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The paper reviews the challenges about haemonchosis—a significant and common parasitic infection of small ruminants. Haemonchus contortus is a highly pathogenic parasite that localises in the abomasum of affected animals and exerts its pathogenicity by blood-sucking activity, adversely affecting the health and productivity of animals. The first challenge is the uneven distribution of the infection globally, this being more prevalent in tropical and subtropical and warm temperate and summer rainfall regions than in cool and cold temperate and arid regions; hence, this leads in differences in the approaches required for its control. Another challenge is the widespread presence of Haemonchus strains resistant to the various anthelmintics available: Benzimidazoles, imidazothiazoles, macrocyclic lactones, closantel and monepantel, which makes the control of the infection difficult. The third challenge refers to the difficulty of diagnosing the disease, given that field evidence can provide suspicion about the infection, which needs to be subsequently confirmed by laboratory tests through parasitological or molecular techniques. The final challenge relates to the difficulties in the control of the infection and the necessity to use pharmaceutical products cautiously and with a planned approach, to avoid further development of anthelmintic resistance, also given that use of a recently licenced vaccine is not widespread. In conclusion, at the moment, we should be concerned, but not worried, about this infection, and apply correctly the appropriate health management plans.

Otodectes cynotis is the agent of otodectic mange, a disease affecting wild and domestic felines, canines and mustelids. It is a highly contagious and pruritic condition and a major cause of otitis externa in dogs. This study was designed to verify the efficacy of NexGard ® PLUS, a combination of afoxolaner, moxidectin and pyrantel pamoate, against natural O. cynotis infestations in dogs. It was a blinded, randomised, single-centre, negative controlled clinical efficacy study. Twenty-four naturally infested dogs were allocated to three groups of eight dogs: an untreated control group, a group treated on Day 0 and a group treated on Days 0 and 29 (both groups were treated per label recommendations). Otoscopic examinations were performed on Days -3, 14, 29, 42 and 55 for clinical otodectic mange evaluation. Ear canal flushings were performed on Day 56 for O. cynotis counts, the primary variable of efficacy, calculated per comparison of live mites in the treated groups, with the untreated control group. The otoscopic examinations revealed treatment effect by Day 14 and a highly significant effect by Day 29, while the conditions remained unchanged or worsened in the untreated control group. The O. cynotis counts revealed reductions of 100% in the group treated once on Day 0 ( p  = 0.0004) and 99.7% ( p  = 0.0006) in the group treated on Days 0 and 29, while 48.8 (geometric mean) live mites were collected in the untreated control group. NexGard ® PLUS was demonstrated to be highly efficacious in dogs naturally infested with O. cynotis following one or two treatments. Otodectes cynotis est l’agent de la gale otodectique, une maladie affectant les félins, les canidés et les mustélidés, sauvages et domestiques. Très contagieuse et prurigineuse, cette affection est une cause majeure d’otite externe chez le chien. Cette étude visait à évaluer l’efficacité de NexGard ® PLUS, une association d’afoxolaner, de moxidectine et de pamoate de pyrantel, contre les infestations naturelles par O. cynotis chez le chien. Il s’agissait d’une étude clinique d’efficacité, monocentrique, randomisée, en double aveugle et contrôlée par un groupe négatif. Vingt-quatre chiens naturellement infestés ont été répartis en trois groupes de huit : un groupe témoin non traité, un groupe traité au jour 0 et un groupe traité aux jours 0 et 29 (les deux derniers groupes ont été traités conformément aux recommandations de l’AMM). Des examens otoscopiques ont été réalisés aux jours -3, 14, 29, 42 et 55 pour l’évaluation clinique de la gale otodectique. Des lavages du conduit auditif ont été effectués au jour 56 pour le dénombrement d’ O. cynotis , principal critère d’efficacité, calculé par comparaison du nombre d’acariens vivants dans les groupes traités avec celui du groupe témoin non traité. Les examens otoscopiques ont révélé un effet du traitement dès le jour 14 et un effet hautement significatif au jour 29, tandis que l’état des chiens du groupe témoin non traité est resté inchangé ou s’est aggravé. Le dénombrement d’ O. cynotis a montré une réduction de 100 % dans le groupe traité une seule fois au jour 0 ( p  = 0,0004) et de 99,7 % ( p  = 0,0006) dans le groupe traité aux jours 0 et 29, tandis que 48,8 acariens vivants (moyenne géométrique) ont été collectés dans le groupe témoin non traité. NexGard ® PLUS s’est avéré très efficace chez les chiens naturellement infestés par O. cynotis après un ou deux traitements.

