scholarly journals Novel High-Throughput Multiplex qPCRs for the Detection of Canine Vector-Borne Pathogens in the Asia-Pacific

2021 ◽  
Vol 9 (5) ◽  
pp. 1092
Author(s):  
Lucas Huggins ◽  
Luca Massetti ◽  
Bettina Schunack ◽  
Vito Colella ◽  
Rebecca Traub
Keyword(s):  
High Throughput ◽  
Clinical Signs ◽  
Asia Pacific ◽  
Hepatozoon Canis ◽  
Babesia Gibsoni ◽  
Babesia Vogeli ◽  
Field Samples ◽  
Vector Borne ◽  
Performance Results ◽  
Next Generation Sequencing Ngs

The Asia-Pacific hosts a large diversity of canine vector-borne pathogens (VBPs) with some of the most common and most pathogenic, generating significant mortality as well as a spectrum of health impacts on local dog populations. The VBPs Anaplasma platys, Babesia gibsoni, Babesia vogeli, Ehrlichia canis, Hepatozoon canis and haemotropic Mycoplasma spp. are all endemic throughout the region, with many exhibiting shifting geographical distributions that warrant urgent attention. Moreover, many of these species cause similar clinical signs when parasitising canine hosts, whilst knowledge of the exact pathogen is critical to ensure treatment is effective. This is complicated by frequent coinfection that can exacerbate pathology. Here, we describe the development, optimisation and validation of two novel quadruplex Taq-Man based real-time PCRs (qPCRs) for the specific and sensitive detection of the aforementioned VBPs. To ensure accurate evaluation of diagnostic performance, results of our qPCRs were evaluated on field samples from Thai dogs and compared with both conventional PCR (cPCR) results and next-generation sequencing (NGS) metabarcoding. Our qPCRs were found to be more sensitive at detecting canine VBP than cPCR and generated results similar to those achieved by NGS. These qPCRs will provide a valuable high-throughput diagnostic tool available to epidemiologists, researchers and clinicians for the diagnosis of key canine VBPs in the Asia-Pacific and further afield.

Critical analysis of vector-borne infections in dogs: Babesia vogeli, Babesia gibsoni, Ehrlichia canis and Hepatozoon canis in Punjab, India

2016 ◽  
Vol 61 (4) ◽  
Author(s):  
Lachhman Das Singla ◽  
Deepak Sumbria ◽  
Ajay Mandhotra ◽  
M.S. Bal ◽  
Paramjit Kaur
Keyword(s):  
Hepatozoon Canis ◽  
Babesia Gibsoni ◽  
Chain Reactions ◽  
Vector Borne Diseases ◽  
Polymerase Chain Reactions ◽  
Relationship Analysis ◽  
Babesia Vogeli ◽  
Blood Smears ◽  
Blood Smear Examination ◽  
Vector Borne

AbstractThere are few published studies on various vector borne diseases of dogs in India and most depict clinical infection in dogs, diagnosed by observation of the haemopathogens in stained blood smears. This study provides the first report regarding molecular confirmation and ancestral relationship analysis of blood smears positive cases of assorted haemopathogens in Punjab province of India. On blood smear examination, haemopathogens were observed in 124 out of 778 (15.95%, 95% CI: 13.53– 18.68) blood smears. Further polymerase chain reactions (PCR) was used on bloods smear positive cases to validate the results. Out of 778 blood samples,

scholarly journals Vector-borne pathogens in dogs of different regions of Iran and Pakistan

2021 ◽  
Author(s):  
Roberta Iatta ◽  
Alireza Sazmand ◽  
Viet-Linh Nguyen ◽  
Farzad Nemati ◽  
Muhammad Mazhar Ayaz ◽  
...  
Keyword(s):  
Control Strategies ◽  
Antibody Test ◽  
Effective Control ◽  
Hepatozoon Canis ◽  
Comprehensive Overview ◽  
Climate Conditions ◽  
Vector Borne Diseases ◽  
Babesia Vogeli ◽  
Vector Borne ◽  
Pathogen Dna

