Morphological and molecular studies on cerebral and non-cerebral coenurosis in sheep and goats

This study explores the causative agent of cerebral and non-cerebral coenurosis in sheep and goats. Cases of non-cerebral coenurosis from a wide geographical range in Africa and western Asia are investigated, and the causative agents are compared to cerebral coenurus cysts from gid cases collected from Greece, where cerebral coenurosis is common but where non-cerebral coenurosis has never been described. The study includes a field and a laboratory-experimental component and provides answers to research questions such as: (i) the presence or absence of non-cerebral coenurosis in sheep; (ii) the description of non-cerebral coenurosis in the intermediate host (sheep and goats); and (iii) the phylogenetic resolution of the T. multiceps cluster and a possible explanation why non-cerebral coenurosis has never been described from some geographical areas. In the field component, a total of 90,415 slaughtered sheep and 2,284 slaughtered goats from abattoirs in the United Arab Emirates (UAE) and Egypt, originating from various tropical and subtropical countries, including India, Pakistan, Iran, Oman, Sudan, Somalia, and Ethiopia, were examined for non-cerebral coenurosis. The field component also included the collection of cerebral coenurus cysts from 20 sheep and 6 goats with cerebral coenurosis and originating from continental Greece and vicinity. Four studies were conducted in the laboratory-experimental component: (1) a morphological study of the characteristics of non-cerebral coenurus cysts and their clusters and protoscolices, (2) a morphological comparison of the rostellar hooks of the collected cerebral and non-cerebral coenurus cysts, (3) a morphological comparison of adult worms produced in dogs experimentally infected with protoscolices from cerebral and non-cerebral cysts of sheep and goats, and (4) a molecular analysis of three partial mitochondrial genes (nad1, cox1, and 12S rRNA) of the above isolates of cerebral and non-cerebral cysts collected in Greece, UAE, and Egypt. The prevalence of non-cerebral coenurosis was 1.75% in goats and only 0.008% in sheep. The only abnormalities observed in the infected goats were large single cysts detected by palpation in thigh muscles and higher serum aspartate aminotransferase (AST) activity. The cysts were found in various muscles and attached to the kidneys, omentum, and heart; they were surrounded by a fibrous, semi-opaque, cloudy-white membrane containing a single coenurus in all cases. The volume of the cysts, the number of the clusters of protoscolices, and the number of protoscolices were similar in sheep and goats. Seventy-six non-cerebral coenurus cysts were collected from goats. The number of protoscolices was significantly positively correlated with the volume of cysts (b = 6.37 > 5; R-Sq = 89.4%; P = 0.000), and the number of clusters was significantly positively correlated with the number of protoscolices (b = 25.13 > 1; R-Sq = 79.8%; P = 0.000), indicating positive allometric growth. The number of clusters was significantly positively correlated with the volume of cysts (b = 0.25 < 0.5; R-Sq = 69.4%; P = 0.000), indicating however negative allometric growth. The biological significance of these allometries is not known, but the parasite may be investing its resources more in the growth of protoscolices, less in the growth of cyst volume, and even less in the number of clusters. Our morphological studies of the rostellar hooks of protoscolices from the cerebral and non-cerebral cysts and the adult worms produced experimentally in dogs did not indicate host-adapted differences (sheep or goats). In contrast, the