Background Canine vector-borne diseases (CVBDs) represent a wide group of diseases of major significance for canine health. In addition to their veterinary importance, many of these diseases are of great zoonotic concern, posing a risk of potential transmission to humans. To date, there has been scant knowledge regarding the prevalence, distribution and risk factors of CVBDs in Greece. Therefore, the objectives of the present study were to update the current knowledge on the seroprevalence of Dirofilaria immitis , Anaplasma spp., Ehrlichia spp. and Borrelia burgdorferi ( sensu lato ) in dogs in Greece and, furthermore, to assess possible environmental and any other risk factors associated with these infections. A total of 1000 apparently healthy and randomly selected dogs, presented in veterinary clinics, were involved at the national level ( n  = 66 municipalities). Serum samples were obtained from each individual dog and were tested using the SNAP ® 4Dx ® Plus kit from IDEXX Laboratories. Possible risk factors were assessed using binary regression models, including dogs’ lifestyle, climatological parameters and the altitude of the region. Results Overall, 21.8% (95% CI: 19.4–24.5%) of the sampled dogs were found to be seropositive to at least one of the four pathogens examined. The most prevalent pathogen was Ehrlichia spp. (12.5%, 95% CI: 10.6–14.7) followed by D. immitis (9.0%, 95% CI: 7.8–11.5) and Anaplasma spp. (6.2%, 95% CI: 4.9–7.9). The lowest prevalence (0.1%) was recorded for B. burgdorferi ( s.l. ) where only one dog was found to be positive. Among the examined risk factors, low mean temperature was found to increase the prevalence of Ehrlichia spp. ( P  ≤ 0.001) and Anaplasma spp. ( P  ≤ 0.001), while low minimum temperature increased the prevalence of D. immitis ( P  ≤ 0.001). In addition, low total annual rainfall had an effect of the prevalence of Ehrlichia spp. ( P  ≤ 0.01). Altitude also had a significant effect on the prevalence of D. immitis ( P  ≤ 0.05) and Anaplasma spp. ( P  ≤ 0.01). Conclusions To our knowledge, this is the first large-scale seroepidemiological study of CVBDs in Greece. It has been evidenced that environmental factors such as temperature, rainfall and altitude can influence the prevalence and distribution of CVBDs.

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.

Canine sarcoptic mange, caused by Sarcoptes scabiei , is a highly contagious and intensely pruritic skin disease in dogs. It is prevalent worldwide and has zoonotic potential. Therefore, effective treatment is important to safeguard animal welfare and public health. The present clinical field study aimed to confirm the efficacy of NexGard ® Plus, an oral combination of afoxolaner, moxidectin and pyrantel pamoate, in treating dogs naturally infested with S. scabiei . It was a blinded, randomised, single-centre, negative-controlled efficacy study. Twenty naturally infested dogs were allocated into two groups: a group treated on Day 0 and Day 26/28 at the label dose, and an untreated control group. Skin scrapings were conducted similarly, once between Day −6 to 0, then on Days 26/28 and 56 for mite counts. Assessments of clinical signs were conducted at the same time intervals. In the treated group, mite infestations were reduced by 97% after the first treatment and were eliminated (100%) after the second treatment ( p  < 0.0005), while all dogs in the untreated control group remained infested for the whole study. Treated dogs had no pruritus, papules or crusts and clear evidence of hair regrowth by Day 56, unlike the dogs in the control group. This study demonstrated the elimination of S. scabiei mites and significant improvement of sarcoptic mange clinical signs in naturally infested dogs treated with the oral combination of afoxolaner, moxidectin and pyrantel. La gale sarcoptique canine, causée par Sarcoptes scabiei , est une dermatose très contagieuse et extrêmement prurigineuse chez le chien. Elle est répandue dans le monde entier et présente un potentiel zoonotique. Un traitement efficace est donc essentiel pour préserver le bien-être animal et la santé publique. La présente étude clinique de terrain visait à confirmer l’efficacité de NexGard ® Plus, une association orale d’afoxolaner, de moxidectine et de pamoate de pyrantel, dans le traitement des chiens naturellement infestés par S. scabiei . Il s’agissait d’une étude d’efficacité monocentrique, randomisée, en double aveugle et contrôlée par un placebo. Vingt chiens naturellement infestés ont été répartis en deux groupes : un groupe traité aux jours 0 et 26/28 à la dose recommandée, et un groupe témoin non traité. Des prélèvements cutanés ont été effectués de la même manière, une première fois entre le jour −6 et le jour 0, puis aux jours 26, 28 et 56 pour le dénombrement des acariens. L’évaluation des signes cliniques a été réalisée aux mêmes intervalles. Dans le groupe traité, l’infestation par les acariens a diminué de 97% après le premier traitement et a été totalement éliminée (100 %) après le second ( p < 0,0005), tandis que tous les chiens du groupe témoin non traité sont restés infestés pendant toute la durée de l’étude. Les chiens traités ne présentaient ni prurit, ni papules, ni croûtes, et une repousse des poils était clairement visible au jour 56, contrairement aux chiens du groupe témoin. Cette étude a démontré l’élimination des acariens S. scabiei et une amélioration significative des signes cliniques de la gale sarcoptique chez les chiens naturellement infestés traités par l’association orale d’afoxolaner, de moxidectine et de pyrantel.