AbstractCanine 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.

scholarly journals Molecular detection of pathogens in ticks and fleas collected from companion dogs and cats in East and Southeast Asia

2020 ◽  
Author(s):  
Viet-Linh Nguyen ◽  
Vito Colella ◽  
Grazia Greco ◽  
Fang Fang ◽  
Wisnu Nurcahyo ◽  
...  
Keyword(s):  
Southeast Asia ◽  
Rhipicephalus Sanguineus ◽  
Sensu Stricto ◽  
The Philippines ◽  
Hepatozoon Canis ◽  
Rickettsia Felis ◽  
Companion Dogs ◽  
Babesia Vogeli ◽  
East And Southeast Asia ◽  
Vector Borne

Abstract Background: Ticks and fleas are considered amongst the most important arthropod vectors of medical and veterinary concern due to their ability to transmit pathogens to a range of animal species including dogs, cats and humans. By sharing a common environment with humans, companion animal-associated parasitic arthropods may potentially transmit zoonotic vector-borne pathogens (VBPs). This study aimed to molecularly detect pathogens from ticks and fleas from companion dogs and cats in East and Southeast Asia.Methods: A total of 392 ticks and 248 fleas were collected from 401 infested animals (i.e. 271 dogs and 130 cats) from China, Taiwan, Indonesia, Malaysia, Singapore, Thailand, the Philippines and Vietnam, were molecularly screened for the presence of pathogens. Ticks were tested for Rickettsia spp., Anaplasma spp., Ehrlichia spp., Babesia spp. and Hepatozoon spp. while fleas were screened for the presence of Rickettsia spp. and Bartonella spp.Result: Of the 392 ticks tested, 37 (9.4%) scored positive for at least one pathogen with Hepatozoon canis being the most prevalent (5.4%), followed by Ehrlichia canis (1.8%), Babesia vogeli (1%), Anaplasma platys (0.8%) and Rickettsia spp. (1%) [including Rickettsia sp. (0.5%), Rickettsia asembonensis (0.3%) and Rickettsia felis (0.3%)]. Out of 248 fleas tested, 106 (42.7%) were harboring at least one pathogen with R. felis being the most common (19.4%), followed by Bartonella spp. (16.5%), Rickettsia asembonensis (10.9%) and “Candidatus Rickettsia senegalensis” (0.4%). Furthermore, 35 Rhipicephalus sanguineus ticks were subjected to phylogenetic analysis, of which 34 ticks belonged to the tropical and only one belonged to the temperate lineage (Rh. sanguineus (sensu stricto)). Conclusion: Our data reveals the circulation of different VBPs in ticks and fleas of dogs and cats from Asia, including zoonotic agents, which may represent a potential risk to animal and human health.

scholarly journals Molecular detection of pathogens in ticks and fleas collected from companion dogs and cats in East and Southeast Asia

2020 ◽  
Author(s):  
Viet-Linh Nguyen ◽  
Vito Colella ◽  
Grazia Greco ◽  
Fang Fang ◽  
Wisnu Nurcahyo ◽  
...  
Keyword(s):  
Southeast Asia ◽  
Animal Species ◽  
Ehrlichia Canis ◽  
Hepatozoon Canis ◽  
Anaplasma Platys ◽  
Rickettsia Felis ◽  
Companion Dogs ◽  
Babesia Vogeli ◽  
East And Southeast Asia ◽  
Vector Borne

