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Comparison of prevalence of infectious agents detected by PCR in cats from Cyprus per categorical variable
Source publication
Background
Feline infectious agent studies are lacking in Cyprus. The aims of this study were to determine the prevalence and risk factors for various feline infectious agents, including feline vector-borne pathogens (FVBP), in cats from Cyprus.
Methods
A cross-sectional, descriptive, multicentre study was performed on 174 feline samples [138 owne...
Contexts in source publication
Context 1
... age of these 174 cats ranged from 0.4 to 22.0 years (median 5.6 years, interquartile range 8 years) and only 15 (8.6%) were pedigree including six Ragdolls, six Persians, two Siamese and one Russian Blue. Tables 2 and 3 show de- scriptive statistics as well as data on the prevalence of infectious agents among the population studied.th=tlb= Specificity testing for the novel PCR assay for Hepatozoon spp. ...
Context 2
... cats (26.4%) were positive for haemoplasmas, including 13 (7.5%) for Mhf, 36 (20.7%) for CMhm and 12 (6.9%) for CMt (Table 2). Sixty-six cats (37.9%) were positive for Hepatozoon spp., while nineteen (10.9%) were positive for B. henselae. ...
Context 3
... analysis showed that many variables had a trend toward significance (P < 0.2) for association with the presence of individual, or groups of infectious agent(s) ( Table 5, Table 6, Additional file 2: Table S1, Additional file 3: Table S2), and these were entered into the multivariable logistic regression analysis, together with variables having significant associations (P ≤ 0.05). ...
Similar publications
Owned, free-roaming domestic cats are abundant in the Chilean countryside, having high probability of contact with wildlife and potentially participating as reservoirs of zoonotic pathogens. In the present study, 131 cats from two remote study areas (Valdivia and Chiloe Island) in southern Chile were analyzed for infection/exposure to eight pathoge...
Citations
... However, in recent years, more importance has been attributed to feline TBDs, especially concerning their epidemiology, clinical aspects, and control [7,14,47,49,52]. Surveys have shown that cats are often positive for pathogens transmitted by ticks in various geographical regions [2,3,20,41,54,55] and, among them, Hepatozoon spp. have been repeatedly reported in domestic and wild felids in Europe and elsewhere. ...
... In analogy to other Hepatozoon species, H. felis is likely transmitted only by ticks, while alternative modes of transmission (i.e., vertical infections) should also be considered [5,41]. Studies have shown that hepatozoonosis may occur in different environments, e.g., from highly urbanized settings [20], to dry, wetland, or forested areas [2,41], as in Brazil [9]. This variability is not surprising considering that Rhipicephalus sanguineus sensu lato (s.l.), i.e., the most distributed tick in the world for its capability to live and reproduce in several environments [16,19,33,61], is probably one of the main vectors of H. felis [8,54]. ...
In the last few years, the number of studies on feline hepatozoonosis has increased, but our knowledge on the actual species of Hepatozoon and/or different genotypes affecting felines is still incipient. At least three species, namely Hepatozoon felis, H. canis, and H. silvestris, have been isolated from domestic cats in various countries. Additionally, there are indications that other species and genotypes may affect felines in given geographic areas. This study was carried out to investigate the occurrence of Hepatozoon spp. in cats from Niterói, a municipality within the metropolitan area of Rio de Janeiro, Brazil. Individual blood samples were collected from 28 cats enrolled in a spaying/castration program. DNA was extracted from all samples and subjected to sequencing specific for Hepatozoon spp. DNA of H. felis was found in 21/28 cats (75%), and four genetic polymorphisms never described thus far were detected. This is the first report of H. felis in cats living in the State of Rio de Janeiro, and the present data confirm that H. felis is a species complex encompassing different genotypes circulating within cat populations. Further studies are warranted to investigate whether different genotypes have different biology or pathogenicity for felids.
