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Gametocyte of Hepatozoon spp. in a neutrophil from peripheral blood smear (Giemsa's stain ×1,000)
Source publication
We attempted to determine the prevalence of Hepatozoon spp. infection in Mashhad, northeast of Iran, via blood smear parasitology.
The prevalence was investigated by examination of blood smear parasitology, using blood samples collected from 254 dogs (51 strays and 203 privately owned-dogs).
Two stray dogs (2/51; 3.92%) and two privately-owned dogs...
Context in source publication
Context 1
... smear parasitology, revealed 1.57% (4/254) of the subjects were infected by Hepatozoon spp. (Fig. 1). These included two stray (2/51; 3.92%) and two privately-owned dogs (2/203; 0.98%). The parasitemia ranged from 1 to 3% with the highest parasitemia (3%) found in a 15-year-old male privately- owned dog. ...Citations
... While microscopically examining thin blood smears detected gamonts in neutrophils only in one sample. Hepatozoon infections have been spotted by blood smear examination ranging from 1% in Iran [9] to 39.2% in Brazil [10] and 10% in Cuba [3]. Hepatozoonosis in blood has been detected by the PCR method in Cuba (47.5%), the Czech Republic (50%), Brazil (53.3%), ...
... According to a study from the Northeast of Iran, the prevalence of H. canis detected by PCR was about 8% [12]. In 2012, another study investigated Hepatozoon infections in dogs in Mashhad by cytological examination [9]. Similarly, Khoshnegah et al. demonstrated the Hepatozoon canis gametocytes in peripheral blood and bone marrow smear in an 11-year-old male dog for the first time in Iran [4]. ...
Canine hepatozoonosis is a tick-transmitted apicomplexan infection caused by two species of Hepatozoon, H. canis, and H. americanum. The present research aimed at detection of Hepatozoon spp. in dogs and its effects on hematological alterations. Blood samples were taken from 108 dogs to assess Hepatozoon spp. Phylogenetic analysis was performed based on the 18S rDNA marker by PCR assay and Giemsa-stained blood smear examination. Of the 108 blood samples of dogs tested in the present study, eight (7.40%, 95% CI: 3.25-14.07%) were positive by the Hepatozoon-specific PCR assay. However, in the microscopic examination, only one sample (0.93%) was positive. All of the sequenced samples were H. canis. The Hepatozoon sequences obtained from PCR amplicons in the canine-positive cases exhibited 100% similarity to each other and 98.47-100% similarity to other relevant sequences in GenBank. These findings represent the first molecular evidence of H. canis in dog populations in South Iran. Furthermore, according to the hematological analysis, significantly higher average numbers of neutrophils and lymphocytes were found in the infected group compared to the non-infected dogs. In this study, no statistically significant connection (P<0.05) was observed between H. canis infection and the examined risk factors.
... Semelhante a outros estudos, também foi verificada coinfecção por E. canis (Spolidorio et al., 2011;Sarma et al., 2012;Chhabra et al., 2013) Ao analisarmos apenas os animais que apresentaram anemia (45,63%) em seus hemogramas verifica-se que as maiores relações de alterações envolveram monocitose (50,00%), seguida de neutrofilia (47,36%) e leucocitose (42,10%). Condizente à estes resultados, a anemia e a leucocitose neutrofílica também são descritas como achados comuns por diversos autores P á g i n a | 276 (Aguiar et al., 2004;O'Dwyer et al., 2006;Otranto et al., 2011;Rahmani Amoli et al., 2012;Sarma et al., 2012;Chhabra et al., 2013) assim como a monocitose (Aguiar et al., 2004;O'Dwyer et al., 2006). ...
... As manifestações clínicas em infecções por H. canis são normalmente relacionadas à infecções concomitantes, imunossupressão e grau de parasitismo (Pereira et al., 2011). Normalmente o grau de parasitismo em cães é baixo (Rahmani Amoli et al, 2012). ...
