Prevalence and seasonal variations of canine dirofilariosis in Portugal
Open accessAbstract
Dirofilariosis is a severe vector-borne emergent disease that is spreading worldwide and becoming a serious threat to human and veterinary public health. Portugal, a Mediterranean country, has favorable climate conditions for mosquito development and survival. At present, accurate data on the prevalence and epidemiological pattern of dirofilariosis in Portugal is scarce and outdated. To study these trends, a project was developed to assess the current prevalence of Dirofilaria immitis infection in shelter dogs as well as the prevalence of Dirofilaria species present in three coastal regions in central Portugal: Coimbra, Santarém, and Setúbal. Blood samples were collected from 696 shelter dogs during three consecutive years: 2011, 2012, and 2013. A rapid immunomigration technique was performed to detect female D. immitis antigens. Concurrently, to detect and identify circulating microfilariae, a modified Knott's technique and acid phosphatase histochemical staining were also performed. Of the 696 dogs sampled, 105 were positive for D. immitis, with an overall prevalence of 15.1%. Forty of the 105 dogs were antigen negative but were positive for D. immitis microfilariae. Three animals were co-infected with D. immitis and Acanthocheilonema dracunculoides, and there was also one dog infected only with A. dracunculoides, all confirmed by polymerase chain reaction. The highest prevalence of canine heartworm during the 3-y period was in Setúbal (24.8%), followed by Coimbra (13.8%), and Santarém (13.2%), with significant inter-district differences. Our results demonstrate a higher prevalence of dirofilariosis compared with findings of previous studies and show an increasing rate of infection in the southern areas of Portugal attributed, at least in part, to bioclimatic and ecological factors. The present study updates the epidemiological situation and correlates the risk of dirofilariosis transmission within each region. These findings are highly relevant to both human and veterinary public health, contributing to the general awareness of pet owners and veterinarian practitioners and reinforcing the need for effective control measures against vectors and preventive therapy in companion animals.
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TL;DR
In a 3-year survey of 696 shelter dogs in central Portugal, canine Dirofilaria immitis prevalence was 15.1%, with significant regional variation and highest prevalence in Setúbal (24.8%). Transmission was seasonal (May–November) and all districts had suitable temperature conditions for Dirofilaria development.
Key findings
Overall D. immitis prevalence was 15.1% (105/696) across 2011–2013; antigen-positive dogs were 9.4% (65/696), microfilariae by modified Knott’s were 12.9% (90/696), and acid phosphatase staining was 12.1% (84/696). Regional prevalence differed significantly (χ²=8.661; p=0.013): Setúbal 24.8% (25/101), Coimbra 13.8% (31/224), and Santarém 13.2% (49/371); the highest yearly rate was 33.3% in Setúbal in 2013 and the lowest was 11.5% in Santarém in 2012. Infection was higher in males (18.2% [56/307]) than females (12.6% [49/389]; p=0.039) and in dogs ≥5 years (p=0.026), while coat length was not significant (p=0.178). Three dogs were co-infected with D. immitis and Acanthocheilonema dracunculoides, one dog was infected only with A. dracunculoides, and no D. repens or A. reconditum was detected; the temperature-based model estimated 150.7 transmission days/year in Santarém, 150.3 in Setúbal, and 129.7 in Coimbra, with seasonal risk from May to November.
Methods
Cross-sectional survey of 696 shelter dogs in nine animal shelters in Coimbra, Santarém, and Setúbal, sampled in October/November 2011, April/May 2012, and May/June 2013; dogs younger than 6 months were excluded and only dogs admitted in the previous 6 months were included. Diagnostic testing used the WITNESS Dirofilaria rapid immunomigration antigen test, modified Knott’s technique, acid phosphatase histochemical staining (Leucognost SP), and PCR with panfilarial primers DIDR-F1/DIDR-R1 plus sequencing for confirmation. Analysis used SPSS v21 and EpiTools, Pearson chi-square for gender, coat length, and age groups, Agresti-Coull prevalence estimates, and a degree-day model based on Dirofilaria Development Units with a 14°C threshold, 130 cumulative DDUs, and 30-day mosquito life expectancy.
Limitations
The paper states that all sampled dogs were strays from shelters with no routine heartworm preventive treatment and no information on prior preventive treatments, which may limit generalizability and may explain the higher prevalence found. The authors state that the transmission risk model used temperature-based DDUs only and note that other factors such as precipitation, relative humidity, vegetation indices, population density, and socioeconomic status also influence transmission, recommending an integrated approach. Evident from the text: antigen testing followed manufacturer instructions without heat treatment, and the authors discuss antigen–antibody complex formation as a possible cause of false-negative antigen results; acid phosphatase staining was sometimes dubious due to low microfilariae burden or faint blood smears.
