Dirofilaria immitis and Dirofilaria repens: Investigating the Prevalence of Zoonotic Parasites in Dogs and Humans in a Hyperenzootic Area.

The mosquito-borne zoonotic nematode parasites Dirofilaria immitis and Dirofilaria repens primarily affect dogs. In recent years, their distribution has expanded due to various factors influencing vector-borne pathogens. This study aimed to investigate the comparative prevalence of infection in dogs and humans within a hyperenzootic region of Europe, and to estimate the proportional relationship between infection prevalence in dogs and humans, within the concept of "One Health". To this end, 604 blood samples from dogs and 625 serum samples from humans living in the Thrace region of northeastern Greece were collected. The dog samples were examined for Dirofilaria spp. microfilariae using Kott's test and for D. immitis antigen using a commercial serological test. The human sera were analyzed for both parasites by Western blot. The overall prevalence of infection in dogs was 177 (29.3%), with 173 (28.6%) testing positive for D. immitis and 7 (1.2%) for D. repens, including 6 (1%) cases of mixed infection. Specific IgG antibodies were detected in 42 (6.7%) human samples, with 24 (3.8%) positive for D. immitis and 18 (2.9%) for D. repens. The infection proportion in humans was 23.4% of the corresponding canine infections, indicating a high risk of human infection in this hyperenzootic region.

Geographical isolation and hyperendemicity of Hepatozoon felis: Epidemiological scenario in Skopelos, Greece, and phylogenetic analysis.

Feline hepatozoonosis is a vector-borne disease caused by different species of the genus Hepatozoon, i.e. Hepatozoon felis, Hepatozoon silvestris and Hepatozoon canis. Knowledge on the biology, epidemiology and taxonomy of Hepatozoon spp. is still limited, despite the fact that the number of documented Hepatozoon spp. infections in domestic cats increased in recent years in different countries. This study was carried out to evaluate the prevalence and the genetic profile of Hepatozoon spp. in cats living on the island of Skopelos, Greece. Individual blood samples were collected from 54 owned cats and were subjected to Giemsa-stained blood smear examination to investigate the presence of Hepatozoon spp. gamonts and to a specific PCR protocol targeting the 18S rRNA gene of Hepatozoon. A total of 45 cats (83.3%) were found infected by Hepatozoon spp. by at least one of the methods applied. In particular, 43 (79.6%) of the cats were PCR-positive, and in 6 (11.1%) cats gamonts of Hepatozoon spp. were found in the blood smears. A total of 26 H. felis sequences were obtained and the presence of three undescribed single nucleotide polymorphisms were detected. The present results indicate that H. felis species complex may be hyperendemic in isolated/confined areas. In such contexts, geographical isolation may favor the origin of new genotypes or haplotypes or even new species.

Comparison of diagnostic methods for laboratory diagnosis of the zoonotic tapeworm Dipylidium caninum in cats.

The tapeworm Dipylidium caninum is the most widely distributed cestode infecting dogs, cats, and sometimes humans, worldwide. The diagnosis of the infection caused by D. caninum is achieved via the visualization of proglottids in feces or with traditional microscopic tests, but both lack sensitivity. The present study has evaluated and compared the diagnostic performance of a PCR protocol on different feline biological samples to detect D. caninum. A sample of feces, a Scotch tape test from the perianal area, and a rectal swab were collected from a total of 100 privately owned cats from Italy and Greece. All fecal samples were subjected to macroscopic examination and to floatation. Based on the results of the above tests the cats were divided in three groups, i.e. (i) cats positive for D. caninum (regardless of positivity for other endoparasites (Group A; n = 50 cats), (ii) cats negative for D. caninum but infected by other helminths (Group B; n = 25 cats), and (iii) cats negative for intestinal endoparasites (Group C; n = 25 cats). For each sample, the DNA was extracted from feces, floatation supernatant, Scotch tape test and rectal swabs and subjected to PCR. For 33 cats from Group A, at least one sample type scored positive at PCR. Of these, all were PCR-positive in the floatation aliquot, while nine and one cats were positive by PCR on feces and Scotch tape test, respectively. Swabs were negative by PCR for all the cats. None of the samples from cats of Groups B and C was positive by any PCR. Sequences obtained from amplicons generated from samples of cats enrolled in Italy had 99-100 % identity with the recently described D. caninum feline genotype. The data presented here suggest that PCR could be a useful tool for diagnosing D. caninum infections, under certain circumstances, e.g. when proglottids are unidentified, unseen or overlooked, even though it has limitations, e.g. false negative results due to fecal PCR inhibitors, uneven distribution of parasitic elements, or to intermittent proglottid and/or egg shedding. Thus, it may not be, currently, the best diagnostic choice for dipylidiosis.

