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.

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.

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.

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.

Haemonchus contortus Parasitism in Intensively Managed Cross-Limousin Beef Calves: Effects on Feed Conversion and Carcass Characteristics and Potential Associations with Climatic Conditions.

The objectives of the study were: (a) to study the effect of Haemonchus spp. on the growth performance characteristics of fattening calves, (b) to assess any potential effects on carcass characteristics and (c) to investigate the potential role of climatic conditions in the process of the infection. The study was conducted for 201 days in an intensively managed cross-Limousin herd. The animals were divided into two equal groups: those receiving anthelmintic treatment (AT) and the untreated (C) controls. The same nutritional regime was applied to both groups and the feed consumption was calculated daily. Standard parasitological examinations were performed at weekly intervals. At slaughter, carcasses were weighed and assessed for conformation and fat cover classes. Climatic variables were obtained for the location of the farm and a temperature-humidity index was calculated. Before anthelmintic treatment with ivermectin, there was no difference in parasitic burdens between the two groups: 544 (AT) vs. 554 (C) epg, whilst after it, counts were 0 and 450-700 epg, respectively, with over 96% of larvae identified as Haemonchus spp. and, later, confirmed as Haemonchus contortus. It was concluded that treated animals had a higher average total bodyweight gain, higher feed conversion ratio and carcass yield of superior quality than controls. There was a difference between the two groups in the pattern of reduction of dry matter intake as the temperature-humidity index increased.

Serological Assessment of Ascaris suum Exposure in Greek Pig Farms and Associated Risk Factors Including Lawsonia intracellularis.

The effects of nematodes and bacteria on intestinal health are of primary importance in modern swine production. The aim of the present study was to assess the seropositivity status of Ascaris suum infection in fatteners in intensive swine farms in Greece and address possible risk factors, including Lawsonia intracellularis as a predisposing factor to swine ascariosis. In total, 360 blood serum samples from pigs in the late fattening period, from 24 Greek swine farrow-to-finish farms (15 samples/farm) were collected and tested with Svanovir® A. suum antibody ELISA and Ileitis antibody ELISA. The results demonstrated 34.4% seropositive samples for A. suum and 42.2% for L. intracellularis. The analysis of predisposing risk factors suggested that the frequency of application of anthelminthic treatment to sows more than two times per year was significantly associated with the lower likelihood of A. suum infection, whereas a greater likelihood of A. suum infection was observed in pigs with concurrent L. intracellularis exposure. The results highlight the importance of proper anthelminthic metaphylaxis of the breeding stock, as well as the likely outcome of concurrent exposure to two intestinal pathogens in pigs, implying a possible association between intestinal nematodes and bacteria in swine.

High Milk Somatic Cell Counts and Increased Teladorsagia Burdens Overshadow Non-Infection-Related Factors as Predictors of Fat and Protein Content of Bulk-Tank Raw Milk in Sheep and Goat Farms.

This paper presents the results of an extensive countrywide investigation performed in 325 dairy sheep flocks and 119 goat herds throughout Greece. The objectives of the study were (a) to investigate fat and protein content in the bulk-tank raw milk of small ruminant farms in Greece and (b) to identify factors potentially influencing that content and factors that can contribute to increased content. The mean fat/protein contents in bulk-tank raw milk of sheep and goats were 6.16 ± 0.05%/4.43 ± 0.01% and 4.77 ± 0.44%/3.23 ± 0.30%, respectively. Significant differences were seen in protein content between farms in the various parts of the country. For sheep, multivariable analyses revealed breed and age of lamb removal from dams as significant for fat content, and somatic cell counts, management system in the farm, administration of anthelmintic treatment during the last stage of pregnancy, and farmer education as significant for protein content. For goats, significant factors were month into lactation period, age of kid removal from dams, and breed for fat content, and somatic cell counts, month into lactation, grazing duration, and % Teladorsagia larvae in faecal samples for protein content. For concurrently high fat and protein content, in multivariable analyses, the following emerged as significant factors: somatic cell counts in milk, numbers of parasite eggs in faeces, and veterinary collaboration (sheep), and month into lactation and somatic cell counts in milk (goats). The results indicate that high somatic cell counts in milk (reflecting the presence of mastitis) and gastrointestinal parasitic infections (mainly Teladorsagia infection) appear to exert a more significant influence on fat and protein content of milk, in comparison to non-infection-related factors.