AbstractTicks and fleas are considered amongst the most important arthropod vectors of medical and veterinary concern due to their ability to transmit pathogens to a range of animal species including dogs, cats and humans. By sharing a common environment with humans, companion animal-associated parasitic arthropods may potentially transmit zoonotic vector-borne pathogens (VBPs). This study aimed to molecularly detect pathogens from ticks (n = 392) and fleas (n = 248) collected from companion dogs and cats in East and Southeast Asia. Of the 392 ticks tested, 37 (9.4%) scored positive for at least one pathogen with Hepatozoon canis being the most prevalent (5.4%), followed by Ehrlichia canis (1.8%), Babesia vogeli (1%), Anaplasma platys (0.8%) and Rickettsia spp. (1%) [including Rickettsia sp. (0.5%), Rickettsia asembonensis (0.3%), Rickettsia felis (0.3%)]. Out of 248 fleas tested, 106 (42.7%) were harboring at least one pathogen with R. felis being the most common (19.4%), followed by Bartonella spp. (16.5%), Rickettsia asembonensis (10.9%) and Candidatus Rickettsia senegalensis (0.4%). Furthermore, 35 Rhipicephalus sanguineus sensu lato ticks were subjected to phylogenetic analysis, of which 34 ticks belonged to the tropical and only one belonged to the temperate lineage (Rh. sanguineus sensu stricto). Our data reveals the circulation of different VBPs in ticks and fleas of dogs and cats from Asia, including zoonotic agents, which may represent a potential risk to animal and human health.Author summaryTicks and fleas are among the most important vectors of pathogens infesting many animal species including humans worldwide. Although a number of vector-borne pathogens have been detected in dogs and cats from East and Southeast Asia, investigation in ticks and fleas collected from them are scant. In order to provide an overview of the pathogens circulating in ticks and fleas from companion dogs and cats in Asia, ticks (n = 392) and fleas (n = 248) were collected from privately-owned dogs and cats from China, Taiwan, Indonesia, Malaysia, Singapore, Thailand, Philippines and Vietnam and molecularly screened for the presence of pathogens. Overall, multiple pathogens were found in ticks (i.e., Babesia vogeli, Hepatozoon canis, Ehrlichia canis, Anaplasma platys and Rickettsia spp.) and fleas (i.e., Rickettsia spp. and Bartonella spp.) from the sampling areas. Of the ticks tested, 9.4% scored positive for at least one pathogen and of fleas 42.7% harbored at least one pathogen with Rickettsia felis being the most common (19.4%). Overall, of the detected pathogens, R. felis stood out as the most important due to its zoonotic potential. The result of this study should increase awareness among pet owners and veterinary practitioners regarding the importance of ticks and fleas, and their transmitted pathogens.

scholarly journals Molecular Detection of Pathogens in Ticks and Fleas Collected from Companion Dogs and Cats in East and Southeast Asia

2020 ◽  
Author(s):  
Viet-Linh Nguyen ◽  
Vito Colella ◽  
Grazia Greco ◽  
Fang Fang ◽  
Wisnu Nurcahyo ◽  
...  
Keyword(s):  
Southeast Asia ◽  
Animal Species ◽  
Rhipicephalus Sanguineus ◽  
Sensu Stricto ◽  
Hepatozoon Canis ◽  
Rickettsia Felis ◽  
Companion Dogs ◽  
Babesia Vogeli ◽  
East And Southeast Asia ◽  
Vector Borne

Abstract Background: Ticks and fleas are considered amongst the most important arthropod vectors of medical and veterinary concern due to their ability to transmit pathogens to a range of animal species including dogs, cats and humans. By sharing a common environment with humans, companion animal-associated parasitic arthropods may potentially transmit zoonotic vector-borne pathogens (VBPs). This study aimed to molecularly detect pathogens from ticks and fleas from companion dogs and cats in East and Southeast Asia.Methods: A total of 392 ticks and 248 fleas were collected from 401 infested animals (i.e., 271 dogs and 130 cats) to molecularly screened for the presence of pathogens. Ticks were tested for Rickettsia spp., Anaplasma spp., Ehrlichia spp., Babesia spp. and Hepatozoon spp. while fleas were screened for the presence of Rickettsia spp. and Bartonella spp.Result: Of the 392 ticks tested, 37 (9.4%) scored positive for at least one pathogen with Hepatozoon canis being the most prevalent (5.4%), followed by Ehrlichia canis (1.8%), Babesia vogeli (1%), Anaplasma platys (0.8%) and Rickettsia spp. (1%) [including Rickettsia sp. (0.5%), Rickettsia asembonensis (0.3%), Rickettsia felis (0.3%)]. Out of 248 fleas tested, 106 (42.7%) were harboring at least one pathogen with R. felis being the most common (19.4%), followed by Bartonella spp. (16.5%), Rickettsia asembonensis (10.9%) and Candidatus Rickettsia senegalensis (0.4%). Furthermore, 35 Rhipicephalus sanguineus ticks were subjected to phylogenetic analysis, of which 34 ticks belonged to the tropical and only one belonged to the temperate lineage (Rh. sanguineus sensu stricto). Conclusion: Our data reveals the circulation of different VBPs in ticks and fleas of dogs and cats from Asia, including zoonotic agents, which may represent a potential risk to animal and human health.

