrina / Control of fleas in captive marsh deer (Blastocerus dichotomus) with imidacloprid and flumetrin collar / Control de pulgas en ciervos de los pantanos (Blastocerus dichotomus) cautivo con uso de collar con imidacloprida e flumetrina

O cervo-do-pantanal (Blastocerus dichotomus) é o maior cervídeo do continente sul-americano e figura na lista de animais ameaçados de extinção. Dentre os principais problemas dessa espécie em ambiente artificial está a puliciose. Essa infestação pode causar anemia expressiva, inapetência e óbito. O controle desses parasitos é realizado de diversas formas, com combate das pulgas nos animais e no ambiente, sendo que o controle conjunto é o mais eficaz. No entanto, o controle das pulgas em cervídeos é bastante restrito, tanto pelo temperamento dos animais, quanto pela inexistência de um produto eficaz e de longa ação. Esse trabalho testou uma coleira à base de imidacloprida e flumetrina, preconizada para uso em cães domésticos, por ser de aplicação única e de longa duração. A coleira se mostrou eficiente por seis meses na eliminação das pulgas, associada à aspersão de deltametrina no recinto; sem ocorrência de efeitos adversos.

The marsh deer (Blastocerus dichotomus) is the largest deer in the South American continent and is listed in the red list of endangered animals. Among the main problems of this species “ex situ” we have puliciosis. This infestation can cause significant anemia, weariness and even death. The control of these parasites is carried out in several ways, with flea combat in the hosts and in the environment, and joint control is the most effective method. However, the control of fleas in animals is quite restricted, both by the behavior of the animal, and by the nonexistence of an effective and long-acting anti parasitic product. This work tested a collar impregnated with imidacloprid and flumethrin recommended for use in domestic dogs, because it is of single application and long duration. The collar showed to be efficient for six months, in the elimination of fleas, associated with spraying of deltamethrin in the enclosure; without occurrence of adverse effects.

El ciervo de los pantanos (Blastocerus dichotomus), es el ciervo más grande del continente sudamericano. Entre los principales problemas dela especie "ex situ" está la puliciosis. Esta infestación puede causar anemia significativa, debilidad, pérdida de apetito y incluso la muerte. El control de los parásitos se lleva de varias maneras, con el combate de las pulgas en el huésped y el medio ambiente, en el que el conjunto de control es más eficaz. Sin embargo, el control de pulgas en animales salvajes es muy restringido, por el comportamiento agresivo del animal, como la ausencia de un producto eficaz y de acción prolongada. Este estudio probó un collar impregnado con imidacloprida y flumetrina, com uso recomiendado en perros domésticos, en una sola aplicación y con largo plazo. El producto ha demostrado eficaz durante seis meses en la eliminacion de pulgas, asociado con la pulverizacion de deltametrina en la habitación; sin efectos adversos en los animales salvajes.

Control de pulgas en ciervos de los pantanos (Blastocerus dichotomus) cautivo con uso de collar con imidacloprida e flumetrina / Control of fleas in captive marsh deer (Blastocerus dichotomus) with imidacloprid and flumetrin collar / Controle de pulgas em cervo-do-pantanal (Blastocerus dichotomus) de cativeiro com uso de coleira com imidacloprida e flumetrina

El ciervo de los pantanos (Blastocerus dichotomus), es el ciervo más grande del continente sudamericano y figura en la lista de animales en peligro de extinción, de acuerdo con el IBAMA y la UICN. Tiene una población bajo el cuidado humano muy limitado, estimada en un centenar de individuos. Entre los principales problemas de la especie "ex situ" está la puliciose. Esta infestación puede causar anemia significativa, debilidad, pérdida de apetito y incluso la muerte. El control de los parásitos se lleva de varias maneras, con el combate de las pulgas en el huésped y el medio ambiente, en el que el conjunto de control es más eficaz. Sin embargo, el control de pulgas en animales salvajes es muy restringido, por el comportamiento agresivo del animal, como la ausencia de un producto eficaz y de acción prolongada. Este estudio probó un collar impregnado con imidacloprid y flumetrina, com uso recomiendado en perros domésticos, em una sola aplicación y con largo plazo. El producto ha demostrado muy eficaz durante dieciocho meses en la eliminacion de pulgas, asociado con la pulverizacion de deltametrina en la habitación; sin efectos adversos em los animales salvajes.