Deltamethrin Application on Pre-Weaned Calves Improves Feed Consumption, Stress and Fatigue Status under Heat Stress Conditions.

Fly infestation remains a universal problem for dairy cattle herds, affecting the animals' health and welfare status. Pre-weaned dairy calves are significantly challenged by the direct and indirect consequences of severe fly infestation, heat-stress and their interaction, which contribute to a stressful and fatiguing environment. Among several physiological, behavioral, clinical and biochemical traits, serum cortisol (SC) and creatine kinase (CK) levels, as well as feed consumption can be used as valid indicators of potential stressful and fatiguing conditions and, therefore, can be efficiently used for stress analysis studies. Hence, the objective of the study was to assess the fly-repellency effect of deltamethrin on pre-weaned dairy calves exposed to heat stress conditions, as well as its association with SC, CK concentrations and feed consumption. Two commercial dairy cattle herds of the Holstein breed in Central Macedonia (Greece) were involved in the study during summer months and under heat stress conditions. Deltamethrin administration resulted in (i) a decreased fly population (100% Musca domestica) landing on pre-weaned dairy calves, (ii) a reduced SC (stress indicator) and CK (fatigue indicator) concentration, and (iii) an increased consumption of feedstuff in deltamethrin treated animals compared to the untreated ones.

Will Fly Repellency Using Deltamethrin Reduce Intramammary Infections, Stress and Fatigue Indicators of Dairy Ewes under Intensive Management?

Intramammary infections (IMIs) caused by various pathogens may lead to clinical or subclinical mastitis, challenging the health and welfare status of infected animals and decreasing the quantity and quality of the produced milk. Additionally, the zoonotic potential of some of the pathogens isolated from IMI cases, the emergence of antibiotic resistance due to the extensive antibiotic use for IMI treatment, and the accumulation of antibiotic residues in milk and meat represent significant concerns for public health. Therefore, the investigation of IMI risk factors and the proposal of efficient measures to mitigate their effects on animal health and welfare is crucial. Although fly infestation is considered to play a significant role in the transmission of IMI pathogens, its adverse effects on udder health and the overall comfort status of dairy ewes have not been quantified and assessed on an evidential basis. Hence, the objectives of this study were to assess, for the first time, the fly repellent effect of deltamethrin and link it to: (i) the occurrence of common bacterial IMI; (ii) the somatic cell counts in milk; and (iii) the serum cortisol and creatine kinase levels (stress and fatigue indicators). The study was carried out in an intensive dairy sheep farm in northern Greece, during peak fly season. Deltamethrin treatment was associated with a reduced (i) number of flies (mostly Musca domestica) landing on treated ewes, compared to untreated ones (p < 0.05); (ii) colony-forming units in the case of Non-aureus Staphylococci IMIs (p < 0.05); and (iii) number of somatic cells in the milk (p < 0.001). Finally, serum cortisol and creatine kinase levels were significantly lower in deltamethrin-treated ewes (p < 0.001), indicating a less stressful environment for them.

Haemonchosis: A Challenging Parasitic Infection of Sheep and Goats.

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.

Rapid on-site diagnosis of canine giardiosis: time versus performance.

