Spatial Distribution of Dermanyssus gallinae Infestations in Greece and Their Association with Ambient Temperature, Humidity, and Altitude.

Dermanyssus gallinae, the poultry red mite (PRM), is the most prevalent and harmful ectoparasite of laying hens globally. Although prevalence and risk factor studies can help veterinarians make decisions regarding farm treatments, relevant data are scarce. The present study investigated the prevalence and infestation severity of PRM in poultry farms across Greece and examined potential risk factors. AviVet traps were used to sample 84 farms (51 backyard, 33 industrial) over three years. Farm altitude, temperature, humidity, region, and production systems were assessed as potential risk factors with chi-square tests, initially for all the studied farms and then exclusively for backyard farms. The overall prevalence was 75.0% and was higher in backyard farms (80.4%) compared with industrial ones (66.7%), varying regionally from 66.7 to 90.9%. Altitude and temperature were not significant risk factors, but farms with humidity <60% had a lower infestation risk. Infestation severity did not significantly differ by risk factors. The poultry red mite is highly prevalent across Greek poultry production systems and regions. In the future, global warming, reduced acaricide options, and a ban on cage systems will all threaten a wider spatio-temporal distribution of the PRM, justifying the urgent need for effective monitoring and control methods to protect hen production and welfare and workers' health.

Diet replacement with whole insect larvae affects intestinal morphology and microbiota of broiler chickens.

Insect-based diets are gaining interest as potential ingredients in improving poultry gut health. This study assessed the dietary treatment with whole dried Tenebrio molitor larvae (TM) on broiler chickens' gut microbiota and morphology. 120 Ross-308 broilers received treated diets with 5% (TM5) and 10% (TM10) replacement ratio in a 35-day trial. Intestinal histomorphometry was assessed, as well as claudin-3 expression pattern and ileal and caecal digesta for microbial community diversity. Null hypothesis was tested with two-way ANOVA considering the intestinal segment and diet as main factors. The TM5 group presented higher villi in the duodenum and ileum compared to the other two (P < 0.001), while treated groups showed shallower crypts in the duodenum (P < 0.001) and deeper in the jejunum and ileum than the control (P < 0.001). Treatments increased the caecal Firmicutes/Bacteroidetes ratio and led to significant changes at the genus level. While Lactobacilli survived in the caecum, a significant reduction was evident in the ileum of both groups, mainly owed to L. aviarius. Staphylococci and Methanobrevibacter significantly increased in the ileum of the TM5 group. Results suggest that dietary supplementation with whole dried TM larvae has no adverse effect on the intestinal epithelium formation and positively affects bacterial population richness and diversity.

Ecological niche modeling analysis (Cx. pipiens), potential risk and projection of Dirofilaria spp. infection in Greece.

Vector-borne diseases continue to increase worldwide. Dirofilariosis is one of the most common vector-borne zoonotic diseases, mainly caused by Dirofilaria spp. (D. immitis and D. repens) and spread by culicid mosquitoes of different species. Greece is one of the countries in southern Europe where it is traditionally endemic, and its distribution is not homogeneous. The aim of this study was to develop an environmental model for Greece that reflects the suitability of the ecological niche for Dirofilaria spp. infection risk and its projection until 2080. For this purpose, we used the potential distribution of suitable habitats for Culex pipiens calculated using an ecological niche model (ENM) and the potential number of generations of Dirofilaria spp. The ecological niche model of Cx. pipiens in Greece showed good predictive power (AUC=0.897) with the parasite at a resolution of 1 km2. The variables that contributed most to the model were mean annual temperature, rivers and human footprint. The highest risk of infection was found in coastal areas and in riverside areas of the main river basins, as well as in irrigated areas of the mainland and peninsular regions and in the whole territory of island areas, and the lowest risk was found in areas of higher altitude. A positive relationship was found between the risk of dirofilariosis and the location of infected dogs, with 86.65% located in very high and high risk areas. In 2080, the percentage of territory gained by Cx. pipiens will increase by 261.52%. This model provides a high predictive value, predicted presence, and risk of Dirofilaria spp. infection and can serve as a tool for the management and control of this disease.

