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Background Vector-borne pathogens are the subject of several investigations due to the zoonotic concern of some of them. However, limited data are available about the simultaneous presence of these pathogens in cats and their ectoparasites. The aim of the present study was to define the species of ectoparasites found on cats as well as to investigate vector-borne pathogens in cats and their ectoparasites in southern Italy. Methods Blood from 42 cats and fleas or flea pools ( n  = 28) and ticks ( n  = 73) collected from them were investigated by quantitative PCR for the detection of vector-borne pathogens. Feline serum samples were tested by IFAT to detect IgG antibodies against Leishmania infantum, Bartonella henselae , Rickettsia conorii, Rickettsia felis, Rickettsia typhi , Babesia microti , Ehrlichia canis and Anaplasma phagocytophilum antigens. Results Only one flea species ( Ctenocephalides felis ) and four tick species belonging to the genera Rhipicephalus and Ixodes were identified on cats from southern Italy. Molecular evidence of Bartonella spp., Rickettsia spp., hemoplasmas, Babesia vogeli and L. infantum was found in ectoparasites (fleas and/or ticks) while DNA from Hepatozoon felis and Ehrlichia/Anaplasma spp. was not detected. Likewise, DNAs from Bartonella, hemoplasma and Leishmania were the only pathogens amplified from feline blood samples. Cats had also antibodies against all the investigated pathogens with the exception of Rickettsia typhi . Agreement between serological and molecular results in individual cats and their ectoparasites was not found. The only exception was for Bartonella with a fair to moderate agreement between individual cats and their ectoparasites. Bartonella clarridgeiae was the species most frequently found in cats and their fleas followed by B. henselae . Conclusions In conclusion, cats harboring ticks and fleas are frequently exposed to vector-borne pathogens. Furthermore, ticks and fleas harbored by cats frequently carry pathogens of zoonotic concern therefore appropriate feline ectoparasiticide preventative treatments should be used in cats.

Background Limited information is available about the species of ticks infesting the cat and the pathogens that they harbor. The aims of the present study were to identify the species of ticks removed from cats living in Sicily and Calabria (Italy) and to detect DNA of vector-borne pathogens in the same ticks. Findings Morphological identification of 132 adult ticks collected throughout the year from cats was carried out. Real-time PCRs for Hepatozoon felis , Piroplasmid, Ehrlichia/Anaplasma spp., Rickettsia spp., Bartonella spp., Mycoplasma spp. and Leishmania infantum were performed from each individual tick. Ticks belonging to Rhipicephalus ( R. sanguineus sensu lato, R. pusillus ) and Ixodes ( I. ricinus , I. ventalloi ) genera were identified. Ixodes ventalloi was the most frequently found tick species (47 %). The positivity rate to at least one pathogen was 14.4 % (19/132 ticks). Leishmania infantum , Rickettsia spp. ( R. monacensis and R. helvetica ), Bartonella spp. ( B. clarridgeiae ), Piroplasmid ( Babesia vogeli ), and Ehrlichia/Anaplasma spp. ( E. canis ) DNAs were amplified in 8.3, 5.3, 1.5, 0.75 and 0.75 % of ticks, respectively. Hepatozoon felis , Anaplasma spp. and hemotropic Mycoplasma spp. DNAs were not detected. Four (21.1 %) out of nineteen positive ticks were co-infected. Conclusions This study provides novel data about ticks infesting cats and the DNA of pathogens that they harbor. In Southern Italy, anti-tick prophylaxis should be implemented throughout the year in cats without neglecting winter time.