Tick paralysis induced by Ixodes gibbosus: enigmatic cases in domestic mammals from Cyprus.

Tick paralysis is a potentially fatal condition caused by toxins produced and secreted by tick salivary glands. This survey presents clinical and epidemiological observations of tick paralysis cases in domestic animals in Cyprus. Local veterinarians report typical tick paralysis cases occurring in goats, sheep, dogs, and cats. The animals suffering from paralysis are free from other neurological diseases, have blood and biochemical parameters within normal ranges, and recover fast by simply removing the ticks found predominantly on the head and around the neck. Tick paralysis cases occur in a specific geographic area of Cyprus (Akamas peninsula), from September through March, but not every year. Instead, the phenomenon has 2 periodic cycles of occurrence, a 3- and a 7-year cycle. The 2 cycles are differentiated by severity based on the number of affected animals and the resulting losses. As described for other tick-borne diseases, these cyclic patterns may be attributed to external factors, self-oscillations of the disease system, or the combined action of these mechanisms. Ticks collected from a recent paralysis case in a goat were morphologically and molecularly identified as Ixodes gibbosus. Efforts should be made to characterize the specific toxins involved in tick paralysis and to develop a vaccine, which could prevent significant losses of small ruminants, especially in free-ranging farming systems, a prevalent management approach observed in Cyprus and various regions worldwide.

Microscopic detection and molecular characterization of Sarcocystis miescheriana in wild boars (Sus scrofa): first report from Greece.

The genus Sarcocystis includes protozoan parasites with an indirect life cycle. Sarcocystis spp. can infect various animal species and humans, causing sarcocystosis, a parasitosis of economic importance and zoonotic concern. Wild boars can act as intermediate hosts for Sarcocystis miescheriana and the zoonotic Sarcocystis suihominis that infects humans by consumption of raw or undercooked infected swine meat. In the present study, the diaphragmatic muscle tissue of 123 wild boars hunted in Greece was examined to determine the frequency of Sarcocystis spp. The samples were examined by tissue compression and molecular techniques. Under light microscopy, 34 out of 123 (27.6%) wild boars tested positive for Sarcocystis spp., while a higher infection prevalence (75%) was revealed by multiplex PCR performed in 100 of the samples. The partial mtDNA cox1 gene (~ 1100 bp) of 20 samples tested positive for S. miescheriana by multiplex PCR was amplified and sequenced. Sarcocystis miescheriana was identified as the only species involved in these infections. This is the first study on the prevalence of Sarcocystis spp. in wild animals in Greece. Further, large-scale surveys are needed to assess the prevalence and species of this parasite in Greece and to design efficient control and preventive measures in a One Health perspective.

First case of macrocyclic lactone-resistant Dirofilaria immitis in Europe - Cause for concern.