... Map was created utilizing mapchart.net. Prevalence is derived from the following manuscripts: Albania [48], Australia [22,49], Brazil [40,[50][51][52][53][54][55], Canada (British Columbia [56], Ontario [18], Saskatchewan [57], Prince Edward Island [58]), Chile [59], China [60], Cyprus [61], Denmark [62], Ecuador [63], Egypt [64], Germany [17,65], Greece [66,67], Iran [68], Ireland [69], Italy [25,45,[70][71][72], Kenya [29] Japan [46,73,74], New Zealand [75], Portugal [32,76], Qatar [77], Romania [78], Russia [79], Saudi Arabia [80], South Africa [81], South Korea [82], Spain [40,83,84], Switzerland [19], Tanzania [28], Thailand [28,[85][86][87], Turkey [88,89], Trinidad and Tobago [90], England [91], USA (Arizona [92], California, Colorado, and Florida [93], Louisiana [94], and Virginia and North Carolina [23]), Scotland [95], and Singapore [93] or average number of fleas per host on reported hemoplasma prevalence. Experimental work completed in this manuscript is limited by the observational nature with a clear need for experimental transmission studies to fully understand the extent of flea involvement in hemoplasma transmission. ...
Background
Feline-associated hemotropic Mycoplasma (hemoplasmas) are believed to be transmitted by two primary mechanisms: (1) direct transmission via fighting and (2) vector-borne transmission by the cat flea (Ctenocephalides felis). While the efficiency of transmission by C. felis appears low, most manuscripts focus on the prevalence of hemoplasmas in wild-caught fleas and report either a very low (< 3%) or a high (> 26%) prevalence. Therefore, we aimed to assess the influence of sample processing and PCR methods on C. felis hemoplasma infection prevalence.
Methods
A systemic review of PubMed articles identified 13 manuscripts (1,531 fleas/flea pools) that met the inclusion criteria (performed PCR for >1 hemoplasma on C. felis collected from cats). Risk of bias was assessed utilizing the ROBINS-E tool. Meta-analysis performed in R of these manuscripts found that not washing samples and a common set of 16S rRNA primers first published in Jensen et al. 2001 were associated with increased hemoplasma prevalence. To evaluate the influence of washing on newly collected fleas, we assessed the hemoplasma status of 20 pools of 5 C. felis each, half of which were washed and half not washed.
Results
Flea washing did not influence the detection of hemoplasma but instead amplified Spiroplasma. To assess non-specific amplification with the Jensen et al. 2001 primers, 67 C. felis samples (34% previously reported hemoplasma infected) were subject to PCR and sequencing. By this method, hemoplasma was detected in only 3% of samples. In the remaining “hemoplasma infected” fleas, PCR amplified Spiroplasma or other bacteria.
Conclusions
Therefore, we concluded that hemoplasma infection in C. felis is rare, and future flea prevalence studies should sequence all positive amplicons to validate PCR specificity. Further investigation of alternative methods of feline-associated hemoplasma transmission and the ability of C. felis to maintain hemoplasma infection is necessary.
Graphical Abstract
... The pathogen, is transmitted by Dermacentor variabilis and the brown dog tick Rhipicephalus sanguineus [5]. The clinical signs and severity of the disease depend upon the bacterial strain, the host immunity, and co-infection with other pathogens such as Babesia canis, Rickettsia rickettsii, and Hepatozoon canis [6,7]. CME has an acute subclinical and chronic phase. ...
Introduction: Canine monocytic ehrlichiosis (CME) is a disease caused by the Gram-negative bacteria Ehrlichia canis, a bacterium that affects domestic dogs but can also infect humans. The diagnosis implies a challenge due to its diversity in clinical manifestations. Methodology: The frequency of E. canis infection, risk factors, and clinical-pathological parameters associated with seropositivity were calculated with the PROC FREQ TABLES and PROC LOGISTIC procedures of the SAS statistical software. Results: The study showed a seroprevalence of 26.62% (156/586). Association between seropositivity and risk factors was found. The age and the presence of ticks including clinical signs such as anorexia, seizures, cough, petechiae, epistaxis, and hematochezia, as well as multiple blood and biochemical alterations were analyzed. The logistic regression analysis showed a high predictive power (c = 0.98) for CME for thrombocytopenia, leukopenia, and anemia. Conclusions: The high prevalence of E. canis in endemic areas makes its diagnosis difficult. Thus, clinical signs must be considered, along with blood and biochemical alterations, as a possible predictor of the disease.