BACKGROUND: Infections by protozoans of the genus Giardia are a common cause of diarrhea in dogs. Canine giardiosis constitutes a disease with a zoonotic potential; however, it is often underestimated due to its challenging diagnosis. The objective of the study was to assess the diagnostic performance of an immunochromatographic strip test (SpeedTM Giardia, Virbac, France) comparing it with microscopy (zinc sulfate flotation) by utilizing the combination of an enzyme immunoassay (ProSpecTTM Giardia EZ Microplate Assay, Oxoid Ltd., UK) and the PCR as the gold standard. A positive result in both ELISA and PCR was set as the gold standard. METHODS: Initially, fecal samples from dogs with clinical signs compatible with giardiosis were tested with the SpeedTM Giardia test and separated into two groups of 50 samples each: group A (positive) and group B (negative). Thereafter, all samples were examined by zinc sulfate centrifugal flotation technique and assayed by the ProSpecTTM Giardia Microplate Assay and PCR. The performance of the SpeedTM Giardia and zinc sulfate centrifugal flotation tests were calculated estimating sensitivity, specificity, and positive and negative likelihood ratio; the chi-square and McNemar tests were used for the comparison of the two methods. RESULTS: Giardia cysts were not detected by microscopy in 16 out of the 50 samples (32%) of group A and in none of group B samples. Eight out of 50 samples in group B (16%) were tested positive both with the ProSpecTTM Giardia Microplate Assay and PCR. Fecal examination with the SpeedTM Giardia test was more sensitive (86.2%) than the parasitological method (58.6%, P < 0.001) while the specificity of both methods was 100%. CONCLUSIONS: The SpeedTM Giardia test is an easy-to-perform diagnostic method for the detection of Giardia spp., which can increase laboratory efficiency by reducing time and cost and decrease underdiagnosis of Giardia spp. infections. This immunochromatographic strip test may be routinely exploited when a rapid and reliable diagnosis is required, other diagnostic techniques are unavailable and microscopy expertise is inefficient. In negative dogs with compatible clinical signs of giardiosis, it is recommended either to repeat the exam or proceed with further ELISA and PCR testing.

Frequency of Resistance to Benzimidazoles of Haemonchus contortus Helminths from Dairy Sheep, Goats, Cattle and Buffaloes in Greece.

The study investigated the presence of resistance to benzimidazoles in Haemonchus contortus helminths from ruminant species in Greece through the detection of the Phe/Tyr polymorphism in the amino acid at position 200 of the ß-tubulin protein. In total, 288 adult female H. contortus helminths collected from the abomasum of various ruminant animals in Greece were tested. Of these, 96 were collected from sheep, 96 from goats, 48 from cattle, and 48 from buffaloes. The frequencies of the homozygous and heterozygous resistant genotypes at the position 200 of the ß-tubulin gene of helminths recovered from sheep were 96.9% and 3.1%, respectively. The frequencies of the homozygous and heterozygous resistant genotypes, respectively, were 100.0% and 0.0% in helminths from goats, 25.0% and 75.0% in helminths from cattle and 8.3% and 91.7% in helminths from buffaloes. In all parasitic populations, no homozygous susceptible genotypes were detected. The present study highlighted, for the first time, the emergence of benzimidazole-resistant H. contortus in goats, cattle, and buffaloes in Greece, using an allele-specific PCR. It is postulated that benzimidazole-resistant alleles were transferred from sheep or goats to cattle and buffaloes at the commonly grazing pastures in Greece.

Anthelmintic activity of injectable eprinomectin (eprecis® 20 mg/mL) in naturally infected dairy sheep.

The anthelmintic activity of an injectable eprinomectin formulation (Eprecis® 20 mg/mL) was evaluated in 150 naturally infected dairy sheep raised in 3 semi-intensive flocks. All ewes were at the same stage of lactation and grazed on natural pastures. Ewes did not receive any anthelmintic treatment for at least 4 months prior to the experiment. In each flock, 50 ewes were selected and randomly allocated to control (C) or treatment (T) groups (n = 25 per group). Groups were balanced according to the ewes' bodyweight (BW) and fecal egg count (FEC) measured seven days before eprinomectin administration (day-7). On study day 0, ewes in group T, received 0.2 mg/kg BW of eprinomectin subcutaneously (Eprecis® 20 mg/mL, Ceva). Ewes in group C were left untreated. Fecal samples were collected on day 0, 7, 14, 21 and 28 post-treatment to assess FEC and for coprocultures. Ewes were weighed on day 0 and 28. Overall and within-flock efficacy of eprinomectin was calculated throughout the experimental period. No local or general adverse reaction after injection was observed. The most prevalent parasite genera were Teladorsagia, Haemonchus and Trichostrongylus. Following treatment, the overall mean FEC of C and T groups differed significantly (P < 0.001). Overall and within-flock efficacy of eprinomectin was 99.8%-100.0% and 99.7%-100.0%, respectively. Contrary to C group, ewes treated with injectable eprinomectin increased their BW during the study (-0.5 kg vs. + 1.5 kg, P < 0.001). In this field study, a single subcutaneous injection of eprinomectin to dairy sheep, at 0.2 mg/kg BW, resulted in excellent curative anthelmintic activity; egg counts remain low for at least 28 days after treatment.