Model-written summary. Check it against the paper before relying on it.
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Claims the paper makes
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- Findingown data
The antigen detection test was positive for D. immitis in 9.4% (65/696) of shelter dogs tested over the 3-year period.
“Of the 696 shelter dogs tested over the 3-y period, 9.4% (65/696) were positive for D. immitis based on the antigen detection test (Table 1).”
Quote in the paperSubstantial claim - Findingown data
Dirofilaria was detected in all three districts in each of the years surveyed.
“Dirofilaria was detected in all three districts in each of the years surveyed.”
Quote in the paperNot a substantial claim - Findingown data
Prevalence of D. immitis in Setúbal over the 3-year study was 24.8% (25/101), 95% CI 17.3-34.0%.
“with the highest prevalence found in Setúbal (24.8% [25/101]) [95% CI: 17.3-34.0%]”
Quote in the paperSubstantial claim- Rests on an assumption: The sampled shelter dogs are representative of the shelter dog population in the three districts.
- Rests on an assumption: The same nine animal shelters tested each year are representative of the three districts.
- Findingown data
The lowest infection rate, 11.5%, was recorded in 2012 in the Santarém area.
“and the lowest 11.5% was recorded in 2012, in the Santarém area (Table 1; Fig. 3).”
Quote in the paperNot a substantial claim - Findingown data
In 49 (46.7%) of the 105 D. immitis-infected dogs, antigen testing and acid phosphatase staining both produced positive results.
“In 49 (46.7%) of the 105 D. immitis-infected dogs, antigen testing and acid phosphatase staining both produced positive results.”
Quote in the paperNot a substantial claim
- Findingown data
Microfilariae were detected in blood samples of 12.9% (90/696) dogs by the modified Knott's technique.
“Microfilariae were detected in blood samples of 12.9% (90/696) dogs by the modified Knott's technique.”
Quote in the paperSubstantial claim - Findingown data
The infection rate was higher in males (18.2% [56/307]) than in females (12.6% [49/389]) (χ2 = 4.268; p = 0.039).
“The infection rate was higher in males (18.2% [56/307]) than in females (12.6% [49/389]) ( 2 = 4.268; p = 0.039).”
Quote in the paperNot a substantial claim - Findingown data
The highest infection rate, 33.3%, was recorded in 2013 in the Setúbal area.
“The highest infection rate 33.3% was recorded in 2013, in the Setúbal area”
Quote in the paperNot a substantial claim - Findingown data
Discordant results between antigen testing and acid phosphatase staining were observed in 16 samples (15.2%) due to occult infections, only detectable by antigen testing.
“Discordant results between the two tests were observed in 16 samples (15.2%) due to occult infections, only detectable by antigen testing.”
Quote in the paperNot a substantial claim - Findingown data
Dirofilaria infection was significantly higher in dogs ≥5 y (χ2 = 7.328; p = 0.026).
“Dirofilaria infection was significantly higher in dogs ≥5 y ( 2 = 7.328; p = 0.026).”
Quote in the paperSubstantial claim - Findingown data
Prevalence of D. immitis in Santarém over the 3-year study was 13.2% (49/371), 95% CI 10.1-17.1%.
“and Santarém (13.2% [49/371]) [CI 95%: 10.1-17.1%]”
Quote in the paperNot a substantial claim- Rests on an assumption: The sampled shelter dogs are representative of the shelter dog population in the three districts.
- Rests on an assumption: The same nine animal shelters tested each year are representative of the three districts.
- Findingown data
No other filarids (i.e., D. repens or A. reconditum) were found during the study.
“No other filarids (i.e., D. repens or A. reconditum) were found during the study (Fig. 2).”
Quote in the paperNot a substantial claim - Findingown data
Prevalence of D. immitis in Coimbra over the 3-year study was 13.8% (31/224), 95% CI 9.9-19.0%.
“followed by Coimbra (13.8% [31/224]) [95% CI: 9.9-19.0%]”
Quote in the paperNot a substantial claim- Rests on an assumption: The sampled shelter dogs are representative of the shelter dog population in the three districts.
- Rests on an assumption: The same nine animal shelters tested each year are representative of the three districts.
- Findingown data
Acid phosphatase staining demonstrated D. immitis-specific microfilariae in 12.1% (84/696) of the animals.
“Acid phosphatase staining demonstrated microfilariae with two distinct bright red dots, representing the excretory and anal pores specific for D. immitis, in 12.1% of the animals (84/696) (Fig. 1).”