parasites produced by cerebral and non-cerebral cysts showed clear morphological differences. Most measurements of the hooks and proglottids differed significantly, but the shape of the small hooks, the distribution of the testes in the mature proglottids, and the appearance of the coils of the vas deferens were the most distinct characters. These morphological differences, albeit between the parasites produced by cerebral and non-cerebral cysts, fell within the range of variation of T. multiceps. The phylogenetic analysis of the mitochondrial haplotypes produced three distinct clusters: one cluster including both cerebral isolates from Greece and non-cerebral isolates from tropical and subtropical countries, and two clusters including only cerebral isolates from Greece. The majority of the non-cerebral specimens clustered together but did not form a monophyletic group. No monophyletic groups were observed based on geography, although specimens from the same region tended to cluster. The recorded clusters indicated high intraspecific diversity. The results of this study support the association between T. multiceps variability and the geographical origin of the isolates and lead us to propose a reformulated hypothesis for the existence of cerebral and non-cerebral forms. Our phylogenetic analysis suggests that the development of cerebral coenuri in sheep may be an ancestral property of T. multiceps and the main mode the parasite uses to complete its life cycle. All variants are therefore able to cause cerebral coenurosis in sheep, but only some variants, predominantly from one genetic cluster, acquired the additional capacity to affect the brain of other species (goats and cattle) or to produce non-cerebral forms, mostly in goats and more rarely in sheep. Our phylogenetic analysis therefore clearly indicates a molecular basis for non-cerebral pathogenicity in some variants of T. multiceps. Specific biosecurity actions should therefore be enforced in areas where non-cerebral coenurosis does not occur to prevent the introduction of T. multiceps variants with such pathogenicity.In dieser Forschungsarbeit wurden Isolate von Taenia multiceps als Erreger der zerebralen und nicht-zerebralen Coenurose bei Schafen und Ziegen vergleichend untersucht. Erreger von nicht-zerebraler Coenurose aus einem breiten geographischen Bereich in Afrika und Westasien wurden verglichen mit Erregern zerebraler Coenuruszysten von gesammelten Drehkrankheitsfällen aus Griechenland, wo zerebrale Coenurose häufig vorkommt, wo aber nicht-zerebrale Coenurose noch nie beschrieben wurde. Diese Forschungsarbeit enthält eine feld- und eine laborexperimentelle Komponente und beschäftigt sich mit Aspekten wie: (i) das Vorhandensein oder Fehlen von nicht-zerebraler Coenurose bei Schafen; (ii) die Beschreibung von nicht-zerebraler Coenurose im Zwischenwirt (Schafe und Ziegen); und (iii) die phylogenetische Auflösung des T. multiceps-Clusters sowie eine mögliche Erklärung, warum nicht-zerebrale Coenurose in bestimmten geographischen Gebieten unbekannt ist. Im Rahmen der Felduntersuchung wurde eine Gesamtzahl von 90,415 geschlachtete Schafe und 2,284 geschlachtete Ziegen aus Schlachthöfen der Vereinigten Arabischen Emirate (UAE) und Ägypten untersucht, die aus verschiedenen tropischen und subtropischen Ländern wie Indien, Pakistan, Iran, Oman, Sudan, Somalia und Äthiopien stammten. Die Feldarbeit umfaßte außerdem die Sammlung von zerebralen Coenurosezysten von 20 Schafen und sechs Ziegen, die vom griechischen Festland und Umgebung stammten. Im Labor wurden vier Teilstudien durchgeführt: (1) eine morphologische Untersuchung der Merkmale von nicht-zerebralen Coenurosezysten, deren Cluster und der Protoskolizes, (2) ein