Caretta caretta is the most common sea turtle species in the Mediterranean Sea. The species is threatened by anthropomorphic activity that causes thousands of deaths and hundreds of strandings along the Mediterranean coast. Stranded turtles are often cared for in rehabilitation centres until they recover or die. The objective of this study was to characterize the gut microbiome of nine sea turtles stranded along the Sicilian coast of the Mediterranean Sea using high-throughput sequencing analysis targeting V3-V4 regions of the bacterial 16S rRNA gene. Stool samples were collected from eight specimens hosted in the recovery centre after a few days of hospitalization (under 7) and from one hosted for many weeks (78 days). To better explore the role of bacterial communities in loggerhead sea turtles, we compared our data with published fecal microbiomes from specimens stranded along the Tuscan and Ligurian coast. Our results highlight that, despite the different origin, size and health conditions of the animals, Firmicutes, Bacteroidetes and Proteobacteria constitute the main components of the microbiota. This study widens our knowledge on the gut microbiome of sea turtles and could be helpful for the set up of rehabilitation therapies of stranded animals after recovery in specialized centres.

Infection with Leishmania parasites can lead to severe disease in humans and dogs, which act as a reservoir in zoonotic transmission. An increasing number of reports suggest that leishmaniosis is not restricted to dogs, but also affects many other mammalian and avian species. Consequently, this expands the potential reservoir and is of great public and veterinary health concern. The present study reviews, based on a comprehensive search of scientific literature published from 1 January 2001 to 31 December 2020, the currently available information on animal leishmaniosis in vertebrates in Europe, other than dogs and humans. This review provides an exhaustive list of mammals and birds in which infections with or exposure to Leishmania parasites have been detected in European countries. Most cases are reported from the Mediterranean region. Domestic animals, in particular cats, pose a concern because of close contact with humans. The wildlife reservoir is less likely to contribute to zoonotic transmission, with the exception of hares. This potentially large reservoir needs to be taken into account when developing control measures for zoonotic leishmaniosis. From a veterinary point of view, it is important that veterinarians are better aware of leishmaniosis and trained in its management.

Background Cats can be carriers of infected arthropods and be infected with several vector-borne pathogens (VBP) but there is limited knowledge about their pathogenic role in cats. Results A cross-sectional controlled study investigated the clinical status and antibody ( Bartonella henselae , Rickettsia conorii , Ehrlichia canis , Anaplasma phagocytophilum , Babesia microti and Leishmania infantum ) and/or blood PCR ( Mycoplasma spp., Bartonella spp., Rickettsia spp., Ehrlichia/Anaplasma spp., piroplasmids, L. infantum , Hepatozoon felis ) prevalence in 197 cats. Outdoor cats lacking ectoparasiticide treatment or hosting ectoparasites (study group [SG], n = 134) and indoor cats treated against ectoparasites (control group [CG], n = 63) were enrolled. Clinical data and retroviral co-infections were compared between the two groups. Multivariable analysis tested associations between variables and VBP exposure. Lymphadenia, stomatitis, and various haematological abnormalities were statistically more frequent in SG. Antibodies against R. conorii , B. henselae , A. phagocytophylum , B. microti , E. canis and L. infantum were detected. Bartonella henselae , Bartonella clarridgeiae , Mycoplasma haemofelis , “ Candidatus Mycoplasma haemominutum” and “ Candidatus Mycoplasma turicensis” DNA were identified. Very high antibody (87.8%) and PCR (40.1%) positivity to at least one pathogen were detected and were significantly higher in SG. Co-infections were confirmed in about one-third of the cats and were more frequent in SG cats. Molecular and overall (antibody and PCR) positivity to Bartonella and antibody positivity to R. conorii were higher in SG. Multivariable analysis found significant associations of Bartonella spp. infection with Feline Immunodeficiency Virus (FIV) infection and increased globulins, and of Mycoplasma spp. infection with adult age, FIV infection, anaemia, and increased creatinine. Conclusions A very high prevalence of exposure to zoonotic VBP was found in cats, with Rickettsia and Bartonella infections being most prevalent. Some risk factors were documented namely for Mycoplasma spp. and Bartonella spp. The lifestyle of cats is clinically relevant and requires specific preventative measures to protect their health.