Extensive Countrywide Molecular Identification and High Genetic Diversity of Haemonchus spp. in Domestic Ruminants in Greece.

The gastrointestinal nematode parasite Haemonchus spp. is one of the most pathogenic parasites of ruminants, due to its blood-sucking activity, which causes large economic losses in the ruminant industry. The latest epizootiological data recorded an increase in the infection, not only in Greece but also in other countries, mainly attributed to climatic changes. The study of the population structure and the investigation of the phylogenetic relationships of Haemonchus spp. are essential for the understanding of its biology and epizootiology to implement appropriate control and prevention strategies. In addition, the molecular approach allows the determination of evolutionary relationships between different species of this parasite, the diverse hosts they infect, as well as the different geographic compartments from which they originate. Therefore, the aim of the present study was to identify the species of the sympatric populations of the genus Haemonchus, a nematode parasite infecting ruminants (sheep, goats, cattle, and buffaloes) from different regions of Greece (continental and insular) using molecular methods. At the same time, an attempt was made to identify the possible subpopulations of Haemonchus spp. in Greece, to investigate their phylogenetic relationships, as well as to determine the genetic diversity of each population. A total of 288 worms of the genus Haemonchus were processed using molecular methods; of these, 96 were collected from sheep, 96 from goats, 48 from cattle, and finally, 48 from buffaloes. A fragment of 321 base pairs of the second internal transcribed spacer (ITS2) sequence of nuclear DNA was amplified for species identification, and, after basic local alignment search tool (Blast) analysis, it was revealed that they belonged to H. contortus. A fragment of 820 base pairs of subunit 4 of the nicotinamide dehydrogenase (ND4) gene of mitochondrial DNA was amplified for genetic diversity analysis. The Greek mitochondrial ND4 sequences of H. contortus were classified into 140 haplotypes, and the values of the average nucleotide and haplotype diversity were lower compared to the respective values derived from Italy, Malaysia, the USA, and China. The phylogenetic analysis of the ND4 gene revealed a clear grouping of the Greek haplotypes when compared with Asian ones, and, at the same time, there was no profound grouping of the same haplotypes with regard to their different hosts and geographical origin within different regions of Greece. The aforementioned findings confirmed that H. contortus prevails in our country and can infect all species of ruminants, without geographical boundaries, when the right conditions (i.e., common grazing) are created.

Diagnostic performance of a rapid immunochromatographic test for the simultaneous detection of antibodies to Theileria equi and Babesia caballi in horses and donkeys.

BACKGROUND: Equine piroplasmosis is caused by two tick-borne protozoan parasites, Theileria equi and Babesia caballi,, which are clinically relevant in susceptible horses, donkeys, and mules. Moreover, equine piroplasmosis significantly constrains international trading and equestrian events. Rapidly diagnosing both parasites in carrier animals is essential for implementing effective control measures. Here, a rapid immunochromatographic test for the simultaneous detection of antibodies to T. equi and B. caballi was evaluated using samples from horses and donkeys collected in Greece, Israel, and Italy. The results were compared with an improved competitive enzyme-linked immunosorbent assay (cELISA) for detecting antibodies to both parasites using the same panel of samples. METHODS: Blood samples were collected from 255 horses and donkeys. The panel consisted of 129 horses sampled at four locations in northern Greece, 105 donkeys sampled at four locations in Sicily, and 21 horses sampled at two locations in Israel. The rapid test and the cELISA were performed according to the manufacturer's instructions, and the results were subjected to a statistical analysis to determine the sensitivity and specificity of both tests and their association. RESULTS: The immunochromatographic test provided a result within 15 min and can be performed in the field, detecting both pathogens simultaneously. The overall coincidence rate between the rapid test and the cELISA for detecting antibodies against T. equi was 93% and 92.9% for B. caballi. The rapid test's sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) for T. equi were above 91.5%. Sixteen samples were positive for both parasites in the rapid test and eight in the cELISA. Either test had no significant association between T. equi and B. caballi detection. The detection rates of both parasites were significantly higher in Italy than in Greece or Israel and in donkeys than in horses. The agreement for T. equi between the results of both tests was high in Greece (93.8%) and Italy (95.2%) and moderate in Israel (76.2%). For B. caballi, the specificity and NPV of the rapid test were high (94.2% and 98.3%, respectively), although the sensitivity and PPV were moderate (69.2% and 39.1%, respectively) due to the small sample size. However, for B. caballi, the sensitivity was higher with the rapid test. CONCLUSIONS: The rapid test detected T. equi and B. caballi simultaneously in the field, potentially replacing laborious cELISA testing and is recommended for import/export purposes. The test can also be helpful for the differential diagnosis of clinical cases, since seropositivity may rule out equine piroplasmosis since it does not indicate current or active infection.