... However, higher haemotropic Mycoplasma spp. prevalence has been reported in Cyprus (26.4%) [40] and Portugal (27.1%) [41]. Haemotropic Mycoplasma sp. are frequent in European cats, with slight differences within the countries. ...
Background: In Europe, feline vector-borne infections are gaining importance due to the changing climate, expanding habitats of potential vectors, and expanding pathogen reservoirs. The main objective of this study was to assess the prevalence of vector-borne pathogens (VBPs) in stray cats in Zaragoza, Spain, and to investigate potential risk factors for infection, including feline leukaemia virus
(FeLV) and feline immunodeficiency virus (FIV).
Methods: Blood samples from stray cats presented to the veterinary faculty in Zaragoza between
February 2020 and 2022 were tested by polymerase chain reaction (PCR) for the presence of Anaplasma phagocytophilum, Anaplasma platys, Bartonella henselae, Ehrlichia canis, Rickettsia spp., haemotropic Mycoplasma spp., Hepatozoon spp., Leishmania infantum, piroplasms, and microfilariae at the LABOKLIN laboratory. The cats were also tested for FeLV and FIV by PCR.
Results: Nearly half of the cats (158/332, 47.6%) were positive for at least one VBP. Hepatozoon spp. were detected in 25.6%, haemotropic Mycoplasma spp. in 22.9%, B. henselae in 9.3%, and L. infantum in 2.1% of the cats. Male sex had a statistically significant association with test results for haemotropic Mycoplasma spp. (odds ratio 1.38 [1.21;1.57]); regionality with Hepatozoon spp., B. henseale, and FIV; and seasonality with Hepatozoon spp., haemotropic Mycoplasma spp., L. infantum, and FeLV (P ≤ 0.05 each). A strong positive correlation was reported for the amount of
rainfall and the number of cats that tested positive for Hepatozoon spp. (ρ = 753, P = 0.05). None of the cats tested positive for A. phagocytophilum, A. platys, Ehrlichia canis, Rickettsia spp., piroplasms,
or microfilariae. Co-infections with multiple VBPs were detected in 56 out of 332 cats (16.9%). Thirty- one of the 332 cats included in the study (9.3%) tested positive for FeLV (6.9%) and for FIV (3.6%). In 20/31 cats (64.5%) that tested positive for FeLV/FIV, coinfections with VBP were detected (P = 0.048, OR 2.15 [0.99; 4.64]).
Conclusions: VBPs were frequently detected in stray cats in Zaragoza. In particular, regionality and seasonality had a statistically significant association with PCR results for most VBPs included in the study.
... In particular, three Hepatozoon species have been described and reported in wild and domestic felids in Europe: H. felis, H. silvestris, and H. canis. Hepatozoon infections are widespread and have been reported in Spain (3), France (4,21), Portugal (22), Cyprus (23), Germany (24), Austria (25), Greece (26), and Italy (15,27,28). ...
... The literature has described conventional polymerase chain reaction (cPCR) assays for Cytauxzoon and Hepatozoon detection, usually targeting 18S rRNA, a highly conserved gene for both protozoa (8,23,(33)(34)(35)(36)(37)(38), and cytochrome B and cytochrome C oxidase subunit I (COI), mitochondrial protein-coding genes, which are more specific for Cytauxzoon species identification (10). Real-time polymerase chain reaction (real-time PCR) may be a reasonable alternative for the rapid screening of large numbers of samples. ...