morphologischer Vergleich der Rostellarhaken der zerebralen und nicht-zerebralen Coenurosezysten, (3) ein morphologischer Vergleich von erwachsenen Würmern aus Hunden, die experimentell mit Protoskolizes von zerebralen und nicht-zerebralen Zysten von Schafen und Ziegen infiziert worden waren, und (4) eine molekulare Analyse von Teilsequenzen dreier mitochondrialer Gene (nad1, cox1 und 12S rRNA) der oben genannten Isolate von zerebralen und nicht-zerebralen Zysten. Die Prävalenz von nicht-zerebraler Coenurose betrug bei Ziegen 1,75% und nur 0,008% bei Schafen. Die Zysten wurden in verschiedenen Muskeln sowie an den Nieren, am Mesenterium und am Herz gefunden. Sechsundsiebzig nicht-zerebrale Coenuruszysten von Ziegen wurden gesammelt und statistisch ausgewertet. Die Anzahl der Protoskolizes korrelierte signifikant positiv mit dem Volumen von Zysten und die Anzahl von Clustern korrelierte signifikant positiv mit der Anzahl der Protoskolizes, was auf positives allometrisches Wachstum hinweist. Die Anzahl von Clustern korrelierte mit der Anzahl von Zysten, was auf ein negatives allometrisches Wachstum hinweist. Die biologische Bedeutung dieser Allometrie ist nicht bekannt, aber der Parasit investiert offenbar seine Ressourcen mehr in das Wachstum von Protoskolizes und weniger in das Wachstum von Zysten und Clustern. Die morphologischen Untersuchungen der Rostellarhaken von Protoskolizes aus zerebralen und nicht-zerebralen Zysten und die in Hunden experimentell erzeugten adulten Würmer zeigten keine Unterschiede in Bezug auf die Wirtstierart (Schafe oder Ziegen). Im Gegensatz dazu wiesen adulte Würmer, die aus der Inokulation zerebraler und nicht-zerebraler Zysten hervorgingen, deutliche morphologische Unterschiede auf. Die meisten Meßwerte der Haken und Proglottiden unterschieden sich signifikant, aber die Form der kleinen Haken, die Verteilung der Hoden in den reifen Proglottiden und das Aussehen der Spiralen des Vas deferens waren die Merkmale mit den auffälligsten Unterschieden. Diese morphologischen Unterschiede fallen allerdings in den Bereich der Variationen von T. multiceps. Die phylogenetische Analyse der mitochondrialen Haplotypen ergab drei distinkte Cluster: eines, das sowohl zerebrale Isolate aus Griechenland als auch nicht-zerebrale Isolate aus tropischen und subtropischen Ländern umfaßte, und zwei Cluster, die ausschließlich aus zerebralen Isolaten aus Griechenland bestanden. Die meisten der nicht-zerebralen Proben gruppierten zusammen, bildeten aber keine monophyletische Gruppe. Dasselbe gilt für geographische Aspekte, obwohl Proben aus derselben Region zu Clustern tendierten. Die Daten zeigten eine hohe intraspezifische Diversität. Die Ergebnisse dieser Studie unterstützen den Zusammenhang zwischen genetischer Identität der T. multiceps-Isolate und der geographischen Herkunft, und führten zum Vorschlag einer neuen Hypothese zum Vorkommen von zerebralen und nicht-zerebralen Formen. Unsere phylogenetische Analyse legt nahe, daß die Entwicklung der zerebralen Coenuri bei Schafen eine ursprüngliche Eigenschaft von T. multiceps sein könnte. Alle Varianten wären damit in der Lage, zerebrale Coenurose bei Schafen zu verursachen, wogegen nur einige Varianten, vor allem aus einem genetischen Cluster, sekundär die Fähigkeit erworben haben, das Gehirn von anderen Arten (Ziegen und Rindern) zu befallen, sowie nicht-zerebrale Formen (vor allem in Ziegen) hervorzubringen. Unsere phylogenetische Analyse zeigt somit eindeutig eine molekulare Basis für nicht-zerebrale Pathogenität innerhalb der Art T. multiceps. Spezifische Biosicherheitsmaßnahmen sollten daher erwogen werden, um die Einführung von T. multiceps Varianten mit einer solchen Pathogenität zu verhindern