Quote in the paperNot a substantial claim - Findingown data
Significant differences in Dirofilaria infection rates were observed within gender and age groups.
“Significant differences in Dirofilaria infection rates were observed within gender and age groups.”
Quote in the paperNot a substantial claim - Findingown data
Sequenced samples showed 99% homology with the sequence DQ018785, specific for A. dracunculoides.
“Sequenced samples (reference HG964682-HG964684, European Nucleotide Archive) showed similarities between them and 99% homology with the sequence DQ018785, specific for A. dracunculoides as described by Rishniw et al. (2006).”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites Rishniw et al. (2006) source
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- Findingown data
The overall prevalence of D. immitis infection in 696 shelter dogs over three consecutive years (2011-2013) in three coastal regions of central Portugal was 15.1% (105/696).
“Of the 696 dogs sampled, 105 were positive for D. immitis, with an overall prevalence of 15.1%.”
- Also stated in: § 3. Results · plain
“The overall prevalence of canine D. immitis infection detected over the 3-y study period in the three areas, was 15.1%.”
- Also stated in: § 4. Discussion · plain
“During the 3 y of this survey conducted in Coimbra, Santarém, and Setúbal districts of Portugal, the average global prevalence of canine D. immitis was 15.1%.”
Quote in the paperSubstantial claim- Rests on an assumption: The sampled shelter dogs are representative of the shelter dog population in the three districts.
- Also stated in: § 3. Results · plain
- Findingown data
Significant interdistrict differences of D. immitis infection rates were observed (χ2 = 8.661; p = 0.013).
“Significant interdistrict differences of D. immitis infection rates were observed ( 2 = 8.661; p = 0.013), with the highest prevalence found in Setúbal (24.8% [25/101]) [95% CI: 17.3-34.0%],followed by Coimbra (13.8% [31/224]) [95% CI: 9.9-19.0%],and Santarém (13.2% [49/371]) [CI 95%: 10.1-17.1%].”
Quote in the paperSubstantial claim- Rests on an assumption: The sampled shelter dogs are representative of the shelter dog population in the three districts.
- Rests on an assumption: The same nine animal shelters tested each year are representative of the three districts.
- Findingown data
A 7-month seasonal risk period (May to November) for Dirofilaria transmission was registered in the three meteorological stations.
“A 7-month seasonal risk period (May to November) for Dirofilaria transmission was also registered in the three meteorological stations.”
Quote in the paperSubstantial claim- Rests on an assumption: The temperature data from the three Portuguese meteorological stations are representative of the climatic conditions in the corresponding districts (Coimbra, Santarém, Setúbal).
- Findingown data
All years and all districts showed suitable conditions for Dirofilaria transmission from 2011 to 2013.
“Considering the estimated transmission risk from 2011 to 2013, all years and all districts showed suitable conditions for Dirofilaria transmission.”
Quote in the paperSubstantial claim- Rests on an assumption: The temperature data from the three Portuguese meteorological stations are representative of the climatic conditions in the corresponding districts (Coimbra, Santarém, Setúbal).
- Findingown data
The highest average number of days with conditions for Dirofilaria transmission was registered in Santarém (150.7 days/y, five estimated Dirofilaria generations).
“The highest average number of days with conditions for Dirofilaria transmission was registered in Santarém (150.7 days/y, five estimated Dirofilaria generations);”
Quote in the paperNot a substantial claim- Rests on an assumption: The temperature data from the three Portuguese meteorological stations are representative of the climatic conditions in the corresponding districts (Coimbra, Santarém, Setúbal).
- Findingown data
Dirofilaria transmission is markedly seasonal in Portugal, with peaks in the summer.
“The results obtained by using the forecast model show that Dirofilaria transmission is markedly seasonal in Portugal, with peaks in the summer, confirming data reported by Genchi et al. (2005) and Rinaldi et al. (2007).”
Quote in the paperSubstantial claim0 of 2 sources checked- cites Genchi et al. (2005) source
Not checked in the free run.
- cites Rinaldi et al. (2007)
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- Rests on an assumption: The temperature data from the three Portuguese meteorological stations are representative of the climatic conditions in the corresponding districts (Coimbra, Santarém, Setúbal).
- Findingown data
Prevalence of canine dirofilariosis increased from 15.3% in 2011 to 17.1% in 2013.
“A variation in the prevalence of canine dirofilariosis was observed between years in this survey, with an increase noted from 2011(15.3%) to 2013 (17.1%)”
Quote in the paperSubstantial claim - Findingnone given
Performance of the diagnostic tests revealed higher sensitivity for direct tests (Knott's and acid phosphatase) in combination with PCR compared with serological methods.
