Cats In Italy Research Articles

Detection of Leishmania infantum DNA in conjunctival swabs of cats by quantitative real-time PCR

Although some studies have investigated the potential role of cats as a reservoir for Leishmania, their role in the epidemiology of visceral leishmaniasis (VL) is still poorly understood. Molecular diagnostic techniques are an important tool in VL diagnosis, and PCR shows high sensitivity and specificity for Leishmania spp. detection. Quantitative real-time PCR (qPCR) is a method that permits quantitative analysis of a large number of samples, resulting in more sensitive, accurate, and reproducible measurements of specific DNA present in the sample. This study compared real-time PCR (qPCR) and conventional PCR (cPCR) for detection of Leishmania spp. in blood and conjunctival swab (CS) samples of healthy cats from a non-endemic area in the state of São Paulo, Brazil. Of all CS samples, 1.85% (2/108) were positive for Leishmania spp. by both cPCR as qPCR (kappa index = 1), indicating excellent agreement between the two methods. The DNA from the two CS-cPCR- and CS-qPCR-positive samples was further tested with a PCR test amplifying the Leishmania spp. discriminative rRNA internal transcribed spacer 1 (ITS 1), of which one sample generated a 300–350-bp DNA fragment whose size varies according to the Leishmania species. Following sequencing, the fragment showed 100% similarity to a GenBank L. infantum sequence obtained from a cat in Italy. In conclusion, the association of qPCR and CS proved to be effective for detection of Leishmania in cats. Conjunctival swab samples were shown to be a practical and better alternative to blood samples and may be useful in the diagnosis and studies of feline leishmaniasis.

Genome characterization of feline morbillivirus from Italy

Feline morbillivirus (FeMV) has been recently identified by RT-PCR in the urine sample of a nephropathic cat in Italy. In this report, we describe the whole genome sequence of strain Piuma/2015 obtained by combination of sequence independent single primer amplification method (SISPA) and next generation sequencing (NGS) starting from RNA purified from the infected urine sample. The existence in Germany and Turkey of FeMVs from cats divergent from Piuma/2015, suggests the presence of FeMV heterogeneity in Europe as it has been described previously in Japan and China.

Molecular identity and prevalence of Cryptococcus spp. nasal carriage in asymptomatic feral cats in Italy.

Cryptococcosis is a life-threatening fungal disease that infects humans and animals worldwide. Inhalation of fungal particles from an environmental source can cause primary infection of the respiratory system. As animals can be considered a sentinel for human diseases, the aim of this study was to determine the prevalence and molecular identity of Cryptococcus spp. in the nasal cavity of feral cats. Cats from 162 urban and rural feral cat colonies were sampled over 3 years. Of 766 cats from which nasal swabs were obtained, Cryptococcus spp. were recovered from 95 (12.6%), including 37 C. magnus (4.8%), 16 C. albidus (2.0%), 15 C. carnescens (1.9%), 12 C. neoformans (1.6%), as well as C. oeirensis (n=3), C. victoriae (n=3), C. albidosimilis (n=2), Filobasidium globisporum (n=2), C. adeliensis (n=1), C. flavescens (n=1), C. dimnae (n=1), C. saitoi (n=1), and C. wieringae (n=1) with prevalence <1%. Thirteen Cryptococcus species were identified by polymerase chain reaction and sequencing of internal transcribed spacer amplicons. Statistical analysis did not identify any predisposing factors that contributed to nasal colonization (eg, sex, age, season, or habitat). Results suggest that asymptomatic feral cats may carry C. neoformans and other Cryptococcus species in their sinonasal cavity. Genotyping of the specific cryptococcal isolates provides a better understanding of the epidemiology of these yeasts.

Genetic and phenotypic characterisation of Escherichia coli producing cefotaximase-type extended-spectrum β-lactamases: first evidence of the ST131 clone in cats with urinary infections in Italy.