scholarly journals Canine Vector-Borne Diseases of Working Dogs of the Sri Lanka Air Force, Free-Roaming, and Privately-Owned Dogs

2021 ◽  
Author(s):  
Rupika Subashini Rajakaruna ◽  
PS Jayathilake ◽  
HSU Wijerathna ◽  
ADS Fernando ◽  
KMH Ginarathne ◽  
...  
Keyword(s):  
Sri Lanka ◽  
Air Force ◽  
Clinical Signs ◽  
Hepatozoon Canis ◽  
Infection Prevalence ◽  
Vector Borne Diseases ◽  
Working Dogs ◽  
Vector Borne ◽  
The Military ◽  
Privately Owned

Abstract Background Canine vector-borne diseases (CVBDs) are a significant health problem among dogs globally. The veterinary records show that the military working dogs of Sri Lanka die early at an average age of six years and CVBDs have been listed as one of the speculated causes. This study examined CVBDs in the working dogs of the Sri Lanka Air Force (SLAF) together with free-roaming and privately-owned dogs living close to the SLAF establishments and country-wide. Methods Blood samples were collected from the cephalic vein for a three year period, from July 2016 to July 2019. Giemsa stained thin blood smears were observed under light microscopy. Results A total of 668 dogs were sampled, of which 169 were infected (25.3%) with hemoparasites. The prevalence of infections among the SLAF working dogs (22.5%), free-roaming (26.3%), or privately-owned (26.2%) dogs did not differ significantly (Chi-square test, p > 0.05). A large number of infected dogs were asymptomatic (57.4%), which was significantly higher in free-roaming dogs (96.3%) compared to the SLAF (41.1%) and privately-owned dogs (38.2%; p < 0.0001) but not between SLAF dogs and privately-owned dogs (χ2 = 0.005, p > 0.938). Seven hemoparasites were identified: Babesia gibsoni, B. canis, Ehrlichia canis, Anaplasma platys, Leishmania sp., Hepatozoon canis, and microfilariae. The most common infection was B. gibsoni (13.8%) followed by E. canis (9.9%) and A. platys (4.3%). There was no difference in the prevalence of B. gibsoni in the three categories of dogs (χ2 = 2.61, p = 0.10). Leishmania and H. canis were always presented as single infections, while microfilaria as mixed infections. Conclusions Even though the SLAF dogs are thoroughly quarantined before introducing them to the military units, the infection prevalence was similar compared to other dog categories. They have likely acquired the infections through ticks and other arthropod vectors. This is the first comprehensive and comparative island-wide study of dog CVBDs of Sri Lanka, especially the military dogs at the SLAF. It highlights that most infected dogs, especially the free-roaming ones, did not show clinical signs. Although it is speculated that they may act as reservoirs of infection, asymptomatic dogs' ability to spread these CVBDS should be investigated.

scholarly journals Molecular survey of vector-borne diseases in two groups of domestic dogs from Lisbon, Portugal

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Ana Mafalda Dordio ◽  
Relja Beck ◽  
Telmo Nunes ◽  
Isabel Pereira da Fonseca ◽  
Jacinto Gomes
Keyword(s):  
Clinical Status ◽  
Molecular Methods ◽  
Hepatozoon Canis ◽  
Clinical Approach ◽  
Limited Information ◽  
Vector Borne Diseases ◽  
Wide Range ◽  
Other Information ◽  
Southern Portugal ◽  
Vector Borne

