Antonella Migliazzo

Feline leishmaniosis (FeL) caused by Leishmania infantum is increasingly recognized in endemic areas, but the inflammatory profile during infection remains poorly defined. This case–control study evaluated the acute phase proteins (APPs) serum amyloid A (SAA), haptoglobin (Hp), and ceruloplasmin (Cp) in cats with anti-L. infantum antibodies and/or DNA and compared them with healthy negative controls. A total of 125 cats were enrolled, including 62 cats positive by IFAT and/or qPCR and 63 healthy controls. Total protein, albumin, albumin-to-globulin ratio (A/G), and serum protein electrophoresis were also assessed, and correlations with APPs were investigated in positive cats. Compared with controls, L. infantum-positive cats had significantly higher Hp and Cp concentrations, whereas SAA did not differ significantly. They also showed higher total protein, beta2- and gamma-globulin concentrations and lower albumin and A/G ratio. SAA and Hp were negatively correlated with albumin, whereas Cp was positively correlated with total protein and beta-globulin fractions. No significant correlations were found between APPs and IFAT antibody titer. In exploratory analyses, IFAT seropositivity (cut-off ≥ 1:80) was associated with higher Hp, whereas qPCR positivity was associated with higher SAA and Cp concentrations; however, these results should be interpreted cautiously because qPCR-positive cats were few. These findings indicate a measurable inflammatory and dysproteinemic profile in cats with laboratory evidence of L. infantum exposure and/or infection, but do not establish APPs as disease-specific diagnostic markers. However, diagnostic heterogeneity, limited qPCR testing, lack of standardized clinical staging, and possible confounding diseases limit causal and clinical interpretation.

Feline leishmaniosis is a worldwide infection caused by the parasite of the genus Leishmania transmitted by sandflies. Based on the complexity of epidemiology and diagnosis of this infection, the role of cats in the epidemiology and clinical impact of disease is still under debate. By using serological and molecular methods, this study aimed to update the epidemiology of the infection in different feline populations from various areas of Italy and to study factors associated with the infection. Of 1490 cats tested, 124 (8.3%, 95% CI 6.9–9.9) were infected, 96 had only specific L. infantum IgG, 18 were only positive for parasite DNA and 10 were both IFAT and qPCR positive. Risk factors for infection were sampling in the winter season (OR = 3.2, 95% CI 2.2–4.8), originating from the Sicily region (OR = 2.0, 95% CI 1.3–3.0), male gender (OR = 1.8, 95% CI 1.1–3.2), outdoor lifestyle (OR = 2.3, 95% CI 0.9–5.6) and seropositivity for FIV antibodies (OR = 2.2, 95% CI 1.2–4.2), while sampling in the spring (OR = 0.5, 95% CI 0.3–0.7) and summer (OR = 0.3, 95% CI 0.1–0.7), and originating from the Lazio region (OR = 0.1, 95% CI 0.05–0.4) were protective factors for infection. In endemic areas, Leishmania infection should be investigated by using both serological and molecular methods and cats should be protected from sandfly bites, particularly if they are FIV infected.

Cats are susceptible to coronavirus infections, including infection by human severe acute respiratory syndrome coronavirus (SARS-CoV). In human ABO system blood groups, alloantibodies can play a direct role in resistance to infectious diseases. Individuals with the AB blood type were over-represented in the SARS-CoV-2 infection group. Blood type AB individuals lack both anti-A and anti-B antibodies, and therefore lack the protective effect against SARS-CoV-2 infection given by these antibodies. Starting from this knowledge, this pilot preliminary study evaluated a possible association between feline blood phenotypes A, B, and AB and serostatus for SARS-CoV-2 antibodies in cats. We also investigated selected risk or protective factors associated with seropositivity for this coronavirus. A feline population of 215 cats was analysed for AB group system blood phenotypes and antibodies against the nucleocapsid (N-protein) SARS-CoV-2 antigen using a double antigen ELISA. SARS-CoV-2 seropositive samples were confirmed using a surrogate virus neutralization test (sVNT). Origin (stray colony/shelter/owned cat), breed (DSH/non DSH), gender (male/female), reproductive status (neutered/intact), age class (kitten/young adult/mature adult/senior), retroviruses status (seropositive/seronegative), and blood phenotype (A, B, and AB) were evaluated as protective or risk factors for SARS-CoV-2 seropositivity. Seropositivity for antibodies against the SARS-CoV-2 N-protein was recorded in eight cats, but only four of these tested positive with sVNT. Of these four SARS-CoV-2 seropositive cats, three were blood phenotype A and one was phenotype AB. Young adult age (1–6 years), FeLV seropositivity and blood type AB were significantly associated with SARS-CoV-2 seropositivity according to a univariate analysis, but only blood type AB (p = 0.0344, OR = 15.4, 95%CI: 1.22–194.39) and FeLV seropositivity (p = 0.0444, OR = 13.2, 95%CI: 1.06–163.63) were significant associated risk factors according to a logistic regression. Blood phenotype AB might be associated with seropositivity for SARS-CoV-2 antibodies. This could be due, as in people, to the protective effect of naturally occurring alloantibodies to blood type antigens which are lacking in type AB cats. The results of this pilot study should be considered very preliminary, and we suggest the need for further research to assess this potential relationship.