The marsh deer (Blastocerus dichotomus) is the largest deer in the South American continent and is listed in the red list of endangered animals according to IBAMA and IUCN. It has a very small population in captivity, estimated at 100 individuals. Among the main problems of this species ex situ we have puliciosis. This infestation can cause significant anemia, weariness and even death. The control of these parasites is carried out in several ways, with flea combat in the hosts and in the environment, and joint control is the most effective method. However, the control of fleas in animals is quite restricted, both by the behavior of the animal, and by the nonexistence of an effective and long-acting anti parasitic product. This work tested a collar impregnated with imidacloprid and flumethrin, recommended for use in domestic dogs, because it is of single application and long duration. The collar showed to be very efficient for eighteen months, in the elimination of fleas, associated with spraying of deltamethrin in the enclosure; without occurrence of adverse effects.

O cervo-do-pantanal (Blastocerus dichotomus), é o maior cervídeo do continente sul-americano e figura na lista de animais ameaçados de extinção, segundo o IBAMA e a IUCN. A espécie tem uma população muito reduzida ex-situ, estimada em cem indivíduos. Dentre os principais problemas dessa espécie em cativeiro temos a puliciose. Essa infestação pode causar anemia expressiva, inapetência e óbito. O controle desses parasitas é realizado de diversas formas, com combate das pulgas nos animais e no ambiente, sendo que o controle conjunto é o mais eficaz. No entanto, o controle das pulgas nos animais é bastante restrito, tanto pelo temperamento do animal, quanto pela inexistência de um produto eficaz e de longa ação. Esse trabalho testou uma coleira a base de imidacloprida e flumetrina, preconizada para uso em cães domésticos, por ser de aplicação única e de longa duração. A coleira se mostrou muito eficiente por seis meses na eliminação das pulgas, associada a aspersão de deltametrina no recinto; sem ocorrência de efeitos adversos.

Efficacy of afoxolaner in the flea control in experimentally infested cats / Eficácia do afoxolaner no controle de pulgas em gatos experimentalmente infestados

The aim of this study was to evaluate the efficacy of a single dose of oral afoxolaner in controlling fleas in cats. Fourteen cats were used. The cats were given identification numbers, housed individually, artificially infested with Ctenocephalides felis felis, and treated (or not) with afoxolaner. Were divided into a treatment group and a control group (n = 7/group), on the basis of the fleas count hours after an infestation applied on Day (one-by-one allocation after ordering by count). At the start of the experimental protocol (designated day 0), the treated group received afoxolaner in a single dose of 2.5 mg/kg and the control group animals received a placebo. All animals were infested with 100 C. felis felis fleas two days before day 0, as well as on days 5, 12, 19, 26, 33, 40, 47, 54, and 63, parasite loads being evaluated at 48 h after each infestation. The efficacy of afoxolaner was 100% on day 2 and remained above 98% until day 42, decreasing to 95.3% by day 63. The findings confirm that a single dose of oral afoxolaner was effective in controlling C. felis felis in cats, and there were no observed adverse events.(AU)