... Normalmente o diagnóstico de hematozoários em cães é feito por exame de esfregaços de sangue corados pelo método de Giemsa (Aguiar et al., 2004;O'Dwyer et al., 2006;Otranto et al., 2011;Rahmani Amoli et al., 2012;Sarma et al., 2012) e analisados sob um microscópio de luz, método esse que pode apresentar baixa sensibilidade, especialmente quando as amostras de sangue não são coletadas na fase aguda da infecção (Pereira et al., 2011;Dantas-Torres e Otranto, 2014). Em contrapartida, dispõem-se do uso de ferramentas moleculares para o diagnóstico de protozoários, que apesar se estarem se tornando ferramentas cada vez mais populares, continuam de certa forma, restritos à pesquisas (Dantas-Torres e Otranto, 2014). ...
A hepatozoonose é o nome dado a doença provocada pelo coccídeo Hepatozoon spp., transmitida por artrópodes e que acomete principalmente os carnívoros domésticos e silvestres. A infecção provocada por Hepatozoon canis é transmitida pelo Rhipicephalus sanguíneus e o cão se infecta através da ingestão do carrapato. O presente trabalho tem como objetivo relatar os aspectos gerais da hepatozoonose canina, durante a realização de estágio em um laboratório particular na região de Goiânia-GO, durante o período de quatro meses. Durante o estágio foi feito um levantamento de casos de cães positivos para Hepatozoon canis, a fim de verificar possíveis relações hematológicas. Foram analisados um total de 72.532 hemogramas, dos quais obteve-se 103 (0,14%) resultados positivos para Hepatozoon canis. Dentre os animais positivos, 38 (36,90%) cães eram do sexo masculino e 65 (63,10%) do sexo feminino. Verificou-se também a maior ocorrência em animais de 13 a 96 meses de idade. Durante o estágio no laboratório a análise hematológica para determinação dos valores de hemograma foi a mais realizada, apesar desse número, não foi verificada grande ocorrência do Hepatozoon canis. Verificou-se que ainda são necessários mais estudos em relação à infecção e doença provocadas pelo Hepatozoon canis a fim de facilitar o diagnóstico nos animais.
... This is because genus Hepatozoon is known as a tick-borne protozoan parasite, and the gene is closely related to Plasmodium sp. (Amoli et al., 2012). The fragments in the universal primer may bind with the samples, and H. ophisauri can be detected. ...
Background
Rodent species are well known for their potential as hosts and reservoirs for various zoonotic diseases. Studies on blood parasite infection in small mammals focused on urban cities in Peninsular Malaysia and have been conducted over the years. In contrast, there are information gaps related to molecular detection of blood parasites in urban areas of Sarawak that are associated with veterinary importance and zoonotic spillover potential. Increasing prevalence and transmission of blood parasite diseases is the most crucial public health issue, particularly in developing urban areas of Sarawak. Therefore, molecular identification studies were performed to determine and identify the blood parasites infecting rodents.
Methods
A total of 40 rodent blood samples were analysed for blood parasite infection and a combined approach using polymerase chain reaction‐based technique, and traditional microscopic examination (blood smear test) was conducted. 18s rRNA (Plasmodium spp.) and cytochrome b (Hepatocystis spp.) gene marker were used to identify the blood parasites.
Results
Note that 67.5% (n = 27) blood samples were tested negative for blood parasites, while 32.5% (n = 13) blood samples collected were infected with at least one protozoan parasite. Out of 13 samples, 69.2% (n = 9) were detected with Hepatocystis sp., while 15.4% (n = 2) were positive with Hepatozoon ophisauri. Two individuals had multiple infections from both species. No Plasmodium spp. have been detected throughout this study using universal primer (targeted Plasmodium spp.); however, different parasite species which were H. ophisauri were detected.
Conclusion
Although there is no evidence of human infection from H. ophisauri and Hepatocystis sp. detected from the study, the data show the host species are heavily infected, and the information is essential for future prevention of zoonotic outbreaks and surveillance programmes. Therefore, it is suggested that the surveillance programmes should be incorporated in targeted areas with a high risk of disease emergence.