“Performance of the diagnostic tests revealed higher sensitivity for direct tests (Knott's and acid phosphatase) in combination with PCR compared with serological methods.”
Quote in the paperSubstantial claim - Study design and conductown data
A total of 696 shelter dogs were randomly selected.
“A total of 696 shelter dogs, including 307 males (44.1%) and 389 females (55.9%), were randomly selected (Table 1).”
Quote in the paperSubstantial claim- Rests on an assumption: The sampled shelter dogs are representative of the shelter dog population in the three districts.
- Premisecitation
A low female worm burden (between one and four) easily decreases the sensitivity of the antigen test.
“Other hypotheses could be a low female worm burden (between one and four) that easily decreases the sensitivity of antigen test, which has been demonstrated in several studies (Martini et al., 1996;Klotins et al., 2000;Atkins, 2003).”
Quote in the paperNot a substantial claim0 of 3 sources checked - Premisecitation
In Portugal, the prevalence of D. immitis in red foxes ranges from 3.23% in north central locations, such as Coimbra, to 11.8% in south central districts, such as Santarém and Setúbal.
“In Portugal, the prevalence of D. immitis in red foxes ranges from 3.23% in north central locations, such as Coimbra (Eira et al., 2006) to 11.8% in south central districts, such as Santarém and Setúbal (Carvalho-Varela and Marcos, 1993;Carvalho-Varela et al., 1993).”
Quote in the paperNot a substantial claim0 of 3 sources checked- cites Eira et al., 2006
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- cites Carvalho-Varela and Marcos, 1993
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- cites Carvalho-Varela et al., 1993
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- Premisecitation
Antigen-antibody complex formation can greatly interfere with the detection of D. immitis antigen.
“The main hypotheses that justify these divergent results could be the antigen-antibody complex formation that can greatly interfere with the detection of the antigen (Brunner et al., 1988;Tonelli and Quentin, 1989;Little et al., 2014).”
Quote in the paperSubstantial claim0 of 3 sources checked - Interpretationcitation
The low rate of filarial infection in Culex theileri (4.76%) suggests the involvement of other Culicidae mosquitoes in dirofilariosis transmission.
“The low rate of filarial infection in this species (4.76%) (Ribeiro et al., 1983) suggests the involvement of other Culicidae mosquitoes in dirofilariosis transmission.”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites Ribeiro et al., 1983
Not checked in the free run.
- Interpretationown data
The prevalence of dirofilariosis found in this study is higher than that reported in previous studies conducted in central Portugal.
“Our results demonstrate a higher prevalence of dirofilariosis compared with findings of previous studies”
- Also stated in: § 4. Discussion · plain
“This prevalence was higher than expected when taking into account findings of previous studies conducted in central Portugal (Araújo, 1996;Balreira et al., 2011;Cardoso et al., 2012).”
Quote in the paperSubstantial claim0 of 3 sources checked- cites Araújo, 1996
Not checked in the free run.
- cites Balreira et al., 2011
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- cites Cardoso et al., 2012 source
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- Rests on an assumption: The prevalence estimates from this study can be compared with previous studies conducted in different dog populations and using different diagnostic methods.
- Also stated in: § 4. Discussion · plain
- Interpretationcitation
The higher prevalence of D. immitis detected in the southern region of Portugal relative to the north may be attributed, at least in part, to bioclimatic and ecological factors.
“The higher prevalence detected in the southern region of Portugal relative to the north may be attributed, at least in part, to bioclimatic and ecological factors, since average temperatures tend to be higher in the southern areas, thus providing more favorable conditions for the vectors (Casimiro et al., 2006).”
Quote in the paperSubstantial claim0 of 1 sources checked- cites Casimiro et al., 2006 source
Not checked in the free run.
- Interpretationcitation
Climate changes related to global warming are most likely implicated in the emergence/re-emergence of canine heartworm disease in Portugal.
“Reasons for the emergence/re-emergence of canine heartworm disease in Portugal are still unclear, but climate changes related to global warming are most likely implicated (Casimiro et al., 2006) as well as globalization that has led to an increased movement of pets (Colwell et al., 2011).”
Quote in the paperNot a substantial claim0 of 2 sources checked - Interpretationown data
At least during the period covered by the present survey, D. immitis appeared to be the sole etiological agent of dirofilariosis in Portugal.
“At least during the period covered by the present survey, D. immitis appeared to be the sole etiological agent of dirofilariosis in Portugal.”
Quote in the paperSubstantial claim
- May combine two claims. 17
- Classified as probably not a substantial claim. 9
- The quote only partly states this claim. 1
May combine two claims.