Heartworm disease caused by the nematode Dirofilaria immitis is one of the most important parasitoses of dogs. The treatment of the infection is long, complicated, risky and expensive. Conversely, prevention is easy, safe, and effective and it is achieved by the administration of macrocyclic lactones (MLs). In recent years, D. immitis strains resistant to MLs have been described in Southern USA, raising concerns for possible emergence, or spreading in other areas of the world. The present study describes the first case of ML-resistant D. immitis in a dog in Europe. The dog arrived in Rome, Italy, from USA in 2023. Less than 6 months after its arrival in Italy, the dog tested positive for D. immitis circulating antigen and microfilariae, despite it having received monthly the ML milbemycin oxime (plus an isoxazoline) after arrival. The microfilariae suppression test suggested a resistant strain. Microfilariae DNA was examined by droplet digital PCR-based duplex assays targeting four marker positions at single nucleotide polymorphisms (SNP1, SNP2, SNP3, SNP7) which differentiate resistant from susceptible isolates. The genetic analysis showed that microfilariae had a ML-resistant genotype at SNP1 and SNP7 positions, compatible with a resistant strain. It is unlikely that the dog acquired the infection after its arrival in Europe, while it is biologically and epidemiologically plausible that the dog was already infected when imported from USA to Europe. The present report highlights the realistic risk of ML-resistant D. immitis strains being imported and possibly transmitted in Europe and other areas of the world. Monitoring dogs travelling from one area to another, especially if they originate from regions where ML-resistance is well-documented, is imperative. Scientists, practitioners, and pet owners should be aware of the risk and remain vigilant against ML-resistance, in order to monitor and reduce the spreading of resistant D. immitis.

Genotyping USA laboratory-maintained isolates and European clinical isolates of Dirofilaria immitis to assess macrocyclic lactone susceptibility or resistance at predictive SNP sites using droplet digital PCR.

Dirofilaria immitis is a parasitic nematode that causes cardiovascular dirofilariosis ("heartworm disease") primarily in canids. The principal approach for mitigating heartworm infection involves the use of macrocyclic lactone (ML) for prophylaxis. Recent research has substantiated the emergence of D. immitis displaying resistance to MLs in the USA. Numerous factors, such as the mobility of companion animals and competent vectors could impact the spread of drug resistance. Genomic analysis has unveiled that isolates resistant to ML exhibit unique genetic profiles when compared to their wild-type (susceptible) counterparts. Out of the ten single nucleotide polymorphism (SNP) markers validated in clinical samples of D. immitis from the USA, four have demonstrated their effectiveness in distinguishing between isolates with varying ML efficacy phenotypes. This study explores the potential of these confirmed SNPs for conducting surveillance studies. Genotypic analysis using SNP markers emerges as a valuable tool for carrying out surveys and evaluating individual clinical isolates. Two USA laboratory-maintained isolates (Berkeley, WildCat) and twenty-five random European clinical samples of either adult worms or microfilariae (mf) pools isolated from domestic dogs, were tested by droplet digital PCR (ddPCR)-based duplex assay. This approach elucidates genetic evidence pertaining to the development of drug resistance and provides baseline data on resistance related genotypes in Europe. The data on these clinical samples suggests genotypes consistent with the continued efficacy of ML treatment regimens in Europe. In addition, this assay can be significant in discriminating cases of drug-resistance from those possibly due to non-compliance to the recommended preventive protocols.

First Case of Human Anisakiosis in Greece: Acute Invasive Infection Mimicking Peritoneal Malignancy.

Consumption of raw and mildly processed seafood, in the context of modern Western world eating trends, is recognized as a major driver for human fish-borne infections. However, these zoonoses and their unfamiliar risks remain neglected and underappreciated among European diagnosticians. In contemporary Europe anisakidosis is one of the most important fish-borne zoonoses. It is caused by ingesting the third-stage infective larvae of the nematode parasites that belong to the family Anisakidae. The case described herein, is an intestinal and ectopic form of anisakiosis (Anisakis spp.), causing symptoms of subacute abdomen and masquerading as an intraperitoneal malignancy. It is the first anisakidosis case reported in Greece, affecting a young patient who had been repeatedly exposed to the parasite by consuming homemade raw fish. Right hemicolectomy, omentectomy and excision of a descending colon nodule were uneventfully performed. The pathology report confirmed granulomatous tissue with eosinophilic infiltration and parasites that were morphologically and molecularly identified as Anisakis spp. Although challenging, acquiring an accurate diagnosis of anisakidosis can prevent unnecessary surgery, as the infection typically is self-resolving, and if treatment is deemed necessary, it can be limited to antiparasitic medication. However, in rare cases, extra-gastrointestinal migration of larvae can cause severe damage with practically unknown risks, posing a diagnostic and therapeutic dilemma. In such a clinical case scenario, surgical exploration can decisively contribute to a definitive diagnosis and early identification of intraabdominal complications necessitating surgical intervention.