Efficacy of eprinomectin 5 mg/mL topical solution administered pour on at 1 mg per kg body weight against Oestrus ovis myiasis in sheep and goats.

The larvae of the nasal bot, Oestrus ovis, mainly parasitize sheep and goats and some species of wild Caprinae but other mammals and humans are also vulnerable to infestation. Eprinomectin 5 mg/mL topical solution (EPRINEX® Multi, Boehringer Ingelheim) administered at 1 mg eprinomectin per kg body weight pour on was recently authorized as an anthelmintic for sheep and goats with zero hours milk withdrawal in several countries in Europe. As the product in cattle has claims against a broad range of parasites including insect parasites and activity against O. ovis has previously been reported following extra-label use in sheep, its therapeutic efficacy against ovine and caprine O. ovis myiasis was evaluated in three regulatory compliant, masked clinical studies. Pre-study recovery of O. ovis larvae from five or six of six randomly selected animals per study site (Bulgaria, one site, sheep; Greece, two sites, sheep or goats) supported the inclusion of the animals from those sites into the studies. The study animals (34 animals per study) were ranked based on bodyweight and allocated randomly to remain untreated (control) or to be treated with eprinomectin 5 mg/mL topical solution at 1 mL per 5 kg body weight pour on. Treatment efficacy was determined based on O. ovis larval counts of eprinomectin 5 mg/mL topical solution-treated vs. untreated animals three weeks after treatment administration. Live O. ovis larvae, including all three instars in each study, were recovered from 13 or 16 of the 17 control animals in the sheep studies (range, 1 to 14 or 5 to 18 larvae, respectively) and from all 17 controls in the goat study (range, 7 to 18 larvae). In each study, eprinomectin 5 mg/mL topical solution-treated animals had significantly (p < 0.001) fewer live O. ovis larvae than the controls. Efficacy of the treatment was 100% and 91.3% against the combined parasitic O. ovis larval stages in sheep and in goats, respectively. The treatment was well accepted by all animals and no health problems were observed throughout the studies. The results of these studies demonstrated eprinomectin 5 mg/mL topical solution administered pour on at 1 mL per 5 kg body weight to be an efficacious and safe treatment of ovine and caprine oestrosis.

Is the European badger a new host for Dirofilaria immitis? The first records in Greece.

Dirofilaria immitis is a ubiquitous nematode parasite with zoonotic potential, transmitted by mosquitoes, that causes heartworm disease in various animal species. Dogs are the parasite's typical final host, and wild carnivores represent the parasite's reservoir in nature. Studies on D. immitis infections in wild animals are essential to assess infection pressure for domestic animals, and until now, there has been only one infection case reported in a European badger (Meles meles). The current report describes the first two European badger cases with cardiovascular dirofilariosis in Greece. Two adult male badgers were rescued in Heraklion and Chania, Crete Island, and admitted to "ANIMA -Wildlife Rehabilitation Centre" in Athens. The detailed clinical examination revealed that the first badger suffered from severe broncho-pneumonitis while the second one displayed clinical signs associated with severe brain trauma. Blood samples were taken for haematology and biochemistry analyses during their short hospitalisation period. In addition, different routine diagnostic tests were carried out, including heartworm antigen testing (ELISA) and the modified Knott's test for microfilariae. Both badgers were positive in both tests. The animals died a few hours after their admission and the detailed necropsies followed, revealed the presence of three parasites in each animal's right heart, morphologically identified as adults of D. immitis. These findings add the European badger in the list of additional potential reservoir hosts for D. immitis and highlight the potential role of wildlife for companion animals and human health.

