Serological and Molecular Findings of Leishmania Infection in Healthy Donkeys (Equus asinus) from a Canine Leishmaniosis Endemic Focus in Tuscany, Italy: A Preliminary Report

Leishmania parasites are considered to be emergent zoonotic pathogens, which is a new concept regarding their epidemiology and the identification of novel animal hosts. The present study is the first in Italy to evaluate anti Leishmania seroprevalence, and the first in Europe to detect parasite DNA in donkeys’ blood. The study was performed on jennies living in a Leishmania infantum endemic area of Central Italy. One hundred and ten blood samples were obtained from 67 healthy lactating Amiatina jennies that were semi-extensively reared in Tuscany. When possible, more than one sample was subsequently obtained from the same subject. All samples were processed by immunofluorescence antibody test (IFAT) and by polymerase chain reaction (PCR). For the results, 11 out of 30 animals (36.7%) showed positive scores under IFAT. In addition, 22 out of the other 37 jennies had positive scores, also. The animals showed titers ranging from 40 to 320. Furthermore, 2 subjects that were submitted for 2 and 3 blood samplings, both had more than one positive score. Moreover, 2 seropositive animals were positive for Leishmania DNA. Donkeys are considered to be a preferred source for a sandfly blood meal, even if clinical leishmaniosis has never been reported in Europe for this animal species. In the view of these facts, our preliminary findings would suggest the role of donkey as a potential reservoir for this protozoan agent. Additional studies would be welcome to elucidate the role of the donkey in Leishmania epidemiology of CanL endemic areas and to confirm the preliminary findings and the hypothesis proposed here.

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Detecting antibodies to Leishmania infantum in horses from areas with different epizooticity levels of canine leishmaniosis and a retrospective revision of Italian data

Parasites & Vectors ◽

10.1186/s13071-020-04385-8 ◽

2020 ◽

Vol 13 (1) ◽

Author(s):

Alessia Libera Gazzonis ◽

Filippo Bertero ◽

Iolanda Moretta ◽

Giulia Morganti ◽

Michele Mortarino ◽

...

Keyword(s):

Leishmania Infantum ◽

Linear Models ◽

Central Italy ◽

Direct Methods ◽

Antibody Test ◽

Canine Leishmaniosis ◽

Northern Regions ◽

Vector Borne ◽

Visceral Leishmaniosis

Abstract Background Leishmania infantum is a vector-borne pathogen endemic in countries in the Mediterranean basin, including Italy. Dogs act as the primary reservoir for this parasite, but other animal species may also be infected. Low-to-moderate seroprevalence levels of infection have been reported in apparent healthy equine populations in southern Europe, reinforcing the importance of exploring those species, including horses, that act as a food source for vectors and may thus participate in the epizoological scenario of canine leishmaniosis (CanL) and zoonotic visceral leishmaniosis (ZVL). Since little is known regarding the exposure to L. infantum in horses in Italy, we assessed the seroprevalence in healthy equine populations from different CanL endemic areas. Methods The survey was conducted on 660 apparently healthy horses distributed throughout central and northern regions of Italy between 2016 and 2019. Blood samples were collected and the presence of anti-Leishmania antibodies (IgG) was investigated by the immunofluorescence antibody test. Information on the location and altitude of the stables, along with the horses’ breed, age, sex, and reproductive status was obtained by filling in a questionnaire. This was then used for statistical analysis by generalized linear models to explore risk factors associated with seroreactivity to L. infantum. Results An average seroprevalence of 13.9% was detected for L. infantum in the equine populations investigated, with statistically significant associations between seroprevalence, geographical variables (northern vs central Italy, origin and altitude) and individual factors (i.e. age and breed morphotype). Conclusions Our results highlight that horses are frequently exposed to L. infantum. Further prevalence surveys in horses, also using direct methods (e.g. PCR), are warranted to clarify the role of these hosts in the epidemiology of Leishmania in Italy.

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Canine visceral leishmaniosis in an area of fishing tourism, Bonito Municipality, Mato Grosso do Sul, Central-West Brazil

Ciência Rural ◽

10.1590/0103-8478cr20170332 ◽

2018 ◽

Vol 48 (3) ◽

Author(s):

Andreia Fernandes Brilhante ◽

Maria Elizabeth Cavalheiros Dorval ◽

Eunice Aparecida Bianchi Galati ◽

Vivianne de Oliveira Landgraf de Castro ◽

Maria Elizabeth Ghizzi Rocca ◽

...