Tick-transmitted apicomplexans of the genera Cytauxzoon and Hepatozoon affect a wide range of felids worldwide, but little is known about them. Recently, several studies addressed the species circulating in Europe, their distribution, and their hosts. Molecular assays are the method of choice for their detection. Unfortunately, conventional PCRs already described are time- and cost-consuming and specific for either Hepatozoon or Cytauxzoon detection. This study was developed to evaluate (i) the occurrence of Cytauxzoon and Hepatozoon in felids using a fast and cost-saving real-time PCR capable of detecting both protozoa simultaneously, (ii) the distribution of Cytauxzoon and Hepatozoon species in north-eastern Italy, and (iii) the involvement of other susceptible felid hosts in the same area. An SYBR® Green-based real-time PCR with primers targeting the 18S-rRNA was validated and applied to 237 felid samples, i.e., whole blood from 206 domestic cats and 12 captive exotic felids, and tissues from 19 wildcats. Positive results were obtained by melting temperature curve analysis due to the specific melting peak (i.e., 81°C Cytauxzoon spp.; 78–78.5°C Hepatozoon spp.). Positive samples were subjected to conventional PCR, followed by sequencing for species identification. Phylogenetic analyses were performed to assess relatedness among European isolates. Data on domestic cats (age class, sex, origin, management, and lifestyle) were recorded, and statistical analyses were performed to identify potential risk factors. A total of 31 (15%) domestic cats were positive for Hepatozoon spp. (i.e., 12 for H. felis, 19 for H. silvestris), while six (2.9%) for C. europaeus. The prevalence of Hepatozoon felis was significantly (p < 0.05) higher in domestic cats, while H. silvestris was higher in strays and animals from the Eastern region (i.e., Friuli-Venezia Giulia). Cytauxzoon europaeus was detected only in stray cats from Friuli-Venezia Giulia (province of Trieste). Among captive felids, one tiger was infected with H. felis and another with H. silvestris; eight out of 19 (42%) wildcats were positive for Hepatozoon spp. (i.e., six with H. felis, two with H. silvestris) and four out of 19 (21%) for Cytauxzoon europaeus. Outdoor lifestyle and origin (i.e., Friuli-Venezia Giulia region) were the most relevant risk factors for H. silvestris and C. europeus infections. Conversely, H. felis was most frequently isolated from domestic cats, suggesting different modes of transmission.
... Albeit the prevalence of feline infection is generally lower than that in dogs from the same localities, the percentage of infected cats is not negligible [56,59,60]. Several epizootiological studies have demonstrated that the parasite circulates among feline populations in Mediterranean countries [59][60][61][62][63][64][65]. Studies in Greece have evaluated the exposure to L. infantum and have recorded a seroprevalence rate of 2.0% in Mykonos island [66], 3.8% in Thessaloniki [59], 8.3% in Athens and 14.7% in Crete island [63]. ...
Leishmaniosis caused by the protozoon Leishmania infantum that is transmitted through the bites of infected phlebotomine sandflies is of major veterinary concern in Greece. The country is endemic with particularly favourable environmental conditions for the spread of this infection. Moreover, Greece remains a popular touristic destination, and the continuous travel of pets raises concern regarding the possible dissemination of infection from endemic to non-endemic areas. Dogs are the main reservoir host, although other animal species, including humans, may also be infected. Canine leishmaniosis manifests as a visceral disease that can result in death if left untreated. Sero-logical and molecular epizootiological studies have confirmed circulation of the parasite in Greek canine and feline populations as well as in other mammals. As a result, constant surveillance and identification of high-risk localities are necessary to establish chemoprophylactic protocols for travelling animals to safeguard animal and public health.
... Reports of feline leishmaniosis most often concern adult cats (median 7 years) from endemic countries, especially in the Mediterranean area [18,19], fitting well with our 6-year-old cat imported from Spain which also tested positive for FeLV and FIV. Clinical leishmaniosis in cats is reportedly associated with an impaired immune response, which may be caused by infections such as FeLV or FIV or by immunosuppressive treatment [2,4,7,[38][39][40][41]. Both the beginning of clinical signs (i.e. ...