Findingown dataDiscordant results were found in 41 samples (38.7%), with positive results detected by acid phosphatase staining, which were later confirmed by PCR analysis.
“Similarly, discordant results were found in 41 samples (38.7%), with positive results detected by acid phosphatase staining, which were later confirmed by PCR analysis (Table 1).”
Quote in the paperNot a substantial claimMay combine two claims.
Findingown dataInfected animals were identified in all age groups: <5 y = 11.7% (44/376); 5-10 y = 19.2% (48/250); and >10 y = 18.6% (13/70).
“Infected animals were identified in all age groups: <5 y = 11.7% (44/376); 5-10 y = 19.2%(48/250), and [>10 y old = 18.6% (13/70)].”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataShort-haired dogs were more frequently infected (16.7%; 65/389) than long-haired dogs (13.0%; 40/307), but the difference was not statistically significant (χ2 = 1.814; p = 0.178).
“Additionally, short-haired dogs were more frequently infected (16.7%; 65/389) than long-haired (13.0%; 40/307), although the length of dog coat was not a statistically significant factor ( 2 = 1.814; p = 0.178).”
Quote in the paperNot a substantial claimMay combine two claims.
Findingown dataOf the 41 microfilaria-positive antigen-negative samples, 37 were identified as single infection with D. immitis, one as single infection with A. dracunculoides, and three were A. dracunculoides and D. immitis coinfections.
“Of these 41 samples, 37 were identified as single infection with D. immitis, one as single infection with A. dracunculoides; the remaining three were A. dracunculoides and D. immitis coinfections.”
Quote in the paperNot a substantial claimMay combine two claims.
Findingown dataInfected animals were found in eight of the nine shelters assessed (88.9%), with prevalence ranging from 5.8% to 31.9% in each shelter.
“Infected animals were found in eight of the nine shelters assessed (88.9%) with prevalence ranging from 5.8% to 31.9% in each shelter.”
Quote in the paperNot a substantial claimMay combine two claims.
Findingown dataThe earliest and latest period for Dirofilaria transmission during the 3 years ranged from May 21 to November 14 in Santarém, from May 20 to November 6 in Setúbal, and from May 29 to November 2 in Coimbra.
“The earliest and latest period for Dirofilaria transmission registered during the 3 y ranged from May 21st to November 14th in Santarém, from May 20th to November 6th in Setúbal, and from May 29th to November 2nd in Coimbra.”
Quote in the paperSubstantial claim- Rests on an assumption: The temperature data from the three Portuguese meteorological stations are representative of the climatic conditions in the corresponding districts (Coimbra, Santarém, Setúbal).
May combine two claims.
Findingown dataThe average number of days with conditions for Dirofilaria transmission in Coimbra was 129.7 days/y, with four estimated Dirofilaria generations.
“and Coimbra (129.7 days/y, four estimated Dirofilaria generations).”
Quote in the paperNot a substantial claim- Rests on an assumption: The temperature data from the three Portuguese meteorological stations are representative of the climatic conditions in the corresponding districts (Coimbra, Santarém, Setúbal).
May combine two claims.
Findingown dataThe average number of days with conditions for Dirofilaria transmission in Setúbal was 150.3 days/y, with five estimated Dirofilaria generations.
“followed by Setúbal (150.3 days/y, five estimated Dirofilaria generations);”
Quote in the paperNot a substantial claim- Rests on an assumption: The temperature data from the three Portuguese meteorological stations are representative of the climatic conditions in the corresponding districts (Coimbra, Santarém, Setúbal).
May combine two claims.
Findingown dataThe maximum number of potential transmission days/y (n = 169.7) occurred in 2011, followed by 2013 (n = 133.3) and 2012 (n = 132.7).
“The maximum number of potential transmission days/y (n = 169.7)occurred in 2011, followed by 2013 (n = 133.3)and 2012 (n = 132.7).”
Quote in the paperNot a substantial claim- Rests on an assumption: The temperature data from the three Portuguese meteorological stations are representative of the climatic conditions in the corresponding districts (Coimbra, Santarém, Setúbal).
May combine two claims.
Interpretationown dataDirofilaria infection is increasing and expanding in Portugal.
“suggesting that Dirofilaria infection is increasing and expanding in Portugal.”
Quote in the paperSubstantial claim- Rests on an assumption: The prevalence estimates from this study can be compared with previous studies conducted in different dog populations and using different diagnostic methods.
May combine two claims.
Premisenone givenCanine dirofilariosis is endemic in mainland Portugal as well as in Madeira Island.
“Canine dirofilariosis is endemic in mainland Portugal as well as in Madeira Island.”