Keyword(s):

Antibody Test ◽

Diagnostic Techniques ◽

Leishmania Infection ◽

Kappa Index ◽

Serological Tests ◽

Mato Grosso ◽

Polymerase Chain ◽

Visceral Leishmaniosis ◽

Immunofluorescence Antibody Test ◽

Immunoenzymatic Assay

ABSTRACT: The study aimed to investigate Leishmania infection in a population of dogs (92 animals) in a fishing area of Bonito Municipality, and evaluate comparatively the serological methods used, immunoenzymatic assay (ELISA), indirect immunofluorescence antibody test (IFAT), and rapid Dual Path-Platform test (DPP®). Blood and aspirate of bone marrow samples were used and parasitological investigation was also performed, such as parasite isolation in Neal, Novy, Nicolle (NNN) medium culture, Woo technique, Giemsa stained smears and specific identification by polymerase chain reaction (PCR). IFAT revealed 56/92 seropositive, the ELISA 8/92 and the DPP®41/92. Regarding the agreement between the serological tests by the Kappa index, there was a slight agreement between ELISA/IFAT and ELISA/DPP®, and fair agreement in IFAT/DPP®. The NNN culture was positive in three out of 21 dogs, and identified by PCR as Leishmania infantum chagasi. No samples were positive by the Woo technique. Our results showed low agreements between the serological tests recommended by the Brazilian Ministry of Heath, and it is necessary to associate other diagnostic techniques, such as parasitological tests and PCR, to increase the sensitivity for canine visceral leishmaniosis diagnosis, mainly regarding asymptomatic dogs in endemic areas.

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Detection of Neospora caninum Infection in Aborted Equine Fetuses in Israel

Pathogens ◽

10.3390/pathogens9110962 ◽

2020 ◽

Vol 9 (11) ◽

pp. 962

Author(s):

Monica Leszkowicz Mazuz ◽

Lea Mimoun ◽

Gili Schvartz ◽

Sharon Tirosh-Levy ◽

Igor Savitzki ◽

...

Keyword(s):

Neospora Caninum ◽

Fluorescent Antibody ◽

Fetal Loss ◽

Antibody Test ◽

Transplacental Transmission ◽

Caninum Infection ◽

Parasite Detection ◽

Polymerase Chain ◽

Equine Abortion

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

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An outbreak of caprine toxoplasmosis - investigation and case report

Ciência Rural ◽

10.1590/0103-8478cr20170790 ◽

2018 ◽

Vol 48 (5) ◽

Author(s):

José Mauricio Ferreira Neto ◽

Fernanda Pinto Ferreira ◽

Ana Carolina Miura ◽

Jonatas Campos de Almeida ◽

Felippe Danyel Cardoso Martins ◽

...

Keyword(s):

Management Practices ◽

Clinical Signs ◽

Antibody Test ◽

Buffy Coat ◽

Soil Samples ◽

Dairy Goat ◽

Blood Samples ◽

Lactating Goats ◽

Polymerase Chain ◽

Immunofluorescence Antibody Test

ABSTRACT: The present study aimed to investigate an abortion outbreak in a dairy goat herd in the municipality of Arapoti, Parana, Brazil. At the beginning of the outbreak, blood samples were collected from 33 goats with clinical signs; later, of the whole goat herd, two cats and two dogs. Milk samples were collected from 78 lactating goats. Four environmental soil samples and four samples of feed residue from goat feeders were collected too. Immunofluorescence antibody test (IFA) was used for serodiagnosis, the molecular analysis was conducted by means of the polymerase chain reaction (PCR), for the isolation of the etiological agent the bioassay was used. The results of the IFA revealed that 76.53% (137/179) of the goats, two dogs and two cats were seropositive for Toxoplasma gondii. Bioassay revealed one buffy coat and two milk sample having viable T. gondii. In the PCR, 11 whole blood samples, eight milk, three feeder troughs, and all soil samples were positive. The findings of the present study confirmed an outbreak caused by environmental contamination (of soil and feed) with T. gondii oocysts that could have been shed by kittens that lived on the farm and had access to the stock of goat food, facilitating this contamination, which reinforces the need for veterinary assistance and good management practices on farms.