... The high prevalence of H. canis in all the provinces of Iran as well as from Bahawalpur in Pakistan suggests that Rh. sanguineus s.l. is highly prevalent in these areas. After the first detection of H. canis gametocytes in a blood smear of an Iranian dog (Khoshnegah et al. 2009), the prevalence of this infection was reported only in dogs from northern half of Iran ranging from 1.6 to 23% (Amoli et al. 2012;Dalimi et al. 2017;Soltani and Dalimi 2018;Barati and Razmi 2018). Conversely, the prevalence herein recorded for H. canis in Pakistan (i.e., 41.4%) was similar to that of farm dogs recorded in Punjab districts (45.5%, Ahmad et al. 2018a). ...
Canine vector-borne diseases (CVBDs) are highly prevalent in tropical and subtropical countries, mainly due to favorable climate conditions and reduced adoption of preventive measures. This study aimed to provide a comprehensive overview on the prevalence of CVBDs in Iran and Pakistan where limited data are available. Blood samples were collected from 403 dogs from six provinces in Iran and Pakistan to assess the presence of pathogen DNA (i.e., Anaplasma spp., Coxiella burnetii , Ehrlichia spp., Rickettsia spp., Babesia spp . , Hepatozoon spp., filarioids, and Leishmania spp.). Sera were also screened by an immunofluorescence antibody test for the detection of antibodies against Leishmania infantum . In total, 46.9% of dogs scored positive to Hepatozoon canis being the most frequently detected (41.4%), followed by Anaplasma platys (6.4%), Ehrlichia canis (3.4%), Rickettsia spp. (2.2%), Babesia vogeli (1.0%), and L. infantum (0.3%). A seroprevalence of 9.6% to anti- L. infantum IgG was also recorded. Data reported herein demonstrate that dogs from Iran and Pakistan are at a high risk of CVBDs, particularly of canine hepatozoonosis. Effective control strategies are advocated for minimizing the risk of infection in animals and humans, also in consideration of the zoonotic potential of some pathogens detected.
... Hepatozoonosis is an emerging vector borne disease (VBD) of both dogs and cats, caused by different species belonging to the genus Hepatozoon [1,2]. Canine hepatozoonosis is caused by Hepatozoon canis that infects dogs worldwide, and Hepatozoon americanum that has been reported, to date, only in the Americas [1,[3][4][5]. Dogs are intermediate hosts of Hepatozoon spp. and become infected via the ingestion of infected ticks (definitive hosts) harboring mature oocysts. ...
Canine hepatozoonosis caused by Hepatozoon canis is an emerging disease in Europe. Clinical pictures vary from subclinical to life-threatening and non-specific clinical signs are predominantly reported. A 2-month-old female puppy originating from Southern Italy was adopted and moved to Northern Italy. Then, the dog was brought to a local veterinary practice for gastrointesti-nal signs, migrating lameness and pruritic dermatitis, and then tested positive for Hepatozoon spp. gamonts at the blood smear. After treatment with imidocarb dipropionate and doxycycline, the dog showed an initial clinical improvement. However, gastrointestinal signs recurred, and diffuse superficial pyoderma appeared on the thoracolumbar region, along with fever, lethargy, and weight loss. Eight months from the first onset of clinical signs, the dog was referred to a veterinary clinic and subjected to complete blood count, urine and fecal analysis, along with abdominal ultrasonog-raphy, whole-body CT and gastroduodenal endoscopy. Skin biopsies and blood samples were subjected to a PCR-coupled sequencing protocol, which scored both positive for H. canis. Alterations were consistent with a pre-existing cholangiohepatitis and multiple acquired extrahepatic shunts secondary to portal hypertension. The dog was euthanatized due to a clinical worsening two months later. The potential role of H. canis in the systemic disease observed, clinic-pathological findings and epizootiological implications are discussed.