Abstract Background Canine vector-borne diseases (CVBDs) are caused by a wide range of pathogens transmitted by arthropods. They have been an issue of growing importance in recent years; however, there is limited information about the vector-borne pathogens circulating in Portugal. The aim of the present study was to detect canine vector-borne bacteria and protozoa of veterinary and zoonotic importance using molecular methods. Methods One hundred and forty-two dogs from Lisbon, southern Portugal, were tested: 48 dogs from a veterinary hospital clinically suspected of vector-borne diseases and 94 apparently healthy dogs from shelters. Anaplasma spp./Ehrlichia spp., Babesia/Theileria spp., Hepatozoon spp., and Mycoplasma spp. infections were detected by PCR from blood samples and examined under light microscopy. Other information including clinical status and diagnostic test results were collected for each animal. Results Infections were detected by PCR in 48 (33.80%) dogs. Single infections were found in 35 dogs (24.64%), and co-infections were found in 13 (9.15%) dogs. Twenty-nine (20.42%) dogs were positive for Hepatozoon spp., 15 (10.56%) for Mycoplasma spp., 11 (7.75%) for Anaplasma spp./Ehrlichia spp., and six (4.21%) for Babesia spp. DNA sequencing was used to identify Babesia vogeli (2.81%), Babesia canis (1.40%), Hepatozoon canis (20.42%), Mycoplasma haematoparvum (2.11%), Mycoplasma haemocanis (8.45%), Anaplasma platys (7.04%), and Ehrlichia canis (0.70%). Conclusions This is the first molecular identification of B. canis and M. haematoparvum in dogs from southern Portugal. This study highlights the importance of molecular methods to identify CVBD pathogens in endemic areas and helps to guide the clinical approach of veterinarians in practice.

scholarly journals Virus Adaptation Following Experimental Infection of Chickens with a Domestic Duck Low Pathogenic Avian Influenza Isolate from the 2017 USA H7N9 Outbreak Identifies Polymorphic Mutations in Multiple Gene Segments

Viruses ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 1166
Author(s):  
Klaudia Chrzastek ◽  
Karen Segovia ◽  
Mia Torchetti ◽  
Mary Lee Killian ◽  
Mary Pantin-Jackwood ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Experimental Infection ◽  
Clinical Signs ◽  
Interspecies Transmission ◽  
Clinical Samples ◽  
Domestic Duck ◽  
Receptor Binding Site ◽  
Synonymous Mutations ◽  
Field Samples ◽  
Virus Adaptation

In March 2017, highly pathogenic (HP) and low pathogenic (LP) avian influenza virus (AIV) subtype H7N9 were detected from poultry farms and backyard birds in several states in the southeast United States. Because interspecies transmission is a known mechanism for evolution of AIVs, we sought to characterize infection and transmission of a domestic duck-origin H7N9 LPAIV in chickens and genetically compare the viruses replicating in the chickens to the original H7N9 clinical field samples used as inoculum. The results of the experimental infection demonstrated virus replication and transmission in chickens, with overt clinical signs of disease and shedding through both oral and cloacal routes. Unexpectedly, higher levels of virus shedding were observed in some cloacal swabs. Next generation sequencing (NGS) analysis identified numerous non-synonymous mutations at the consensus level in the polymerase genes (i.e., PA, PB1, and PB2) and the hemagglutinin (HA) receptor binding site in viruses recovered from chickens, indicating possible virus adaptation in the new host. For comparison, NGS analysis of clinical samples obtained from duck specimen collected during the outbreak indicated three polymorphic sides in the M1 segment and a minor population of viruses carrying the D139N (21.4%) substitution in the NS1 segment. Interestingly, at consensus level, A/duck/Alabama (H7N9) had isoleucine at position 105 in NP protein, similar to HPAIV (H7N9) but not to LPAIV (H7N9) isolated from the same 2017 influenza outbreak in the US. Taken together, this work demonstrates that the H7N9 viruses could readily jump between avian species, which may have contributed to the evolution of the virus and its spread in the region.

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