Efficacy of afoxolaner or the combination of afoxolaner with milbemycin oxime against Otodectes cynotis in naturally infested dogs.

Otodectes cynotis, commonly known as "the ear mite," is a highly contagious ectoparasite and a significant cause of otitis externa in canines. The objective of the current study was to determine the efficacy of the isoxazoline afoxolaner (Nexgard®), and the combination of afoxolaner with milbemycin oxime (Nexgard Spectra®), in dogs naturally infested with O. cynotis. In total, 32 infested client-owned dogs from two different sites in Greece were included. The animals were randomly divided into four equal groups based on their infestation score. Group 1 served as the negative control, group 2 received one oral administration of Nexgard (Day 0), group 3 received two monthly oral administrations of Nexgard (Days 0, 30), and group 4 received two monthly oral administrations of Nexgard Spectra (Days 0, 30), according to label instructions. Otoscopic examinations for mites and observations on debris/cerumen in the ears were carried out on Days 0, 15, 30, and 45. A quantitative assessment of ear mites by ear duct flushing and live mite counts was performed on Day 45. The results demonstrated that a single oral dose of afoxolaner and two monthly doses of afoxolaner or afoxolaner with milbemycin oxime resulted in a 99.9% reduction in live mite counts compared to the untreated control group by Day 45. Additionally, treated dogs showed improved clinical symptoms, such as ear cerumen/debris decrease, while untreated dogs experienced worsening symptoms over the study duration. No adverse events were reported. Overall, these results support the use of afoxolaner-based products to treat O. cynotis infestation in dogs.

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.

A Review of Toxoplasma gondii in Animals in Greece: A FoodBorne Pathogen of Public Health Importance.

Toxoplasma gondii is a zoonotic protozoon with a complex life cycle and the second most important foodborne pathogen in Europe. Surveillance of toxoplasmosis is based on national considerations since there are no mandatory controls along the food chain in the European Union, and underreporting of meat is still a problem in many countries like Greece. The current review provides an overview of T. gondii prevalence, associated risk factors, and surveillance in animals in Greece, focusing on the transmission role of meat and highlighting the control measures that should be adopted by consumers. Sows, wild boars, hares, equines, and cats had lower, while sheep and goats generally had higher seroprevalence than their respective pooled European and global values. Seroprevalence in chickens was similar between Greece and Europe, while there was high variation in cattle studies, with no data regarding dairy products. Though a comprehensive meat safety assurance system is the most effective approach to control the principal biological hazards associated with meat, such as T. gondii, the prerequisite risk categorisation of farms and abattoirs based on EFSA's proposed harmonised epidemiological indicators has not materialised as yet in Greece. Therefore, comprehensive control strategies are still required to ensure food safety and safeguard public health.

First clinical case of tick-borne encephalitis (TBE) in a dog in Greece.

Tick-borne encephalitis virus (TBEV) causes tick-borne encephalitis (TBE), affecting human health in Europe and Asia. Reports on canine clinical cases of TBE are rare, although dogs are used as sentinels for assessing human health risks. The first canine clinical TBE case in Greece is reported in this case report. The dog had a history of tick infestation and displayed neurological symptoms, particularly tetraparesis, neck hyperalgesia, and a sudden behavior change. Serum samples were obtained and examined in a commercial ELISA to detect anti-TBEV specific IgG and IgM antibodies. The dog tested seropositive for both IgG and IgM, and based on its history and compatible clinical signs, the diagnosis of TBE infection was reached. The prognosis was initially poor, and treatment included the administration of fluids, corticosteroids, and antibiotics, followed by physical therapy. After a 10-day hospitalization, the dog had a much better prognosis. This case highlights that TBEV does emerge in new locations, increasing human and animal infection risk. Veterinarians should include TBE in their differential diagnosis of canine patients with a history of tick infestations, progressive neurological symptoms, and abnormal behavior.

Leishmaniosis in Greece: The Veterinary Perspective.

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. Serological 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.

Hepatozoon species infecting domestic cats from countries of the Mediterranean basin.