EPIZOOTIOLOGIC RESEARCH OF SELENIUM AND VITAMIN E CONCENTRATIONS IN CATTLE OF THESSALONIKI'S COYNTY

Η ΕΡΓΑΣΙΑ ΠΕΡΙΛΑΜΒΑΝΕΙ ΜΙΑ ΕΠΙΖΩΟΤΙΟΛΟΓΙΚΗ ΚΑΙ ΜΙΑ ΠΕΙΡΑΜΑΤΙΚΗ ΕΡΕΥΝΑ. ΑΠΟ ΤΗΝΕΠΙΖΩΟΤΙΟΛΟΓΙΚΗ ΠΡΟΕΚΥΨΕ ΟΤΙ: 1) ΣΗΜΑΝΤΙΚΟ ΠΟΣΟΣΤΟ ΤΩΝ ΒΟΟΕΙΔΩΝ ΤΗΣ ΧΩΡΑΣ ΜΑΣΚΙΝΔΥΝΕΥΕΙ ΑΠΟ ΣΕΛΗΝΙΟΠΕΝΙΚΑ ΝΟΣΗΜΑΤΑ. ΑΝΤΙΘΕΤΑ, Η ΠΕΡΙΠΤΩΣΗ ΑΒΙΤΑΜΙΝΩΣΗΣ Ε ΠΡΕΠΕΙ ΝΑ ΘΕΩΡΕΙΤΑΙ ΑΠΙ ΘΑΝΗ ΓΙΑ ΤΑ ΖΩΑ ΑΥΤΑ. 2) ΟΙ ΑΓΕΛΑΔΕΣ ΠΟΥ ΒΡΙΣΚΟΝΤΑΝ ΣΤΟΝ 9Ο ΜΗΝΑ ΤΗΣ ΚΥΟΦΟΡΙΑΣ ΚΑΙ ΣΤΟΝ 1Ο ΜΗΝΑ ΜΕΤΑ ΤΟΝ ΤΟΚΕΤΟ ΠΑΡΟΥΣΙΑΖΑΝ ΜΙΚΡΟΤΕΡΕΣ ΣΥΓΚΕΝΤΡΩΣΕΙΣ ΣΕΛΗΝΙΟΥ ΣΕ ΣΧΕΣΗ ΜΕ ΤΙΣ ΑΓΕΛΑΔΕΣ ΠΟΥ ΒΡΙΣΚΟΤΑΝ ΣΤΑ ΥΠΟΛΟΙΠΑ ΣΤΑΔΙΑ ΑΝΑΠΑΡΑΓΩΓΙΚΗΣ ΠΕΡΙΟΔΟΥ. 3) ΤΟ ΜΑΥΡΟ ΤΡ ΙΧΩΜΑ ΤΩΝ HOLSTEIN ΠΑΡΟΥΣΙΑΖΕΙΜΕΓΑΛΥΤΕΡΗ ΣΥΓΚΕΝΤΡΩΣΗ ΣΕΛΗΝΙΟΥ ΣΕ ΣΧΕΣΗ ΜΕ ΤΟ ΚΟΚΚΙΝΟ ΤΡΙΧΩΜΑ ΤΩΝ SIMMENTAL ΚΑΙ ΤΟ ΦΑΙΟ ΤΩΝ SWISS. 4) ΣΤΑ HOLSTEIN ΥΠΑΡΧΕΙ ΓΡΑΜΜΙΚΗ ΣΥΣΧΕΤΙΣΗ ΤΗΣ ΣΥΓΚΕΝΤΡΩΣΗΣ ΣΕΛΗΝΙΟΥ ΣΤΟ ΑΣΠΡΟ ΤΡΙΧΩΜΑ. 5) ΥΠΑΡΧΕΙ ΓΡΑΜΜΙΚΗ ΣΥΣΧΕΤΙΣΗ ΤΗΣ ΣΥΓ ΚΕΝΤΡΩΣΗΣ ΣΕΛΗΝΙΟΥ ΣΤΟ ΑΙΜΑ ΤΩΝ ΒΟΟΕΙΔΩΝ ΚΑΙ ΤΗΣ ΣΥΓΚΕΝΤΡΩΣΗΣ ΤΟΥ ΣΤΟΙΧΕΙΟΥ ΣΤΟ ΑΙΜΑ. ΑΠΟ ΤΗΝ ΠΕΙΡΑΜΑΤΙΚΗ ΕΡΕΥΝΑ ΠΡΟΕΚΥΨΕ ΟΤΙ Η ΔΡΑΣΤΗΡΙΟΤΗΤΑ ΤΟΥ ΕΝΖΥΜΟΥ 5'-ΑΠΟΙΩΔΙΑΣΗ ΤΗΣ ΘΥΡΟΖΙΝΗΣ, ΤΥΠΟΥ Ι ΔΕΝ ΑΥΞΑΝΕΙ, ΟΤΑΝ ΕΝΙΣΧΥΘΟΥΝ ΜΕ ΣΕΛΗΝΙΟ, ΜΟΣΧΑΡΙΑHOLSTEIN ΠΟΥ ΗΔ Η ΕΧΟΥΝ ΣΥΓΚΕΝΤΡΩΣΗ ΣΕΛΗΝΙΟΥ ΑΙΜΑΤΟΣ ΠΕΡΙΠΟΥ 0.05ΜG/ML.THE RESEARCH INCLUDES BOTH AN