Quote in the paperSubstantial claimMay combine two claims.
Interpretationown dataD. immitis infection is endemic and continuously transmitted in the three districts studied (Coimbra, Santarém, and Setúbal) from 2011 to 2013.
“D. immitis was detected in all three districts studied from 2011 to 2013, confirming the endemicity and continuous transmission of this life-threatening disease.”
Quote in the paperSubstantial claimClassified as probably not a substantial claim.
Study design and conductnone givenThe number of animals required was estimated using WinEpi program (stratified sample approach), taking into account the global prevalence of canine dirofilariosis in each district and the average number of animals in the shelters.
“The number of animals required was estimated using WinEpi program (stratified sample approach), taking into account the global prevalence of canine dirofilariosis in each district as well as the average number of animals in the shelters.”
Quote in the paperSubstantial claimMay combine two claims.
PremisecitationBased exclusively on microfilariae detection, without species differentiation, prevalence rates of 16.7% in Ribatejo, 16.5% in Alentejo, 12% in Algarve, and 30% in Madeira were found.
“Based exclusively on microfilariae detection, without species differentiation, prevalence rates of 16.7% in Ribatejo, 16.5% in Alentejo, 12% in Algarve, and 30% in Madeira were found (Araújo, 1996).”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites Araújo, 1996
Not checked in the free run.
Classified as probably not a substantial claim.
Study design and conductreasoningOnly dogs admitted to the kennels in the previous 6 months were included in the survey to preclude evaluating the same animal in subsequent years.
“Only dogs admitted to the kennels in the previous 6 months were included in the survey to preclude evaluating the same animal in subsequent years.”
Quote in the paperSubstantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
PremisecitationD. immitis also has been reported in three European otters Lutra lutra in Portuguese natural freshwater ecosystems.
“Additionally, D. immitis also has been reported in three European otters Lutra lutra in Portuguese natural freshwater ecosystems (Torres et al., 2004;Saraiva et al., 2012).”
Quote in the paperNot a substantial claim0 of 2 sources checked- cites Torres et al., 2004
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- cites Saraiva et al., 2012
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May combine two claims. The quote only partly states this claim.
InterpretationcitationThe lower sensitivity of the antigen test was unexpected, although similar findings have been reported by Tarello (2001) and Vezzani et al. (2008).
“The lower sensitivity of the antigen test was unexpected, although similar findings have also been reported by Tarello (2001), who found seven dogs in Italy harboring D. immitis microfilariae that tested negative for D. immitis antigen.”
Quote in the paperNot a substantial claim0 of 2 sources checked- cites Tarello (2001)
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- cites Vezzani et al. (2008)
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Classified as probably not a substantial claim.
Study design and conductreasoningAll microfilaremic samples that either tested positive for D. immitis or were in doubt by the modified Knott's technique and acid phosphatase staining but were negative for D. immitis circulating antigens were subjected to molecular techniques to confirm diagnosis.
“Some microfilaremic samples that either tested positive for D. immitis or were in doubt by the modified Knott's technique and acid phosphatase staining were negative for D. immitis circulating antigens.For this reason and to ensure an accurate analysis, all of these samples were subjected to molecular techniques to confirm its diagnosis.”
Quote in the paperSubstantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
PremisecitationIn Portugal, only Culex theileri was found to be naturally infected with Dirofilaria spp. larvae.
“In Portugal, only Culex theileri was found to be naturally infected with Dirofilaria spp.larvae.”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites Ribeiro et al., 1983
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Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.
PremisecitationRecent data from Cardoso et al. (2012) reported the national average prevalence of D. immitis in apparently healthy dogs to be 3.6% and 8.9% in dogs suspected to have clinical signs associated with canine vector-borne disease.
“Recent data from Cardoso et al. (2012) using a commercial ELISA kit (SNAP ® 4Dx ® , IDEXX Laboratories, Inc., Westbrook, Maine, USA) reported the national average prevalence of D. immitis in apparently healthy dogs to be 3.6% and 8.9% in dogs suspected to have clinical signs associated with canine vector-borne disease.”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites Cardoso et al. (2012) source
Not checked in the free run.
Classified as probably not a substantial claim.
Study design and conductreasoningDogs younger than 6 months were excluded due to the long life cycle of D. immitis.
“Dogs younger than 6 months were excluded due to the long life cycle of D. immitis.”
Quote in the paperSubstantial claimProbably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.
PremisecitationIn 2011, Balreira et al. reported a 2.1% seroprevalence of canine Dirofilaria infection in northern and north central Portugal, with higher rates in Aveiro (6.8%) and Coimbra (8.8%).