Background Anti- Leishmania antibodies are increasingly investigated in cats for epidemiological studies or for the diagnosis of clinical feline leishmaniosis. The immunofluorescent antibody test (IFAT), the enzyme-linked immunosorbent assay (ELISA) and western blot (WB) are the serological tests more frequently used. The aim of the present study was to assess diagnostic performance of IFAT, ELISA and WB to detect anti- L. infantum antibodies in feline serum samples obtained from endemic ( n  = 76) and non-endemic ( n  = 64) areas and from cats affected by feline leishmaniosis ( n  = 21) by a Bayesian approach without a gold standard. Methods Cut-offs were set at 80 titre for IFAT and 40 ELISA units for ELISA. WB was considered positive in presence of at least a 18 KDa band. Statistical analysis was performed through a written routine with MATLAB software in the Bayesian framework. The latent data and observations from the joint posterior were simulated in the Bayesian approach by an iterative Markov Chain Monte Carlo technique using the Gibbs sampler for estimating sensitivity and specificity of the three tests. Results The median seroprevalence in the sample used for evaluating the performance of tests was estimated at 0.27 [credible interval (CI) = 0.20–0.34]. The median sensitivity of the three different methods was 0.97 (CI: 0.86–1.00), 0.75 (CI: 0.61–0.87) and 0.70 (CI: 0.56–0.83) for WB, IFAT and ELISA, respectively. Median specificity reached 0.99 (CI: 0.96–1.00) with WB, 0.97 (CI: 0.93–0.99) with IFAT and 0.98 (CI: 0.94–1.00) with ELISA. IFAT was more sensitive than ELISA (75 vs 70%) for the detection of subclinical infection while ELISA was better for diagnosing clinical leishmaniosis when compared with IFAT (98 vs 97%). Conclusions The overall performance of all serological techniques was good and the most accurate test for anti- Leishmania antibody detection in feline serum samples was WB.

Neutrophil-to-lymphocyte (NLR), monocyte-to-lymphocyte (MLR), and platelet-to-lymphocyte (PLR) ratios have been proposed as diagnostic and prognostic markers for neoplastic and inflammatory diseases in dogs and cats. The aim of this retrospective preliminary study was to evaluate the relationship between these ratios and markers of inflammation routinely measured in cats. A total of 275 cats were enrolled. Complete blood count, serum amyloid A (SAA), albumin, globulin, and albumin-to-globulin ratio (AGR) data were analyzed, as well as the presence of leukocyte alterations considered suggestive of inflammation (LAI: neutrophils left shift, toxic neutrophils, and reactive lymphocytes) evaluated in blood smears. The NLR and MLR correlated positively with SAA and globulins and negatively with albumin and AGR. Higher NLR and MLR were found in cats with increased SAA and globulins and decreased albumin and AGR. The PLR correlated negatively with albumin and AGR. A higher PLR was found in cats with hypoalbuminemia. Cats with LAI had higher NLR, MLR, and PLR. In cats with no changes in parameters indicative of inflammation, 11.25, 0.42, and 528.3 were identified as upper limits for NLR, MLR, and PLR, respectively. In conclusion, the NLR, MLR, and PLR act as good inflammatory markers easily evaluated by routine hematology.