The incidence of cefotaximase (CTX-M)-type extended-spectrum β-lactamase (ESBL)-producing Escherichia coli has increased dramatically in humans and animals since the middle of the last century. E coli that produce CTX-M β-lactamase represent a major cause of urinary tract infections, and pose a significant therapeutic challenge to both human and veterinary medicine. As data on uropathogenic CTX-M-producing strains in cats are limited, the aim of this study was to describe the genetic character and antibiotic resistance phenotypes of CTX-M-producing E coli isolated from cats with cystitis. Seven of 15 E coli bacteria isolated from 138 urine samples had the CTX-M gene and were therefore included in this study. These isolates were screened by polymerase chain reaction for the presence of 14 extra-intestinal virulence factors, class 1 and class 2 integrons, and to identify their phylogenetic groups. Multi-locus sequence typing (MLST) of the strains and susceptibility testing (disc diffusion method) were also performed. Virulence factor iutA was the most frequent determinant identified (86.7%), and the majority of CTX-M-producing strains (n = 5) carried class 1 integrons. MLST allowed us to discriminate four known sequence types (ST131, ST555, ST602, ST155) and three novel sequence types (ST3847, ST3848, ST4181). To the best of our knowledge, this is the first study to report uropathogenic CTX-M-producing E coli ST131 in cats in Italy. Accurate diagnostics and prudent use of antimicrobials are recommended to avoid the spread of multidrug-resistant pathogens in veterinary medicine and to prevent their transmission to humans.

Virulence factors and genetic variability of uropathogenicEscherichia coliisolated from dogs and cats in Italy

In this study, the association between virulence genotypes and phylogenetic groups among Escherichia (E.) coli isolates obtained from pet dogs and cats with cystitis was detected, and fingerprinting methods were used to explore the relationship among strains. Forty uropathogenic E. coli (UPEC) isolated from dogs (n = 30) and cats (n = 10) in Italy were analysed by polymerase chain reaction (PCR) for the presence of virulence factors and their classification into phylogenetic groups. The same strains were characterized by repetitive extragenic palindromic (REP)- and enterobacterial repetitive intergenic consensus (ERIC)-PCR techniques. We found a high number of virulence factors such as fimbriae A, S fimbriae (sfa) and cytotoxic necrotizing factor 1 (cnf1) significantly associated with phylogenetic group B2. We demonstrated a high correlation between α-hemolysin A and pyelonephritis C, sfa, and cnf1 operons, confirming the presence of pathogenicity islands in these strains. In addition, UPEC belonging to group B2 harboured a greater number of virulence factors than strains from phylogenetic groups A, B1, and D. REP- and ERIC-PCR grouped the UPEC isolates into two major clusters, the former grouping E. coli strains belonging to phylogenetic group B2 and D, the latter grouping those belonging to groups A and B1. Given the significant genetic variability among the UPEC strains found in our study, it can be hypothesized that no specific genotype is responsible for cystitis in cats or dogs.

Isolation and Characterization of Bartonella Strains in Cats in Italy

Bartonella henselae and Bartonella clarridgeiae were isolated in two cats in Sardinia, Italy. Infection by B. clarridgeiae was characterized by fever and submandibular lymph nodes enlargement while B. henselae infection was asymptomatic. This is the first report of B. clarridgeiae in a cat in Italy and the first isolation of B. henselae and B. clarridgeiae in Sardinia.