EPIZOOTIOLOGIC AND AN EXPERIMENTAL RESEARCH: FROM THE EPIZOOTIOLOGIC RESEARCH RESULTED THAT: 1) IN OUR COUNTRY, A SIGNIFICANT PERCENTAGE OF THE CATTLE RUN THE RISK OF SE DEFICIENCY DISEASES, ON CONTRARY, THE CASE OF VIT. E DEF ICIENCY, SHOULD BE REGARDED AS IMPROBABLE FOR THESE ANIMALS. 2) THE COWS WHICH WERE IN THE NINTH MONTH OF PREGNANCY AND THE FIRST MONTH AFTER PARTURITION PRESENTED LOWER SE CONCENTRATION COMPARED TO THE COWS INTHE OTHER STAGES OF THE REPRODUCTIVE PERIOD. 3) THE BLACK HAIR OF HOLSTEIN PRESENTED HIGHER CONCENTRATION OF SE IN COMPARISON WITH THE RED HAIR OF SIMMENTAL AND THE GREYISCH BROWN OF SWISS. 4) IN THE HOLSTEIN CATTLE, THERE IS A LINEAR COORELATION BETWEEN THE SE CONCENTRATION IN BLOOD AND THE SE CONCE NTRATION IN BLACK COLOURED HAIR, ALSO BETWEEN THE CONCENTRATION OF SE IN BLACK HAIR AND THAT IN WHITE HAIR. 5) THERE IS A LINEAR COORELATION BETWEEN THE CONCENTRATION OF SE IN THE BLOOD OF CATTLE AND THE CONCENTRATION IN THE LIVER TISSUE. FROM THE EXPERIME NTAL RESEARCH RESULTED THAT THE ACTIVITY OF THE ENZYME TYPE I IODOTHYRONINE 5'-DEIODINASE DOES NOT INCREASE WHEN HOLSTEIN CALVES, WHICH ALREADY HAVE A SE CONCENTRATION IN BLOOD ABOUT 0,05ΜG/ML, ARE SUPPLEMENTED WITH SE