Unusual parasitoses in urban dogs: Urinary capillariosis and linguatulosis, cases report.

Parasitic diseases caused by uncommonly diagnosed parasites may pose a threat to companion animals' health in urban environments where they are least expected. The pentastomid Linguatula serrata (tongue worm) and the capillarid Pearsonema plica (syn. Capillaria plica) are parasites with an indirect life cycle, infecting both domestic and wild carnivores. The present report describes two cases: the first one of urinary capillariosis and the other of linguatulosis, in two dogs living in the urban environment of Athens, Greece. In the case of capillariosis, the dog never lived out of the city, so it was presumably infected in the urban environment. On the contrary, in the case of linguatulosis, the dog was adopted at a young age from a rural environment but remained asymptomatic for several months while living in the city. Both dogs had mild symptoms, compatible with these infections. Urinary capillariosis and linguatulosis are uncommon in owned, pet dogs, living in cities due to epizootiological characteristics, i.e. need for wildlife reservoir for P. plica and consumption of raw infected viscera for L. serrata. Different factors contribute to the fact that such infections may occur in scenarios where they are least expected. Recent studies show a progressive worldwide increase in the number of uncommon parasitoses in pet animals, that in some cases, such as linguatulosis, are of zoonotic relevance. Regular parasitological examinations and preventive antiparasitic schemes are necessary in order to treat and prevent infections in pet animals and safeguard the health of both animals and humans under the concept of One Health.

Biting Back: Advances in Fighting Ticks and Understanding Tick-Borne Pathogens.

Ticks are blood-feeding arthropods and obligate ectoparasites of virtually all animal species (except fish) and humans [...].

Pets, Wildlife and Parasites.

In our dynamic world, borders of different sorts are being rapidly altered or even erased [...].

Diversity and geographic distribution of haplotypes of Dirofilaria immitis across European endemic countries.

BACKGROUND: Dirofilaria immitis, also known as heartworm, is one of the most important parasitic nematodes of domestic dogs, causing a potentially serious disease, cardiopulmonary dirofilariosis, which can be lethal. This species seems to be less 'expansive' than its sister species Dirofilaria repens, and it is believed that climate change facilitates the spread of this parasite to new non-endemic regions. METHODS: In total, 122 heartworm isolates were analysed from nine endemic countries in Europe (Portugal, Spain, Italy, Greece, Hungary, Romania, Slovakia, and Ukraine) and a single isolate from Bangladesh by amplification and sequencing of two mitochondrial (mt) DNA markers: cytochrome c oxidase subunit 1 (COI) and dehydrogenase subunit 1 (NADH). The main aim of the current study was to determine the genetic diversity of D. immitis and compare it with D. repens haplotype diversity and distribution. DNA was extracted from adult heartworms or microfilariae in blood. Most isolates originated from dogs (Canis lupus familiaris) while 10 isolates originated from wildlife species from Romania, including eight isolates from golden jackals (Canis aureus), one isolate from a Eurasian otter (Lutra lutra) and one isolate from a red fox (Vulpes vulpes). RESULTS: Median spanning network analysis was based on the combined sequence (1721 bp) obtained from two mt markers and successfully delineated nine haplotypes (Di1-Di9). Haplotype Di1 was the dominant haplotype encompassing 91 out of the 122 sequences (75%) from all nine countries and four host species. Haplotype Di2 was the second most common haplotype, formed solely by 13 isolates from Italy. The remaining sequences were assigned to Di3-Di9 haplotypes, differing by 1-4 SNPs from the dominant Di1 haplotype. There was evidence for geographical segregation of haplotypes, with three unique haplotypes associated with Italy and four others associated with certain countries (Di4 and Di7 with Slovakia; Di8 with Greece; Di6 with Hungary). CONCLUSION: Diversity in D. immitis mt haplotypes was lower by half than in D. repens (9 vs. 18 haplotypes in D. immitis and D. repens, respectively), which may be associated with the slower expansion of heartworm in Central and NE Europe. NADH gene appears to be conserved in Dirofilaria sp. by showing lower genetic diversity than the analysed COI gene.