... This species transmits several zoonoses in Europe such as anaplasmosis, babesiosis, bartonellosis, borreliosis (including Lyme), ehrlichiosis, louping ill, tick-bite fever and different rickettsioses, tick-borne encephalitis and tularemia (Salman and Teres-Call 2013). Rhipicephalus sanguineus is known the vector of Crimean-Congo hemorrhagic fever (CCHF) (Telmadarraiy et al. 2015), anaplasmosis and ehrlichiosis (Khazeni et al. 2013), babesiosis (Razmi et al. 2002) and hepatozoonosis (Rahmani Amoli et al. 2012) in Iran. Also, piroplasmosis (theileriosis) is reported in Iran which R. sanguineus is one of its vectors (Bigdeli et al. 2012). ...
The raccoon, Procyon lotor, is native to North and Central America but has been introduced to several Old Word countries including Iran. The aim of this study was to determine the prevalence of ectoparasites of raccoons in Iran. During 2015–2017, a total of 30 feral raccoons were collected from Guilan Province, northern Iran. The fur of raccoons was examined for the presence of ectoparasites. The collected ectoparasites were preserved in 70% ethanol and microscopically and morphologically identified. In total, 11 fleas and 244 ixodid ticks were collected from 18 (60%) raccoons for the first time in Iran. Two raccoons (6.7%) harbored just fleas, five raccoons (16.7%) both fleas and ticks and 11 raccoons (36.7%) just ticks. Two species of fleas: Ctenocephalides felis (cat flea) and Pulex irritans (human flea) were collected from seven raccoons (23.3%). Four species of ixodid ticks: Haemaphysalis concinna, Ixodes ricinus, Rhipicephalus sanguineus and R. turanicus were collected from 16 raccoons (53.3%). All ectoparasites, except for cat flea, were found on raccoons for the first time. The prevalence, mean abundance and mean intensity of ectoparasites and the indices of affinity, association and diversity were calculated. The pair of species H. concinna/ I. ricinus showed significant association and the infestation with I. ricinus in cubs was significantly higher than in adults (P ˂ 0.05). The medical importance of zoonotic infections transmittable by raccoons and their ectoparasites is discussed. The investigation of different possible vector-borne infections in raccoons and their ectoparasites in the study area is suggested.
... Received 1 April 2020; Received in revised form 9 July 2020; Accepted 15 July 2020 parasitemia [4]. However, microscopic examination of blood smears is not a reliable method of detection of H. canis, particularly for asymptomatic carriers, where the level of parasitemia is low or intermittent, such that there is a low number of circulating gamonts in the host's blood [8]. Furthermore, this is also a time-consuming process to perform. ...
... infections (10%) in the population of stray dogs examined in this study. This result is consistent with the prevalence of Hepatozoon infections inferred by blood smear examination in other studies, where values range from 1% in Iran [8] to 39.2% in Brazil [27]. In contrast, the overall prevalence determined by molecular analysis using a real-time PCR assay, revealed that 47.5% of sampled dogs were infected with H. canis. ...
Canine hepatozoonosis caused by Hepatozoon canis is a worldwide distributed tick-borne disease of domestic and wild canids that is transmitted by ingestion of Rhipicephalus sanguineus sensu lato (s.l.) ticks. The present study was aimed to determine the prevalence of Hepatozoon infections in 80 stray dogs from Havana Province in Cuba, and to confirm the species identity and phylogenetic relationships of the causative agent. Samples were screened by microscopical examination of thin blood smears for the presence of Hepatozoon spp. gamonts and by genus-specific SYBR green-based real-time PCR assay targeting the 18S rRNA gene. Direct microscopy examination revealed Hepatozoon gamonts in the peripheral blood of 8 dogs (10.0%; 95% CI: 4.80–18.0%), while 38 animals (47.5%; 95% CI: 36.8–58.4%) were PCR-positive, including all microscopically positive dogs. Hence, the agreement between the two detection methods was ‘poor’ (κ = 0.20). Hematological parameters did not differ significantly between PCR-positive and PCR-negative dogs (p > 0.05). The DNA sequences of the 18S rRNA gene of the Hepatozoon spp. from Cuban dogs showed a nucleotide identity >99% with those of 18S rRNA sequences of Hepatozoon canis isolates from Czech Republic, Brazil and Spain. Phylogenetic analysis revealed that obtained sequences clustered within the Hepatozoon canis clade, different from the Hepatozoon felis or Hepatozoon americanum clades. The present study represents the first molecular characterization of Hepatozoon canis in stray dogs within Cuba.