Rumen development is critical for optimal growth, feed efficiency and health in pre-weaned dairy calves. Butyric acid, a short-chain fatty acid produced in the rumen, has been shown to stimulate epithelial development and modulate immune responses. This study evaluated the effects of butyric salt supplementations, as metabiotics, on rumen morphometry and investigated the potential associations with feed intake, growth performance and health indicators in pre-weaned dairy calves. Thirty calves were randomized into three groups: controls (G1), 25 g (G2) or 50 g (G3) sodium butyrate from birth until weaning. The gross and histological rumen morphometry, dry matter intake, growth performance, complete blood count, total protein determination and fecal (oo)cyst counts for Giardia spp. and Eimeria spp. were analyzed using one-way ANOVA. When significant effects were detected, treatment means were compared using Tukey’s HSD at p ≤ 0.05. Sodium butyrate supplementation produced dose-dependent responses. Compared with controls (p ≤ 0.05), G3 calves had a greater mean papillae height (3.20 vs. 1.36 mm) and rumen wall thickness (7.08 vs. 3.62 mm), higher final body weight (93.9 vs. 80.1 kg), increased daily weight gain (798 vs. 550 g/day), increased total dry matter intake (69.4 vs. 55.1 kg) and improved feed conversion ratio (1.45 vs. 1.67). In addition, G3 calves showed reduced Giardia and Eimeria (oo)cyst excretions and favorable hematological shifts at weaning (i.e., higher hematocrit, hemoglobin and mean corpuscular hemoglobin concentration and altered leukocyte differentials) consistent with improved health. In conclusion, sodium butyrate supplementation in pre-weaned dairy calves resulted in dose-dependent improvements in rumen morphometry, voluntary dry matter intake and growth performance. These physiological benefits were accompanied by decreased Giardia spp. and Eimeria spp. (oo)cyst shedding and by positive shifts in hematological health indicators, suggesting a potential enhancement of the animals’ immune response.

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.

Background There is a demand for broad-spectrum products effective against internal and external parasites to simplify dog-owner compliance with veterinary parasiticide treatment recommendations. A field study investigated the safety and efficacy of a broad-spectrum formulation of fluralaner, moxidectin, and pyrantel pamoate compared with a licensed afoxolaner–milbemycin oxime combination product in treating canine intestinal nematode infections in client-owned dogs at multiple sites in Europe. Methods Dogs with positive pre-treatment fecal egg counts (FEC) were randomized to treatment on day 0 with the investigative veterinary product (IVP), at dose rates of: IVP, fluralaner 10–20 mg/kg, moxidectin 0.025–0.050 mg/kg, pyrantel 5–10 mg/kg ( n  = 172); or a control product (CP), afoxolaner 2.5–5.2 mg/kg, milbemycin oxime 0.5–1.0 mg/kg ( n  = 86). On day 14, FEC were completed on all study dogs. Efficacy against any intestinal nematode species was claimed if the FEC reduction in ≥ 10 dogs initially positive for that species was ≥ 90%, and if the geometric mean difference between pre- and post-treatment FEC was statistically significant ( P  ≤ 0.05). Results Study dogs were enrolled at veterinary clinics in Albania ( n  = 60), Bulgaria ( n  = 111), Greece ( n  = 27), and Italy ( n  = 60). Where a sufficient number of infected dogs allowed for statistical comparison, geometric mean FEC reductions on day 14 versus day 0 in the IVP group were significant ( P  < 0.0001) for hookworms, Toxocara canis , Toxascaris leonina, Trichuris vulpis , and capillarid-type eggs. On the basis of geometric mean reductions in FEC relative to day 0, the efficacy of the IVP exceeded 99% against all intestinal nematode parasites, and FEC reductions in the CP exceeded 98%. The percentage of nematode-free dogs in the IVP group was statistically noninferior ( P  < 0.0001) to the CP group and even statistically superior at the α  = 0.025 level of significance (one-sided). There were no treatment-related adverse events. Conclusions In client-owned dogs, the combination of fluralaner–moxidectin–pyrantel is safe and effective for the treatment of intestinal infections with hookworms, ascarids, including T. canis , and whipworms. On the basis of the complete elimination of parasite ova from the feces evaluated in this study, this fluralaner–moxidectin–pyrantel combination demonstrated to be significantly superior ( P  < 0.025) to an afoxolaner–milbemycin combination in the treatment of canine intestinal nematode infections. Graphical Abstract

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.

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.