Fly repellency using deltamethrin may reduce intramammary infections of dairy cows under intensive management.

The aim of this study was to assess the possible effect of the fly repellent deltamethrin on the full microbiological profile of the intramammary infections as well as on the somatic cell count in one Greek dairy cattle farm under intensive management, during peak fly season. Fifty five multiparous cows, stabled in the same farm, were randomly divided in three groups; cows of Group A were dressed on their back with deltamethrin, cows of Group B remained untreated within the same pen and cows of Group C remained untreated in a separate pen. Clinical records of the herd showed a history of clinical and subclinical mastitis (especially during spring and early summer) and fly infestation. Deltamethrin reduced fly population, landing on the cows of Group A, facilitating a significant decrease of S. aureus, coagulase negative staplylococci, E. coli intramammary infections and somatic cell count, throughout the study period. Consequently, there is a relationship between fly abundance and transmission of bacteria resulting in intramammary infections and mastitis in dairy cows. The use of the pyrethroid deltamethrin reduced fly abundance and therefore, the possibility of transmitting pathogenic bacteria in the mammary gland.

Human taeniasis/cysticercosis: a potentially emerging parasitic disease in Europe.

Taenia saginata (T. saginata)/Taenia solium (T. solium) taeniasis/cysticercosis disease complexes remain a significant challenge for food safety and public health. Human taeniasis is an infectious disease caused by the ingestion of the metacestode larval stage, the cysticerci of T. saginata in beef or T. solium in pork. Humans can also become infected via the ingestion of T. solium eggs. In this case, the cysticerci can establish in the central nervous system, causing the infection called neurocysticercosis. T. solium is of higher importance than T. saginata because the former species can cause neurocysticercosis in humans, a major cause of neurological morbidity in the world. The taeniasis/cysticercosis complex is included in the list of neglected zoonotic diseases by the World Health Organization and Food and Agriculture Organization, with T. solium being the number one foodborne parasite; it occurs mostly in developing countries, such as regions of Asia, Africa and Latin America, where the disease remains endemic. Long absent in Western Europe and other developed countries, cysticercosis has been recently re-emerged as a result of immigration, travel and commerce. In this review, cysticercosis is presented with special emphasis on some aspects of this neglected disease: the main clinical manifestations, risk factors and epidemiology. In addition, any recent advances in diagnostic approaches and treatment are discussed. Finally, the complexities involved in the control of the disease and the need to revise current management strategies are highlighted.

Feline gastrointestinal parasitism in Greece: emergent zoonotic species and associated risk factors.