Tick-borne diseases (TBDs) are caused by pathogens of human and veterinary concern representing a major public health issue worldwide. Although feline medicine has progressed much in the recent decades, data on feline TBDs (FeTBDs) remain scant. Therefore, this study aimed to assess the prevalence of apicomplexan parasite infections, associated risk factors and clinical-hematological abnormalities in domestic feline populations from countries of the Mediterranean basin. Blood and serum samples from cats (n = 600) living in France, Greece, Israel, Italy, Portugal and Spain were collected along with animal data (i.e., age, sex, breed, housing conditions and geographical origin), clinical signs and laboratory blood test parameters. Cats were grouped according to their age as kitten (up to one year), young (between one and six years), mature (between seven and ten years) and senior (older than ten years). Blood samples were tested for Hepatozoon spp. and piroplasmids by conventional PCR targeting 18S rRNA gene. The overall prevalence of Hepatozoon spp. infection was 14.5%, being significantly higher in cats from Greece (30%) and Portugal (23%), followed by Spain (15%), Israel (15%) and France (4%). Cats from Italy scored negative. Hepatozoon felis was identified in 86 animals, with three different sequence types and H. silvestris was detected in one shelter cat from Portugal. No piroplasmid DNA was amplified. The risk of Hepatozoon spp. infection was related to feline geographical provenience, housing condition and age. No statistical correlation was reported with any clinical signs, while increased alanine aminotransferase (ALT) activity was the only laboratory abnormality significantly associated (p = 0.03) with the infection. Data suggest a high circulation of H. felis, and only occasionally of H. silvestris, within domestic feline populations in the Mediterranean basin, mainly in shelter or free roaming and young cats with asymptomatic or subclinical infection.

Growth performance, welfare traits and meat characteristics of broilers fed diets partly replaced with whole Tenebrio molitor larvae.

The role of insects in animal nutrition has been reconsidered during recent years, paving the way for an increasing market for edible insects. Their protein and amino acid balance make them a promising source of protein for replacing high value proteins. Yellow mealworm, Tenebrio molitor L. (TM; Coleoptera: Tenebrionidae) larvae, have shown positive effects on broiler performance in several research studies and have a strong potential as a sustainable alternative protein source for monogastric animals. This study aimed to assess the effect of replacing various ratios of basal diets with T. molitor larvae on broiler performance as well as on several meat and welfare characteristics. For the study, 120 one-day-old male chicks (Ross 308) were randomly allocated in 3 treatments and 4 replications (10 birds per pen). Birds of the control group (basal diet) were fed with typical commercial maize and soymeal-based rations in mash form. The other 2 groups were treated with the same diet, after replacing 5% and 10% with dried TM larvae, respectively. On d 35 (end of trial), meat samples were collected and analysed. Body weight, feed intake, body weight gain and feed conversion ratio during the periods of 1 to 10 d (starter period), 11 to 24 d (grower period), 25 to 35 d (finisher period) and 1 to 35 d (total period) were assessed. Pododermatitis, diarrhoea, feather score and litter conditions were also assessed during the trial. The results indicated that TM larvae inclusion in the broilers' diet positively affected body weight gain values, as well as the carcass yield, the meat composition and various welfare traits. Additionally, the dietary treatments with TM larvae favourably affected meat composition and colour parameters, whereas there were also some positive effects on lipid and protein oxidation. Saturated fatty acids were decreased by the dietary supplementation whereas the polyunsaturated fatty acids to SFA ratio increased. In general, the study showed that whole TM larvae addition can provide a promising alternative to soybean meal in the diet of broilers, demonstrating a positive impact on growth, welfare and meat characteristics.

Helminth Infections in Dairy Sheep Found in an Extensive Countrywide Study in Greece and Potential Predictors for Their Presence in Faecal Samples.