Use of In-Clinic Diagnostic Kits for the Detection of Seropositivity to Leishmania infantum and Other Major Vector-Borne Pathogens in Healthy Dogs.

Canine Vector-Borne Diseases (CVBDs) are widespread in Europe and enzootic in many other countries. Though severe illnesses may occur, dogs living in enzootic areas often show vague or no clinical signs of CVBDs. Undiagnosed infections/co-infections in subclinically infected animals favor the spread of CVBDs and increase the risk of transmission to other animals and, in some cases, humans. This study has evaluated the exposure of dogs living in key enzootic countries, i.e., Italy and Greece, to major CVBDs via the use of in-clinic diagnostic kits. Overall, 300 privately owned dogs without/with single mild clinical signs living in different regions of Italy (n. 150) and Greece (n. 150) were included in the study. As part of a clinical examination, a blood sample was collected from each dog and subjected to two serological rapid tests, i.e., the SNAP® 4Dx®Plus (IDEXX Laboratories Inc.) for the detection of antibodies against Ehrlichia spp., Anaplasma spp., Borrelia burgdorferi s.l. and Dirofilaria immitis antigen and the SNAP®Leishmania (IDEXX Laboratories Inc.) for the detection of antibodies against Leishmania infantum. In all, 51 dogs (17%; 95% CI 12.9-21.7) were seropositive to at least 1 pathogen, i.e., 4 in Italy (2.7%; 95% CI 1.4-13.1) and 47 in Greece (31.3%; 95% CI 24-39.4). Dirofilaria immitis antigens were found in 39 dogs (13%; 95% CI 9.4-17.3), while antibodies against Ehrlichia, Anaplasma and Leishmania were detected in 25 (8.3%; 95% CI 5.5-12.1), 8 (2.7%; 95% CI 1.2-5.2) and 5 (1.7%; 95% CI 0.5-3.8) dogs, respectively. None of the dogs tested seropositive for B. burgdorferi s.l. Statistical analyses were performed to evaluate associations between exposure to CVBDs and possible risk factors. The present results indicate that dogs living in enzootic areas may be seropositive for one or more CVBDs in absence of clinical signs. Rapid kits are among first line tools for the detection of CVBDs in clinical settings, as they are cost-effective, straightforward and quick to use. Also, in-clinic tests used herein allowed detection of co-exposure to CVBDs investigated.

Wild Mesocarnivores as Reservoirs of Endoparasites Causing Important Zoonoses and Emerging Bridging Infections across Europe.

Mesocarnivores are small- or mid-sized carnivore species that display a variety of ecologies and behaviours. In Europe, wild mesocarnivores are represented by the red fox (Vulpes vulpes), the golden jackal (Canis aureus), the European wildcat (Felis silvestris), the Mustelidae of the genera Meles, Martes, Mustela, Lutra, the invasive species of raccoon dog (Nyctereutes procyonoides), raccoons (Procyon lotor), and American mink (Neogale vison). These abundant animals thrive in various habitats and often develop their activity close to human settlements. Thus, they may play an important role in the introduction, maintenance, and transmission of major parasitic zoonoses and promote bridging infections with domestic animals. Against this background, this article reports and discusses some of the most important endoparasites of wild mesocarnivores living in Europe, on the basis of their actual role as reservoirs, spreaders, or sentinels. The data derived from epizootiological studies in different European countries, and the proven or speculated implications of the detected endoparasites in human and domestic animals' health, are discussed. Through older and recent literature review, the state-of-the-art knowledge on the occurrence and prevalence of the parasites under consideration is presented, showing further, warranted investigations and the need for surveillance and vigilance.