Heterologous Challenge with PRRSV-1 MLV in Pregnant Vaccinated Gilts: Potential Risk on Health and Immunity of Piglets

The objective of the present study was to evaluate the potential risks of the four commercial PRRS-1 MLV vaccines in pregnant vaccinated gilts at the last stage of gestation under field conditions. The study was conducted at four pig farms, including 25 gilts from each farm (25 × 4 = 100 gilts), which were equally allocated to five different study groups. A PRRS-1 MLV vaccination was applied on the 100th day of their pregnancy with the different commercial vaccines that are available in the Greek market. The results indicated virus congenital infection and viremia in piglets (20/200 = 10% PRRSV infected piglets), and detection of PRRSV-specific antibodies (181/200 = 90.5% piglets found with PRRSV antibodies). The subsequent phylogenetic analyses revealed high percentages of similarity between the PRRSV-1 strain detected in infected litters and the PRRSV-1 vaccine strain to which the study gilts had been previously exposed to. Health status analyses of trial piglets resulted in differences between litters from vaccinated sows and litters from non-vaccinated sows at 110th day of gestation as regards the number of weak-born piglets, mummies, and piglets with splay-leg and/or respiratory symptoms. The current study’s results indicate several potential dangers of the PRRS MLV vaccination in late gestation

Heterologous Challenge with PRRSV-1 MLV in Pregnant Vaccinated Gilts: Potential Risk on Health and Immunity of Piglets

The objective of the present study was to evaluate the potential risks of the four commercial PRRS-1 MLV vaccines in pregnant vaccinated gilts at the last stage of gestation under field conditions. The study was conducted at four pig farms, including 25 gilts from each farm (25 &times; 4 = 100 gilts), which were equally allocated to five different study groups. A PRRS-1 MLV vaccination was applied on the 100th day of their pregnancy with the different commercial vaccines that are available in the Greek market. The results indicated virus congenital infection and viremia in piglets (20/200 = 10% PRRSV infected piglets), and detection of PRRSV-specific antibodies (181/200 = 90.5% piglets found with PRRSV antibodies). The subsequent phylogenetic analyses revealed high percentages of similarity between the PRRSV-1 strain detected in infected litters and the PRRSV-1 vaccine strain to which the study gilts had been previously exposed to. Health status analyses of trial piglets resulted in differences between litters from vaccinated sows and litters from non-vaccinated sows at 110th day of gestation as regards the number of weak-born piglets, mummies, and piglets with splay-leg and/or respiratory symptoms. The current study&rsquo;s results indicate several potential dangers of the PRRS MLV vaccination in late gestation

Evaluation of Intradermal PRRSV MLV Vaccination of Suckling Piglets on Health and Performance Parameters under Field Conditions

Porcine reproductive and respiratory syndrome virus (PRRSV) causes respiratory disease in weaning and growing pigs. A vaccination against PRRSV is one of the most important control measures. This trial aimed to evaluate the effect of the intradermal (ID) administration of a PRRSV-1 modified live virus (MLV) vaccine in comparison to the intramuscular (IM) administration on the piglets’ health and performance. A total of 187 suckling piglets of a PRRSV-positive commercial farrow-to-finish farm were assigned to four groups: group A—PRRSV ID, group B—PRRSV IM, group C—control ID, and group D—control IM. At 2 weeks of age, all the study piglets were either vaccinated with a PRRSV-1 MLV vaccine or injected with the vaccine adjuvant (controls). The collected blood serum samples were tested by ELISA and qRT-PCR. The side effects, body weight (BW), average daily gain (ADG), mortality rate, and lung and pleurisy lesions scores (LLS, PLS) were also recorded. The ELISA results indicated that the vaccination induced an important seroconversion at 4 and 7 weeks. Significant differences in the qRT-PCR results were noticed only at 10 weeks in group A vs. group C (p p < 0.05). High viral loads, as evidenced by the qRT-PCR Ct values, were noticed in animals of both non-vaccinated groups at 7, 10, and 13 weeks. An ID vaccination has a positive impact on the BW at the piglets’ slaughter, while both an ID and IM vaccination had a positive impact on the ADG. The mortality rate was lower in vaccinated groups at the finishing stage. The LLS and PLS were significantly lower in the vaccinated groups. In conclusion, our study demonstrated that the ID vaccination of suckling piglets with a PRRSV-1 MLV vaccine has a positive effect on the piglets’ health and performance, including an improved BW and a lower LLS and PLS index at their slaughter, as well as a decreased mortality rate at the growing/finishing stage

Angiotensin II blood serum levels in piglets, after intra-dermal or intra-muscular vaccination against PRRSV