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Application of different techniques to detect Toxoplasma gondii in slaughtered sheep for human consumption

Revista Brasileira de Parasitologia Veterinária ◽

10.1590/s1984-29612015076 ◽

2015 ◽

Vol 24 (4) ◽

pp. 416-421 ◽

Cited By ~ 5

Author(s):

Annelise Castanha Barreto Tenório Nunes ◽

Edna Maria Vieira da Silva ◽

José Aelson de Oliveira ◽

Elise Myuki Yamasaki ◽

Pomy de Cássia Peixoto Kim ◽

...

Keyword(s):

Toxoplasma Gondii ◽

Antibody Test ◽

Human Consumption ◽

Cerebral Tissue ◽

Serum Samples ◽

Kappa Test ◽

Polymerase Chain ◽

Immunofluorescence Antibody Test ◽

Perivascular Cuff ◽

Immunofluorescence Antibody

Abstract The aim of this study was to investigate occurrence of Toxoplasma gondii in sheep slaughtered in the state of Alagoas, Brazil, by means of different diagnosis techniques. Serum samples and tissues from 100 slaughtered sheep were used. To detect antibodies, the indirect immunofluorescence antibody test (IFAT) was used, and tissues from seropositive animals (cut-off ≥1:64) were submitted to Polymerase Chain Reaction (PCR) and immunohistochemistry (IHC). To assess the concordance between the direct techniques, the kappa test was used. In the IFAT, it was observed that 14% (14/100) of the ovine samples were serum-positive. In the PCR, 21.43% (3/14) of the animals were positive and in IHC, it was observed that 7.14% (1/14) were positively stained for T. gondii in cerebral tissue. Histopathologically, the predominant finding was the presence of mononuclear cell infiltrate in the heart and a perivascular cuff in the cerebrum and cerebellum. The concordance between the direct diagnosis techniques was moderate (k=0.44). Thus, it is important to use different direct techniques in diagnosing toxoplasmosis in naturally infected sheep.

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Toxoplasma gondii, Neospora caninum and Leishmania spp. serology and Leishmania spp. PCR in dogs from Pirassununga, SP

Revista Brasileira de Parasitologia Veterinária ◽

10.1590/s1984-29612015046 ◽

2015 ◽

Vol 24 (4) ◽

pp. 454-458 ◽

Cited By ~ 6

Author(s):

Nathália Mendonça de Seabra ◽

Vanessa Figueredo Pereira ◽

Marcos Vinícius Kuwassaki ◽

Julia Cristina Benassi ◽

Trícia Maria Ferreira de Sousa Oliveira

Keyword(s):

Toxoplasma Gondii ◽

Neospora Caninum ◽

Antibody Test ◽

Seropositivity Rate ◽

Three Samples ◽

Leishmania Spp ◽

Polymerase Chain ◽

The City ◽

Immunofluorescence Antibody Test ◽

Immunofluorescence Antibody

Abstract We examined the presence of antibodies against the parasites Toxoplasma gondii, Neospora caninum, and Leishmania spp., as well the presence of DNA from Leishmania spp., in dogs from Pirassununga - SP. The seropositivity rate was compared with the animals’ originating location. Three hundred seventy-three blood samples from the county’s kennel and local veterinary clinics were collected and analyzed. A total of 300 samples were tested for T. gondii and N. caninum using an indirect immunofluorescence antibody test (IFAT); 45% (135/300) were positive for T. gondii and 24.3% (73/300) were positive for N. caninum. Three hundred seventy-three samples were tested for Leishmania spp. using the IFAT. Of these, 4.6% (17/373) were positive. Additionally, 145 samples were tested using a polymerase chain reaction (PCR); of these samples, 0.7% (1/145) was positive. Considering the results, we conclude that these parasites are present in the city of Pirassununga - SP and that the animals have contact with the protozoan. It is therefore necessary to create methods for disease prevention to maintain both animal and human health in regard to leishmaniasis and toxoplasmosis.