O objetivo do estudo foi avaliar a eficácia de uma dose única de afoxolaner oral no controle de pulgas em gatos. Foram utilizados 14 gatos. Os animais foram identificados, alojados individualmente, infestados artificialmente com C. felis felis e tratados (ou não) com afoxolaner. Foram divididos em um grupo de tratamento e um grupo controle (n = 7/ grupo), com base na contagem de pulgas, horas após a infestação aplicada no dia (alocação de um por um após o período por contagem). No início do protocolo experimental (dia 0), o grupo tratado recebeu afoxolaner em dose inicial de 2,5 mg / kg e os animais do grupo controle receberam um placebo. Todos os animais foram infestados com 100 pulgas C. felis felis dois dias antes do dia 0, assim como nos dias 5, 12, 19, 26, 33, 40, 47, 54 e 63, sendo avaliadas as cargas parasitárias às 48 h após cada infestação. A eficácia do afoxolaner foi de 100% no dia 2 e permaneceu acima de 98% até o dia 42, diminuindo para 95,3% no dia 63. Os resultados confirmam que uma dose única de afoxolaner oral foi eficaz no controle de C. felis felis em gatos, e não houve eventos adversos observados.(AU)

Control de pulgas en ciervos de los pantanos (Blastocerus dichotomus) cautivo con uso de collar con imidacloprida e flumetrina. / Control of fleas in captive marsh deer (Blastocerus dichotomus) with imidacloprid and flumetrin collar / Controle de pulgas em cervo-do-pantanal (Blastocerus dichotomus) de cativeiro com uso de coleira com imidacloprida e flumetrina

El ciervo de los pantanos (Blastocerus dichotomus), es el ciervo más grande del continente sudamericano y figura en la lista de animales en peligro de extinción, de acuerdo con el IBAMA y la UICN. Tiene una población bajo el cuidado humano muy limitado, estimada en un centenar de individuos. Entre los principales problemas de la especie "ex situ" está la puliciose. Esta infestación puede causar anemia significativa, debilidad, pérdida de apetito y incluso la muerte. El control de los parásitos se lleva de varias maneras, con el combate de las pulgas en el huésped y el medio ambiente, en el que el conjunto de control es más eficaz. Sin embargo, el control de pulgas en animales salvajes es muy restringido, por el comportamiento agresivo del animal, como la ausencia de un producto eficaz y de acción prolongada. Este estudio probó un collar impregnado con imidacloprid y flumetrina, com uso recomiendado en perros domésticos, em una sola aplicación y con largo plazo. El producto ha demostrado muy eficaz durante dieciocho meses en la eliminacion de pulgas, asociado con la pulverizacion de deltametrina en la habitación; sin efectos adversos em los animales salvajes.

The marsh deer (Blastocerus dichotomus) is the largest deer in the South American continent and is listed in the red list of endangered animals according to IBAMA and IUCN. It has a very small population in captivity, estimated at 100 individuals. Among the main problems of this species ex situ we have puliciosis. This infestation can cause significant anemia, weariness and even death. The control of these parasites is carried out in several ways, with flea combat in the hosts and in the environment, and joint control is the most effective method. However, the control of fleas in animals is quite restricted, both by the behavior of the animal, and by the nonexistence of an effective and long-acting anti parasitic product. This work tested a collar impregnated with imidacloprid and flumethrin, recommended for use in domestic dogs, because it is of single application and long duration. The collar showed to be very efficient for eighteen months, in the elimination of fleas, associated with spraying of deltamethrin in the enclosure; without occurrence of adverse effects.

O cervo-do-pantanal (Blastocerus dichotomus), é o maior cervídeo do continente sul-americano e figura na lista de animais ameaçados de extinção, segundo o IBAMA e a IUCN. A espécie tem uma população muito reduzida ex-situ, estimada em cem indivíduos. Dentre os principais problemas dessa espécie em cativeiro temos a puliciose. Essa infestação pode causar anemia expressiva, inapetência e óbito. O controle desses parasitas é realizado de diversas formas, com combate das pulgas nos animais e no ambiente, sendo que o controle conjunto é o mais eficaz. No entanto, o controle das pulgas nos animais é bastante restrito, tanto pelo temperamento do animal, quanto pela inexistência de um produto eficaz e de longa ação. Esse trabalho testou uma coleira a base de imidacloprida e flumetrina, preconizada para uso em cães domésticos, por ser de aplicação única e de longa duração. A coleira se mostrou muito eficiente por seis meses na eliminação das pulgas, associada a aspersão de deltametrina no recinto; sem ocorrência de efeitos adversos.