Free-roaming dogs and cats in central Italy: Public perceptions of the problem

A cross-sectional telephone survey of randomly selected households examined the extent and types of problems associated with free-roaming dogs and cats in the Teramo province of Italy. The households were sampled randomly within each municipality; municipalities were combined into coastal, central hills and mountain regions for analysis. The survey was conducted in May and June of 2004 with a response rate of 74% (397/536). Ninety percent of respondents ( N = 356) believed that free-roaming dogs and cats were a problem. They were most commonly concerned about personal safety, followed by animal welfare, public health and environmental sanitation. Sixty-nine percent of respondents (274) actually saw free-roaming dogs or cats where they live. While dogs were most commonly seen, cats were seen in greater numbers. Overall, 10% (39/297) and 5% (21/397) of respondents cared for free-roaming cats and dogs, respectively. Two-thirds of the respondents (251/397) believed that animals were abandoned because the owners lost interest. About 2/3 of respondents (251/397) reported that the community government should have the responsibility for dealing with free-roaming dogs and cats. The respondents supported the idea of building more shelters and controlling the birth rate as control measures rather than euthanizing dogs and cats. The results suggest that free-roaming dogs and cats are a very common sight in this part of Italy with substantial concerns by the residents. However, concerns about the animals’ welfare were clearly raised, supporting the laws that make it illegal to euthanize a healthy dog or cat in Italy. Using the information from this study, research on the underlying causes of abandonment of dogs and cats or failing to sterilize them should be undertaken to begin to address this problem. Further, national and regional funding must be provided to support existing laws which should help protect and eventually decrease the numbers of free-roaming and homeless dogs and cats.

Prevalence of the polycystic kidney disease and renal and urinary bladder ultrasonographic abnormalities in Persian and Exotic Shorthair cats in Italy

The ultrasonographic findings of kidneys, liver and urinary bladder of 288 Persian and 44 Exotic Shorthair clinically normal cats that underwent screening for polycystic kidney disease (PKD) between July 2003 and December 2005 were reviewed. Cats were divided into two groups, one including cats aged <9 months (group 1) and one cats aged >/=9 months (group 2). Cats were classified as PKD-positive when at least one renal cyst was found. One hundred and thirty-six cats (41.0%) had more than one cyst in at least one kidney. The prevalence of PKD was similar in both groups. Eight PKD-positive cats had cystic livers (5.9%). Other renal abnormalities included a pelvic calculus and a medullary rim sign (MRS). The difference in prevalence of an MRS in group 2 compared to group 1 and the difference between PKD-positive and -negative cats in group 2 were not significant. There was no difference in mean kidney length between PKD-positive and -negative cats in group 2. Urinary bladder anomalies were principally represented by urinary sediment, with prevalence significantly higher in group 2. No difference was detected in group 2 between PKD-positive and -negative cats. In conclusion feline PKD is common in Italy. The ultrasonographic findings of MRS and urinary bladder sediment did not correlate with feline PKD. Urinary bladder sediment is common in Persians and Exotic Shorthairs and more likely in adults.

Characterization of the first Bartonella henselae strain isolated from a cat in Italy

Bartonella henselae has been identified and characterized for the first time in Italy. A strain, designed Pavia-1, was isolated from the blood of a cat whose owner developed cat scratch disease (CSD). Pavia-1 and two American B. henselae strains (Houston-1, ATCC 49882, type I and strain 269608, UC Davis, type II) were compared by whole-cell fatty analysis (CFA), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for protein profiles, Western immunoblotting (WB) for reactivity with polyclonal antibodies, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), type-specific 16S rRNA PCRs, and pulsed-field gel electrophoresis (PFGE). Bartonella clarridgeiae (ATCC 51734) was also included for comparison. Pavia-1 was identified as a B. henselae type I. PFGE allowed differentiation between B. clarridgeiae and B. henselae and furthermore, between all the B. henselae strains. The fingerprints of PFGE observed for Pavia-1 were distinct from those of B. henselae type II and also of Houston-1, suggesting that the two type I strains derived from two different clones. These results show the capability of B. henselae to develop genotypic variability between genetically related strains.

Genetic identification of wild and domestic cats (Felis silvestris) and their hybrids using Bayesian clustering methods.