Background
In Europe, feline leishmaniosis is commonly caused by Leishmaniainfantum. There is little knowledge regarding pathogenesis, ocular manifestations and long-term follow-ups in cats with leishmaniosis.
Findings
A 6-year-old female, spayed European Shorthair cat was imported from Spain to Germany 2 years prior to its first clinical presentation. The cat showed lethargy, weight loss, ulcerative lesions on the front limbs and high-grade chronic uveitis. The diagnosis of L. infantum infection was based on the cytological finding of amastigotes in skin lesions, positive qPCR of EDTA-blood and positive PCR of a cyto-brush sample from the conjunctiva. Supportive findings included positive serology by IFAT, serum protein capillary electrophoresis with peaks in alpha2- and gamma-globulin sections and marked elevation of SAA. Enucleation had to be performed on day 288 on both eyes because of blindness, glaucoma and high-grade uveitis. Histologically, high numbers of Leishmania spp. amastigotes were found in histiocytes. IFAT and PCR were positive in the aqueous humor in both eyes, respectively. Feline leukemia virus antigen and feline immunodeficiency virus antibody testings were positive. Hematological and biochemical results revealed mild leukocytosis with lymphocytosis, monocytosis and eosinopenia as well as marked elevation of SAA and hyperglobulinemia. The cat was treated with allopurinol, responded well and was still alive at follow-up on day 288 after first presentation. However, enucleation was necessary because of refractory glaucoma and uveitis.
Conclusion
For the first time, ocular evidence of Leishmania IgG antibodies was demonstrated in the aqueous humor of both eyes in cats. There is limited knowledge about the pathogenesis, treatment options and outcomes in cats infected with L. infantum. This case report supports the hypothesis that immunosuppression increases the risk of clinical signs of leishmaniasis in cats. Alpha2- and gamma-globulin peaks in serum protein capillary electrophoresis are supportive criteria for the diagnosis of L. infantum infection. SAA is valuable for monitoring. Regarding ophthalmology, uveitis and glaucoma may have a poor prognosis.
Graphical abstract
... The isolate showed 99.37 per cent identity with H. felis isolate from wild (Jungle) cat collected from Wayanad, Kerala and 99.05 per cent similarity towards H. felis isolate of a tiger from Trivandrum Zoo. Malangmei et al. (2021), Morelli et al. (2021) in Greece and Attipa et al. (2017) in cats of Cyprus. ...
Vector-borne diseases are an important cause of morbidity and mortality in domestic cat population and hepatozoonosis is one such infection. Incidence of hepatozoonosis has been described in a variety of animal species but information on cats is scarce. To investigate the occurrence of this pathogen in domestic cats, blood samples were collected from domestic cats from three districts of Kerala (Thrissur, Kannur and Wayanad). Field-stained blood smears were examined to detect the presence of gamonts of Hepatozoon spp. and the samples were subjected to molecular analysis by PCR amplification. Out of 122 blood samples screened, none of the sample revealed the presence of gamonts of Hepatozoon on microscopic examination. Polymerase chain reaction targeting 18S rRNA gene of Hepatozoon spp. demonstrated seven positive cases with a prevalence of 5.74 per cent. Results of this study indicate that hepatozoonosis is established within the domestic cats in Kerala and warrant the adoption of control measures.
... Since then, Hepatozoon infections have been found in domestic cats and various wild felids worldwide [22]. In Europe, Hepatozoon felis is recognized as the main agent infecting domestic and wild felines [15,16,[23][24][25][26][27][28][29][30][31][32][33][34][35][36]. Hepatozoon canis has also been reported in domestic cats in Europe [28,29,[37][38][39]. ...
... The geographical distribution of Hepatozoon spp. in domestic cats is apparently wide in Europe, with reports originating mainly from Mediterranean countries, such as Spain [23,24,28,37], Portugal [15,16], Italy [29,32,35], Cyprus [27], Greece [36], and France [39], but also from Central Europe: Austria [33] and Switzerland [40]. In our study, the exact identity of the species involved could not be established due to the low quality of the obtained sequences. ...