... Based on the microscopic examination of blood smears, Hepatozoon spp. infection in sampled dogs in the present study (2.03%) is higher than the previous study conducted in Iran (1.6%) [29] which it may be related to the paraseitemia rate in sampled dogs [14]. There were some similar studies conducted in different countries which used microscopic examination of dog's blood smears and they showed Hepatozoon spp. ...
... This study is the rst in Iran to record H. canis in R. sanguineus (s.l) ticks from dogs. It should be noted that the previous studies have mentioned R. sanguineus s.l. as the most prevalent specie of tick to infest the dogs in Iran [29], but the vector competence of R. sanguineus s. l. for Hepatozoon protozoa from Iran has never been demonstrated. In the present study, R. sanguineus (s.l.) ticks was the only species found in sampled dogs of the study area which could be related to its huge capability to adapt [49][50][51][52]. ...
Background
Hepatozoon canis is a protozoan that is transmitted by the ixodid ticks. Ingesting the tick or a section of the tick organ which is infected by the mature oocysts containing infectious sporozoite is the main source of infection in dogs. Canine hepatozoonosis infects dogs in Iran, but the vector for Hepatozoon protozoa from Iran has never been demonstrated. The present study aims to detect H. canis in dogs and vector ticks in Iran.
Methods
During the period of 2018–2019, Blood samples and ixodid ticks were collected and examined using microscopical, molecular, and serological methods. A total of 246 blood samples were collected from the cephalic vein of pet, stray, and shelter dogs (103 stray, 99 shelter, and 44 pets) of both genders and varying ages in Northwest of Iran.
Results
Microscopically, infected neutrophils with Hepatozoon spp. were detected in 5 of 246 (2.03%) thin stained blood smears with low parasitemia. Indirect immunofluorescent antibody test (IFAT) was used to test the serum samples and antibodies against H. canis were detected in 31 (12.6%) of the serum samples. Molecularly, 23 out of 246 (9.34%) blood samples were found to be infected with H. canis. A comparison of the results of 3 diagnostic methods demonstrated a good agreement between IFAT and PCR and a poor agreement between microscopical examination with IFAT and PCR. There was no significant difference in different age groups and sex of sampled dogs. However, stray dogs had significant infection rate of than pets and shelter ones. In body inspection, 141 adult ticks (31 partially engorged females, 26 fully engorged female and 84 fed males) were collected from examined dogs and all ticks were belonging to species of Rhipicephalus sanguineus (sensu lato). Positive reaction to H. canis was observed in the genomic DNA of the 7 ticks (4.96%). A BLAST analysis of obtaining sequences isolated from both dogs and ticks indicated a 99–100% similarity with H. canis 18S rRNA gene sequences in GenBank. This is the first study in Iran to detect H. canis in R. sanguineus tick.
... Farkas et al. (2014), tested the blood samples of 334 red foxes and 15 golden jackals to investigate H. canis parasite by PCR method; the obtained results indicated that 26 red foxes (8%) and 9 golden jackals (60%) had been infected by H. canis. Amoli et al. (2012), tested 254 blood samples of 51 stray dogs and 203 pet dogs and only 4 cases were diagnosed infected by H. canis. In Dalimi et al. (2017) study, twenty-four out of the 104 (23.07%) samples were found to be positive for H. canis by molecular detection in dogs of Ardabil province. ...