BACKGROUND: Feline gastrointestinal parasitism constitutes an issue of concern for veterinarians since parasites are widespread and affect animals' health and welfare. Furthermore, some of these pathogens have zoonotic potential. To provide detailed data on the current epizootiology of feline endoparasitism, a multicentric survey was conducted during 2016. METHODS: Faeces from 1150 cats were collected from all regions of Greece and examined by sedimentation and flotation techniques. Possible risk factors including gender, age, ownership status, living conditions and co-infections with other parasites were assessed using binary regression models for each one of the most prevalent parasites. RESULTS: The overall gastrointestinal parasite prevalence in cats was 50.7%. The study population consisted of cats of both sexes, different age groups, ownership status and living conditions. A total of 10 gastrointestinal parasitic species were detected and up to 5 different parasites were isolated in the same faecal sample. The most frequently identified parasites were Toxocara cati (n = 278; 24.2%), followed by Cystoisospora spp. (n = 189; 16.4%), Ancylostomatidae (n = 186; 16.2%), Aelurostrongylus abstrusus (n = 40; 3.5%), Giardia spp. (n = 26; 2.3%), Joyeuxiella pasqualei (n = 14; 1.2%), Capillaria aerophila (n = 8; 0.7%), Dipylidium caninum (n = 3; 0.2%), Toxascaris leonina (n = 2; 0.1%) and Troglostrongylus brevior (n = 2; 0.1%). The occurrence of co-infections was 11.6%. Concerning risk factors, the likelihood of T. cati infection was higher for female cats living outdoors and for cats being infected with Cystoisospora spp. In the same frame, young, stray, male and free of A. abstrusus cats were more likely to be infected with Cystoisospora spp. Correspondingly, stray, infected with Giardia spp. but free of Cystoisospora spp. cats were more likely to be infected with Ancylostomatidae. Regarding A. abstrusus infection, a higher probability was reported for cats living outdoors and for cats free of Cystoisospora spp., while Giardia spp. infections were more common in young and co-infected with Ancylostomatidae animals. CONCLUSIONS: The prevalence of parasitized cats in Greece was high and thus consideration should be paid to control the risk factors, to implement targeted preventive antiparasitic treatments and educate cat owners on the value of prevention for the health and welfare of their cats.

Evaluation of the in-field efficacy of oregano essential oil administration on the control of neonatal diarrhea syndrome in calves.

The objective of this study was to evaluate under field conditions, whether daily administration of oregano essential oil is effective in preventing and/or diminishing the severity of neonatal diarrhea syndrome in calves aged less than 15days. Ninety-one newborn calves from three dairy farms were assigned into two groups; "Eco" group (n=46) calves were drenched with Greek oregano (Origanum vulgare ssp. Hirtum) essential oil (ECODIAR® liquid 5%) at the dose of 12.5mg/kg body weight once per day until the age of 10days. "Conts" group (n=45) calves were left untreated and served as controls. All animals were monitored daily for the incidence of diarrhea until the age of 15days and their fecal score was recorded. Fecal samples were collected on days 3, 6 and 10 for microbiological and parasitological evaluation. Average fecal score throughout the experiment, incidence of diarrhea, duration and severity of diarrhea episodes were significantly lower in Eco group compared to the controls. Daily administration of oregano essential oil in calves for the first 10days of their life effectively diminishes the severity of naturally acquired diarrhea under field conditions and, under certain hygiene practices, possess a preventive effect against neonatal diarrhea syndrome.

Antioxidant effect of a polyphenol-rich grape pomace extract on motility, viability and lipid peroxidation of thawed bovine spermatozoa.

BACKGROUND: Grape extracts of the Greek species Vitis vinifera possess potent antioxidant properties in vitro. The freeze/thaw process and the preparation of semen during assisted reproductive techniques can adversely affect the functional integrity of spermatozoa. The objective was to assess the effect of three different concentrations (1 µg ml(-1), 2 µg ml(-1) and 5 µg ml(-1)) of a polyphenol-rich grape pomace extract on motility, viability, acrosomal and lipid peroxidation status of thawed bovine spermatozoa after 2 and 4 hrs of incubation. RESULTS: The results indicate that the percentage of "Rapid" spermatozoa remained significantly increased (p <0.05) in the presence of 5 µg ml(-1) of the extract, compared to the control after 2 hrs of incubation. Additionally, the incubation of spermatozoa with 2 µg ml(-1) and 5 µg ml(-1) of the extract for 2 hrs resulted in a significantly better maintenance of viable spermatozoa with intact acrosome (p <0.05). The other parameters did not show statistically significant changes. Moreover, the presence of 2 µg ml(-1) and 5 µg ml(-1) of the extract kept the levels of Malondialdehyde (MDA) production in significantly lower level, compared to the other groups, after 2 hrs and 4 hrs of incubation (p <0.05). Particularly, a dose-dependent effect was noticed after 2 hrs of incubation. CONCLUSIONS: Our results suggest that the grape pomace extract exerts a powerful antioxidant role, by suppressing lipid peroxidation, and provides protection in terms of motility and acrosomal integrity, which are correlated with in vivo fertility. The optimal extract concentration is 5 µg ml(-1).