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. 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. 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: Leishmaniasis is a widespread infectious disease, but there is not much information about its prevalence in high risk occupational categories. Objectives: The aim of this study is to assess the prevalence of Leishmania immunological positivity in human skin tissues collected from subjects living in Western Sicily, with suspected cutaneous Leishmania infection, in order to explore the risk possibly related to occupation. Methods: 318 consecutive subjects (M/F ratio=1.0, mean age=40±25.4 years), attending the Dermatology Department of the University of Palermo Hospital from 2013 to 2015, without any previous history of Leishmania infection and performing various occupations, were included. Parasite isolation and PCR-RT test on skin scrapings were performed to evaluate the immunological status; all data were analyzed by the chi square test, comparing all positive results from the different provinces. Results: 81 (50.9%) out of 159 females and 79 (49.7%) out of 159 males were found PCR-RT positive to Leishmania infantum, with a higher risk in the Agrigento district (p<0.001) and in subjects living in rural areas (p=0.0038), regardless of the type of work performed. The observed animal leishmaniasis prevalence in the same areas shows the endemic status of the disease in Sicily. Conclusions: Although based on a relatively small sample, our study shows that cutaneous leishmaniasis represents a health care problem with a medical and social impact in Western Sicily. An active surveillance system and the establishment of diagnosis and treatment centres could be useful in controlling this public health problem.

Background: Identification ofLeishmania infantum‐infected dogs is crucial for control of canine leishmaniosis. In particular, in areas where access to specialized laboratories is limited, the availability of reliable and rapid in‐clinic serologic tests may support immediate diagnosis in suspected cases and permit detection of asymptomatic canine carriers ofL infantuminfection.Objective: The purpose of the study was to validate the immunochromatographic test (ICT) Anigen Rapid Leishmania Ab Test kit for detection ofL infantumantibodies in naturally exposed dogs in comparison with the immunofluorescence antibody test (IFAT).Methods: Serum samples from 66 dogs, including 20 healthy control dogs and 46 dogs suspected or confirmed with canine leishmaniosis, were measured by both tests. Anti‐LeishmaniaIgG titers ≥ 1:40 byIFATwere considered positive. Kappa statistic with a 95% CI was calculated to evaluate agreement between the 2 testing methods, and sensitivity, specificity, and positive and negative likelihood ratio were calculated.Results: Anti‐L infantumIgG antibodies were found in 35 of 66 samples using theIFATtest (titers 1:40–1:5120). Thirty‐one out of 66 samples tested positive with the qualitativeICT. FourIFAT‐positive (titers < 1:80) samples wereICT‐negative. The Kappa value of 0.853 demonstrated very good agreement between the 2 tests.Conclusion: The Anigen Rapid Leishmania Ab Test kit reliably identified canine sera with anti‐L infantumIgG antibody titers ≥ 1:40. TheICTrequires neither special preparation of the serum nor specialized equipment and can be stored at ambient temperature. The test is applicable as a field test because it is easy to use and provides rapid results.

Pentadecane is an organic compound that is made up of primarily carbon and hydrogen atoms and can be found as floral volatile in different plants. Many essential oils and crude extracts of some plants, containing pentadecane, show an interesting antimicrobial activity. On the basis of absence of the toxicity of pentadecane directed toward eukaryotic cells, the authors investigated in vitro effects of pentadecane in Leishmania infantum parasites. Pentadecane (300 µM) was found to decrease the growth by 86 % both in promastigotes (IC50: 65.3) and amastigotes (IC50: 60.5 µM). This compound also caused a reduction of macrophage infection showing a growth inhibition of 77% at 300 µM.Analysis of propidium iodide incorporation in Leishmania, treated with pentadecane at 48 hours, suggested that cells were arresting in the sub G0/G1 and G1 phase of the cell cycle.Cytotoxicity assay of pentadecane in immortalized cells lines (DH82 and U937) and in primary epithelial cells of Cercopiteco (CPE) showed that i...

Leishmaniasis is a disease with a worldwide distribution affecting both humans and animals. There is a need to identify and develop new drugs for the treatment of leishmaniasis. This study showed that crude ethanolic extracts of the red alga Asparagopsis taxiformis have a powerful effect against L. infantum, the prevalent species of the genus Leishmania in the Mediterranean basin. L. infantum demonstrated decreased vitality with increasing concentration of the algal extracts. At a concentration of 40 μg/mL, the extracts achieved 100% mortality of the parasite and the LD₅₀ value was 25 μg/mL for promastigotes and 9 μg/mL for amastigotes. Algal extracts caused morphological alterations and apoptosis in Leishmania cells. The potential cytotoxic action of crude extracts was investigated by a MTT viability assay on DH82 and Vero cell lines but there was no cytotoxic effect. The potential of red alga A. taxiformis metabolites as anti-leishmanial agents merits further pharmacological investigation.