Crossbreeding with free-ranging domestic cats is supposed to threaten the genetic integrity of wildcat populations in Europe, although the diagnostic markers to identify "pure" or "admixed" wildcats have never been clearly defined. Here we use mitochondrial (mt) DNA sequences and allelic variation at 12 microsatellite loci to genotype 128 wild and domestic cats sampled in Italy which were preclassified into three separate groups: European wildcats (Felis silvestris silvestris), Sardinian wildcats (Felis silvestris libyca), and domestic cats (Felis silvestris catus), according to their coat color patterns, collection localities, and other phenotypical traits, independently of any genetic information. For comparison, we included some captive-reared hybrids of European wild and domestic cats. Genetic variability was significantly partitioned among the three groups (mtDNA estimate of F(ST) = 0.36; microsatellite estimate of R(ST) = 0.30; P < 0.001), suggesting that morphological diversity reflects the existence of distinct gene pools. Multivariate ordination of individual genotypes and clustering of interindividual genetic distances also showed evidence of distinct cat groups, partially congruent with the morphological classification. Cluster analysis, however, did not enable hybrid cats to be identified from genetic information alone, nor were all individuals assigned to their populations. In contrast, a Bayesian admixture analysis simultaneously assigned the European wildcats, the Sardinian wildcats, and the domestic cats to different clusters, independent of any prior information, and pointed out the admixed gene composition of the hybrids, which were assigned to more than one cluster. Only one putative Sardinian wildcat was assigned to the domestic cat cluster, and one presumed European wildcat showed mixed (hybrid) ancestry in the domestic cat gene pool. Mitochondrial DNA sequences indicated that three additional presumed European wildcats might have hybrid ancestry. These four cats were sampled from the same area in the northernmost edge of the European wildcat distribution in the Italian Apennines. Admixture analyses suggest that wild and domestic cats in Italy are distinct, reproductively isolated gene pools and that introgression of domestic alleles into the wild-living population is very limited and geographically localized.

Simultaneous occurrence of spongiform encephalopathy in a man and his cat in Italy

follow only simple commands. Repeat EEGs showed periodic triphasic complexes. 2 weeks after admission, he was mute, akinetic, and unable to swallow. He died in early January, 1994. His 7-year-old, neutered, female shorthaired cat presented in November, 1993, with episodes of frenzy, twitching of its body, and hyperaesthesia. The cat was usually fed on canned food and slept on its owner’s bed. No bites from the cat were recalled. In the next few days, the cat became ataxic, with hindquarter locomotor dysfunction; the ataxia got worse and there was diffuse myoclonus. The cat was killed in mid-January, 1994. No pathogenic mutations in the patient’s PrP gene were found. The patient and the cat were methionine homozygous at codon 129. Histology of the patient’s brain showed neocortical and cerebellar neuronal loss, astrocytosis, and spongiosis (figure A). PrP immunoreactivity showed a punctate pattern and paralleled spongiform changes (figure B). The cat’s brain showed mild and focal spongiosis in deeper cortical layers of all four lobes (figure C), vacuolated cortical neurons (figure D), and mild astrogliosis. The cerebellar cortex and the dentate nucleus were gliosed. Immunoreactive PrP showed a punctate pattern in neocortex, allocortex, and caudate nucleus (figure E). Western blot analysis of control and affected human and cat brain homogenates showed 3 PrP bands of 27–35 kDa. After digestion with proteinase K and deglycosylation, only samples from the affected patient and cat showed type-1 PrP, with PrP glycoform ratios comparable to those observed in sporadic CJD (details available from author). This study shows a spatio-temporal association between Simultaneous occurrence of spongiform encephalopathy in a man and his cat in Italy

Prevalence of feline immunodeficiency virus and other retroviral infections in sick cats in Italy

Two hundred and seventy-seven sick pet cats living in Italy were tested for antibodies to feline immunodeficiency virus (FIV) and for feline leukemia virus (FeLV) antigen. Overall, 24% of the cats resulted positive for anti-FIV antibody and 18% for FeLV antigen. FIV was isolated from the peripheral mononuclear blood cells of ten out of 15 seropositive cats examined and from one out of eight saliva samples. No FIV isolations were obtained from six serum samples cultured. Feline syncytium forming virus (FeSFV) could be isolated from blood and/or saliva in ten out of 11 FIV seropositive cats examined, in six out of nine FeLV antigen positive cats, in two cats found positive for both infection markers, and in three out of 11 cats negative for both markers. Thus, the probability of isolating FeSFV was enhanced by infection with other exogenous retroviruses.