Hepatozoonosis is a protozoal disease caused by various species of Hepatozoon. This parasite is transmitted from tick; the main vector of Hepatozoon canis is usually the brown dog tick (Rhipicephalus sanguineus). However, several species of ticks are disposed as the alternative vectors. Dogs are usually infected by eating the tick or a part of the tick organ infected by the mature oocysts containing infectious sporozoite. In the current study, a total of 145 blood samples were collected from the cephalic vein of pet, stray, and shelter dogs in Tehran. To conduct this study, first thin blood smears were prepared from all the samples and stained with the Giemsa method. Then, after extraction of DNA from the blood samples, in order to trace Hepatozoon canis, the 18S rRNA gene segment of the parasite was amplified using polymerase chain reaction (PCR). To confirm the PCR-positive results, five randomly selected PCR-positive samples were sequenced. According to the results, through direct observation of microscopic slides, no infection of H. canis parasite was observed, but according to the PCR results, 32 out of the 145 blood samples were found to be infected by H. canis. In this study, infection to H. canis in older dogs was higher than in young dogs, and more male dogs were found to be infected by the parasite compared to female dogs; but no significant difference was observed in this regard (P > 0.05). Moreover, stray dogs showed a significantly higher rate of infection, compared to the pet and shelter ones (P < 0.05).
... H. canis infections occur when mature oocysts contained within the definitive host, the brown dog tick, Rhipicephalus sanguineus, are ingested by the dog. Infections may also be transmitted transplacentally from an infected dam to the foetus (Forlano et al., 2007;Amoli et al., 2012). Potential vectors of canine hepatozoonosis include Amblyomma ovale, Rhipicephalus turanicus, Rhipicephalus spp. ...
... (Forlano et al., 2007;Ivanov and Tsachev, 2008;Giannelli et al., 2013;Latrofa et al., 2014). The parasite produces gamonts in the peripheral blood and schizonts in the lungs, heart, skeletal muscle, liver, spleen and lymph nodes (Ivanov and Tsachev, 2008;Amoli et al., 2012). The life cycle of H. canis is approximately 81 days with meronts observed in the bone marrow of the dog 13 days post inoculation and gamonts in the peripheral blood 28 days after infection (Baneth, 2011). ...
... Cases involving low parasitaemias of H. canis are usually asymptomatic, however, clinical signs such as fever, weight loss, lethargy, pale mucous membranes, cachexia, paralysis, hind limb weakness, muscle hyperaesthesia and anorexia are observed in cases with high parasitaemias (Ivanov and Tsachev, 2008;Baneth, 2011;Chhabra et al., 2013). Diagnosis can be made by observation of gamonts in the cytoplasm of neutrophils or monocytes in peripheral blood or buffy coat smears (Amoli et al., 2012). The gamonts appear as a clear oblong capsule with a faintly staining nucleus within the cytoplasm of the neutrophil or monocyte. ...
A five-year old mixed breed bitch was presented to the veterinary clinic in lateral recumbency with a history of anorexia and muscle hyperaesthesia. Examination of the blood smear of this animal revealed the presence of Hepatozoon spp. gamonts in the neutrophils and monocytes with a parasitaemia level of approximately 2%. Complete blood count (CBC) revealed a neutrophilia, and a normocytic normochromic non-regenerative anaemia which were consistent with Hepatozoon infections. Diagnosis was confirmed by polymerase chain reaction (PCR) amplification of the 18S rRNA followed by DNA sequencing of the amplicon. Although the other dog in the household appeared asymptomatic, Hepatozoon canis infections was confirmed by both microscopic examination of blood smear and PCR. Both dogs were infested with Rhipicephalus sanguineus ticks. Phylogenetic analysis revealed that the H. canis sequences from these two dogs were similar to those from Venezuela and St Kitts but not Brazil. This is the first reported case of Hepatozoon canis infections in dogs in Trinidad that were confirmed by molecular techniques.