Ticks, Fleas, and Harboured Pathogens from Dogs and Cats in Cyprus.

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 Echidnophaga 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.

Multicenter randomized, and blinded European field study evaluating the efficacy and safety of Felpreva®, a novel spot-on formulation containing tigolaner, emodepside and praziquantel, in treating cats with mixed infection with intestinal nematodes, cestodes and/or lungworms.

This paper describes a multicentric field study which has evaluated the safety and efficacy of a novel spot on formulation containing emodepside 2.04% w/v, praziquantel 8.14% w/v and tigolaner 9.79% w/v (Felpreva®, Vetoquinol) when administered at the intended commercial dose of 0.15 ml/kg body weight to privately owned cats infected with major intestinal nematodes (Toxocara cati, Toxascaris leonina, Ancylostoma tubaeforme, Uncinaria stenocephala) and/or cestodes (Dipylidium caninum, Taenia taeniaeformis) and/or lungworms (Aelurostrongylus abstrusus, Troglostrongylus brevior). A total of 219 cats from 26 veterinary clinics located in Albania, Greece, Hungary, Italy and Portugal were included in the study. Feces from the cats were examined on a single occasion between Study Day -7 and Day 0 (baseline) and post-treatment (i) twice between Day 7 and Day 14 (± 2) (for intestinal helminths) or (ii) twice between Day 21 (± 2) and Day 28 (± 2) (for lungworms). Cats were allocated into two groups at a ratio of 2:1 (Felpreva®: Profender®, i.e. a commercial control product containing emodepside and praziquantel). Cats infected with intestinal helminths were treated once on Day 0 (i) with Felpreva® (Group 1) or (ii) with Profender® (Group 2). Animals infected with lungworms received a second treatment with Profender® on Day 14 (± 2) regardless of group allocation. Faecal egg or larval count reduction for Felpreva® was 97.47% for intestinal nematodes and 96.80% for lungworms. No cats infected with cestodes at baseline resulted positive after treatment with Felpreva®. However, the low number of cats (n = 10) did not allow for a statistical analysis to be performed. Non-inferiority of Felpreva® compared to Profender® was statistically demonstrated for all target intestinal and respiratory parasites. No adverse events nor application site reactions were observed. These results show that the new topical combination product Felpreva® is highly safe and efficacious in treating infections caused by major species of feline intestinal nematodes, cestodes and lungworms under field conditions.

Endoparasites in wild populations of Cyprus mouflon (Ovis gmelini ophion).

The Cyprus mouflon (Ovis gmelini ophion) is a subspecies of wild sheep, endemic to Cyprus and strictly protected by national and international legislation. In the present study, 107 Cyprus mouflon faecal samples were collected from locations within the species geographical range and examined by standard parasitological methods, i.e. flotation, sedimentation and Ziehl-Neelsen stained smears. Parasites were found in 104 (97.39%) samples. First stage larvae of the lungworms Muellerius capillaris and Cystocaulus ocreatus were found in 93 (86.9%) and in 65 (60.7%) of the samples, respectively, strongylid eggs in 46 (43%), Eimeria spp. oocysts in 44 (41.1%) and Nematodirus spp. Trichuris spp. and Moniezia spp. eggs in 10 (9.3%), 7 (6.5%) and 1 (0.9%) sample, respectively. To the best of the authors' knowledge, this is the first survey on the endoparasites of Cyprus mouflon. The possible role of the parasites found on the health status of the animals and the risk of endoparasite transmission from domestic ruminants to Cyprus mouflon are discussed.

The role of the sequential organ failure assessment score in evaluating the outcome in dogs with parvoviral enteritis.