Background Feline leishmaniosis caused by Leishmania infantum is often associated with feline immunodeficiency virus (FIV) infection; however, the role and clinical significance of this coinfection remain unknown. This study aimed to assess whether FIV is associated with L. infantum infection in cats from canine leishmaniosis endemic areas and to report the clinical signs and hematological alterations associated with coinfection. Methods A retrospective matched case-control study (ratio 1:2) was conducted. Data of clinical examination and complete blood count (CBC) were selected from a cohort of 705 cats examined for epidemiological studies on feline leishmaniosis conducted between 2012 and 2019. Ninety-one FIV seropositive cases and 182 FIV seronegative control cats were selected. Matching was done according to age, sex, lifestyle and geographic provenience of case cats. Rapid ELISA devices were mainly used to detect anti-FIV antibodies. Anti- Leishmania IgG antibodies were detected by indirect-immunofluorescence test (IFAT). Leishmania DNA was searched in blood, oral and conjunctival swabs by quantitative real-time PCR. Results Feline immunodeficiency virus seropositive cats had no hematological abnormalities suggestive of an advanced stage of FIV infection and were statistically more frequently IFAT positive, and their risk of being L. infantum antibody positive was 2.8 greater than in the FIV seronegatives. The association of FIV seropositivity with L. infantum antibody positivity was confirmed in the univariable model of logistic regression. A multivariate model found FIV infection and L. infantum PCR positivity as predictors of a positive L. infantum IFAT result. Male outdoor cats from rural or suburban areas were at risk for FIV and L. infantum antibody positivity. Clinical signs more frequently associated with the coinfection were oral lesions, pale mucous membranes and low body condition score (BCS). Conclusions This study documents that FIV seropositive cats with no hematological abnormalities suggestive of an advanced stage of FIV infection are more prone to be L. infantum seroreactive by IFAT in endemic areas. Therefore, FIV seropositive cats should be tested for L. infantum antibodies and treated for preventing sand fly bites. Pale mucous membranes, low BCS and oral lesions but no CBC abnormalities were significantly associated with the coinfection. Graphical abstract

It has been estimated that 44,000 Caretta caretta turtles die every year due to anthropomorphic activity in the Mediterranean Sea, and that longline fishing is one of the most significant causes of mortality. A total of 482 specimens of C. caretta were rescued from different parts of the Sicilian coast (Mediterranean Sea) from 2014 to 2016. The most numerous stranding was recorded during the spring and summer seasons, mainly along the north and eastern coasts of Sicily. The curved carapace length for all the specimens ranged from between 19 and 95 cm and most of these were young or sub adults. The highest number of strandings was recorded in 2014 and 2015, with 206 and 169 individuals, respectively. A total of 66 live specimens out of 239 were successfully rehabilitated and released after surgery or drug therapy; fishing hooks were found in 129 specimens in different parts of the digestive tract with greater frequency in the oesophagus (47.3%) followed by the gut (24.8%), stomach (14.7%), and mouth (13.2%). This paper will highlight the incidence of the incidental catch by longline fishing of C. caretta along the Sicilian coasts and also relate the size of ingested hooks to the size of examined specimens.

In feline Leishmania infantum (Li) infection and in clinical cases of feline leishmaniosis, co-infection with feline immunodeficiency virus (FIV) has been reported. However, the role of the retroviral co-infection in the impairment of feline clinical health is still controversial. The aim of this study was to evaluate hemogram changes in cats from regions endemic for both Li and FIV infection. Four hundred and ninety-six cats tested for Li (EDTA blood polymerase chain reaction and immunofluorescence antibody test) and for FIV infection (enzyme-linked immune assay) were retrospectively evaluated. Hemogram results including blood smear morphological evaluation were statistically compared considering four infection patterns: Li+FIV+, Li+FIV−, Li−FIV+, and Li−FIV−. Significantly lower values of erythrocytes (Li+FIV−: p = 0.0248; Li−FIV+: p = 0.0392) and hemoglobin (Li+FIV: p = 0.0086; Li−FIV+: p = 0.0249) were found in both infections when compared to Li−FIV− cats, and severity of anemia was more frequently moderate in Li-positive cats (p = 0.0206) and severe in FIV infection (p = 0.024). Li infection was associated with monocytosis (p = 0.0013) and morphologically activated monocytes (p = 0.0209). Moreover, FIV infection was associated with the presence of inflammatory leukogram (p = 0.023), and an association between thrombocytosis and the co-infection was found (p = 0.0347). Li infection in cats induces hematological changes compatible with chronic inflammation, some of which are due to co-infection with FIV.