“In 2011, Balreira et al. reported a 2.1% seroprevalence of canine Dirofilaria infection in northern and north central Portugal, with higher rates in Aveiro (6.8%) and Coimbra (8.8%).”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites Balreira et al.
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Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.
InterpretationcitationOccult infections may be due to several factors, such as prepatency, unisex infection (by female worms), drug-induced sterility of adult filariae, or immune-mediated clearance of microfilariae.
“According to Genchi et al. (2007), occult infections may be due to several factors, such as prepatency, unisex infection (by female worms), drug-induced sterility of adult filariae, or immune-mediated clearance of microfilariae.”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites Genchi et al. (2007)
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The paper relies on this without support.
There are no published data on seasonal dirofilariosis transmission trends in Portugal.
“There are no published data on seasonal dirofilariosis transmission trends in Portugal.”
The paper relies on this without support.
The temperature data from the three Portuguese meteorological stations are representative of the climatic conditions in the corresponding districts (Coimbra, Santarém, Setúbal).
“Minimum and maximum daily temperatures registered in the three respective Portuguese meteorological stations were obtained from Instituto Português do Mar e da Atmosfera (IPMA).”
The paper relies on this without support.
The sampled shelter dogs are representative of the shelter dog population in the three districts.
“A total of 696 shelter dogs, including 307 males (44.1%) and 389 females (55.9%), were randomly selected (Table 1).”
The paper relies on this without support.
The prevalence estimates from this study can be compared with previous studies conducted in different dog populations and using different diagnostic methods.
“This prevalence was higher than expected when taking into account findings of previous studies conducted in central Portugal (Araújo, 1996;Balreira et al., 2011;Cardoso et al., 2012).”
The paper relies on this without support.
The same nine animal shelters tested each year are representative of the three districts.
“The same nine animal shelters were tested each year (October/November 2011, April/May 2012, May/June 2013) to determine trends in the prevalence of Dirofilaria spp. in these regions.”
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
In Portugal, the prevalence of canine heartworm reported in the past has remained at similar percentages and has even increased in some regions.
“In Portugal, the prevalence reported in the past has remained at similar percentages and has even increased in some regions.”
From What has happened to heartworm disease in Europe in the last 10 years? (2022) · cites this paper as [12]
In central coastal regions of Portugal, canine heartworm prevalence has been reported at 15.1%, while in the central region it ranged from 8.8 to 27.3%.
“Other studies have reported prevalence of 15.1% in the central coastal regions of Portugal, while in the central region, prevalence ranged from 8.8 to 27.3% [12,13,14]”
From What has happened to heartworm disease in Europe in the last 10 years? (2022) · cites this paper as [12]
In neighbouring Portugal, canine dirofilariosis is endemic with an overall national seroprevalence of 2.1%-15.1%.
“In neighbouring Portugal, canine dirofilariosis is endemic with an overall national seroprevalence of 2.1%- 15.1% ( Alho et al., 2014 and Vieira et al., 2015).”
From Culex pipiens as a potential vector for transmission of Dirofilaria immitis and other unclassified Filarioidea in Southwest Spain (2016) · cites this paper as Alho et al., 2014
Rice field abundance was acknowledged as one of the possible elements contributing to the higher Dirofilaria infection rates in certain areas of Portugal.
“Rice field abundance was acknowledged as one of the possible elements contributing to the higher Dirofilaria infection rates in certain areas of Portugal [24].”
From Dirofilaria infections in dogs in different areas of Greece (2016) · cites this paper as [24]
Canine dirofilariosis due to D. immitis is known to be endemic and widely distributed in Portugal, with prevalence ranging between 0.9 and 27.3% in mainland regions to over 30% in Madeira Island.
“Canine dirofilariosis due to D. immitis is known to be endemic and widely distributed in Portugal, with prevalence ranging between 0.9 and 27.3% in mainland regions to over 30% in Madeira Island [22–25]”
From Molecular characterization of Dirofilaria spp. circulating in Portugal (2017) · cites this paper as [25]
Molecular results based on ITS2-PCR also confirmed previous findings of D. immitis infection in dogs related to sex, age, regional distribution and prevalence.
“Molecular results based on ITS2-PCR also confirmed previous findings of D. immitis infection in dogs related to sex, age, regional distribution and prevalence [25]”
From Molecular characterization of Dirofilaria spp. circulating in Portugal (2017) · cites this paper as [25]
In 2011-2013, the overall parasitological prevalence of canine dirofilariasis in three districts of Centre-South Portugal was 15.1%.