The aims of the present study were: (a) to describe the prevalence of helminth infections from pooled faecal samples from sheep flocks across Greece and (b) to evaluate flock-related factors potentially associated with the presence of these infections in the flocks. An extensive countrywide study was performed on 325 sheep farms throughout Greece; faecal samples were collected from ewes and processed for the identification of helminth parasites. Helminths were detected in samples from 92.9% of flocks; these included Dicrocoelium dendriticum (16.7% of flocks), Fasciola hepatica (0.6%), Paramphistomum cervi (2.2%), Moniezia spp. (18.8%), Trichostrongylidae (85.5%), Nematodirus spp. (18.8%), Strongyloides papillosus (7.1%), Trichuris spp. (20.0%) and lungworms (17.8%). Mean Trichostrongylidae counts across all flocks in the study were 215 epg. Specifically, for Trichostrongylidae, there were differences between flocks that had or had not received anthelmintics during the two months prior to sampling, as well as between flocks located in various areas of the country. In multivariable analyses, for the outcome 'high (>300) epg counts in faecal samples', the month into the lactation period at sampling and the application of reproductive control practices on the farm emerged as significant factors; for the outcome 'high proportion (>63%) of Teladorsagia spp. in faecal samples', the availability of straw bedding emerged as a significant factor; and for the outcome 'high proportion (>63%) of Haemonchus contortus in faecal samples', the age of the farmer emerged as a significant factor. For the outcome 'presence of Trichuris spp. in faecal samples', the provision of finished feed (concentrate) to animals emerged as a significant factor, whilst, finally, for the outcomes 'presence of D. dendriticum in faecal samples' and 'presence of lungworms in faecal samples,' no significant factors emerged.

In Vitro Acaricidal Activity of Silver Nanoparticles (AgNPs) against the Poultry Red Mite (Dermanyssus gallinae).

Dermanyssus gallinae (PRM) is the most common blood-sucking ectoparasite in laying hens and is resistant against numerous acaricides. Silver nanoparticles (AgNPs) represent an innovative solution against PRM. The current study aimed to assess the in vitro acaricidal activity of AgNPs against PRM and describe their potential mechanism of action. Nanoparticles were produced using a wet chemistry approach. Mites were collected using AviVet traps from 18 poultry farms in Greece. Contact toxicity bioassays were carried out for 24 h with negative controls, 20, 40, 60, or 80 ppm AgNPs. Analysis of variance was used to compare the mortality rates of PRM between the control and treatment groups, while LC50, LC90, and LC99 values were estimated using probit regression analysis for the total farms jointly and separately. Nanoparticles displayed strong acaricidal activity, and mortality rates were significantly different between groups and increased by AgNPs concentration. Overall mean LC50, LC90, and LC99 values were 26.5, 58.8, and 112.3 ppm, respectively. Scanning electron microscopy on mites treated with 80 ppm AgNPs revealed cracks in their exoskeleton and limb detachments, presumably resulting from the interaction between AgNPs and the mites' chitin. Future studies should focus on assessing AgNPs residues in chicken tissues before moving into field trials.

Isolation of Listeria monocytogenes from poultry red mite (Dermanyssus gallinae) infesting a backyard chicken farm in Greece.

The poultry red mite (PRM), Dermanyssus gallinae, is arguably the most harmful, ubiquitous haematophagous ectoparasite infesting egg-laying hens. PRM is a vector of various microorganisms, with some being important for food microbiology and public health. The present study aimed to investigate the presence of specific pathogens, including Escherichia coli, Salmonella spp. and Listeria spp., carried by PRM infesting a chicken farm in Greece. Mites were caught using cardboard traps (Avivet), and 100 unwashed PRM were homogenized and used for microbiological cultures. Microbiological cultures were carried out on general and selective substrates to detect the above-mentioned bacteria. Specifically for Listeria spp., DNA was extracted from bacteria grown in Tryptone Soya Yeast Extract Agar using a commercial kit. The hly gene encoding the Listeriolysin O protein was amplified by PCR. Mites were identified as D. gallinae using morphological keys as well as by COI DNA barcoding. Microbiological cultures and PCR assays were positive for Listeria monocytogenes. No other bacteria were detected. The current study constitutes the first molecular isolation of L. monocytogenes from D. gallinae, confirming that PRM can carry this food-borne pathogen. PRM control measures and hygiene practices should be applied to minimize any possible contamination risk of poultry products with L. monocytogenes and safeguard public health.

Intracellular IFN-γ and IL-4 levels of CD4 + and CD8 + T cells in the peripheral blood of naturally infected (Leishmania infantum) symptomatic dogs before and following a 4-week treatment with miltefosine and allopurinol: a double-blinded, controlled and cross-sectional study.