Simple Summary Porcine reproductive and respiratory syndrome virus (PRRSV) infection causes massive financial losses in pig production worldwide. Vaccination is still the most cost-effective tool to handle PRRSV infection. PRRSV induces apoptosis in different organs. Angiotensin II (Ang II) participates in the inflammatory response, cell proliferation, migration, and apoptosis. The objective of the current study was to assess the concentration of Ang II in the serum of piglets following immunization against PRRSV through intradermal (ID) or intramuscular (IM) vaccination with a commercial PRRS modified live virus (MLV) vaccine. The results indicated differences in viremia of tested piglets at 7 weeks of age, while piglets at 10 weeks of age were all found qRT-PCR positive for PRRSV. Moreover, significant differences were noticed in Ang II in 7-week-old piglets. In conclusion, our study provides evidence that ID vaccination induces less tissue damage, based on the lower measurements of Ang II in the serum of ID vaccinated piglets. The Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) induces apoptosis in different organs. Angiotensin II (Ang II) is the main effector of the renin-angiotensin system and participates in apoptosis. Thus, this study aimed to investigate changes in piglet serum Ang II levels following intradermal (ID) and intramuscular (IM) vaccination with a commercial PRRS modified live virus (MLV) vaccine. The trial was conducted in a commercial pig farm, including 104 piglets which were randomly allocated to four groups: Group A-Porcilis PRRS ID, Group B-Porcilis PRRS IM, Group C-Diluvac ID and Group D-Diluvac IM. The study piglets were either vaccinated or injected at 2 weeks of age and they were tested by qRT-PCR for PRRSV and by ELISA for Ang II. The results indicated differences in viremia of tested piglets at 7 weeks of age, while piglets at 10 weeks of age were all found qRT-PCR positive for PRRSV. In addition, significant differences were noticed in Ang II in 7-week-old piglets. In conclusion, the present study provides evidence that ID vaccination induces less tissue damage, based on the lower measurements of Ang II in the serum of ID vaccinated piglets

The role of lactate dehydrogenase, alkaline phosphatase and aspartate aminotransferase in the diagnosis of subclinical intramammary infections in dairy sheep and goats

The objective was to investigate the changes occurring in the activities of the enzymes lactate dehydrogenase (LDH), alkaline phosphatase (ALP) and aspartate aminotransferase (AST) in sheep and goat milk as a result of subclinical intramammary infections (IMI) and to evaluate the use of these enzymes for the diagnosis of subclinical IMI in dairy sheep and goats. A total of 206 samples of sheep milk and 162 samples of goat milk, obtained from equal udder halves, were used in the study. For each species they were divided into two groups: a no-infection group and a subclinical infection group. Activities of LDH, ALP and AST were significantly higher in the subclinical infection group than in the no-infection group (P<0.05) in both sheep (LDH: 350.42 +/- 11.25 v. 120.91 +/- 4.41; ALP: 2773.43 +/- 105.18 v. 2189 +/- 94.24; AST: 29.57 +/- 0.74 v. 17.32 +/- 0.46) and goats (LDH: 354.07 +/- 13.33 v. 103.79 +/- 3.75; ALP: 311.13 +/- 25.74 v. 137.24 +/- 19.62; AST: 27.59 +/- 6.42 v. 15.87 +/- 0.45). The activity of LDH was identified as indicator for subclinical IMI in both sheep and goats. The Optimum cut-off values for LDH activity, offering the highest diagnostic sensitivity (DSn) and diagnostic specificity (DSp), determined by receiver operating characteristic (ROC) analysis, were at 197 U/l, 185 U/l and 197 U/l for sheep, goats and both species, respectively. DSn for sheep, goats and both species at these cut-off values was 92.8%, 98.2% and 94.0%, whereas DSp was 95.4%, 96.3% and 96.3%, respectively. It was concluded that the determination of LDH activity in milk serum is a sensitive and reliable method for the detection of subclinical IMI in dairy sheep and goats

Investigation of Fas (APO-1)-related apoptosis in piglets intradermally or intramuscularly vaccinated with a commercial PRRSV MLV