... In recent decades, Hepatozoon infection in felids has been increasingly reported worldwide, usually with low infection rates, but ranging up to 37.9% in Cyprus [27]. The overall prevalence of Hepatozoon spp. in the present study was 0.5%, similar to that found in Spain [37] or Italy [32]. ...
Background
Apicomplexan haemoparasites are protozoans that infect a variety of domestic and wild animal species, as well as humans. Data regarding haemoprotozoans in domestic cats are limited; therefore, the aim of this study was to assess the occurrence of Babesia spp., Cytauxzoon spp., and Hepatozoon spp. in domestic cats in Romania using molecular tools.
Methods
Blood samples from 371 domestic cats were screened for the presence of piroplasmids. All samples that yielded a visible band in agarose gels were subsequently tested by specific assays targeting the 18S rDNA of Babesia spp., Cytauxzoon spp., and Hepatozoon spp. Moreover, nested PCR assays targeting mitochondrial genes of Babesia spp. were used for screening of all Babesia spp. 18S rDNA-positive samples.
Results
From the total number of sampled cats, 19.4% were positive in the PCR assay targeting piroplasmids. Babesia spp. were identified in 15.1% of cats, while 0.5% were positive for Hepatozoon spp. Molecular analyses confirmed the presence of Babesia canis. No samples were positive for Cytauxzoon spp.
Conclusions
The high infection rates of domestic cats with Babesia spp. and the need for species differentiation highlight the importance of mitochondrial genes as targets for molecular protocols.
... Both H. canis and H. felis were found in ticks, albeit at a low prevalence. In Cyprus, H. canis has been previously reported in dogs [45], while H. felis has been detected with a high prevalence (37.9%) in cats [46]. Similarly, H. felis occurs with a high prevalence in cats in other European enzootic areas, reaching 25.5% in Greece [47]. ...
... Ctenocephalides felis is the vector of important pathogens, including B. henselae detected herein and previously reported in rats and cats of Cyprus [6,46,52]. On the other hand, to the best of the authors' knowledge, the present report of B. koehlerae and B. clarridgeiae is the first in the country. Bartonella spp. ...
Ticks and fleas are blood-sucking ectoparasites that cause irritation and anaemia to their hosts and act as vectors of pathogens (vector-borne pathogens, VBPs) of relevance for animal and human health. In the present study, tick and flea species in dogs and cats from Cyprus were recorded and VBPs were detected in the collected specimens. Ectoparasites were collected from 220 animals (161 dogs and 59 cats), and a questionnaire including demographic, clinical, and other information was filled out for each animal. The ectoparasites were morphologically identified and the detection of VBPs was performed by PCR-coupled sequencing. Rhipicephalus sanguineus sensu lato was found on 108 dogs and 13 cats, and Ixodes gibbosus on 2 dogs. Ctenocephalides felis was the predominant flea species (on 62 dogs and 45 cats), while one dog and one cat were infested by Ctenocephalides canis and Echid-nophaga gallinacea, respectively. The VBPs in ticks were Anaplasma platys, Rickettsia massiliae, Rickettsia conorii, Rickettsia felis, Hepatozoon felis and Hepatozoon canis, while Rickettsia felis, Rickettsia sp., Bartonella koehlerae, Bartonella clarridgeiae, and Bartonella henselae were recorded in fleas. Statistical analysis (chi-square test and multiple univariate generalized linear model) showed that animals up to 6 months of age were less likely to be infested with ticks than older animals, but more likely to be infested with fleas. Ticks were more prevalent in sheltered than in owned animals, while the odds ratio of flea presence was higher in owned animals than those living in shelters. The present study is the first investigation on the occurrence of ticks and fleas in dogs and cats from Cyprus, showing the presence of different VBPs in these important ectoparasites. The results point out the importance of systematic ectoparasite control in dogs and cats.