BACKGROUND: Canine leishmaniosis (CanL) is a systemic disease caused by the protozoan parasite Leishmania infantum with a wide spectrum of clinical signs, with cutaneous, ocular, renal and lymphoreactive conditions prevailing in the clinical setting. The immune system plays a pivotal role in the evolution of Leishmania infection and its response to antileishmanial treatment. Cytokines are important immune response mediators that are released by activated lymphocytes and less so by other immunocytes. In dogs with leishmaniosis, IFN-γ and IL-4 have been recognized as the main activators of cellular and humoral immunity, respectively. The objective of this study was to investigate intracellular IL-4 and IFN-γ expression by CD4 + and CD8 + lymphocytes in the peripheral blood of symptomatic dogs before and after combined antileishmanial treatment with miltefosine and allopurinol. RESULTS: Postantileishmanial treatment CD4 + IL-4 + and CD8 + IL-4 + cell counts were significantly decreased, although no similar changes were observed in the comparisons made between the pre- and posttreatment CD4 + IFN-γ + and CD8 + IFN-γ + counts and ratios. CONCLUSION: The findings indicate that IL-4 production by T cells may facilitate the symptomatic phase of CanL, whereas IFN-γ production by CD4 + and CD8 + cells may indicate its negligible role in the evolution of natural CanL and perhaps the equivocal positive influence of antileishmanial treatment.

Feline Toxoplasmosis in Greece: A Countrywide Seroprevalence Study and Associated Risk Factors.

Toxoplasma gondii is a ubiquitous zoonotic parasite, with felines being the only definitive hosts. Cats shed oocysts with their faeces, and seroprevalence studies can be used to indirectly assess the environmental contamination. The current study aimed to evaluate T. gondii seroprevalence in Greek cats and identify possible risk factors. In total, 1554 blood samples were analyzed from different cats across all nine geographic regions of Greece, and a short questionnaire was completed for each cat. A rapid immunochromatographic test was used to detect anti-T. gondii antibodies, IgG type, and 21.8% of cats were seropositive. Regarding risk factors, when chi-square tests were applied, seropositivity was significantly higher (p < 0.05) in rural cats, cats with outdoor access, and hunting cats. Gender, age, ownership, and raw feeding were not significant risk factors, although female, adult, stray, and raw-feeding cats had a higher seroprevalence than their counterparts. Binary logistic regression models were developed to adjust for the confounding effects of the initially recognized risk factors, and only hunting in urban areas remained a significant risk factor. Greek cats had lower seropositivity than the average European value, and the present research highlights the importance of updated seroprevalence and risk factor studies within the context of One-Health.

Protozoan Parasites in Adult Dairy Small Ruminants and Potential Predictors for Their Presence in Faecal Samples.

There is a scope to study protozoan infections in adult ewes and does, as these animals can act as reservoirs of infection for lambs and kids, for which these pathogens are harmful. The objectives of this study were to describe the prevalence of protozoan infections in faecal samples from adult sheep and goats on dairy farms across Greece and to evaluate farm-related factors potentially associated with the presence of protozoan infections in these animals. A cross-sectional study was performed on 325 sheep and 119 goat farms throughout Greece; faecal samples were collected from ewes and does and processed for the identification of protozoan parasites. Eimeria oocysts were found in faecal samples from 69% of farms (72% of sheep farms and 61% of goat farms), Giardia cysts in samples from 33% of farms (33% of sheep farms and 34% of goat farms) and Cryptosporidium oocysts in samples from 8% of farms (7% of sheep farms and 11% of goat farms). In a multivariable analysis, for the presence of Eimeria in samples from sheep farms, the lack of a designated building for lambs emerged as a significant factor; for the presence of Giardia in samples from goat farms, the availability of a main building for animals emerged as a significant factor; for the presence of Cryptosporidium, the lack of grazing and the management system emerged as the main significant factors in sheep and goat farms, respectively. Protozoa were found significantly more frequently in samples collected from farms on which farmers considered diarrhoea as an important health problem in their lambs/kids.