Comparative Morphology of Ancylostoma tubaeforme (Zeder, 1800) and Ancylostoma caninum (Ercolani, 1859)

As a result of studies of Ancylostoma from European, African, and Australian dogs and cats, it was found that dogs harbored A. caninum and cats A. tubaeforme. A detailed study of these hookworms from dogs and cats in the United States confirmed the observations made by other investigators that A. caninum is not the common species in cats. A. tubaeforme differs from A. caninum in the following features: smaller size, larger ventral and esophageal teeth, shorter esophagus, thicker cuticle, smaller bursa, different arrangement of lateral bursal rays, smaller costa dorsalis, longer spicules, and shorter, narrower female tail. The two species apparently have world-wide distribution and A. caninum is seldom or never found as a natural infection of cats or A. tubaeforme as a natural infection of dogs. In 1800 Zeder (Biocca, 1954) described hookworms he found in a cat in 1783, giving them the name Strongylus tubaeformis. Then in 1859 Ercolani (Biocca, 1954) published a description of the dog hookworm, naming it Sclerostoma caninum. Later S. caninum was transferred to the genus Ancylostoma, but S. tubaeformis apparently was overlooked by most investigators. For nearly 100 years the common hookworm of both dogs and cats was referred to as Ancylostoma caninum. A number of investigators (Scott, 1928, 1929; McCoy, 1931; Foster and Cort, 1932; and others) found that it was difficult to infect dogs with larvae from cats and cats with larvae from dogs. However, the differences were believed to be physiological rather than morphological, and the two were considered different strains of the same species. In 1954 Biocca published the results of his detailed study of Ancylostoma from dogs and cats in Italy. He found that there were two species rather than two strains of one species. In his paper he gave the original descriptions of both Zeder and Ercolani, a redescription of A. tubaeforme, and the major morphological differences between the two species. Bohm (1955) in Austria, Rohde (1959) in Germany, and Fitzsimmons (1961a, b), who studied hookworms from both African (Nyasaland) and Australian dogs and cats, agreed with the findings of Biocca. In the United States the writer has noticed in chemotherapeutic trials of several active compounds that the activity is not always the Received for publication 4 June 1962. same against Ancylostoma from dogs and those from cats. After consulting the papers by Biocca and Rohde, a detailed examination was made of hookworms obtained from dogs and cats in this country. As the morphological differences agree with those found by other investigators in Europe, Africa, and Australia, it is evident that we are dealing with two differ nt species, each of which appears to have a world-wide distribution. The major differences are described and illustrated below. MATERIALS AND METHODS Ancylostoma were obtained from 12 dogs and 9 cats used in chemotherapeutic trials in this laboratory. Numerous measurements were made from relaxed worms obtained either as a result of treatment or at autopsy. Numbers of worms were fixed; the anterior and posterior ends cut off; the middle sections discarded; and the ends dehydrated, cleared, and mounted for detailed study. The best of the numerous mounted ends were photographed to illustrate the major differences between the two species. Ancylostoma from a number of other dogs and cats were examined to determine whether natural infections of A. caninum occurred in cats or A. tubaeforme in dogs. DIFFERENTIATION OF THE TWO SPECIES (All measurements in millimeters, unless otherwise stated) Sizes of worms: Measurements of lengths and widths of 20 mature males and 20 mature females of each species gave the following ranges and averages: A. tubaeforme: males, 9.5 to 11.0 by 0.30 to 0.35 (avg. 10.3 by 0.32); females, 12.0 to 15.0 by 0.38 to 0.43 (avg. 13.2 by 0.41); A. caninum: males, 11.0 to 13.0 by