Porcine reproductive and respiratory syndrome virus (PRRSV) induces apoptosis through the activation of death receptors, including cell-surface Fas receptor. The aim of this study was to investigate the impact of intradermal (ID) and intramuscular (IM) vaccination with a commercial PRRSV-modified live vaccine in piglets on Fas-related apoptosis. The study included 104 suckling piglets from a commercial farrow-to-finish pig farm, suffering from positive unstable PRRSV status. Animals were assigned in four groups: group A-Porcilis PRRS ID-vaccinated pigs, group B-Porcilis PRRS IM-vaccinated pigs, group C-Diluvac ID adjuvant-administered pigs, and group D-Diluvac IM adjuvant-administered pigs. Vaccines were administered at 2 weeks of age. Blood samples were collected from the same pigs at 4, 7, and 10 weeks of age. Sera were examined by quantitative real-time reverse transcription-PCR (qRT-PCR) for PRRSV and by ELISA for soluble Fas (sFas). At 4 weeks of age, all groups were negative qRT-PCR for PRRSV; at 7 weeks only group A was negative; and at 10 weeks all groups were positive. sFas was significantly increased in groups C (4 vs. 7, 4 vs. 10, and 7 vs. 10 weeks) and D (7 vs. 10 weeks). Significant differences among groups were noticed only at 10 weeks (A vs. C, A vs. D, B vs. C, B vs. D). A significant positive and moderate correlation between PRRSV viral load and Fas level was observed. In unvaccinated piglets, increased serum sFas levels reveal apoptotic suppression compared with vaccinated piglets. In the latter, vaccine-derived antibodies limit the infection and may attribute to the reduced Fas expression, suggesting a weak induction of lymphocyte-mediated cytotoxicity

Cross-Sectional Serosurvey and Factors Associated with Exposure of Dogs to Vector-Borne Pathogens in Greece

Blood samples from 2620 dogs living in 7 different areas were used for the detection of antibodies against Ehrlichia canis and Borrelia burgdorferi and Dirofilaria immitis antigen. Previously published seropositivity data against Leishmania infantum from the same samples were also used to identify multiple-pathogen seropositivity. The associations between seropositivity against each pathogen and gender, age, utility, hair length, and region of residence were evaluated by multivariate logistic regression analyses. The prevalence of seropositivity against E. canis, B. burgdorferi, and D. immitis was 12.25%, 2.23%, and 5.96%, respectively. Double- and triple-pathogen seropositivity was detected in seven different combinations. Age and region of residence were strongly associated with seropositivity against all pathogens. The association between seropositivity and the area of residence highlights the need for regular testing of dogs for vector-borne pathogens in areas with similar conditions to define control measures

Investigation of Fas (APO-1)-Related Apoptosis in Piglets Intradermally or Intramuscularly Vaccinated with a Commercial PRRSV MLV

Porcine reproductive and respiratory syndrome virus (PRRSV) induces apoptosis through the activation of death receptors, including cell-surface Fas receptor. The aim of this study was to investigate the impact of intradermal (ID) and intramuscular (IM) vaccination with a commercial PRRSV-modified live vaccine in piglets on Fas-related apoptosis. The study included 104 suckling piglets from a commercial farrow-to-finish pig farm, suffering from positive unstable PRRSV status. Animals were assigned in four groups: group A-Porcilis PRRS ID-vaccinated pigs, group B-Porcilis PRRS IM-vaccinated pigs, group C-Diluvac ID adjuvant-administered pigs, and group D-Diluvac IM adjuvant-administered pigs. Vaccines were administered at 2 weeks of age. Blood samples were collected from the same pigs at 4, 7, and 10 weeks of age. Sera were examined by quantitative real-time reverse transcription-PCR (qRT-PCR) for PRRSV and by ELISA for soluble Fas (sFas). At 4 weeks of age, all groups were negative qRT-PCR for PRRSV; at 7 weeks only group A was negative; and at 10 weeks all groups were positive. sFas was significantly increased in groups C (4 vs. 7, 4 vs. 10, and 7 vs. 10 weeks) and D (7 vs. 10 weeks). Significant differences among groups were noticed only at 10 weeks (A vs. C, A vs. D, B vs. C, B vs. D). A significant positive and moderate correlation between PRRSV viral load and Fas level was observed. In unvaccinated piglets, increased serum sFas levels reveal apoptotic suppression compared with vaccinated piglets. In the latter, vaccine-derived antibodies limit the infection and may attribute to the reduced Fas expression, suggesting a weak induction of lymphocyte-mediated cytotoxicity