Parasites of small Indian mongoose, Herpestes auropunctatus, on St. Kitts, West Indies.

Herpestes auropunctatus, the small Indian mongoose, is an invasive omnivore introduced to the Caribbean, including the island of St. Kitts over 150 years ago. It has played a role in changing native fauna and can carry zoonotic pathogens of public health importance. The aim of the current study was to estimate the prevalence of parasites harbored by mongooses. In total, 87 mongooses trapped from April to July 2015 were examined for parasites using (1) hair plucks (N = 79), ear swabs (N = 79), and general coat and skin examination (N = 87) for mites, ticks, lice, and fleas; (2) dissection of the trachea, bronchi, and lungs for lungworms and flukes (N = 76); (3) a double centrifugation fecal flotation method for parasites of the gastrointestinal tract (N = 75); and (4) PCR of heart homogenates for Toxoplasma gondii (N = 60). The only ectoparasite seen was Ctenocephalides felis (79.3%; 69/87), with most mongooses having > 10 fleas (based on a subjective assessment) but insufficient numbers to result in signs of pruritus or anemia. On fecal flotation, coccidial oocysts were found with a prevalence of 69.3% (52/75). Neither T. gondii, lungworm, nor fluke infections were detected with the methods used. The high number of C. felis-infested mongooses and the infestation level of the individual mongooses suggest that they could serve as a reservoir for these potential vectors of pathogens. No evidence was found to support that mongooses are a component of T. gondii cycles on St. Kitts, although this finding needs to be confirmed with a larger sample size from other geographic locations.

Fipronil/(S)-methoprene spot-on to control fleas on cats in a field trial in Spain

The study was conducted in order to evaluate the effect of a fipronil/(S)-methoprene formulation against fleas on naturally infested cats. The study involved a population of 89 cats distributed among 24 veterinary practices in 9 regions of Spain. The product was applied according to label instructions on days 0, 30 and 60. Animals underwent parasitological and clinical assessments on day 0 and thereafter in monthly intervals (every 30 days) until day 90. Ctenocephalides felis was the most abundant species (98.9% of all fleas collected), and flea abundance on Day 0 was associated with the hair type, the location of the household, and the time elapsed from the last anti-flea treatment. Fipronil/(S)-methoprene demonstrated high efficacy and induced the reduction of clinical signs related to the presence of fleas. Clinical signs and flea abundance decreased significantly throughout time (P=0.001) with an efficacy rate of 72.6% at Day 30, 88.4% at Day 60 and 93.9% at Day 90. A high level of flea control and a remission of the clinical signs related to presence of fleas were observed on cats following 3 monthly applications a fipronil/(S)-methoprene formulation.(AU)

Fipronil/(S)-methoprene spot-on to control fleas on cats in a field trial in Spain

The study was conducted in order to evaluate the effect of a fipronil/(S)-methoprene formulation against fleas on naturally infested cats. The study involved a population of 89 cats distributed among 24 veterinary practices in 9 regions of Spain. The product was applied according to label instructions on days 0, 30 and 60. Animals underwent parasitological and clinical assessments on day 0 and thereafter in monthly intervals (every 30 days) until day 90. Ctenocephalides felis was the most abundant species (98.9% of all fleas collected), and flea abundance on Day 0 was associated with the hair type, the location of the household, and the time elapsed from the last anti-flea treatment. Fipronil/(S)-methoprene demonstrated high efficacy and induced the reduction of clinical signs related to the presence of fleas. Clinical signs and flea abundance decreased significantly throughout time (P=0.001) with an efficacy rate of 72.6% at Day 30, 88.4% at Day 60 and 93.9% at Day 90. A high level of flea control and a remission of the clinical signs related to presence of fleas were observed on cats following 3 monthly applications a fipronil/(S)-methoprene formulation.(AU)