“A recent survey, 2011–2013, in three districts of Centre-South, has revealed an overall parasitological prevalence rate of 15.1%, the highest in Setúbal (24.8%), followed by Coimbra (13.8%) and Santarém (13.2%) [24].”
From First molecular identification of mosquito vectors of Dirofilaria immitis in continental Portugal (2015) · cites this paper as [24]
D. repens was not found in parallel animal surveys in the same districts of Portugal.
“D. repens was not found in parallel animal surveys in the same districts of Portugal [24]”
From First molecular identification of mosquito vectors of Dirofilaria immitis in continental Portugal (2015) · cites this paper as [24]
Coimbra and Santarém had similar prevalence rates of canine dirofilariasis.
“which had similar prevalence rates of CD.”
From First molecular identification of mosquito vectors of Dirofilaria immitis in continental Portugal (2015) · cites this paper as [24]
Setúbal had the highest canine dirofilariasis prevalence rate.
“Setúbal, which had the highest CD prevalence rate”
From First molecular identification of mosquito vectors of Dirofilaria immitis in continental Portugal (2015) · cites this paper as [24]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
Acid phosphatase-positive groups exceed the stated total
MinorThe results describe 49 dogs positive on both antigen testing and acid phosphatase staining and 41 discordant dogs positive on acid phosphatase staining. These disjoint groups total 90, exceeding the stated 84 animals with positive acid phosphatase staining.
Reported 84 · recomputed 90
“Acid phosphatase staining demonstrated microfilariae with two distinct bright red dots, representing the excretory and anal pores specific for D. immitis, in 12.1% of the animals (84/696) (Fig. 1).”
The citation lists haven’t been fetched yet
References 29 · Cited by 45
Research works printed but not linked: 7
- Araújo, 1996, Canine and human Dirofilaria immitis infections in Portugal. A review, Parassitologia, 38, 366
- Carvalho-Varela, 1993, A helmintofauna da raposa (Vulpes vulpes silacea Miller, 1907) in Portugal, Acta Parasitol. Port., 1, 73
- Carvalho-Varela, 1993, Some ecological aspects of the helminthic fauna of the red fox (Vulpes vulpes L.) of the Palearctic Zone. II – Vulpine Iberian populations, Acta Parasitol. Port., 1, 81
- Gomes, 2003, Acta Parasitol. Port., 10, 10
- Guerrero, 1989, Estudio de la incidencia de la enfermedad del gusano del corazón en la población canina Española, Med. Vet., 6, 217
- Ribeiro, 1983, Contribuição para o Estudo dos Vectores das Filaríases Animais em Portugal, J. Soc. Cienc. Med., 147, 143
- Tarello, 2001, Importance in the dog of concentration tests for the diagnosis of heartworm disease in non-endemic areas, Vet On-Line, 2
- Authors
- Alho, A. M., Landum, M., Ferreira, C., Meireles, J., Gonçalves, L., de Carvalho, L. M., & Belo, S.
- Affiliations
- University of Lisbon (PT)
- Universidade Nova de Lisboa (PT)
- Corresponding author
- Alho, Ana Margarida
- ORCID
- Venue
- Veterinary Parasitology
- Publisher
- Elsevier BV
- Published
- 2014-11
- Volume
- 206
- Issue
- 1-2
- Pages
- 99-105
- ISSN
- 0304-4017 (ISSN-L)
- OpenAlex
- W1966437261
- Semantic Scholar
- e6d90cdb2bf712af59a831ce5301c74211d6f7b2
- Cited by
- 45
- References
- 29
- Citations by year
- 2026: 1 · 2025: 2 · 2024: 2 · 2023: 4 · 2022: 4 · 2021: 3 · 2020: 4 · 2019: 6 · 2018: 3 · 2017: 4 · 2016: 10 · 2015: 2
- FWCI
- 3.74
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Veterinary Parasitology (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Funders
- Fundação para a Ciência e Tecnologia (FCT) (SFRH/BD/85427/2012)
- Topics
- Health Sciences › Medicine › Infectious Diseases › Parasitic Diseases Research and Treatment
- Health Sciences › Veterinary › Small Animals › Helminth infection and control
- Health Sciences › Medicine › Public Health, Environmental and Occupational Health › Research on Leishmaniasis Studies
- Keywords
- canine dirofilariasis, Dirofilaria immitis, heartworm disease, prevalence, seasonal variation, shelter dogs, microfilariae, antigen detection, co-infection, vector-borne diseases, Portugal, coastal areas, bioclimatic factors
- MeSH terms
- Animals, Dirofilaria immitis, Dirofilariasis, Dog Diseases, Dogs, Female, Male, Portugal, Seasons, Prevalence
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Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
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Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-09.
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