SCIENTIFIC BACKGROUND: The aim of this prospective study was to assess whether the Sequential Organ Failure Assessment (SOFA) score could be indicative of outcome (survival to discharge) in dogs with parvoviral enteritis. METHODS: In 35 naturally infected dogs, the SOFA score and clinical score were calculated and the presence of systemic inflammatory response syndrome was verified on admission and during the first four days of hospitalization. RESULTS: 26 dogs survived, and out of the 9 non-survivors, 6 dogs had positive blood cultures. Mean SOFA scores and clinical scores between survivors and non-survivors and between septic and non-septic dogs on admission and on each hospitalization day were significantly different. Trends in SOFA score indicated that in non-survivors and septic dogs there was an increase in SOFA score during the first four days of hospitalization and a decrease occurred in survivors and non-septic dogs. The area under the curve (ROC curve analysis) for SOFA score predicting the outcome was 0.797 and predicting sepsis was 0.834. The best cut-off point of SOFA score for predicting the final outcome was 3.5 and the best cut-off of SOFA score for predicting sepsis was also 3.5. CONCLUSIONS: Either single values or trends in SOFA score can assist in suspecting sepsis and reaching prognosis in parvoviral enteritis.

Risk factors for Cryptosporidium infection in small ruminants in northern Greece.

The knowledge of risk factors for Cryptosporidium spp. infection in small ruminants is based on limited data. Therefore, the current research aimed to describe the prevalence and risk factors associated with the occurrence of Cryptosporidium infection in sheep and goat herds in northern Greece. Hence, 530 fresh fecal samples from 59 sheep and goat farms were collected and examined for Cryptosporidium oocysts using microscopy of fecal smears stained by the modified Ziehl-Neelsen technique. The overall prevalence of Cryptosporidium infection for both host species was 34% (180/530; 95% confidence interval (CI): 29.9-38). Specifically, the prevalence for sheep and goats was 33.5% (112/334; 95% CI: 28.4-35.6) and 34.7% (68/196; 95% CI: 28-41.4), respectively. Additionally, standardized questionnaires were filled-in to collect data regarding animals' health status, feeding, and other management practices in each farm. In total 22 risk factors hypothesized to be associated with Cryptosporidium infection were investigated. Multiple logistic regression analysis showed that farms with stagnant water were 11.78 (95% CI: 66-61.5) times more likely to be infected with Cryptosporidium than farms without stagnant water (p < 0.05). Furthermore, farms with more than 25% of their animals suffering from diarrhea were 17.39 (95% CI: 3.43-88.3) times more likely to be infected with Cryptosporidium than farms with ≤ 25% of the animals having diarrhea (p < 0.05). These results suggest that the animal health status and the prevailing environmental conditions play an important role in transmitting Cryptosporidium spp. infection.

A Comparison of Copromicroscopic and Molecular Methods for the Diagnosis of Cat Aelurostrongylosis.

The gold standard method for the diagnosis of cat aelurostrongylosis is the detection of Aelurostrongylus abstrusus first stage larvae with the Baermann's examination. Nevertheless, molecular assays have shown higher diagnostic performances compared to copromicroscopy. This study evaluated the usefulness of an A. abstrusus species-specific PCR on different biological samples collected in clinical settings from 100 privately-owned cats in Italy (n. 60) and Greece (n. 40). A fecal sample was collected from each animal and a pharyngeal swab was also obtained for cats from Italy. All stool samples were subjected to flotation and Baermann's test. The cats were categorized in three groups based on the results of copromicroscopy, i.e., Group A (n. 50 cats with A. abstrusus infection regardless of positivity for other helminths), Group B (n. 25 cats negative for A. abstrusus but positive for at least one of any other helminth), Group C (n. 25 cats negative for any helminth). DNA was extracted from individual aliquots of feces, flotation supernatant, Baermann's sediment and the pharyngeal swab and then subjected to a PCR specific for A. abstrusus. At least one fecal aliquot or the pharyngeal swab scored positive by the A. abstrusus-specific PCR for 48/50 (96%) cats enrolled in Group A; in particular, 38/50 (76%), 35/50 (70%), 41/50 (82%) and 21/25 (84%) DNA extracts from feces, flotation supernatant, Baermann's sediment and pharyngeal swabs were positive by PCR. These results confirm that molecular tools are highly sensitive and specific and indicate that pharyngeal swabs are the most suitable sample for molecular analysis in clinical settings.