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Naturally acquired Lawsonia intracellularis infection in pigs studied from weaning to slaughter by indirect immunofluorescence antibody test and polymerase chain reaction on faeces

Research in Veterinary Science ◽

10.1016/j.rvsc.2004.08.001 ◽

2005 ◽

Vol 79 (2) ◽

pp. 93-98 ◽

Cited By ~ 8

Author(s):

T.K. Jensen ◽

H. Vigre ◽

V. Sørensen ◽

K. Møller

Keyword(s):

Polymerase Chain Reaction ◽

Indirect Immunofluorescence ◽

Antibody Test ◽

Lawsonia Intracellularis ◽

Chain Reaction ◽

Polymerase Chain ◽

Immunofluorescence Antibody Test ◽

Immunofluorescence Antibody

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Transplacental Transmission of Theileria equi Is Not a Common Cause of Abortions and Infection of Foals in Israel

Animals ◽

10.3390/ani10020341 ◽

2020 ◽

Vol 10 (2) ◽

pp. 341 ◽

Cited By ~ 3

Author(s):

Sharon Tirosh-Levy ◽

Yuval Gottlieb ◽

Lea Mimoun ◽

Monica L. Mazuz ◽

Amir Steinman

Keyword(s):

Antibody Test ◽

Transplacental Transmission ◽

Tick Feeding ◽

Theileria Equi ◽

Source Of Infection ◽

Route Of Transmission ◽

Immunofluorescence Antibody Test ◽

High Parasitemia ◽

Immunofluorescence Antibody

Although the main route of transmission of Theileria equi is through tick feeding, transplacental transmission is also possible and may lead to abortion, or to the birth of a sick or carrier foal. The aim of this study was to evaluate the role of T. equi as a cause of abortions in Israel and the risk of foals being infected at a young age. Eight aborting mares were serologically evaluated for exposure to T. equi via the immunofluorescence antibody test (IFAT) and their aborted fetuses were evaluated using PCR and qPCR. In addition, five mares and their foals (aged 4–6 months) from a highly endemic farm were tested for T. equi infection using IFAT, PCR and qPCR. Five of the eight aborting mares were seropositive for T. equi; however, none of the aborted fetuses was infected. All five mares from the endemic farm were subclinically infected with T. equi. Of their five foals, one was infected, with relatively high parasitemia and different parasite genotype than its dam’s, suggesting another source of infection. The results of this study suggest that transplacental transmission of T. equi is not common and does not appear to be a prominent cause of abortion in chronically infected mares.

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Antigenic and genetic characterization of the first rabies virus isolated from the bat Eumops perotis in Brazil

Revista do Instituto de Medicina Tropical de São Paulo ◽

10.1590/s0036-46652008000200006 ◽

2008 ◽

Vol 50 (2) ◽

pp. 95-99 ◽

Cited By ~ 12

Author(s):

Juliana Galera Castilho ◽

Flávia Marchizeli Canello ◽

Karin Corrêa Scheffer ◽

Samira Maria Achkar ◽

Maria Luiza Carrieri ◽

...

Keyword(s):

Rabies Virus ◽

Antibody Test ◽

Nucleoprotein Gene ◽

Adult Mice ◽

Vampire Bats ◽

The City ◽

Immunofluorescence Antibody Test ◽

Immunofluorescence Antibody

Although the main transmitters of rabies in Brazil are dogs and vampire bats, the role of other species such as insectivorous and frugivorous bats deserves special attention, as the rabies virus has been isolated from 36 bat species. This study describes the first isolation of the rabies virus from the insectivorous bat Eumops perotis. The infected animal was found in the city of Ribeirão Preto, São Paulo. The virus was identified by immunofluorescence antibody test (FAT) in central nervous system (CNS) samples, and the isolation was carried out in N2A cell culture and adult mice. The sample was submitted to antigenic typing using a panel of monoclonal antibodies (CDC/Atlanta/USA). The DNA sequence of the nucleoprotein gene located between nucleotides 102 and 1385 was aligned with homologous sequences from GenBank using the CLUSTAL/W method, and the alignment was used to build a neighbor-joining distance-based phylogenetic tree with the K-2-P model. CNS was negative by FAT, and only one mouse died after inoculation with a suspension from the bat's CNS. Antigenic typing gave a result that was not compatible with the patterns defined by the panel. Phylogenetic analysis showed that the virus isolated segregated into the same cluster related to other viruses isolated from insectivorous bats belonging to genus Nyctinomops ssp. (98.8% nucleotide identity with each other).

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